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Simultaneous identification and quantitation of 11 flavonoid constituents in Kaempferia parviflora by gas chromatography
Abstract
Kaempferia parviflora (Krachaidum; KD) is a Thai herb, the rhizomes of which have been used in folk medicine and ritual ceremonies. The increasing use of KD has led to concerns regarding the variation of quality, potency and efficacy of KD products. A gas chromatographic method was developed and validated using 11 flavonoids that had been fully characterized as reference. Limits of detection ranged from a low of 0.1 ppm to a high of 1.0 ppm. The limits of quantitation were a low of 0.5 ppm (5-hydroxy-3,7-dimethoxyflavone) to a high of 3.0 ppm (5,7,4'-trimethoxyflavone and 5,7,3',4'-tetramethoxyflavone). Precision of intra- and inter-day analyses gave a RSD range of 3.02-8.25 and 2.84-12.37, respectively. The diversity of flavonoid content and their distribution profiles in KD samples from 12 different origins was investigated using the validated method. Total flavonoid content in these samples ranged from 23.86 to 60.98 mg/g. Two of the compounds, 5,7-dimethoxyflavone and 5,7,4'-trimethoxyflavone, emerged as major constituents. Samples contained as much as 21.68 and 9.88 mg/g, respectively. Two distinct patterns of the distribution of the flavonoids, as characterized by the ratio of these two compounds in the KD rhizome samples, were observed. This method is expected to be useful in the quantitative and qualitative analyses of the flavonoid content of KD samples and as a quality control assessment of KD raw materials and products.
... These included nanosuspensions, selfmicroemulsifying and self-nanoemulsifying drug delivery systems, complexation with cyclodextrin (4,5), and solid dispersion (6). Nonetheless, studies focusing on improving the oral delivery of pure methoxyflavones are quite scarce (7). It is generally understood that poor oral bioavailability of bioflavonoids arising from inadequate solubility in the gastrointestinal tract coupled with the presystemic metabolism plays a major role in their lack of clinical success (3,8). ...
... DSC thermograms of KAPE methoxyflavones powder, liquid glycols, and mixtures of the methoxyflavones in glycols are presented in Fig. 3. Clearly-defined endothermic peaks were present in the solid methoxyflavones suggestive of their crystalline nature (Fig. 3a). The thermal events occurred between 145 to 155 °C and were comparable to the melting peaks of the methylated flavones (7,24). The melting temperatures of PG and PEG 400 have been reported in the literature as -60 °C and 5.8 °C, respectively (25). ...
... The 1 H NMR spectrum of PG also displayed the 1 H vibrational signals representative of C 2 and C 3 at around 3 ppm and 4 ppm, respectively (Fig. 5a). Examination of KAPE methoxyflavones such as DMF revealed a band of vibrational signals at about 3 to 4 ppm and another at 6 to 8 ppm which had been attributed to protons of the methoxy and the 2-phenylchromone units, respectively (7). Surprisingly, the signals from the 2-chromone core of the methoxyflavones were completely absent in solutions of PEG400 and PG containing the solutes (Fig. 5b). ...
Herein, thermal and non-thermal techniques were used to elucidate the putative physical and chemical interactions between poorly water-soluble Kaempferia methoxyflavones and PEG400/ propylene glycol. Additionally, the biocompatibility of methoxy-flavone-glycol solutions was evaluated using Caco-2 cells whereas the absorptive transport was investigated by measuring the apparent permeability coefficient (P app) of the methoxyflavones and transepithelial electrical resistance (TEER) of the Caco-2 cell monolayer. Data from differential scanning calorimetry, Fourier-transform infrared (FTIR), and proton nuclear magnetic resonance (1 H NMR) spectroscopic analysis revealed physico-chemical compatibility between the three methoxyflavones and PEG400/propylene glycol. Furthermore, PEG400 and propy-lene glycol solutions of the methoxyflavones were shown to be compatible with Caco-2 cells at pharmacologically effective concentrations. In vitro transport studies across the Caco-2 cell monolayer revealed high P app values of 24.07 × 10-6 to 19.63 × 10-6 cm s-1 for PEG400 solutions of the methoxyflavones. The TEER values of the Caco-2 cell monolayers indicated that the increased drug transport was partly due to increased tight junction openings, but without compromising the epithelial barrier integrity. The good pharmaceutical and biocompatibility profiles , as well as improved transport of the methoxyflavones in PEG400 and propylene glycol solutions, are suggestive of the worthiness of this approach for further consideration pertaining to the development of these drugs into oral liquid dosage forms.
... The major components of K. parviflora rhizomes are methoxyflavones, structurally identified as 5,7-dimethoxyflavone, 5,7,4 -trimethoxyflavone, and 3,5,7,3 ,4 -pentamethoxyflavone [18,22,23], the pharmacokinetic characteristics of which have been investigated [20,24]. In a previous study, 5,7-dimethoxyflavone was shown to reduce the viability of HepG2 cancer cells with an IC 50 of 25 µM by generating reactive oxygen species and significantly reducing the mitochondrial membrane potential, suggesting that it might be considered to be an anti-liver cancer lead compound [25]. ...
... Column chromatography and semi-preparative HPLC separation were efficiently applied, leading to the isolation of six structurally related methoxyflavones ( Figure 1). These methoxyflavones were determined to be 3,7-dimethoxy-5hydroxyflavone (1) [34], 5-hydroxy-7-methoxyflavone (2) [35] [22], and 5-hydroxy-3,7,3 ,4tetramethoxyflavone (6) [38] (Figure 2) by comparing their 1D nuclear magnetic resonance (NMR) spectroscopic data (Figures S1, S3, S5, S7, S9 and S11) with those previously reported and MS data obtained from LC-MS analyses ( Figures S2, S4, S6, S8, S10 and S12). separation and structural elucidation of Compounds 1-6, the evaluation of their anti-melanogenic activity, and SARs. ...
... ,4′-trimethylkaempferol (5) [22], and 5-hydroxy-3,7,3′,4′-tetramethoxyflavone (6) [38] (Figure 2) by comparing their 1D nuclear magnetic resonance (NMR) spectroscopic data (Figures S1, S3, S5, S7, S9, and S11) with those previously reported and MS data obtained from LC-MS analyses ( Figures S2, S4, S6, S8, S10, and S12). ...
Kaempferia parviflora Wall. ex Baker (Zingiberaceae), commonly known as Thai ginseng or black ginger, is a tropical medicinal plant in many regions. It has been traditionally used to treat various ailments, including ulcers, dysentery, gout, allergies, abscesses, and osteoarthritis. As part of our ongoing phytochemical study aimed at discovering bioactive natural products, we investigated potential bioactive methoxyflavones from K. parviflora rhizomes. Phytochemical analysis aided by liquid chromatography–mass spectrometry (LC-MS) led to the isolation of six methoxyflavones (1–6) from the n-hexane fraction of the methanolic extract of K. parviflora rhizomes. The isolated compounds were structurally determined to be 3,7-dimethoxy-5-hydroxyflavone (1), 5-hydroxy-7-methoxyflavone (2), 7,4′-dimethylapigenin (3), 3,5,7-trimethoxyflavone (4), 3,7,4′-trimethylkaempferol (5), and 5-hydroxy-3,7,3′,4′-tetramethoxyflavone (6), based on NMR data and LC-MS analysis. All of the isolated compounds were evaluated for their anti-melanogenic activities. In the activity assay, 7,4′-dimethylapigenin (3) and 3,5,7-trimethoxyflavone (4) significantly inhibited tyrosinase activity and melanin content in IBMX-stimulated B16F10 cells. In addition, structure–activity relationship analysis revealed that the methoxy group at C-5 in methoxyflavones is key to their anti-melanogenic activity. This study experimentally demonstrated that K. parviflora rhizomes are rich in methoxyflavones and can be a valuable natural resource for anti-melanogenic compounds.
... From the inflorescences of Pseudognaphalium liebmannii were isolated seven known flavones: 5-hydroxy-3,7-dimethoxyflavone 1 (Jaipetch et al. 1983;Sutthanut et al. 2007), 5,8-dihydroxy-3,7-dimethoxyflavone 2 (Norbedo et al. 1984), 5,7-dihydroxy-3, 8-dimethoxyflavone 3 (gnaphaliin A), 3,5-dihydroxy-7,8-dimethoxyflavone 4 (gnaphaliin B) (Rodríguez-Ramos and Navarrete 2009), 3,5-dihydroxy-6,7,8trimethoxyflavone 5 (Torrenegra et al. 1982;Van Puyvelde et al. 1989;Yong et al. 2011), 3,5,7-trimethoxyflavone (Jaipetch et al. 1983;Sutthanut et al. 2007) (Figure 1). Unequivocal structure of 6 was confirmed by X-ray diffraction analysis (These data can be obtained from the Cambridge Crystallographic Data Center, DOI: 10.5517/ccdc.csd.cc2bqbt7. ...
... From the inflorescences of Pseudognaphalium liebmannii were isolated seven known flavones: 5-hydroxy-3,7-dimethoxyflavone 1 (Jaipetch et al. 1983;Sutthanut et al. 2007), 5,8-dihydroxy-3,7-dimethoxyflavone 2 (Norbedo et al. 1984), 5,7-dihydroxy-3, 8-dimethoxyflavone 3 (gnaphaliin A), 3,5-dihydroxy-7,8-dimethoxyflavone 4 (gnaphaliin B) (Rodríguez-Ramos and Navarrete 2009), 3,5-dihydroxy-6,7,8trimethoxyflavone 5 (Torrenegra et al. 1982;Van Puyvelde et al. 1989;Yong et al. 2011), 3,5,7-trimethoxyflavone (Jaipetch et al. 1983;Sutthanut et al. 2007) (Figure 1). Unequivocal structure of 6 was confirmed by X-ray diffraction analysis (These data can be obtained from the Cambridge Crystallographic Data Center, DOI: 10.5517/ccdc.csd.cc2bqbt7. ...
Abstract
The inflorescences of Pseudognaphalium liebmannii are used as folk medicine to treat various respiratory diseases. In this work, we report the isolation of seven known flavones: 5-hydroxy-3,7-dimethoxyflavone 1, 5,8-dihydroxy-3,7-dimethoxyflavone 2, 5,7-dihydroxy-3,8-dimethoxyflavone 3 (gnaphaliin A), 3,5-dihydroxy-7,8-dimethoxyflavone 4 (gnaphaliin B), 3,5-dihydroxy-6,7,8-trimethoxyflavone 5, 3,5,7-trimethoxyflavone 6 and 3-O-methylquercetin 7. All these flavones except 1 and 6 showed a relaxant effect on guinea pig tracheal preparation with EC50 between 69.91 ± 15.32 and 118.72 ± 7.06 µM. Aminophylline (EC50 = 122.03 ± 7.05 µM) was used as a relaxant reference drug. The active flavones shifted the concentration-response curves of forskolin and nitroprusside leftward, and significantly reduced the EC50 values of these drugs. Furthermore, these flavones dose-dependently inhibited phosphodiesterase (PDE) in an in vitro assay. This reveals that the inflorescences of P. liebmannii contain several flavones with relaxant effect on airway smooth muscle and with PDEs inhibition that contribute to supporting the anti-asthmatic traditional use.
... Da, was conclusively identified as 5,7,4 ′ -trimethoxyflavone. This determination was based on its NMR spectroscopic data, which sufficiently matches the literature report [41] and is further supported by mass fragmentation analysis, which revealed the presence of a fragment with an m/z of 287 Da [M−OCH 3 ] + , as shown in Figure 2A. ...
... Da, was conclusively identified as 5,7,4′-trimethoxyflavone. This determination was based on its NMR spectroscopic data, which sufficiently matches the literature report [41] and is further supported by mass fragmentation analysis, which revealed the presence of a fragment with an m/z of 287 Da [M−OCH3] + , as shown in Figure 2A. An analysis of the fragmentation of 7 suggested that it contained a methoxy unit due to the observation of a m/z 30 Da (-OCH3) loss. ...
The plant Allium hookeri, a member of the Allium genus, has a rich history of culinary and medicinal use. Recent studies have unveiled its potent antioxidant and anti-inflammatory properties. While research on A. hookeri has demonstrated its neuroprotective and anti-neuroinflammatory effects, the specific bioactive compounds responsible for these effects remain unidentified in prior research. This study utilized an untargeted metabolomic approach, employing HRESI-qTOF MS/MS-based molecular networking, to comprehensively profile the chemical composition of metabolites in A. hookeri and identify new compounds within the plant. As a result, ten compounds, comprising one novel flavonoid (2) and nine known compounds (1 and 3–10), were isolated and identified through NMR analysis. The inhibitory effects of all isolated compounds on the senescent cell-associated secretory phenotype (SASP), which is pivotal in neuroprotective actions, were evaluated. Biological activity testing revealed N-trans-feruloyltyramine (7) to be the most potent compound, effectively inhibiting SASP markers and contributing to the senomorphic activities of A. hookeri. These findings underscore the potential of phenolamides from A. hookeri as a promising source of bioactive compounds for mitigating senescence-associated diseases.
... The content of methoxyflavones isolated from KP was previously analyzed by many researchers (Sutthanut et al., 2007;Burapan et al., 2014;Gelila Asamenew et al., 2019). However, the relationship between the contents of bioactive compounds in different extracts from KP with their pharmacological activities was not much explored. ...
... Particularly, among isolated compounds, 5,7,4 ′ -trimethoxyflavone (compound 2) was presented as the most abundant component in the total crude extract ME (31.10 mg/g or 41% of all compounds), which agrees with previous reports (Sutthanut et al., 2007;Ninomiya et al., 2016;Gelila Asamenew et al., 2019). In addition, compound 2, the most highlighted component in KP with various pharmacological activities such as anticancer (Walle and Walle, 2007), vasorelaxant effect (Tep-areenan and Sawasdee, 2010), anti-inflammatory effect (Sae-Wong et al., 2011), anti-Alzheimer action (Youn et al., 2016), was currently found to be at a much higher amount than previous reports. ...
Scientific evidence on the antithrombotic activity of Kaempferia parviflora (KP) remains limited. The present study investigated the antiplatelet aggregation effect and anticoagulant activity of this plant on human blood. Among four tested fractions, the ethyl acetate (EA) fraction exhibited the strongest inhibitory effect on platelet aggregation (percentage of inhibition %I = 95.07% at 0.4 mg/mL and 94.44% at 0.2 mg/mL for ADP and collagen, respectively, p > 0.05 compared with the positive control) and blood coagulation (mean activated partial thromboplastin time - APTT, 43.23 vs. 46.70 s with heparin, p > 0.05). From this extract, 6 known methoxyflavones were isolated: 5-hydroxy-3,7- dimethoxyflavone (1), 5,7,4′- trimethoxyflavone (2), 5-hydroxy-3,7,4′- trimethoxyflavone (3), 5-hydroxy-7-methoxyflavone (4), 3,5,7- trimethoxyflavone (5) and 3,5,7,4′- tetramethoxyflavone (6). Four compounds 1 – 4 were tested for their activities. Compounds 2, and 3 showed a stronger inhibitory effect than compounds 1 and 4 on both agonists used (p < 0.05). For anticoagulant activity, 1, 3 and 4 at 0.4 mg/mL had capacity to extend APTT value (p < 0.05 compared to DMSO 0.1%). Moreover, only 3 possessed a similar anticoagulant effect to the positive control (mean APTT, 45.15 vs. 46.70 s, p > 0.05). No compounds could lengthen prothrombin time and thrombin time parameters. The LC-MS analysis indicated that EA contained the highest content of methoxyflavones (119.85 mg/g, DW) compared to other fractions. Compound 2 (41% of total methoxyflavones) was the most abundant component among 6 isolated compounds. Molecular docking indicated that all 4 tested methoxyflavones were tightly bound to several platelet receptors such as COX-1, P2Y12, GPVI while their interaction with coagulation factors such as ATIII, FIX or VKOR was less efficient. In conclusion, KP is rich in methoxyflavones which are potential antithrombotic agents. Further studies should be performed to elucidate the in vivo activity and mechanism of action of these compounds.
