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Abstract
Mitragyna speciosa, a tropical plant indigenous to Southeast Asia, is well known for its psychoactive properties. Its leaves are traditionally chewed by Thai and Malaysian farmers and manual labourers as it causes a numbing, stimulating effect. The present study aims to evaluate alkaloid yield and composition in the leaf extracts. For this purpose we have compared several non-conventional extraction techniques with classic procedures (room temperature or under heating). Dried M. speciosa leaves belonging to three batches of different origin (from Thailand, Malaysia and Indonesia) were extracted using ultrasound-assisted extraction (UAE), microwave-assisted extraction (MAE) and supercritical carbon dioxide extraction SFE-CO(2), using methanol, ethanol, water and binary mixtures. The extracts were compared using an HPLC/ESI-MS analysis of mitragynine and four other related alkaloids which were present in the alkaloid fraction. The extraction technique influences both the raw product yield and the relative alkaloid content of M. speciosa leaves. Of the several methods tested, MAE in a closed vessel at 110 °C (60 W, methanol/water 1:1) gave the highest alkaloid fraction amount, while UAE with an immersion horn at 25 °C (21.4 kHz, 50 W, methanol) showed the best yield for mitragynine. This work may prove to be a useful contribution to forensic, toxicological and pharmacognosy studies. Although the potential applications of M. speciosa alkaloids clearly need further investigation, these results may facilitate the scaling-up of their extraction.
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... Finally, 41 articles that complied with the requirement of inclusion criteria were retained for this scoping review. Out of the 41 articles screened, 14 articles were focused on the identification of bioactive compounds in the M. speciosa extract (6,7,(29)(30)(31)(32)(21)(22)(23)(24)(25)(26)(27)(28), while 27 articles were focused on the pharmacological properties (3,6,(35)(36)(37)(38)(39)(40)(41)(42)(43)(44)7,45,21,23,24,26,28,33,34). ...
... Methanol was found to be the most common solvent used in the extraction of secondary metabolites from M. speciosa. Out of 11 studies that used methanol, two studies practiced Soxhlet (7,25), three studies with maceration alone (26,28,32), one study with combination of maceration and sonication (21), one study with sonication alone (29), and three studies with modern methods such as UAE, MAE, Supercritical Fluid Extraction (SFE) and/or Accelerated Solvent Extraction (ASE) (6,22,30). ...
... Methanol in both maceration and Soxhlet methods are the predominant conventional practices in many M. speciosa extraction studies (22). It has been suggested that utilizing methanol in the Soxhlet conventional method for the M. speciosa extraction yielded a high amount of mitragynine in addition to 24 unique peaks, as analyzed using Gas Chromatography-Mass Spectrometer (GC-MS) (42). ...
Introduction: This scoping review aimed to provide a comprehensive summary and evaluation of solvents and methods for the extraction of bioactive compounds with pharmacological properties from Mitragyna speciosa (M. speciosa) Korth. Methods: The relevant articles were screened on electronic databases such as Scopus, PubMed, and Science Direct and verified their qualities based on the Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for Scoping Reviews (PRISMA-ScR) guideline. Results: We selected 41 articles according to two features; the extraction of bioactive compounds and pharmacological properties of M. speciosa extract that involved different solvents and methods. Evidence shows that methanol was the commonly used solvent along with the maceration process in the extraction of M. speciosa to obtain valuable bioactive compounds with clinical benefits. Alternatively, Soxhlet provides less exertion to the extraction process with similar value. Conclusion: Despite various potential modern techniques and solvents available, the synergy between traditional maceration and Soxhlet and methanol was found to potentially attain pharmacological values and bioactive substances in M. speciosa.
... In recent decades, the extraction of M. speciosa mitragynine has been carried out in various ways, including ultrasound (US) [4][5][6], maceration [4,7], and accelerated solvent extraction (ASE) [8]. Conventional extractions, such as soaking, maceration, and Soxhlet extraction, have more or fewer deficiencies and limitations, including long processing time, low extraction yields, and poor extraction efficiency [4,9]. ...
... In recent decades, the extraction of M. speciosa mitragynine has been carried out in various ways, including ultrasound (US) [4][5][6], maceration [4,7], and accelerated solvent extraction (ASE) [8]. Conventional extractions, such as soaking, maceration, and Soxhlet extraction, have more or fewer deficiencies and limitations, including long processing time, low extraction yields, and poor extraction efficiency [4,9]. ...
... In recent decades, the extraction of M. speciosa mitragynine has been carried out in various ways, including ultrasound (US) [4][5][6], maceration [4,7], and accelerated solvent extraction (ASE) [8]. Conventional extractions, such as soaking, maceration, and Soxhlet extraction, have more or fewer deficiencies and limitations, including long processing time, low extraction yields, and poor extraction efficiency [4,9]. To overcome these limitations of conventional extraction methods, new 2 and promising extraction techniques have been introduced. ...
The aim of this work was to determine the potential of using a pulsed electric field (PEF) and ultra-sound (US) apparatus to produce mitragynine extracts from Mitragyna speciosa dried leaves. Four modes of the device were tested: PEF, US, US + PEF, and PEF + US. The results were compared with extracts obtained using a conventional technique (maceration, as the control). Changes in the mitragynine content were determined with the liquid chromatography/mass spectrometry (LC-MS/MS) method. The LC-MS/MS analysis showed that the mitragynine contents from kratom extracts using four different modes were different. The highest extraction (106.63 ± 0.85 mg/L) of mitragynine was achieved by the PEF + US procedure, followed by US + PEF (97.27 ± 1.33 mg/L), with increased extraction efficiencies of 45.81 ± 0.59% and 33.00 ± 1.85%, respectively. Moreover, the total energy consumption under the combination technique was 25.0% lower than that with PEF assistance. Furthermore, scanning electron microscopy (SEM) and Fourier Transform Infrared Spectroscopy (FTIR) provided confirmatory evidence for the extraction of M. speciosa by the prototype. This study confirmed that PEF and US devices can be considered a green alternative method and may contribute to the application of agricultural products.
... From previous studies, UAE has been used for the extraction of ketum leaves. [11][12][13][14]. ...
... Collected leaves were air-dried and powdered. Then, 10 g of ground Ketum leaves powder were soaked in 300 mL of solvent and sonicated for 20, 40, and 60 min using a Branson 5510R-MT Ultrasonic extraction chamber with an output of 40 kHz and 135 W, following protocol [13], but with slight modifications. The temperature of the water bath was measured using a thermometer and maintained at 60 °C. ...
... The extraction conditions, the type of solvents, and the techniques used play vital roles in the extraction of phytochemicals and their compositions in studied plants [13,19]. The solvent extraction technique is one of the most crucial steps in the preparation of the sample for natural product analysis, particularly for herbal extracts. ...
Mitragynine and its congeners are one of the major phytoconstituents present in Mitragyna speciosa Korth. (ketum) leaves and a well-known factor that