... Compared with synthetic dyes, natural dyes possess advantageous features such as their easy availability, mass availability, no need for purification, and no pollution, thus significantly reducing the cost of equipment. In natural pigments, there are four kinds of compounds that are mainly used Nanomaterials 2022, 12, 563 3 of 16 as photosensitizers of DSSCs: chlorophylls [20,21]; flavonoids [22,23]; carotenoids [24]; and anthocyanins [25]. In addition, the electrocatalyst in the paired electrodes plays a vital role in determining the overall photovoltaic efficiency of DSSCs since the correct selection of electrocatalysts helps improve the I − /I 3 − reduction reaction and prevent the recombination of oxide cations (cationic form of photosensitizers) with electrons in the semiconductor CB [6]. ...
... synthetic dyes, natural dyes possess advantageous features such as their easy availability, mass availability, no need for purification, and no pollution, thus significantly reducing the cost of equipment. In natural pigments, there are four kinds of compounds that are mainly used as photosensitizers of DSSCs: chlorophylls [20,21]; flavonoids [22,23]; carotenoids [24]; and anthocyanins [25]. In addition, the electrocatalyst in the paired electrodes plays a vital role in determining the overall photovoltaic efficiency of DSSCs since the correct selection of electrocatalysts helps improve the I − /I3 − reduction reaction and prevent the recombination of oxide cations (cationic form of photosensitizers) with electrons in the semiconductor CB [6]. ...
Dye-sensitized solar cells (DSSCs), a powerful system to convert solar energy into electrical energy, suffer from the high cost of the Pt counter electrode and photosensitizer. In this study, the dual application of waste grape skin is realized by employing the grape skin and its extract as the carbon source of the carbon-based counter electrode and photosensitizer, respectively. The ultraviolet–visible absorption and Fourier transform infrared spectroscopy verify the strong binding between the dye molecules (anthocyanins) in the extract and the TiO2 nanostructure on the photoanode, contributing to a high open-circuit voltage (VOC) value of 0.48 V for the assembled DSSC device. Moreover, the waste grape skin was subjected to pyrolysis and KOH activation and the resultant KOH-activated grape skin-derived carbon (KA-GSDC) possesses a large surface area (620.79 m2 g−1) and hierarchical porous structure, leading to a high short circuit current density (JSC) value of 1.52 mA cm−2. Additionally, the electrochemical impedance spectroscopy reveals the efficient electron transfer between the electrocatalyst and the redox couples and the slow recombination of electrolytic cations and the photo-induced electrons in the conduction band of TiO2. These merits endow the DSSC with a high photovoltaic efficiency of 0.48%, which is 33% higher than that of a common Pt-based DSSC (0.36%). The efficiency is also competitive, compared with some congeneric DSSCs based on other natural dyes and Pt counter electrode. The result confirms the feasibility of achieving the high-value application of waste grape skin in DSSCs.
... Kaempferia parviflora (KP) is one of the most popular plants in the family Zingiberaceae. It has many active constituents like 7-dimethoxyflavone and 5,7,40-trimethoxyflavone [17][18][19][20][21][22][23][24]. Traditionally, KP and its compounds are used as a folk medicine for managing a variety of diseases, particularly male infertility, due to its aphrodisiac, antioxidant, and anti-inflammatory activities [17][18][19][20][21][22]. ...
... Previous studies confirmed that KP phytoconstituents like 5,7-dimethoxyflavone and 5,7,40-trimethoxyflavone were reported to have inhibitory activity against phosphodiesterase 5 (PDE5), suppression of NO production, increasing male libido, and improving erectile dysfunction in treated males [17,18,[23][24][25]90]. Also, these polyphenols possess aphrodisiac properties which are being used to improve sexual activities and performance [24][25][26][27][28]. ...
Background. Therapeutic strategies based on herbal plants and diets containing sufficient amounts of antioxidants and essential vitamins are very important factors in treating reproduction and male infertility worldwide. Thus, the aim of this study was to investigate the potential effects of Kaempferia parviflora (KP) on the role of some microRNAs in treated and nontreated infertile rats. In addition, the correlation of expressed microRNAs with sperm count, sperm motility, and sperm viability was identified. The probable use of these microRNAs as a diagnostic marker for predicting the clinical response of infertility to the treatment with KP was also achieved. Methods. In the present study, the potential effects of Kaempferia parviflora (KP) at different doses (140, 280, and 420 mg/kg) for six weeks on male rats with subinfertility were explored. In addition, the effect of KP on the expression of circulating microRNAs and its correlation with the parameters of sexual infertility was identified by performing both in vitro and in vivo assays. In vitro antioxidant activity, sperm functional analysis, serum testosterone, and expression of circulating microRNAs were conducted using colorimetric, ELISA, and real-time RT-PCR analysis, respectively. Results. Kaempferia parviflora (KP) at nontoxic doses of 140–420 mg/kg/day for six weeks significantly improved serum testosterone and epididymal sperm parameters (sperm count, motility, and sperm viability), increased testicular weight, and provided a reduction in the percentage of abnormal spermatozoon in infertile male rats. The expression of miR-328 and miR-19b significantly decreased, and miR-34 significantly increased in infertile rats treated with KP compared to infertile nontreated rats. After six weeks of KP therapy, the change in the expression levels of miRNAs was correlated positively with higher levels of serum testosterone and the measures of epididymal sperm parameters. The respective area under the receiver operating characteristic curve (AUC-ROC) was applied to predict the potential use of miR-328, miR-19b, and miR-34 in the diagnosis of male infertility in treated and nontreated infertile male rats. The data showed that AUC cutoff values of 0.91 for miR-328, 0.89 for miR-19b, and 0.86 for miR34 were the best estimated values for the clinical diagnosis of male rats with infertility. In rats treated with KP for six weeks, AUC cutoff values of 0.76 for miR-328, 0.79 for miR-19b, and 0.81 for miR-34 were the best cutoff values reported for the clinical response of infertility to KP therapy after six weeks. Conclusions. In this study, the improvement of male infertility might proceed via antioxidant and antiapoptotic pathways, which significantly improve spermatogenesis and aphrodisiac properties of males. In addition, the expression of miRNAs, miR-328, miR-34, and miR-19b, in KP-treated and nontreated infertile rats significantly correlated with increased serum testosterone levels and epididymal sperm parameters as well. MicroRNAs, miR-328, miR-34, and miR-19b, might be related to oxidative and apoptotic pathways that proceeded in spermatogenesis. Thus, the use of miRNAs could have a role as diagnostic, therapeutic, and predictive markers for assessing the clinical response of Kaempferia parviflora treatment for six weeks. This may have potential applications in the therapeutic strategies based on herbal plants for male infertility. However, in subsequent studies, the genetic regulatory mechanisms of the expressed miRNAs should be fully characterized.
1. Introduction
In life, infertility is considered one of the most health problems facing 30–50% of males world wide [1, 2]. Previously, defects in male spermatogenesis, reduction in sperm quality, and seminal production were greatly affected by several treating conditions such as hypogonadism, varicocele, infections, and obstructions [1–3]. In addition, spermatogenesis and sperm normal production were shown to be affected by inadequate vitamins intake, chemotherapy, type of drugs used, toxins, and polluted air [3]. Diets containing sufficient amounts of antioxidants and vitamins A, B, C, and E can enhance barrier stability of testis by increasing blood flow and protect sperm DNA from cellular oxidative-free radical activity [4, 5]. Antioxidants were shown to protect DNA and other cellular components from oxidation and damage, improving sperm quality, which in turn raises the rates of fertility among males [6–8].
Therefore, therapeutic strategies based on herbal plants are very important factors in treating reproduction and male infertility. Natural plants are concomitantly used as a possibility traditional medicine for treating male infertility and other human diseases in up to 60% of the world’s population [9–12]. Medicinal plants related to the family Zingiberaceae are used worldwide as spices and are shown to have versatile medical activities, particularly as antioxidative [13], free radical scavenging activities [14, 15], androgenic activity [16], aphrodisiac [17, 18], anticancer [19], and anti-inflammatory [20].
Kaempferia parviflora (KP) is one of the most popular plants in the family Zingiberaceae. It has many active constituents like 7-dimethoxyflavone and 5,7,40-trimethoxyflavone [17–24]. Traditionally, KP and its compounds are used as a folk medicine for managing a variety of diseases, particularly male infertility, due to its aphrodisiac, antioxidant, and anti-inflammatory activities [17–22].
Previously, a number of biological activities of KP were identified, particularly antioxidant, anti-inflammatory, and inhibition of NO production, increasing male libido and erectile dysfunction, having aphrodisiac properties, and being used to improve sexual activities and performance [24–28].
In rabbit semen, KP (Krachaidum, KD) showed previously a quiet tendency to increase ejaculation volume and a subsequent increase of the total number, viability, and progressive motility of spermatozoa [29]. Additionally, the seminal vesicle and spermatogenesis significantly improved in rats, following the use of tea or extracts from KP (Krachaidum, KD) [30].
This might be due to the presence of active components like phenolics and flavonoids present mainly in the KP extracts [24]. In other studies, like other plants (curcuma and ginger) in the Zingiberaceae family, it was reported that KP extracts modulate changes in reproductive function by relaxation of the smooth muscles of the blood vessels [31–33], leading to an increase in blood flow to the reproductive organs and finally an improvement in functions of male reproductive organs. Also, KP in association with physical exercise interventions significantly stimulated both increase in sexual motivation and enhancement of sexual performance as well [34].
However, little is known about the roles of circulating miRNAs in reproductive function and male infertility in cases treated with traditional medicine particularly, KP extracts. It was reported previously in many studies that microRNAs as short noncoding transcripts of up to 22 nucleotides have considerable potential as diagnostic and therapeutic tools against many diseases [35, 36]. At the posttranscriptional level, microRNAs might regulate gene expression. So, it could be used for monitoring diagnosis and for treatment of male infertility with therapeutic or herbal medicine [35–37]. In addition, a set of miRNAs was shown to regulate significantly more biological processes like embryonic development, cell differentiation, cell cycle, cell growth, and apoptosis [38–40]. Thus, dysregulation of miRNA functions can lead to the development of disease. miRNAs are shown to contribute to human spermatogenesis and to be retained after the completion of spermatogenesis, and any changes in the expression of spermatozoal RNAs have been associated with male infertility [41–44].
The role of small noncoding RNAs was significantly reported in male germ cell development [45, 46]. In previous studies, miRNAs were reported to have a role in male and female gametogenesis and the development of the embryo [46, 47]. miRNAs were identified in the male reproductive system and in testis, epididymis, sperm cells, seminal plasma, and extracellular vesicles (i.e., exosomes and microvesicles) were suggested to represent known functions. Thus, any alterations in spermatogenesis and embryogenesis could be attributed to the change in the expression of miRNAs [48–54]. These signs could clearly have the potential association of miRNAs in various forms of infertility [53, 54]. In the testis, the critical role of miRNAs was demonstrated during mitotic proliferation and formation of spermatogonia from germ cells. Additionally, their roles also start during spermatogonial stem cells (SSCs) in the epithelium of seminiferous tubules or during spermatocyte meiosis and spermiogenesis [55].
In normozoospermic controls and in infertile males, miR-19b and other miRNAs were clearly expressed in human seminal plasma from fertile controls; however, they significantly increased in the seminal plasma of the infertile men [56]. Thus, a significant increase in the expression levels of miR-19b may be a possible indicator of the degrees of spermatogenic failure in treated and nontreated cases. In addition, other studies reported the expression of many miRNAs, including miR-34, which were associated with many vital processes of male fertility, particularly the regulation of germ cell function as well as cell differentiation during spermatogenesis [56, 57]. It was reported that lower expression and hypermethylation of the promotor of cellular miR-34 types were significantly identified in infertile males. Thus, it was reported that obvious lower expression with hypermethylation of the promoter region makes miR-34 types be an indicator of the deficiency of spermatogenesis [58].
In animal models, inactivation and lower expression of miR34-b,c along with others miRNAs clusters caused low sperm counts, abnormal sperm morphology with low motility, and subsequent male infertility [57, 59, 60]. This might be due to unsuitable or hypermethylation of CpG in their promoter regions. In n somatic cells, miR-34 types additionally act as tumor suppressor genes aside from the P53 gene [61].
Also, an increase in the expression levels of miR-328 was reported to govern the pathogenesis of male erectile dysfunction (ED) in many ways. It was found that miR-328 might impair stem cell or neuronal survival, control zonation morphogenesis, and affect calcium homeostasis [62–66].
For aforementioned facts [45–67], identifying the vital role of miR-19b, miR-328, and miR-34 enforces studying their expression profile in treated and nontreated infertility male rats with conventional KP herbal medicine. In addition, there are no scientific reports on the effect of therapeutic or herbal-based treatments such as KP on the role of these circulating microRNAs in reproductive function and male infertility. Therefore, the aim of this study was to investigate the potential effects of KP on the role of microRNAs, miR-19b, miR-328, and miR-34, in treated and nontreated infertile rats and also the correlation of expressed microRNAs with sperm count, sperm motility, and sperm viability, as well as its potential use as diagnostic biomarkers in predicting the clinical response of Kaempferia parviflora treatment.
2. Materials and Methods
2.1. Plant Material
The Kaempferia parviflora (KP) rhizomes obtained were purchased from a convenience store (Othaim Markets) in Riyadh, KSA. The plant rhizomes were cut into small pieces and dried in a hot air oven at 55°C [28, 65]. Then, the dried materials were macerated in ethanol twice, for 3 days each, and filtered. To prepare a 1% of fresh KP suspension, the dry KP powder was suspended in distilled water with Tween 80 [28, 65].
2.2. Assessment of Total Phenolic Content (TPC) and Total Flavonoid (TF) Content
2.2.1. Preparation of KP Extract
In this test, a mechanical blender was used to prepare a fine powder of the dried rhizomes of KP. At room temperature, the rhizomes of the plant were dried in the shade and then chopped into small pieces. KP was ground to a fine powder and became ready for the extraction step. By using a Soxhlet apparatus, 20 g of the dried rhizome powder was extracted in 300 mL methanol at 60–65°C for 3-4 h. Then, Whatman filter paper No. 1 was used to filtrate the extract and exposed to pressure at 40°C for the concentration process. Finally, the extract was further dried, weighed (2.6 g), and stored in storage vials at 4°C for reuse in the study [28, 65, 66].
2.2.2. Total Phenolic Content
In this experiment, the total phenolic content of the KP extract was estimated by using Folin–Ciocalteu method as previously reported [68, 69]. “A total of 200 μL of crude KP extract (1 mg/mL/3 mL dH2O) was mixed thoroughly with 0.5 mL of Folin–Ciocalteu reagent for 3 min. To the mixture, 2 mL of 20% (w/v) sodium carbonate was added, and the whole mixture was stored in the dark for 60 min as mentioned previously” [68, 69]. Then, “the absorbance of the produced mixture was measured at 650 nm. Finally, calibration curves were used to calculate the concentrations of the total phenolic contents and expressed as mg of gallic acid equivalent per g dry weight as mentioned before” [68, 69].
2.2.3. Total Flavonoid Content
In this test, the aluminum chloride colorimetric method was used to estimate the total flavonoid content of crude KP extract, as mentioned previously [70]. In this experiment, “50 μL of KP extract (1 mg/mL ethanol) was completed to 1 mL with methanol and mixed with 4 mL of dH2O. Moreover, after 5 min of incubation, 0.3 mL of 5% NaNO2 solution and 0.3 mL of 10% AlCl3 solution were added, and the mixture was allowed to stand for 6 min [70], followed by the addition of 2 mL of 1 mol/L NaOH solution and the final volume of the mixture reached to 10 mL by adding dH2O” [70]. “The absorbance of the mixture was measured after 15 min. Finally, from a calibration curve, the concentration of the total flavonoid content present was calculated, and the result was expressed as mg rutin equivalent per g dry weight” [70].
2.3. Assessment of Antioxidant Activity
2.3.1. DPPH Assay
The 1,1-diphenyl-2-picryl-hydrazyl (DPPH) was used to estimate the antioxidant activity of the KP extract as mentioned before [71]. “In this test, a mixture of the KP extract was prepared, whereas 3.8 mL DPPH solution was added to 200 μL of each extract (100–500 μg/mL) and the whole mixture left in the dark for one hour at room temperature as mentioned before. In the final stage, the KP mixture was subjected to measure the absorbance at 517 nm against ascorbic acid as a positive control” [71]. Finally, “the ability of the sample to scavenge DPPH radical was determined as follows” [71]: {DPPH scavenging effect = Control OD − Sample OD/Control OD × 100}---[71].
2.3.2. Nitroblue Tetrazolium (NBT) Assay
The free radical scavenging activity of KP extract to superoxide anion was identified by nitroblue tetrazolium (NBT) as previously reported [72]. “In this test, a total of 100–500 μg/mL of the KP extract reacted with a mixture of 1.5 mmol/L riboflavin, 50 mmol/L nitroblue tetrazolium (NBT), 10 mmol/L D,l-methionine, and 0.025% (v/v) Triton X-100 in 50 mmol/L phosphate buffer at pH 7.8, respectively” [72]. “Then, the reaction mixture was initiated by the illuminating process to produce a colored formazan compound. The absorbance of formazan was recorded at 560 nm against ascorbic acid as a positive control. Finally, the percentage of the scavenging activity was identified as the inverse of the produced formazan” [72].
2.3.3. FRAP Assay
In this experiment, the antioxidant capacity of the KP extract was identified by estimation of ferric (Fe +3) reducing antioxidant power (FRAP), as mentioned previously [73]. “In this method, 100 μL of the KP extract (100–500 μg/mL) was incubated with 2.5 mL of 200 mmol/L phosphate buffer (pH 6.6) and 2.5 mL of1% potassium ferricyanide for 20 min at 50°C. After that, 2.5 mL of 10% trichloroacetic acid was added to the reaction mixture and was centrifuged at 10,000 rpm for10 min” [73]. “Then, a mixture from the upper layer was performed, whereas 5 mL of this layer was mixed with 5.0 mL dH2O and 1 mL of 0.1% Fe Cl3. The reaction mixtures were subjected to measure the absorbance at 700 nm against ascorbic acid as a positive control” [73].
2.4. Acute Toxicity Test
In this test, although different concentrations of KP (140, 280, and 420 mg/rat) were previously studied as improving agents for sexual performance in streptozotocin- (STZ-) induced diabetic male rats with infertility [28], the cytotoxicity of KP extract at doses of 140, 280, and 420 mg/rat was subjected to measure cellular toxicity in a healthy group of rats (10 rats) as previously reported in many toxicity studies [74, 75]. After the first 4 h of dosing, all animals have been observed for the appearance of any symptoms of toxicity. In addition, the survived animals were recorded following 24 h and maintained under daily observations for two weeks [74–76].
2.5. Animals Care and Experimental Design
Fifty adult male Wistar rats weighing about 180–200 g were included in this study. One week before starting the experiment, all rats were allowed to acclimatize to the laboratory environment like controlled conditions of the light cycle (12 hr : 12 hr, light : dark), room temperature (25 ± 2°C), and relative humidity (60%–70%) with free access water and rat chow. Animals were randomly classified into five groups of 10 animals each. Group 1 (normal group) was administered with the vehicle (distilled water), and group 2 (subinfertility group) received para-amino salicylic acid (PAS) at a dose of 400 mg/kg bw, while groups 3, 4, and 5 were given PAS at a dose of 400 mg/kg bw with an aqueous suspension of KP extract at doses of 140, 280, and 420 mg/kg, respectively [28, 75]. The animals were orally administered KP once a day for 6 weeks. “The experiment and the experimental procedures were performed according to the guidelines of the Experimental Animal Care Center, College of Applied Medical Sciences, and were approved by the Ethics Committee of the Experimental Animal Care Society, RRC, College of Applied Medical Sciences, King Saud Univ., Riyadh, Saudi Arabia, under file number (RRC-2019-021)” [76].
2.6. Sperm Collection and Functional Analysis
An overdose of “pentobarbital sodium was applied for anesthesia; then, all animals were sacrificed to collect the testes and epididymis” [28]. “Before the collection of spermatozoa, the testes and epididymis were weighted. In addition, to collect rat spermatozoa, a cauda part of the epididymis was minced into small pieces and mixed in 1 ml of Hanks’ balanced salt solution prewarmed at 37°C. Also, sperm parameters such as sperm count, motility, and viability were examined by microscope as previously mentioned” [77].
A Neubauer cell counting chamber under 10× magnification was used to collect sperm counts, as mentioned previously [78]. In addition, “the one-step eosin-nigrosin staining technique was applied to assess the percentage of sperm viability and morphology like normality and abnormality” [77, 78]. “In this test, sperm viability and morphology were then evaluated by counting alive and dead cells, whereas nonstained cells were considered alive and orange-red colored cells were considered dead cells” [77].
2.7. Assessment of Serum Testosterone
Serum samples were collected from the blood by “centrifugation at 2200 g for 15 min at 4°C and subjected to testosterone analysis using immunoassay ELISA Kit (Testosterone ELISA Kit, Abcam, Cambridge, UK)” [28, 65]. The level of testosterone in each sample was calculated according to the manufacturer’s instructions, as mentioned before [28, 65].
2.8. Real-Time RT-PCR Analysis of Circulating miRNAs
2.8.1. Extraction of RNA and Synthesis of cDNA
In this experiment, “RNA of all samples was estimated by using a reverse transcription-polymerase chain reaction (RT-PCR) analyses and the miRNease isolation kit (Qiagen, Hilden, Germany) as mentioned previously” [79–82]. Then, “reverse transcription miScriptII RT kits (Qiagen) were applied to generate a complementary DNA (cDNA), and then the levels of miRNAs were evaluated by optical density” [79–82].
2.8.2. Real-Time RT-PCR Analysis
The expression of “miRNAs in the serum was identified by using quantitative real-time RT-PCR analyses and primers of circulating miRNAs, miR-328, miR-34, and miR-19b (Applied Biosystems, Foster City, CA, USA)” [79]. In this test, “GAPDH gene was applied as an internal housekeeping gene to normalize the average copy number of the resultant PCR components as previously stated in the literature” [80–82].
In PCR process, “templets of respective cDNA were subjected to four thermal phases: primary denaturation phase (I) (at 94°C for 2 minutes); denaturation phase (II) (at 94°C for 30 seconds); annealing phase (III) (at 59°C for 30 seconds); and amplification phase (IV) (at 72°C for 30 seconds)” [80–82]. “The PCR phases (II to IV) proceeded for 45 cycles, and all reactions were measured in a triplicate manner” [80–82].
2.8.3. Statistical Analysis
In this study, “the data obtained were analyzed by using an SPSS statistical program (SPSS, IBM Statistics V.17) and the results of the continuous variables were expressed as mean ± SD” [82]. In addition, the “nonparametric test (Mann–Whitney-Wilcoxon test) and the χ2 test were performed to estimate the frequency differences between the groups, respectively” [82]. Moreover, “to compare between the studied variables like serum testosterone levels, sperm viability and morphology, and expression levels of miRNAs, two independent sample t-tests were used for all groups. Additionally, multiple stepwise regressions and Pearson’s correlations analysis were used to estimate the association between the expressed miRNAs and the studied independent variables in KP-treated and nontreated rats” [82]. “The area under the receiver operating characteristic (ROC) curve was used to measure the susceptibility and sensitivity of the studied parameters like testosterone, sperm viability, morphology, and miRNAs, miR-328, miR-34, and miR-19b, for the diagnosis of male infertility in treated and nontreated rats as previously reported” [82]. All tests were two-tailed; because of multiple assessments, results were only considered statistically significant at a value of < 0.05.
3. Results
3.1. Phenolic and Flavonoid Contents
Total phenolic and flavonoids constituents were calculated from the calibration curves at R² = 0.965 for total phenolic content and R² = 0.986 for the total flavonoid content, respectively. The total phenolic content estimated from the methanolic KP extract was 76.8 ± 3.8 gallic acid equivalents/g, and the total flavonoid content was 42.8 ± 2.7 rutin equivalents/g (Table 1).
Phytoconstituents
Quantity
Total phenolics contenta
76.8 ± 3.8 (R² = 0.965)
Total flavonoids contentb
42.8 ± 2.7(R² = 0.986)
Values are means of three biological replicates. amg gallic acid equivalent (GAE)/g DW. bmg rutin equivalent/g DW.
... Và H-2', H-6' tương quan với δ C 130.8 (C-1'), δ C 131.1 (C-4') và δ C 159.3 nên carbon ở δ C 159.3 chính là C-2. Từ các d liệu phổ trên và so sánh với tài liệu tham khảo (Sutthanut et al., 2007) xác định cấu trúc của 1 là 5,7dimethoxyflavone Hợp chất 2 thu được có dạng bột vô định hình màu vàng nhạt. ...
... Trên phổ HMBC cho thấy các nhóm methoxyl được nối vào C-3, C-5 và C-7 do có s tương quan của proton này với carbon của chúng. Từ các d liệu phổ NMR và so sánh tài liệu (Sutthanut et al., 2007) hợp chất 2 xác định là 3,5,7-trimethoxyflavone Hợp chất 3 thu được có dạng bột trắng ngà. Phân tích phổ hai chiều HMBC của hợp chất 3 cho thấy 2 proton ở δ H 6.19 (2H, s) đều cho tương quan với carbon thơm gắn oxy ở δ C 166.6 ( C-7), δ C 165.6 (C-5), δ C 163.2 (C-9) và carbon vòng thơm không mang oxy ở δ C 106.7 (C-10). ...
Five coumpounds including 5,7-dimethoxyflavone (1), 3,5,7-trimethoxyflavone (2), di-O-methylpinocembrin (3), bisdemethoxycurcumin (4), aloe-emodin (5) were isolated from the n hexane extract of Kaempferia parviflora rhizomes. Their structures were elucidated by ESI-MS, 1D & 2D-NMR spectra and compared their spectra with published data. Among them, compounds 4, 5 were reported for the first time from Kaempferia parviflora species.
... in certain countries such as Thailand and Japan. Among the various phytochemical components of the K. parviflora rhizome are polymethoxflavones, including 5,7-dimethoxflavone, 5,7,4'-trimethoxyflavone, 3,5,7,3',4'-pentamethoxyflavone, 5-hydroxy-3,7-dimethoxyflavone and 5-hydroxy-3,7,4'trimethoxyflavone (Gopi et al., 2017;Sutthanut et al., 2007). The biological effects of the major flavonoids, phenolic, tannins, and terpenoids in K. parviflora rhizomes have not yet been clarified through research studies. ...
Medicinal plants are used in traditional medicine to treat various diseases. They have been utilized to prevent and cure numerous diseases, the treatment of which is established in conventional knowledge practices. K. parviflora is a perennial plant with numerous medicinal properties. This study seeks to conduct a preliminary investigation of traditional medical treatment using K. parviflora, which has been employed for treating hypertension and promoting longevity through overall good health. In its results, this study shows that K. parviflora contains potential medicinal compounds such as flavonoids, phenolics, tannins, and terpenoids. The preliminary colorimetric detection of the K. parviflora extracts provides high confidence in the positive assessment of traditional medical practice. The present investigation suggests that using colorimetric detection for initial screening is acceptable. However, it is strongly recommended that chromatography be used in chemical concentration analysis for further study.
... However, it seems plausible that a high dose of oral KP in the rooster diet might have a greater impact on these parameters. However, it is important to note that KP is extracted with alcohol to produce the active components of methoxyflavones (Sutthanut et al., 2007). Despite the alcohol being removed from the KP extract prior to consumption, the taste and flavor are undesirable for the rooster, subsequently declining to eat. ...
Thai indigenous roosters are exposed to unsuitable temperatures and humidity, resulting in a lower reproductive potential. Kaempferia parviflora (KP) extract containing methoxyflavones was fed to roosters to improve their reproductive performance. Thirty-two Thai native roosters were orally administered KP extract at 300, 450, and 600 mg, calculated according to their average body weight, for at least 14 d before semen collection and continued supplementation until the end of the experiment. The nonsupplemented group served as the control. Fresh semen in terms of semen volume, sperm concentration, mass movement score, and sperm viability were evaluated. Semen preservation at 5°C and fertility test were examined for total motility (MOT), progressive motility (PMOT), sperm viability, and lipid peroxidation up to 48 h of storage. Testosterone concentrations and testicular function were also determined. The results showed that the highest sperm concentration and sperm motility of fresh semen were observed in KP extract at 600 mg (P < 0.001). KP extract at 600 mg resulted in higher sperm viability than the control and KP extract at 300 mg (P < 0.05), but was not different from KP at 450 mg (P > 0.05). The highest MOT, PMOT, and viability were found in the roosters that received 600 mg oral KP extract (P < 0.05), while those of the roosters that received oral KP extract 300 mg and the control were the lowest (P < 0.05) at all storage times. Lipid peroxidation was significantly lower in the KP extract up to 24 h (P < 0.05). The fertility and hatchability of the KP extract at 600 mg at T48 showed a minor decrease compared to the control at T0. These results might be inferred as a result of good spermatogenesis, as revealed by the results of histological examination and testosterone activity. In summary, oral administration of 600 mg KP extract improved sperm production and successfully preserved rooster semen for a long duration of up to 48 h of storage.
... Da, was conclusively identified as 5,7,4′-trimethoxyflavone. This determination was based on its NMR spectroscopic data, well matching with the literature report [29], and further supported by mass fragmentation analysis, which revealed the presence of a fragment with an m/z of 287 Da [M -OCH3] + as shown in Figure 3A. An analysis of the fragmentation of 7 suggested that it contained a methoxy unit due to the observation of a m/z 30 Da (-OCH3) loss. ...
The plant Allium hookeri, belonging to the Allium genus, has a history of being used both as a common food ingredient and in herbal medicine. It has recently been reported to have anti-oxidant and anti-inflammatory effects. A. hookeri has been also shown to exhibit neuroprotective and anti-neuroinflammatory activities, but the active compounds responsible for these effects have not been identified in previous studies. This study aimed to perform a metabolite profiling using an HRESI-qTOF MS/MS-based molecular networking approach and identify the active compounds from A. hookeri that target senescent cell-associated secretory phenotype (SASP) inhibitory effects, which contribute to neuroprotective activities. As a result, ten compounds, including one new flavonoid (2) and nine known compounds (1 and 3–10), were identified, and their biological activity was tested. The most potent compound was N-trans-feruloyltyramine (7), which inhibited SASP markers and contributed to the senomorphic activities of A. hookeri. These findings suggest that the phenolamides from A. hookeri could be a promising source of bioactive compounds for preventing senescence-associated diseases.
... Flavonoids isolated from A. bracteosum (A1--A11) were determined as xanthomicrol (A1), [20] gardenin D (A2), [21] (A3), [22] 5-hydroxy-7,8,4'-trimethoxyflavone (A4), [23] 5,6,30-trihydroxy-7,8,40-trimethoxyflavone (A5), [24] 8methoxycirsilineol (A6), [25] cirsilineol (A7), [26] tamarixetin (A8), [27] nobiletin (A9), [28] chrysoeriol (A10), [29] and 5,7-dimethoxyflavone (A11). [30] The 1 H-NMR data of flavones A1-A11 were shown in Table S1. These flavones which had a free H-3 were different from flavonoids isolated from V. negundo. ...
Adenosma bracteosum and Vitex negundo are natural sources of methoxylated flavonoids. Little is known about the α-glucosidase inhibition of multi-methoxylated flavonoid derivatives. Eighteen natural flavonoids were isolated from A. bracteosum and V. negundo. Seven halogenated derivatives were synthesized. Their chemical structures were elucidated by extensive NMR analysis and high-resolution mass spectroscopy as well as comparisons in literature. All compounds were evaluated for their α-glucosidase inhibition. Most compounds showed good activity with IC50 values ranging from 16.7 to 421.8 µM. 6,8-Dibromocatechin was the most active compound with an IC50 value of 16.7 µM. A molecular docking study was conducted, indicating that those compounds are potent α-glucosidase inhibitors.
... in certain countries such as Thailand and Japan. Among the various phytochemical components of the K. parviflora rhizome are polymethoxflavones, including 5,7-dimethoxflavone, 5,7,4'-trimethoxyflavone, 3,5,7,3',4'-pentamethoxyflavone, 5-hydroxy-3,7-dimethoxyflavone and 5-hydroxy-3,7,4'trimethoxyflavone (Gopi et al., 2017;Sutthanut et al., 2007). The biological effects of the major flavonoids, phenolic, tannins, and terpenoids in K. parviflora rhizomes have not yet been clarified through research studies. ...
Medicinal plants are used in traditional medicine to treat various diseases. They have been utilized to prevent and cure numerous diseases, the treatment of which is established in conventional knowledge practices. 𝘒. 𝘱𝘢𝘳𝘷𝘪𝘧𝘭𝘰𝘳𝘢 is a perennial plant with numerous medicinal properties. This study seeks to conduct a preliminary investigation of traditional medical treatment using 𝘒. 𝘱𝘢𝘳𝘷𝘪𝘧𝘭𝘰𝘳𝘢, which has been employed for treating hypertension and promoting longevity through overall good health. In its results, this study shows that 𝘒. 𝘱𝘢𝘳𝘷𝘪𝘧𝘭𝘰𝘳𝘢 contains potential medicinal compounds such as flavonoids, phenolics, tannins, and terpenoids. The preliminary colorimetric detection of the 𝘒. 𝘱𝘢𝘳𝘷𝘪𝘧𝘭𝘰𝘳𝘢 extracts provides high confidence in the positive assessment of traditional medical practice. The present investigation suggests that using colorimetric detection for initial screening is acceptable. However, it is strongly recommended that chromatography be used in chemical concentration analysis for further study.
... 14 compounds, including two polyoxygenated cyclohexanes (1 and 2), eleven flavonoids (3-13), and sitosterol (14), were identified through phytochemical analysis of K. parviflora extract. The primary methoxyflavones in K. parviflora have been identified structurally, according to phytochemical analysis (Kim et al., 2007 andSutthanut et al., 2007). Five coumpounds including 5,7-dimethoxyflavone (1), 3,5,7-trimethoxyflavone (2), di-O-methylpinocembrin (3), bisdemethoxycurcumin (4), aloe-emodin (5) were isolated from the n hexane extract of K. parviflora rhizomes (Tri et al., 2021). ...
... Ehnopharmacologically, the plant rhizomes have long been used as community medicines among the hill tribe people to promote health and increase lifelongevity. Polymethoxyflavones are identified as phytochemicals of the plant rhizomes with 5,7-dimethoxyflavone (DMF); 3,5,7-trimethixyflavone (TMF), and 3,5,7,3',4'-pentamethoxyflavone (PMF) as the main constituents (Sutthanut et al., 2007). Pharmacological data indicate that the health beneficial activities of the plant rhizomes are due to antioxidant, antiinflammatory, antistress, hypoglycemic, hypolipidemic, and antiobesity properties (Banjerdpongchai et al., 2009;Somintara et al., 2019;Wattanathorn et al., 2007). ...
The chapter focuses on current insights into anti-aging strategies, plant bioactives, and drug development. The aging process poses a significant risk of noncommunicable diseases and disorders depending on habitual factors leading to effective therapeutic interventions that are generally based on healthy diets and supplements containing antioxidants and anti-inflammatory compounds. Evidence-based applications of bioactive phytochemicals, Ayurvedic medicinal
herbs and their formulations, traditional foods, and dietary supplements for protecting aging are highly needed to proceed towards plant-based anti-aging supplementations.
... 14,35 The existence of 11 flavonoid constituents have been quantified on Kaempferia parviflora rhizome using gas chromatographic method in which 5,7,4ʹ trimethoxyflavone and 5,7 dimethoxyflavone were considered to be main constituents. 35 36,37 However, to the best of our knowledge, there are no reports on the putative identification of phytoconstituents present in the chloroform extract of Kaempferia parviflora Wall ex Baker rhizome and its evaluation for antioxidant/ antimicrobial/antifungal activities. In general, dissolution of bioactive compounds present in the plant sample is higher with higher polarity of the solvent and thus plant extracts prepared using polar solvents like water, and ethanol contain generally a large number of bioactive compounds with close retention factor (Rf) values. ...
... Compound 8 was previously isolated from Kaempferia parviflora [55]. Compound 9 was reported from H. decumbens and H. graveolens [56], compound 10 was isolated from Halimodendron halodendron [57] whereas, the compound 11 was isolated from Alipinia officinarum and showed pancreatic lipase inhibitory activity [58]. ...
Diabetes mellitus (DM) is a group of systemic metabolic disorders with a high rate of morbidity and mortality worldwide. Due to the detrimental side effects of the current treatment, there is a great need to develop more effective antidiabetic drugs with fewer side effects. Natural products are a well-known source for the discovery of new scaffolds for drug discovery, including new antidiabetic drugs. The genus Helichrysum has been shown to produce antidiabetic natural products. In this investigation, the methanolic extract of H. cymosum and H. pandurifolium resulted in the isolation and identification of eleven known compounds viz 5,8-dihydroxy-7-methoxy-2-phenyl flavanone (1), pinostrobin (2), dihydrobaicalein (3), glabranin (4), allopatuletin (5), pinostrobin chalcone (6), helichrysetin (7), 5-hydroxy-3,7-dimethoxyflavone (8), 3,5-dihydroxy-6,7,8-trimethoxyflavone (9), 3-O-methylquercetin (10), and 3-methylethergalangin (11). The in vitro bio-evaluation of isolated compounds against alpha-glucosidase showed that 10, 5, and 11 demonstrated the highest alpha-glucosidase inhibitory activity with IC50 values of 9.24 ± 0.4, 12.94 ± 0.2, and 16.00 ± 2.4 μM respectively, followed by 7 and 3 with IC50 values of 18.16 ± 1.2 and 44.44 ± 0.2 μM respectively. However, none of these compounds showed a measurable inhibitory effect on alpha-amylase under the experimental conditions used except compound 10 which showed a poor alpha-amylase inhibitory activity with an IC50 value of 230.66 ± 15.8 μM. Additionally, strong total antioxidant capacities were demonstrated by 10, 5 and 7 in ferric-ion reducing antioxidant power assay (374.34 ± 69.7; 334.37 ± 1.7; 279.93 ± 0.8) µmol AAE/mmol. This is the first scientific report to be carried out on alpha-glucosidase inhibitory activities and antioxidant capacities of H. cymosum constituents and a first report on the isolation and identification of methoxyflavanoids from H. pandurifolium. Our findings suggest that these compounds are promising candidates to inhibit alpha-glucosidase as well as oxidative stress related to diabetes. Results from molecular docking provided insight into the observed in vitro alpha-glucosidase inhibitory activities for 5, 7, 10, and 11. It is envisaged that the isolated phytochemicals from these plants may contribute to the development of hypoglycemic lead compounds with anti-diabetic potential.
... Methanol ext. 19 ...
Kaempferia parviflora (Black ginger) is used widely in medical fields as an anti-microorganism and anti-inflammation. In this study, the aim was to evaluate the in vitro and in vivo anti-acne efficacy of black ginger extract. The results indicate that the methanol and ethanol extracts showed the highest total phenolic contents, without a significant difference, whereas the n-hexane extract showed the highest total flavonoid content. Nine flavones were detected using UPLC−QTOF−MS, and the ethyl acetate extract showed the highest amount of 5,7-dimethoxyflavone (DMF) according to HPLC. Antibacterial activity against Staphylococcus aureus, S. epidermidis, and Cutibacterium acnes was observed. All the extracts showed antimicrobial activity against C. acnes, revealing MICs in the range of 0.015 to 0.030 mg/mL, whereas the ethyl acetate extract inhibited the growth of S. epidermidis with a MIC of 3.84 mg/mL. In addition, the ethyl acetate extract showed the highest activity regarding nitric oxide inhibition (IC50 = 12.59 ± 0.35 µg/mL). The ethyl acetate extract was shown to be safe regarding cell viability at 0.1 mg/mL. The anti-acne efficacy was evaluated on volunteers. The volunteers were treated in two groups: one administered a 0.02% ethyl acetate extract gel-cream (n = 9) and one administered a placebo (n = 9) for 6 weeks. The group treated with the gel-cream containing the extract showed 36.52 and 52.20% decreases in acne severity index (ASI) after 4 and 6 weeks, respectively, and 18.19 and 18.54% decreases in erythema, respectively. The results suggest that K. parviflora could be a potent active ingredient in anti-inflammatory and anti-acne products.
... KP rhizomes were reported to contain 5-Hydroxy-3,7-dimethoxyflavone, 5-hydroxy-7-methoxyflavone, 5-hydroxy-3,7,4'-trimethoxyflavone, 5-hydroxy-7,4'-dimethoxyflavone, 5-hydroxy-3,7,3',4'-tetramethoxyflavone, 3,5,7-trimethoxyflavone, 3,5,7,4'-tetramethoxyflavone, 5,7,4'-trimethoxyflavone, 5,7,3', 4'-tetramethoxyflavone, 5,7-dimethoxyflavone, 3,5, 7,3',4'-pentamethoxyflavone, tectochrysin, 5,3'-dihydroxy-3,7,4'-trimethoxyflavone, 5-hydroxy-7,3', 4'-trimethoxyflavone and genkwanin as some of the major flavonoids and phytoconstituents [6][7][8]. ...
... The use of V oc , I sc , FF, and η, DSSC has been evaluated. [26] Sensitizers used in DSSC by various pigments such as chlorophyll, [91,192] carotenoid, [191,193] anthocyanin, [87,98] flavonoid, [194] cyanine, [195,196] betalains [197,198] and tannin. [199] Pigmentary particles and colors derived largely from plants, animals and minerals are known as natural colorants and can be treated chemically. ...
The rapid growth of the population severely influences the supply of energy, accordingly ensuring clean energy has become a big challenge now and will be in the future. Fossil fuels have been satisfying the energy demand until now, but fossil fuels, being non‐renewable sources, will not be able to satisfy the energy demand in the future and will have a negative impact on the environment. Renewable energy sources have become the most demanding topic for researchers in this crisis. The solar cell, which is an abundant renewable energy resource, converts solar power into electrical energy without any environmental damage. Silicon solar cells have higher efficiency, but their high manufacturing cost, complicated procedures and environmental issues restrict their usage. Then dye‐sensitized solar cells (DSSCs) have been introduced as an alternative to silicon solar cells. In DSSC, both natural and synthetic dyes are used. Though synthetic dyes provide higher efficiency, they are environmentally harmful. Afterward, the concept of natural dye‐sensitized solar cells (NDSSC) have been materialized where only natural dyes are used. Researchers and environmentalists are looking for natural dyes as a replacement for synthetic dyes in recent times, as natural dyes are plentiful, can be collected naturally and have no environmental effects. Natural dyes in the form of anthocyanins, carotenoids, flavonoids, chlorophylls, tannins and betalains are extracted from various portions of plants that include leaves, roots, flowers, fruits, seeds, barks, etc. In this review, we investigate natural sources of dyes, natural sensitizers (dyes), shortcomings and remedies, improvements in efficiency and stability, developments, and commercialization. In addition, recent advances and the comparison of natural and synthetic dyes have been discussed in this review.
... flavonol, flavone, and anthocyanin), glycosides, lipophenols, polyphenols, saponins, steroids, tannins, terpenes, but reducing sugar is absent (Putiyanan et al., 2008). Many studies have demonstrated that KP rhizome extract contained various flavonoid constituents which 5,7,4'trimethoxyflavone, 5,7-dimethoxyflavone, and 3,5,7,3',4'-pentamethoxyflavone were major components (Sutthanut et al., 2007;Tuntiyasawasdikul et al., 2014). GC-MS analysis could detect more than 50 compounds from KP rhizome oils, which were mainly classified as monoterpenes and sesquiterpenes (Pojanagaroon and Kaewrak, 2004). ...
... The occurring natural compounds of these groups were isolated by successive CC procedures on silica gel 60, using a CH 2 Cl 2 :acetone gradient as eluent. Gsu1 was gathered in five subgroups: Gsu1A (fractions 1-4, 2.2 g, CC, CH 2 Cl 2 :acetone 95:5) yielded a 6:4 mixture of kaempferol 7,4 ′ -dimethyl ether (26) and kaempferol 3,7,4 ′ -trimethyl ether (27) (277.4 mg, 0.86% yield respect dry extract, Sutthanut et al., 2007), apigenin 7,4 ′ -dimethyl ether (22, 24.6 mg, 0.076%, Chen et al., 2012), kaempferol 7,4 ′ -dimethyl ether (26, 39.6 mg, 0.012%), and acacetin (apigenin 4 ′ -methyl ether, 21, 18.6 mg, 0.058%, Yang et al., 2014); Gsu1B (fractions 5-8, 608 mg, CH 2 Cl 2 :acetone 9:1) yielded precipitation of apigenin (19, 29.8 mg, 0.09%, Liu et al., 2013), and a kaempferol (25, Jørgensen et al., 1998) and ent-dihydrotucumanoic acid (5) mixture in the mother liquor; Gsu1C (fractions 15-17, 300 mg, CH 2 Cl 2 :acetone 9:1) yielded apigenin and the precipitation of kaempferol (25, 21.6 mg, 0.067%, Jørgensen et al., 1998); Gsu1D (fractions 25-28, 158 mg CC, CH 2 Cl 2 :MeOH 8:2) yielded ent-dihydrotucumanoic acid (5, 282.5 mg, 0.87%, Martínez et al., 1994); and Gsu1E (fractions 37-45, 720 mg, CH 2 Cl 2 :acetone 8:2) also yielded 5 (57.8 mg, 0.18%). Additionally, 600 mg of Gsu2 were submitted into two consecutive reverse phase CC procedures (C 18 column, H 2 O:MeOH 3:7) to render ent-dihydrotucumanoic acid (5, 79.2 mg, 0.24%) and impure ent-neoclerodan-2S,3S,4R,15-tetrol [4, (1R,2S,3S,4aS,5R,6S,8aS)-5-[(S)-4-hydroxy-3-methylbutyl]-1,5,6,8a-tetramethyl-decahydronaphthalene-1,2, 3-triol, ent-dihydrotucumanol, 62.2 mg, 0.19%], which also is the major compound the Gsu3 group (uncalculated yield). ...
Chemical analysis of the hydroalcoholic extracts from the aerial parts of the two accepted varieties of Ageratina espinosarum (A.Gray) R.M. King & H. Rob. (Asteraceae) resulted in the identification of ent-dihydrotucumanol (4), ent-dihydrotucumanoic acid (5), apigenin (19), acacetin (21), apigenin 7,4′-dimethyl ether (22), kaempferol (25), kaempferol 7,4′-dimethyl ether (26), and a mixture of 26 and kaempferol 3,7,4′-trimethyl ether (27) from A. espinosarum var. subintegrifolia; and taxifolin (7), naringenin (9), crysoeriol 4′-methyl ether (10), alysifolinone (11), sakuranetin (12), a mixture of sakuranetin 4′-methyl ether (13) and persicogenin (14), homoeriodictyol (15), and apigenin (19) from A. espinosarum var. espinosarum. Compounds 9, 12, 19 and 25 have been previously isolated of the aerial parts from this plant. Compound 19 was present in both varieties herein analyzed. Compounds 4, 5, 7, 10, 11, 13–15, 21, 22, 26, and 27 were isolated for the first time from A. espinosarum, despite this, the chemical content found in both varieties is consistent with that previously described. The flavanones, flavones and flavonols methoxy and dimethoxy derivatives are the chemophenetic markers of A. espinosarum. The structure of the known compounds was determined by spectroscopic and spectrometric techniques and their data compared with those previously reported. This is the first report of the isolation of compound 4, which was characterized as its 2,3,15-triacetyl (4A) and 2,15-diacetyl (4B) derivatives, whose spectroscopic and spectrometric data are described within.
... After filtration, the resulting solution was concentrated under a vacuum to yield a greenish oily crude extract. This extract was fractionated and purified over silica gel columns to afford 12 compounds among which a new flavonoid trivially named flavolippia (1), alongside eleven known compounds: 2,4-dimethylpyridin-3,5-diol (2) (Morton and Gustav 1947), 5-hydroxyl-6,7,4 0 -trimethoxylflavone (3) (Ulubelen et al. 1968, Tabatabaei et al. 2017; 5-hydroxyl-3,7,4 0 -trimethoxylflavone (4) (Sutthanut et al. 2007, Nantapap et al. 2017, Burapan et al. 2017; 7-hydroxyl-5,6,4 0 -trimethoxylflavone (5) (Deng et al. 2009, Momeni et al. 2016Tsopmejio et al. 2019); a-amyrin (6) (Jabeen et al. 2011); lupeol acetate (7) (Silva et al. 2017); lupeol (8) (Silva et al. 2017); betulin (9) (Siddiqui et al. 1988); fridelin (10) (Olmedo et al. 2008); saccharose (11) (Silva et al. 2017) and cosanol (12) (Silva et al. 2017). A new alkaloid derivative named lippiamicin (13) was synthesized from the known 2,4-dimethylpyridin-3,5-diol. ...
The chemical investigation of the leaf extract of a Cameroonian medicinal plant, Lippia rugosa A Chev (Verbenaceae) led to the isolation of a new flavonoid derivative flavolippia (1), alongside eleven known compounds: 2,4-dimethylpyridin-3,5-diol (2), 5-hydroxy-6,7,4'-trimethoxylflavone (3), 5-hydroxy-3,7,4'-trimethoxyflavone (4), 7-hydroxy-5,6,4'-trimethoxyflavone (5), 3β-hydroxy-urs-12-en-3-ol (α-amyrin) (6), lupeol acetate (7), lup-20(29)-en-3β-ol (lupeol) (8), lup-20(29)-en-3β,28-diol (betulin) (9), fridelan-3-one (fridelan) (10), saccharose (11), cosanol (12). In addition, a new semi-synthetic alkaloid derivative named lippiamicin (13) was prepared from 2,4-dimethylpyridin-3,5-diol (2). Their structures were established on the basis of their spectroscopic data, as well as 1 D and 2 D NMR. Compounds 1-13 were evaluated for their antioxidant activities. The results obtained showed that compounds 2 and 12 were the most active with IC50 values of 0.145 ± 0.011 and 0.195 ± 0.017 µM/mL respectively (for DPPH•) and 0.241 ± 0.027 and 0.223 ± 0.024 µM/mL respectively (for FRAP) compared to butylated hydroxyltoluene used as positive control.
... Considering the good chemical properties of HES and NAR, a wide range of analytical methods have been proposed in the literature for the determination of HES and NAR in different matrices including pharmaceutical formulations, biological fluids, citrus fruit peels and pulp, and juice, to name a few. The most widely applied methods for the determination of HES and NAR have included mainly chromatographic techniques [12][13][14][15][16]. Although these techniques offer high sensitivity, good selectivity, and low detection limits, they often require expensive and sophisticated instrumentation, long time of analysis, sample preparation, use of organic chemical solvents, and highlytrained personnel. ...
This paper reports the versatile preparation of three-dimensional nanostructured porous platinum (3DnpPt) directly on the surface of a screen-printed electrode via a simple electrochemical method and its application for the simultaneous voltammetric determination of hesperidin (HES) and narirutin (NAR) in residual water from the citrus industry. The surface morphology of the electrode was characterized by field-emission scanning electron microscopy (FEG-SEM), electron diffraction X-ray (EDX), X-ray photoelectron spectroscopy (XPS), and by the electrochemical impedance spectroscopic (EIS) technique. The results obtained from the voltammetric studies conducted showed that the sensor has good electrocatalytic activity and selectivity for HES and NAR oxidation. The linear scanning voltammetry (LSV) technique employed yielded linear ranges of 10 µmol L⁻¹ to 0.4 mmol L⁻¹ and 10 µmol L⁻¹ to 0.5 mmol L⁻¹, with detection limits of 6.61 µmol L⁻¹ and 0.21 µmol L⁻¹, and amperometric sensitivity of 0.52 A L mol⁻¹ and 0.79 A L mol⁻¹ for HES and NAR, respectively. The proposed 3DnpPt-SPE sensor also exhibited good repeatability and high selectivity, as well as long-term stability. The sensor was successfully applied in residual water sample for the simultaneous quantification of HES and NAR where good recovery rates were obtained.
... 3,5,7,3′,4′-Pentamethoxyflavone (PMF) is one of the major polymethoxyflavones found in black ginger (Kaempferia parviflora). 19 Several in vitro and in vivo studies have reported the health benefits of black ginger extracts, including anti-inflammation, anticancer, cardioprotective, and antimicrobial effects. 20−22 However, the role of polymethoxyflavones in the regulation of the intestinal barrier is poorly understood, and no studies have examined the transcriptional regulation of TJ proteins by PMF to date. ...
... 第 28 卷 黑姜在东南亚地区广泛种植。在泰国，黑姜被 称为"Krachaidum"，也被称为"Thai ginseng 泰国人 参"，民间用其作药物长达几个世纪 [10][11] ，现已成为 该国收入冠军的草药之一，主要用于治疗过敏、胃 肠道疾病和抗菌，其根茎的酒精浸泡液是治疗阳痿 的滋补饮品 [12] 。Yenjai 等 [13] 较早报道黑姜根茎含有 多种黄酮类物质，这些化合物具有抗疟原虫和抗真 菌的活性。此后，黑姜的多种药理作用逐渐受到关 注，如抗肿瘤 [14][15] 、抗炎 [16] 、抗氧化 [17] 、血管扩张 和心血管保护 [18][19][20][21] 、抗骨关节炎 [3] 作用等。另外黑姜 用于增强男性性能力，近年受到较大关注 [22][23][24][25][26][27][28] ，市 场上出现许多利用黑姜制成的相关功能产品。 虽然黑姜在东南亚地区被广泛种植 [3] ，被用于 制造食品、化妆品和医药品 [29] ，但国内鲜有黑姜应 用的报道。因此，本文就近年来黑姜化学成分、药 理作用、毒理学研究等方面进行综述，旨在为该植 物资源的进一步开发利用提供科学依据。 1 化学成分 1.1 黄酮类化合物 黑姜根茎中含有丰富的黄酮类化合物，目前报 道的有 32 种(表 1)，主要活性物质是 5,7-二甲氧基 黄酮及多甲氧基黄酮(PMFs) [30] 。黑姜根茎的黄酮类 化合物含量因测量方法不同而有所不同，不同产地 亦存在差异 [30] 。 Sutthanut 等 [31] 建立了测定黑姜黄酮 类化合物含量的气相色谱法；同时对泰国不同产地 黑姜黄酮类化合物的含量进行了分析，结果表明, 不同产地的含量差异较大，最低为 23.86 mg/g，最 高为 60.98 mg/g。此外，黑姜根茎提取物作为保健 食品，其 5,7-二甲氧基黄烷酮的标准化含量应不低 于 2.5%，总 PMFs 含量不低于 10% [10,30] ...
Kaempferia parviflora is widely cultivated in Southeast Asia as food, cosmetics and medicine. It has also been cultivated as a medicinal plant for a long time in Xishuangbanna Dai Autonomous Prefecture, China. Rhizome is an important medicinal part of K. parviflora, mainly containing flavonoids. Pharmacological studies showed that black ginger had anti-cancer, aphrodisiac, anti-inflammatory, anti-oxidant, anti-fungal, anti-viral, neuroprotective, vascular relaxant and cardioprotective, and transdermal permeable activities, etc. The advances in chemical constituents, pharmacological activities and toxicology of black ginger were reviewed, which would provide help for further research, development and utilization of K. parviflora resource.
... The eudesmane sesquiterpenes are (Figure 1) 4aaH-3,5a,8ab-trimethyl-4,4a,8a,9-tetrahydronaphtho[2,3b]-furan-8-one: (siphonochilone) (2); 9ab-hydroxy-4aaH-3,5a,8ab-trimethyl-4,4a,8a,9-tetrahydronaphtho-([2,3b]-dihydrofuran-2-one)-8-one (3); 4aaH-3,5a,8ab-trimethyl-4,4a,8a,9-tetrahydronaphtho-([2,3b]-dihydrofuran-2-one)-8-one (4); 2-hydroxy-4aaH-3,5a,8ab-trimethyl-4,4a,8a,9tetrahydro-naphtho[2,3b]-furan-8(5H)-one (5); 4aaH-3,5a,8ab-trimethyl-4,4a,8a-trihydronaphtho-([2,3b]-dihydrofuran-2-one)-8-one (6) (Holzapfel et al. 2002;Lategan et al. 2009). Other class of compounds that have been isolated from the South African variety were the diarylheptanoid (2,3-diacetoxy-7-(3 0' ,4 0' -dihydroxy-5 0'methoxyphenyl)-1-(4 0 -hydroxy-3 0 -methoxyphenyl)-5-heptene and flavonoids (3,4 0 ,7trimethyl kaempferol, 3,7-dimethoxyquercetin) (Guerrero et al. 2002;Holzapfel et al. 2002;Sutthanut et al. 2007;Igoli et al. 2020). We, hereby, report on the isolation and characterisation of a new furanoeudesmane-type sesquiterpenoid from the South African variety, as shown in Figure 1, which is characterised as phaeusmane F acetate (1) as a common name and 9ab-Hydroxy-4aaH-3,5a, 8ab-trimethyl-4, 4a, 6, 7, 8a, 9-hexahydronaphtho-([2, 3 b]-dihydrofuran-2-one)-8-one as a systematic name. ...
A new eusdesmane sesquiterpenoid, characterised as 5-acetoxy-9aβ-hydroxy-4aαH-3,5α, 8aβ-trimethyl-4, 4a, 6, 7, 8a, 9-hexahydronaphtho-([2, 3 b]-dihydrofuran-2-one)-8-one or phaeusmane F acetate (1) has been isolated from the rhizomes of the South African variety of wild ginger (iphonochilus aethiopicus (Schweinf) B.L. Burtt). The compound was obtained after a series of column and gel filtration chromatography. Its structure was elucidated by NMR and Mass-Spectrometric analyses, including 1 D-, 2 D-NMR and HR-LCMS. This is an initial report of the compound from a Siphonochilus sp. Previously isolated similar compounds from the plant material were 4aαH-3,5α,8aβ-trimethyl-4,4a,8a,9-tetrahydronaphtho[2,3b]-furan-8-one (siphonochilone) (2), 9aβ-hydroxy-4aαH-3,5α,8aβ-trimethyl-4,4a,8a,9-tetrahydronaphtho-([2,3b]-dihydrofuran-2-one)-8-one (3), 4aαH-3,5α,8aβ-trimethyl-4,4a,8a,9-tetrahydronaphtho-([2,3b]-dihydrofuran-2-one)-8-one (4), 2-hydroxy-4aαH-3,5α,8aβ-trimethyl-4,4a,8a,9-tetrahydro-naphtho[2,3b]- furan-8(5H)-one (5), 4aαH-3,5α,8aβ-trimethyl-4,4a,8a-trihydronaphtho-([2,3b]-dihydrofuran-2-one)-8-one (6).
... It possesses antioxidant [4], anti-allergenic [5], anticancer [6], antimicrobial [1], anticholinesterase [7], anti-inflammatory [8], anti-obesity [9], and antimutagenic [10] properties. Phytochemical analysis of black ginger rhizome extracts has confirmed the presence of flavonoids [1,11], methoxyflavones [5,7,10,[12][13][14], phenolic glycosides [15,16], and terpenoids [17]. Leaf extract of Kaempferia galanga has been reported to exhibit antinociceptive, anti-inflammatory [18], and sedative [19] properties. ...
This study aimed to establish a rapid in vitro plant regeneration method from rhizome buds of Kaempferia parviflora to obtain the valuable secondary metabolites with antioxidant and enzyme inhibition properties. The disinfection effect of silver oxide nanoparticles (AgO NPs) on rhizome and effects of plant growth regulators on shoot multiplication and subsequent rooting were investigated. Surface sterilization of rhizome buds with sodium hypochlorite was insufficient to control contamination. However, immersing rhizome buds in 100 mg L−1 AgO NPs for 60 min eliminated contamination without affecting the survival of explants. The number of shoots (12.2) produced per rhizome bud was higher in Murashige and Skoog (MS) medium containing 8 µM of 6-Benzyladenine (6-BA) and 0.5 µM of Thidiazuron (TDZ) than other treatments. The highest number of roots (24), with a mean root length of 7.8 cm and the maximum shoot length (9.8 cm), were obtained on medium MS with 2 µM of Indole-3-butyric acid (IBA). A survival rate of 98% was attained when plantlets of K. parviflora were acclimatized in a growth room. Liquid chromatography with tandem mass spectrometry (LC-MS/MS) was used to determine the chemical profile of K. parviflora leaf extracts. Results showed that several biologically active flavonoids reported in rhizomes were also present in leaf tissues of both in vitro cultured and ex vitro (greenhouse-grown) plantlets of K. parviflora. We found 40 and 36 compounds in in vitro cultured and ex vitro grown leaf samples, respectively. Greenhouse leaves exhibited more potent antioxidant activities than leaves from in vitro cultures. A higher acetylcholinesterase inhibitory ability was obtained for greenhouse leaves (1.07 mg/mL). However, leaves from in vitro cultures exhibited stronger butyrylcholinesterase inhibitory abilities. These results suggest that leaves of K. parviflora, as major byproducts of black ginger cultivation, could be used as valuable alternative sources for extracting bioactive compounds.
... Kay and Grätzel were the earliest to study the use of Chl products to sensitize nanoporous TiO 2 [105]. Since then, several studies have been performed to investigate the use of Chl as a sensitizer in DSSC [106][107][108]. The electronic combination between the sensitizer and TiO 2 CB is usually established via the carboxylic acid groups in the sensitizer, which helps the dye molecule attach and introduce electrons to the CB TiO 2 . ...
Humankind is currently heavily reliant on fossil fuel for its energy supply. However, fossil fuels are a nonrenewable source. It requires hundreds of millions of years for fossil fuels to develop inside the earth, and with our daily rate of consumption, we will run out of fuel unless we find an alternative source. Solar energy has gained much attention as a solution for the current energy deficit since it converts light energy directly to electrical energy. Dye-sensitized solar cells deliver a cheaper and dependable alternative for numerous photovoltaic devices such as organic, inorganic, and hybrid solar cells. Dye-sensitized-solar-cell technology characteristically depends on photosensitizer (Dye), electrolyte, and the metal oxide semiconductor. This review paper enlightens the working principle, components, and development of DSSCs present development and future prospectus for this novel technology.
... Methoxyflavones in KP have been claimed for physiological and pharmacological activities (Chen et al., 2018). A quality control assessment of KP raw materials and products was performed by a gas chromatography using 11 flavonoids as a reference (Sutthanut et al., 2007). Three main effective methoxyflavones, 3,5,7,3',4'-pentamethoxyflavone ( P M F ) , 5 , 7 -d i m e t h o x y f l a v o n e ( D M F ) , a n d 5,7,4'-trimethoxyflavone (TMF) are always mentioned for pharmacological effects (Chen et al., 2018). ...
Objective:
Aims of this study were to (1) compare anti-proliferative activity between aqueous and ethanol Kaempferia parviflora (KP) extracts in both cancer (Human urinary bladder cancer cell, T24) and normal cell lines (Human umbilical vein endothelial cell, HUVEC). (2) confirm selective cytotoxicity of ethanol KP extract to normal and different cancer cell lines (3) investigate its cellular mechanism through p53 and SIRT1 gene expression.
Methods:
Phytochemical difference between aqueous and ethanol extract was determined by thin layer chromatography (TLC). Screening for cytotoxic activity in human cell lines was performed by cell viability assay using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reagent. P53 and SIRT1 gene expression were quantified using RT-PCR.
Results:
Results from the cell viability assay were shown as follows: (1) ethanol extract possessed higher toxicity to cancerous cells than aqueous extract (2) ethanol extract exhibited higher cytotoxicity to cancerous cells than normal cells (3) ethanol extract also showed cytotoxicity, with different levels, to three prostate cancer cell lines varying in aggressiveness. (4) ethanol KP extract induced cell death in T24 via p53 gene expression and prolonged cell survival in HUVEC through SIRT1 gene expression.
Conclusion:
These findings implied that ethanol KP extract might possibly be an alternative for cancer adjuvant therapy through the mechanism of selective p53 and SIRT1 gene expression.
... Because the growth of medicinal herbs is closely related to the natural environment (e.g., soil, water quality, and climate), their quality and efficacy vary among growing regions [38,39]. Therefore, herbs from different locations need to be clearly distinguished and then used to produce specific medicines. ...
Panax quinquefolium is a perennial herbaceous plant that contains many beneficial ginsenosides with diverse pharmacological effects. 24(R)-pseudoginsenoside F 11 is specific to P . quinquefolium , a useful biomarker for distinguishing this species from other related plants. However, because of its nonconjugated property and the complexity of existing detection methods, this biomarker cannot be used as the identification standard. We herein present a stable 24(R)-pseudoginsenoside F 11 fingerprint spectrum in the terahertz band, thereby proving that F 11 can be detected and quantitatively analyzed via terahertz spectroscopy. We also analyzed the sample by high-performance liquid chromatography-triple quadrupole mass spectrometry. The difference between the normalized data for the two analytical methods was less than 5%. Furthermore, P . quinquefolium from different areas and other substances can be clearly distinguished based on these terahertz spectra with a standard principal component analysis. Our method is a fast, simple, and cost-effective approach for identifying and quantitatively analyzing P . quinquefolium .
Kaempferia species have contained various crucial ethnobotanical features, and are being used as traditional folk medicines in some Southeast Asia. This review tends to highlight important information (phytochemistry, biosynthesis, synthesis, pharmacology, and pharmacokinetics) of Kaempferia principal phytochemical classes diterpenoids and flavonoids. The electronic sources, e.g., Google Scholar, Sci-Finder, and Web of Science, and the most meaningful keywords "Kaempferia", "diterpenoids", and "flavonoids" have been more often utilized for searching the literature. More than 190 phytochemicals type diterpenoids (153 compounds) and flavonoids (42 compounds) have been separated from Kaempferia species. Isopiramanes and flavones are representative compounds. Kaempferia diterpenoids and flavonoids established great attention due to their pharmacological values such as antioxidative, anti-inflammatory, antimicrobial, antiviral, and antimalarial activities, especially anticancer. They also protected against harms to the neuron, skin, and liver. Pharmacokinetic studies revealed that metabolism of Kaempferia flavonoids might be related to the transformation of hydroxyl groups. Advances in chromatographic separations to obtain huge amounts of Kaempferia isolated compounds are expected. Future in vivo and clinical investigations on the components of Kaempferia should be carried out to provide accurate dosage and normative recommendations.
Zingiberaceae, commonly known as ginger, is one of the largest families of the plant kingdom with 52 genera and over 1300 species and distributed widely throughout the tropics, particularly in Southeast Asia. The family consists of a large number of economically and medicinally important plants well known for their use in ethnomedicine. Various plant parts from these plants, such as rhizome and seeds, have been used as ingredients of many herbal preparations in Chinese, Thai, African, and Indian traditional medicinal systems including Ayurveda. Interestingly, many of these plant parts are also used as spices since ancient times, becoming an essential part of ethnic cuisine. Many plants from different genera of Zingiberaceae were studied for their medicinal usages, and a number of compounds were purified with potent bioactivities including antimicrobial, antifungal, anti-inflammatory, anticancer, antioxidant, antiviral, antidiabetic, antiarthiritic, larvicidal, neuroprotective, and heptoprotective activities. Phytochemical analyses of different genera of Zingiberaceae have revealed the presence of a wide range of pharmacologically active phytochemical groups which mainly include terpenoids, diarylheptanoids, phenylpropanoids, and flavanoids. Interestingly, there are a number of essential oils, plant extracts, and compounds from Zingiberaceae species known to possess potent cytotoxic / antiproliferative / anticancer activities. These extracts and compounds are capable of specifically targeting cancer-related proteins and signaling pathways to kill cancer cells without severe damage to normal healthy cells. This chapter attempts to provide a comprehensive review on anticancer potential and mechanistics of action of such plant-derived extracts and bioactive compounds reported from various genera of the family such as Curcuma, Zingiber, Kaempferia, Alpinia, Amomum, and Hedychium as evidenced by in vitro and in vivo studies.KeywordsAnticancerApoptosisChemotherapeuticsPhytochemicalsZingiberaceae
Volatile organic compounds (VOCs) play an important role in the biological activities of the medicinal Zingiberaceae species. In commercial preparations of VOCs from Kaempferia parviflora rhizomes, its leaves are wasted as by-products. The foliage could be an alternative source to rhizome, but its VOCs composition has not been explored previously. In this study, the VOCs in the leaves and rhizomes of K. parviflora plants grown in a growth room and in the field were analyzed using the headspace solid-phase microextraction (HS-SPME) method coupled with gas chromatography and time-of-flight mass spectrometry (GC-TOF-MS). The results showed a total of 75 and 78 VOCs identified from the leaves and rhizomes, respectively, of plants grown in the growth room. In the field samples, 96 VOCs were detected from the leaves and 98 from the rhizomes. These numbers are higher compared to the previous reports, which can be attributed to the analytical techniques used. It was also observed that monoterpenes were dominant in leaves, whereas sesquiterpenes were more abundant in rhizomes. Principal component analysis (PCA) revealed significantly higher abundance and diversity of VOCs in plants grown in the field than in the growth room. A high level of similarity of identified VOCs between the two tissues was also observed, as they shared 68 and 94 VOCs in the growth room and field samples, respectively. The difference lies in the relative abundance of VOCs, as most of them are abundant in rhizomes. Overall, the current study showed that the leaves of K. parviflora, grown in any growth conditions, can be further utilized as an alternative source of VOCs for rhizomes.
Phytochemical study on the rhizomes of Kaempferia parviflora led to the isolation of twenty-three compounds including six phenolic glycosides (1-6), thirteen flavones (7-19), and five phenolic compounds (20-23). Of these, the new compounds were determined to be 2,4-dihydroxy-6-methoxyacetophenone-2-β-D-apiofuranosyl-(1→6)-β-D-glucopyranoside (1), 2-hydroxy-4-propionyl-phenyl O-β-D-glucopyranoside (2), and 4-hydroxy-3,5-dimethoxyacetophenone 8-O-α-L-rhamnopyranosyl-(1→6)-β-D-glucopyranoside (3) and named as kaempanosides A-C, respectively. Their chemical structures were established based on HR-ESI-MS, 1D and 2D NMR spectra. All compounds 1-23 exhibited acetylcholinesterase inhibitory activity with IC50 values ranging from 57.76 to 253.31 µM.
Black ginger is used as an herbal medicine for self-care and health promotion. Black ginger extract has been shown to alter the function of transporters in several cell types. This study demonstrates the interaction between the extract and 5,7-dimethoxyflavone (DMF) on drug efflux mediated by breast cancer resistance proteins (BCRP) and P-glycoprotein (P-gp) in Caco-2 cells and heterologous cell systems [Madin-Darby canine kidney type II (MDCKII) stably transfected with human BCRP (MDCKII/BCRP) or human P-gp (MDCKII/P-gp)]. The transepithelial flux of 3H-Digoxin and 3H-Estrone sulfate, prototypic substrates of P-gp, and BCRP, respectively, across Caco-2 cell monolayers, MDCKII/BCRP, and MDCKII/P-gp cells were determined. The results demonstrate that black ginger extract (10 μg/ml) significantly increases 3H-Digoxin and 3H-Estrone sulfate transport from the apical to basolateral side while decreasing transport from the basolateral to apical side of Caco-2 cells and MDCKII cell overexpression of BCRP or P-gp. The effect of the extract on 3H-Digoxin and 3H-Estrone sulfate transport was related to a decrease in efflux ratio. Likewise, DMF (5 μM) significantly increased 3H-Digoxin and 3H-Estrone sulfate absorption with a decreased efflux ratio compared to the control. Interestingly, the extract also significantly increased absorption of paclitaxel, an anti-cancer drug, which has poor oral absorption. Taken together, co-administration of drugs as substrates of BCRP and P-gp, with the black ginger extract containing DMF, might alter the pharmacokinetic profiles of the medicine.
Kaempferia, a genus of the family Zingiberaceae, is widely distributed with more than 50 species which are mostly found throughout Southeast Asia. These plants have important ethnobotanical significance as many species are used in Ayurvedic and other traditional medicine preparations. This genus has received a lot of scholarly attention recently as a result of the numerous health advantages it possesses. In this review, we have compiled the scientific information regarding the relevance, distribution, industrial applications, phytochemistry, ethnopharmacology, tissue culture and conservation initiative of the Kaempferia genus along with the commercial realities and limitations of the research as well as missing industrial linkages followed by an exploration of some of the likely future promising clinical potential. The current review provides a richer and deeper understanding of Kaempferia, which can be applied in areas like phytopharmacology, molecular research, and industrial biology. The knowledge from this study can be further implemented for the establishment of new conservation strategies.
Flavone has recently been proved as a promising scaffold for the development of a novel drug against dengue fever, one of the major health threats globally. However, the structure–activity relationship study of flavones on the anti-dengue activity remains mostly limited to the natural-occuring analogs. Herein, 27 flavone analogs were successfully synthesized, of which 5 analogs ( 5e , 5h , 5o , 5q , and 5r ) were novel. In total, 33 analogs bearing a diverse range of substituents were evaluated for their efficacy against DENV2-infected LLC/MK2 cells. The introduction of electron-withdrawing groups on ring B such as Br ( 5m ) or NO 2 ( 5n and 5q ) enhanced the activity significantly. In particular, the tri-ester 5d and di-ester 5e exhibited low toxicity against normal cell, and exceptional DENV2 inhibition with the EC 50 as low as 70 and 68 nM, respectively, which is over 300-fold more active compared to the original baicalein reference. The viral targets for these potent flavone analogs were predicted to be NS5 MTase and NS5 RdRp, as suggested by the likelihood ratios from the molecular docking study. The great binding interaction energy of 8-bromobaicalein ( 5f ) confirms the anti-dengue activity at atomistic level. The physicochemical property of all the synthetic flavone analogs in this study were predicted to be within the acceptable range. Moreover, the QSAR model showed the strong correlation between the anti-dengue activity and the selected molecular descriptors. This study emphasizes the great potential of flavone as a core structure for further development as a novel anti-dengue agent in the future.
Rapid and on-site qualitative and quantitative analysis of small molecules (including bioflavonoids) in biofluids are of great importance in biomedical applications. Herein, we have developed two deep learning models based on the 3D fluorescence spectra of gold nanoclusters as a single probe for rapid qualitative and quantitative analysis of eight bioflavonoids in serum. The results proved the efficiency and stability of the random forest-bidirectional long short-term memory (RF-BLSTM) model, which was used only with the most important features after deleting the unimportant features that might hinder the performance of the model in identifying the selected bioflavonoids in serum at very low concentrations. The optimized model achieves excellent overall accuracy (98-100%) in the qualitative analysis of the selected bioflavonoids. Next, the optimized model was transferred to quantify the selected bioflavonoids in serum at nanoscale concentrations. The transferred model achieved excellent accuracy, and the overall determination coefficient (R2) value range was 99-100%. Furthermore, the optimized model achieved excellent accuracies in other applications, including multiplex detection in serum and model applicability in urine. Also, LOD in serum at nanoscale concentration was considered. Therefore, this approach opens the window for qualitative and quantitative analysis of small molecules in biofluids at nanoscale concentrations, which may help in the rapid inclusion of sensor arrays in biomedical and other applications.
BACKGROUND: The rhizomes of Kaempferia parviflora (KP), have been traditionally used for treating various ailments with 5,7-dimethoxyflavone (DMF) as a prominent compound. OBJECTIVE: To investigate the anti-aging and neuroprotective properties of KP and DMF in Caenorhabditis elegans. METHODS: C. elegans (wild-type (N2), transgenic and mutant strains) were treated with KP and DMF and were monitored for lifespan and neuroprotection through physiological assays, fluorescence microscopy and qPCR analysis. Molecular docking studies were employed to identify the interaction mode of DMF with DAF-16 and SKN-1. RESULTS: KP and DMF significantly increased the lifespan of N2 along with modulating pharyngeal pumping and lipofuscin accumulation. They also exhibited neuroprotection in Aβ transgenic strains by improving lifespan and delaying paralysis. Further, they reduced ROS accumulation significantly in worms exposed to UV-A, thereby exhibiting anti-photoaging potential. KP and DMF could activate SKN-1, DAF-16 which was evident from molecular docking and qPCR analysis. The DAF-2 and DAF-16 mutants did not exhibit any variations in lifespan upon treatment with KP and DMF suggesting the involvement of the DAF-16 mediated pathway in regulating the anti-aging and neuroprotective effects. CONCLUSION: Our findings suggest that KP with DMF as an active ingredient is a potential nutraceutical for aging and associated disorders.
This research was aimed to determine the effects of extraction methods on antioxidant properties and methoxyflavone contents of Kaempferia parviflora (KP) rhizome extracts. The KP rhizomes were extracted by maceration using ethanol at the concentrations of 25, 50, 75 and 95% V/V for 7 days and 95% V/V ethanolic extraction with sonication-assisted extraction (SAE) for 15-45 mins. Antioxidant components (phenolics, flavonoids and anthocyanins) and antioxidant activities (DPPH and FRAP) were examined. Methoxyflavones of the KP extracts were identified by a GC-MS technique. It was found that extraction methods affected the antioxidant properties of the extracts. Increasing ethanol concentrations enhanced anthocyanins but not phenolics and flavonoids. Ethanol concentration at 75% V/V exhibited the greatest DPPH while 25% V/V ethanol showed the greatest FRAP values. In this study, 10 methoxyflavones from KP extracts were separated and identified by GC chromatograms. The content of 5,7-dimethoxyflavone increased from 1.1 g/100 mL extract to 48.10 g/100 mL extract as the ethanol concentrations increased from 25% to 95% V/V. SAE for 15-45 mins had little impact on antioxidant properties as well as methoxyflavone contents. In general, SAE enhanced the extraction of KP rhizomes by increasing 5,7-dimethoxyflavone contents.
Introduction:
Kaempferia parviflora or black ginger is abundantly cultivated because its rhizomes contain methoxyflavones that have many pharmacological properties. K. parviflora can be divided into two types, based on morphological characteristics, but differences in their chemical compositions have never been explored.
Objectives:
This research aims to find chemical markers that can be used to differentiate between the two types of K. parviflora, the red-leaf and green-leaf types, by quantifying the amounts of methoxyflavones.
Material and methods:
K. parviflora samples were collected from 39 locations in Thailand. Their genetic diversity was assessed by a genotyping-by-sequencing (GBS) technique to construct the population structure. Their chemical compositions were analyzed by high performance liquid chromatography-photodiode array detection to determine the methoxyflavone contents.
Results:
The population structure based on >3,000 single nucleotide polymorphism (SNP) markers showed that the samples can be divided into two groups, which were consistent with the classification by leaf margin color (red-leaf and green-leaf types). HPLC analysis revealed 3,5,7,3',4'-pentamethoxyflavone (PMF), 5,7-dimethoxyflavone (DMF), 5,7,4'-trimethoxyflavone (TMF), 3,5,7-trimethoxyflavone and 3,5,7,4'-tetramethoxyflavone as major methoxyflavones that can be used as chemical markers. The red-leaf type showed higher amounts of PMF, TMF and 3,5,7,4'-tetramethoxyflavone than the green-leaf type, while the green-leaf type showed higher amounts of DMF and 3,5,7-trimethoxyflavone than the red-leaf type.
Conclusion:
These results provide another approach to discriminate the two types of K. parviflora using chemical profiles alongside genetic and morphological analyses. Therefore, a specific type of K. parviflora can be selected over the other based on preferences for a certain methoxyflavone.
Kaempferia spp. have traditionally played a role in treating various ailments and are attracting increasing economic and scientific interest. This review evaluates the traditional use, phytochemical compounds, and biological properties of Kaempferia parviflora, Kaempferia galanga, Kaempferia rotunda, Kaempferia pulchra, and Kaempferia angustifolia. The health benefits associated with consuming Kaempferia spp. and their active components, namely flavonoids and other volatile compounds, include antibacterial, anticancer, antidiabetic, cardiovascular protective, immunoregulatory, neuroprotective, and skin-whitening effects, suggesting the use of Kaempferia spp. as a potential health aid for the aging population. As agricultural production is important to ensure consistent quality of Kaempferia produce, resource management techniques related to species identification, cultivation, storage, and processing were also highlighted. It was suggested that the value-added concept in agricultural production may be limited. Investigations into plant biology and physiology are required to provide a solid theoretical basis for the sustainable production of Kaempferia.
Background
Ovarian cancer is hard to diagnose and its survival rates dramatically decrease according to the stage progression. Anthocyanidins derived from Kaempferia parviflora has been studied for anti-inflammatory, antioxidant, and anticancer effects in various disease both in vitro and in vivo. However, no studies have evaluated the molecular mechanism of Kaempferia parviflora anthocyanidin fractions in ovarian cancer cells.Objectives
To study the effects of Kaempferia parviflora anthocyanidins on the progression of ovarian cancer, we extracted anthocyanidins from Kaempferia parviflora and analyzed it using HPLC. Then, we determined the change of cell characteristics by treatment of anthocyanidins derived from Kaempferia parviflora.ResultsBased on HPLC analysis, the most abundant anthocyanidins of Kaempferia parviflora was peonidin, followed by cyanidin, delphinidin, and pelargonidin. We selected high-grade serous ovarian cancer and clear cell cancer cell lines to investigate the anticancer effects of anthocyanidin extracts. Treatment with anthocyanidin extracts decreased ES-2 and OV-90 cell proliferation and increased late apoptosis with cell cycle arrest at sub G0/G1 phase. In addition, anthocyanidin extracts hampered the mitochondrial membrane permeability in both cell lines. Moreover, the cytosolic calcium accumulation was detected in ES-2 cells, and the overproduction of reactive oxygen species was estimated in OV-90 cells, respectively.Conclusion
Collectively, these results showed the anticancer effects of Kaempferia parviflora anthocyanidin extracts against the progression of ovarian cancer.
Kaempferia parviflora and Curcuma longa have been widely reported to have a potent anti-inflammatory effect. However, both extracts have been shown to have low bioavailability and high-level first pass metabolism upon oral administration. This study aimed to develop transdermal delivery of a combination of both extracts in matrix-patch formulations with five different volatile oils which possess analgesic and anti-inflammatory properties. Fourteen formulations exhibited good physicochemical properties and stability. The release of methoxyflavones increased significantly when the concentration of PVP (polyvinylpyrrolidone) in the patches increased. The drug release kinetics were best-fitted with the zero-order and Higuchi models, depending on the amount of PVP. When combined with Curcuma longa, the patch provided the highest accumulation of methoxyflavones and curcuminoids within the skin. In the clinical, randomized controlled trial, a significant pain improvement was found after seven days of application on the pain area. The pain scores decreased from 5.84 ± 1.57 to 2.74 ± 1.37 and the mean pain pressure threshold increased significantly from 1.79 ± 0.51 N to 2.55 ± 0.41 N. Our results indicated that the developed herbal patches were useful for transdermal application and can be considered as an alternative treatment for pain relief, highlighted by a decrease in pain intensity and an increase in pain tolerance, without skin irritation.
Skin aging is accompanied by an increase in the number of senescent cells, resulting in various pathological outcomes. These include inflammation, impaired barrier function, and susceptibility to skin disorders such as cancer. Kaempferia parviflora (Thai black ginger), a medicinal plant native to Thailand, has been shown to counteract inflammation, cancer, and senescence. This study demonstrates that polymethoxyflavones (5,7-dimethoxyflavone, 5,7,4′-trimethoxyflavone, and 3,5,7,3′,4′-pentamethoxyflavone) purified from K. parviflora rhizomes suppressed cellular senescence, reactive oxygen species, and the senescence-associated secretory phenotype in primary human dermal fibroblasts. In addition, they increased tropocollagen synthesis and alleviated free radical-induced cellular and mitochondrial damage. Moreover, the compounds mitigated chronological aging in a human ex vivo skin model by attenuating senescence and restoring expression of essential components of the extracellular matrix, including collagen type I, fibrillin-1, and hyaluronic acid. Finally, we report that polymethoxyflavones enhanced epidermal thickness and epidermal-dermal stability, while blocking age-related inflammation in skin explants. Our findings support the use of polymethoxyflavones from K. parviflora as natural anti-aging agents, highlighting their potential as active ingredients in cosmeceutical and nutraceutical products.
Phytochemical investigations of the ethanolic extract of leaves of Crateva religiosa (Family: Capparaceae) resulted in the isolation of four compounds viz. (E)-ethyl-4-((3-(4-hydroxyphenyl)acryloyl)oxy)-2-methyl-5-oxotetrahydrofuran-2-carboxylate (Cg-1) named as Cratin, Cg-2, Cg-3, and Cg-4. Cg-1 has been reported for the first time from any natural source and has not been synthesized so far. Cg-2, Cg-3, and Cg-4 were also isolated first time from this plant source. Their structures were elucidated based on chemical and physical data viz. (elemental analysis, FT-IR, UV, ¹H NMR, ¹³C NMR, and mass). Single crystal X-ray analysis was further used for the authentication of the structure of the isolated compounds in the case of Cg-1, including chemical modification of Kaempferol Cg-2 to Cg-2-Me, to get suitable crystals for X-ray diffraction study. The isolated novel compound Cg-1 and the methyl ether of kaempferol (Cg-2-Me) were screened for DNA binding studies with ct-DNA using UV-Visible spectroscopy, fluorescence, circular dichroism studies. Molecular docking was performed to understand the interaction between DNA and the compound studied and showed groove binding interaction (non-intercalation).
Flavonoids are closely related to human health. The identification and determination of flavonoids is an important and difficult issue. In view of this, this work puts forward a colorimetric sensing array for flavonoid discrimination based on gold nanoparticle (AuNP) aggregation. Two substances (acetylthiocholine iodide (Atc) and S-propionylthiocholine iodide (Ptc)) are hydrolyzed to choline (Chi) under the action of acetylcholinesterase (AcChE). Choline can agglomerate AuNPs by forming Au-S bond with its own sulfhydryl functional group. The presence of flavonoids inhibits the AcChE activity, resulting in the formation of different amounts of Chi and different degrees of AuNP aggregation. Controlled by Atc and Ptc as array’s receptors, the five flavonoids, including proanthocyanidin (Pro), naringenin (Nar), quercetin (que), flavone (Fla), and curcumin (Cur), were differentiated through their unique colorimetric “fingerprint” pattern-based recognition with the aid of linear discriminant analysis (LDA). The assay is validated under phosphate buffered saline (PBS) buffer conditions as well as in human serum. In addition, identification for diverse concentrations of single flavonoid and different binary and ternary mixtures of flavonoids were also demonstrated.
Nghiên cứu được thực hiện nhằm đánh giá ảnh hưởng của trạng thái nguyên liệu (làm choáng và không làm choáng), môi trường xả tiết (nước và không khí) và điều kiện xả tiết (nhiệt độ và thời gian) đến chất lượng của sản phẩm phi lê cá lóc. Các chỉ tiêu hóa lý được đánh giá gồm màu sắc (L*, a* và b*), hiệu suất thu hồi sau gia nhiệt, trạng thái cấu trúc, hàm lượng sắt heme, sắt non-heme và oxy hóa lipid (chỉ số peroxide-PV và TBARS). Kết quả cho thấy cá được làm choáng trước khi cắt tiết có giá trị L* cao hơn và giá trị a*, b* thấp hơn và hiệu quả loại máu tốt hơn (hàm lượng sắt heme và sắt non-heme thấp hơn) so với cá không được làm choáng. Nước là môi trường phù hợp để xả tiết cá lóc. Nhiệt độ và thời gian xả tiết là hai yếu tố quan trọng ảnh hưởng đến hiệu quả loại máu, điều kiện phù hợp để xả tiết cá lóc là nhiệt độ nước 23-25°C trong thời gian 20 phút.
Sweet cherry (Prunus avium L.) is a tree widely cultivated in temperate regions for its tasty and healing fruits. Pruning works on the tree give each year considerable amounts of woody wastes that hardly have any utility. The aims of this work were to detect the most active antioxidants present in a cherry pruning wood sample, to isolate and characterize them, and study the antimicrobial and antibiofilm activities of components found in the wood ethyl acetate extract against a selection of foodborne microorganisms. The online HPLC–DPPH technique allowed the detection of two active antioxidant peaks that, after being isolated by a combination of countercurrent chromatography (FCPC) and conventional preparative techniques, and subsequent structural characterization by NMR, MS and polarimetry, resulted to be (‒)-catechin (1) and (‒)-taxifolin (4). Other components of the cherry wood extract were also isolated, among which compounds 1, 4, and (+)-dihydrowogonin (12) have never been reported in P. avium. A selection of the isolated flavonoids was submitted to antimicrobial and antibiofilm activity evaluations against strains from type culture collections, as well as on multi-resistant strains previously isolated in our laboratory. Those compounds with antimicrobial activity detected in preliminary screenings by standard agar diffusion tests ‒the flavan-3-ol 1, the flavanonols 4 and (+)-aromadendrin (5), the flavanone (+)-pinocembrin (15), and the flavone tectochrysin (17)‒ were subjected to the minimal inhibitory concentration (MIC) test, showing all of them MIC values of 100 μg/mL. Compound 4 also induced a significative inhibition on the formation of biofilms by Enterobacter sp. UJA37p at a concentration of 1 μg/mL and a significative disruption of preformed biofilm by this strain at 0.1 µg/mL. Similar results on biofilm disruption were observed with compound 17.
Nine flavonoids (1-9) have been isolated from Kaempferia parviflora. Among these, 5,7,4'-trimethoxyflavone (8) and 5,7,3',4'-tetramethoxyflavone (9) exhibited antiplasmodial activity against Plasmodium falciparum, with IC50 values of 3.70 and 4.06 microg/ml, respectively. 3,5,7,4'-Tetramethoxyflavone (7) and compound 8 possessed antifungal activity against Candida albicans with respective IC50 values of 39.71 and 17.63 microg/ml, and also showed mild antimycobacterial activity with the minimum inhibitory concentrations (MIC) of 200 and 50 microg/ml, respectively. However, none of the isolated compounds demonstrated cytotoxicity against KB, BC and NCI-H187 cell lines.
There is an increasing interest in the usage of chromatographic methods on the analysis of chemical compounds present in Hippophae rhamnoides L. In this paper, the chromatographic techniques applied for the determination, separation and identification of chemical compounds of H. rhamnoides L. are reviewed. We examined the existing chromatographic methods based on separations by paper and thin-layer chromatography, gas chromatography, high-performance liquid chromatography and capillary electrophoresis and also methods of detection by ultraviolet absorption, fluorescence, refractive index, electrochemical and mass spectrometry. Biological properties of the plant and its pharmacological effects and use in traditional medicine have also been reviewed.
The biologically active secondary metabolites of Ginkgo biloba extract EGb 761 in phytopharmaceuticals were analyzed using two simple, rapid, accurate and sensitive HPLC methods. The proposed methods were successfully applied in the determination of terpenes and flavonoids in four phytopharmaceutical preparations selected from the Egyptian market. The terpenes; ginkgolide A, ginkgolide B, and bilobalide were analyzed using RP 18 column with a mobile phase consisting of water/methanol/isopropanol (72.5:17.5:10, v/v) at a flow rate of 1 ml min-1 and UV detection at 220 nm. The flavonoids; quercetin and kaempferol were analyzed using RP 18 column in a step gradient elution with acetonitrile and water at pH 3.3 and flow rate of 1.5 ml min-1 with UV detection at 370 nm. The two HPLC methods were completely validated.
5-Hydroxy-7-methoxyflavanone, 5,7-dimethoxyflavanone, 5-hydroxy-7-methoxyflavone 5-hydroxy-7,4′-dimethoxyflavone, 5,7-dimethoxyfiavone, 5,7,4′-trimethoxyflavone, 5,7,3′,4′-tetramethoxyflavone, 5-hydroxy-3,7-dimethoxyflavone, 5-hydroxy-3,7,4′-trimethoxyflavone, 3,5,7-trimethoxyflavone and 5-hydroxy-3,7,3′,4′-tetramethoxyflavone have been isolated from the black rhizomes of Boesenbergia pandurata.
Flavanoids exist as secondary plant metabolites, which displays a wide variety of biological effects. The polymethoxylated flavones, such as nobiletin and tangeretin, present in Citrus reticulata peels are of great interest due to their pharmacological effects. Separation and isolation of these structurally very similar flavones has been achieved by using commercially available ion exchange resins. The strong cation exchange resin [H+] selectively adsorbs the tangeretin and other phenolic compounds present in the matrix in comparison to the hexamethoxylated flavone, i.e. nobiletin. The cation exchange resin can be used successfully to isolate nobiletin and tangeretin from a plant extract. Purity of the isolated compounds was monitored by HPLC using a C-18 column with photometric detection at 280 nm. The structures of the isolated compounds have been confirmed by NMR and tandem mass spectrometry.
The anti-inflammatory activity of 5,7-dimethoxyflavone (5,7-DMF) has been assessed. It was found to possess a comparable effect to aspirin on the rat paw edema model, and it showed no inhibition on cotton pellet-induced granuloma formation. On the rat pleurisy model, 5,7-DMF exhibited an antiexudative effect, interfered with leukocyte migration, and markedly inhibited prostaglandin biosynthesis. In addition, 5,7-DMF caused marked lowering of the rectal temperature of rats. The results obtained from Hippocratic screening revealed that 5,7-DMF possessed a weak CNS depressant activity.
Six compounds were isolated from fresh rhizomes of fingerroot (Boesenbergia pandurata Schult.) as strong antimutagens toward 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1) in Salmonella typhimurium TA98. These compounds were 2',4',6'-trihydroxychalcone (pinocembrin chalcone; 1), 2',4'-dihydroxy-6'-methoxychalcone (cardamonin; 2), 5,7-dihydroxyflavanone (pinocembrin; 3), 5-hydroxy-7-methoxyflavanone (pinostrobin; 4), (2,4,6-trihydroxyphenyl)-[3'-methyl-2'-(3' '-methylbut-2' '-enyl)-6'-phenylcyclohex-3'-enyl]methanone (5), and (2,6-dihydroxy-4-methoxyphenyl)-[3'-methyl-2'-(3' '-methylbut-2' '-enyl)-6'-phenylcyclohex-3'-enyl]methanone (panduratin A; 6). Compound 5 was a novel compound (tentatively termed 4-hydroxypanduratin A), and 1 was not previously reported in this plant, whereas 2-4 and 6 were known compounds. The antimutagenic IC(50) values of compounds 1-6 were 5.2 +/- 0.4, 5.9 +/- 0.7, 6.9 +/- 0.8, 5.3 +/- 1.0, 12.7 +/- 0.7, and 12.1 +/- 0.8 microM in the preincubation mixture, respectively. They also similarly inhibited the mutagenicity of 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). All of them strongly inhibited the N-hydroxylation of Trp-P-2. Thus, the antimutagenic effect of compounds 1-6 was mainly due to the inhibition of the first step of enzymatic activation of heterocyclic amines.
The cyclohexenyl chalcone derivative [(-)-hydroxypanduratin A], together with the previously known panduratin A, sakuranetin, pinostrobin, pinocembrin, and dihydro-5,6-dehydrokawain were isolated from the chloroform extract of the red rhizome variety of Boesenbergia pandurata (Robx.) Schltr. [currently known as Boesenbergia rotunda (L.) Mansf., Kulturpfl.]. Their structures were assigned on the basis of their spectroscopic data. (-)-Hydroxypanduratin A and (-)-panduratin A showed significant topical anti-inflammatory activity in the assay of TPA-induced ear edema in rats.
A gas chromatography-mass spectrometry (GC-MS) method was developed and validated for the simultaneous determination of seven major chemical markers (bilobalide, ginkgolides A, B, C, kaempferol, quercetin and isorhamnetin) in phytopharmaceuticals of Ginkgo biloba L. The intra-day relative standard deviations (RSD) and inter-day RSD's were based on the analysis of the standardized Ginkgo biloba L. extract on the same day and on the following 3 consecutive days. The intra-day RSD's ranged from 1.21% (bilobalide) to 6.20% (kaempferol). The inter-day RSD's ranged from 2.10% (bilobalide) to 10.42% (isorhamnetin). Mean recoveries ranged from a low of 63.0 +/- 5.3% (isorhamnetin) to a maximum of 103.5 +/- 6.0% (ginkgolide A). Calibration curves were linear in ranges between 2.73 and 36.36 microg/ml for the markers. Limits of detection ranged from a low of 0.5 microg/ml (bilobalide) to a high of 2.5 microg/ml (quercetin). The limits of quantitation were a low of 1.1 microg/ml (gingkolides A, B, C) to a high of 7.5 microg/ml (kaempferol). The method was applied to a standard extract (>6% total terpenoids and >24% total flavonoids) and six ginkgo capsule phytopharmaceuticals.
Flavonoids of nine Australian monofloral Eucalyptus honeys have been analyzed and related to their botanical origins. The mean content of total flavonoids varied from 1.90 mg/100 g of honey for stringybark (E. globoidia) honey to 8.15 mg/100 g of honey for narrow-leaved ironbark (E. crebra) honey, suggesting that species-specific differences occur quantitatively among these Eucalyptus honeys. All of the honey samples analyzed in this study have a common flavonoid profile comprising tricetin (5,7,3',4',5'-pentahydroxyflavone), quercetin (3,5,7,3',4'-pentahydroxyflavone), and luteolin (5,7,3',4'-tetrahydroxyflavone), which, together with myricetin (3,5,7,3',4',5'-hexahydroxyflavone) and kaempferol (3,5,7,4'-tetrahydroxyflavone), were previously suggested as floral markers for European Eucalyptus honeys. Thus, flavonoid analysis could be used as an objective method for the authentication of the botanical origin of Eucalyptus honeys. Moreover, species-specific differences can also be found in the composition of honey flavonoid profiles. Among these honeys, bloodwood (E. intermedia) honey contains myricetin and tricetin as the main flavonoid compounds, whereas there is no myricetin detected in yapunyah (E. ochrophloia), narrow-leaved ironbark (E. crebra), and black box (E. largiflorens) honeys. Instead, these types of Eucalyptus honeys may contain tricetin, quercetin, and/or luteolin as their main flavonoid compounds. Compared to honeys from other geographical origins, the absence or minor presence of propolis-derived flavonoids such as pinobanksin, pinocembrin, and chrysin in Australian honeys is significant. In conclusion, these results demonstrate that a common flavonoid profile exists for all of the Eucalyptus honeys, regardless of their geographical origins; the individual species-specific floral types of Eucalyptus honey so common in Australia could be possibly differentiated by their flavonoid profile differences, either qualitatively or quantitatively or both.
Phenolics are ubiquitous compounds found in all plants as their secondary metabolites. These include simple phenols, hydroxybenzoic acid and cinnamic acid derivatives, flavonoids, coumarines and tannins, among others. The extraction of phenolics from source materials is the first step involved in their analysis. While chemical methods are used for determination of total content of phenolics, chromatographic and spectrometric analyses are employed for identification and quantification of individual compounds present. This paper provides a summary of background information and methodologies used for the analysis of phenolics in foods and nutraceuticals.
Reversed-phase high-performance liquid chromatography (RP-HPLC) with ultraviolet diode array detection (UV-DAD) was used for the simultaneous determination of four flavone C-glucosides, i.e. orientin, homoorientin, vitexin and isovitexin in several food systems fortified by the antioxidant of bamboo leaves (AOB), such as high temperature sterilized milk, sunflower seed oil and extruded rice cake for the first time. The method included extraction of flavone C-glucosides from AOB-fortified foods by methanol aqueous solution, deproteinating with saturated lead acetate solution and potassium oxalate, defatting with n-hexane and clean-up by solid-phase extraction (SPE) with Phenomenex C18 cartridges. Analytes were separated with Luna C18 5 microm 250mm x 4.6mm column using acetonitrile and 1% (v/v) acetic acid (pH 3.0) as mobile phase. Good results were obtained with respect to repeatability (relative standard deviation (RSD)< 2.2%) and recovery (81.4-91.8%) which fulfilled the requirements defined by European Union (EU) legislation. The total amounts of four flavone C-glucosides were 12.56 microg/100 mL, 881.08 microg/100 mL and 1420.83 microg/100 g dry weight in AOB-fortified sterilized milk, sunflower seed oil and extruded rice cake, respectively. The method was successfully applied to the analysis of flavone C-glucosides in AOB-fortified samples. The optimized procedure could also be referenced for the separation of flavone C-glucosides in other fortified foodstuffs.
Quercetin and catechin are among the major flavonoids in plant foods and their intake has been associated to a risk reduction in several degenerative diseases. The aim of the present study was to bring data on the bioavailability of quercetin and catechin when administered simultaneously. The study was performed on rats adapted to diets containing (i) 0.25% quercetin, or (ii) 0.25% catechin, or (iii) 0.25% quercetin+0.25% catechin. Quercetin, catechin and their metabolites were determined in plasma, urine and liver by HPLC with UV or coulometric detection. When quercetin and catechin were fed in association, their respective plasma concentration significantly decreased (-35% and -28% respectively), whereas the urinary and hepatic concentrations were only affected for quercetin (-36%). These data may be explained by a competitive interaction between quercetin and catechin at the digestive level, leading to a reduction of the intestinal absorption of quercetin and a possible delaying of catechin absorption over time. The simultaneous administration of quercetin and catechin had no effect on the formation of their glucurono and sulfo conjugates, indicating the absence of competition between quercetin and catechin for the corresponding conjugative enzymes.
Kaempferia parviflora is a Zingiberaceous plant, which has been reputed for its beneficial medicinal effects. The present study was undertaken to evaluate the Kaempferia parviflora ethanolic extract (KPE) for its anti-gastric ulcer activity by experimental models. Oral administration of the KPE at 30, 60 and 120 mg/kg significantly inhibited gastric ulcer formation induced by indomethacin, HCl/EtOH and water immersion restraint-stress in rats. In pylorus-ligated rats, pretreatment with the KPE had no effect on gastric volume, pH and acidity output. In ethanol-induced ulcerated rats, gastric wall mucus was significantly preserved by the KPE pretreatment at doses of 60 and 120 but not at 30 mg/kg. The findings indicate that the ethanolic extract of Kaempferia parviflora possesses gastroprotective potential which is related partly to preservation of gastric mucus secretion and unrelated to the inhibition of gastric acid secretion.
Sugarcane (Saccharum officinarum L., Gramineae) bagasse and leaves were investigated for their flavonoid content and transgenic sugarcane ("Bowman-Birk" and "Kunitz") was compared with non-modified ("control") plants. Analyses were carried out by high-performance liquid chromatography coupled to diode array UV detection (LC/UV), also using post-column addition of shift reagents, and tandem MS (atmospheric pressure chemical ionization-MS/MS and collision-induced dissociation-MS). On-line UV and MS data demonstrated the presence of methoxyflavone glycosides and aglycones in a total of seven compounds. Three naturally occurring flavones glycosides and two unusual erythro- and threo-diastereoisomeric flavolignan 7-O-glucosides were identified together with their aglycones.
An overview mapping recent trends in the determination of polyphenols of natural origin (mostly flavonoids) and their synthetic derivatives by electromigration methods is presented. The overview (covering the period of the recent 5 years and comprising 61 references) is focused on capillary zone electrophoresis (CZE) and micellar electrokinetic chromatography (MEKC) with various detection methods. Techniques comprising on-line pre-separation such as isotachophoresis (ITP)-CZE and flow-injection-CZE, chiral separations and CZE evaluation of antioxidation activity are also discussed.
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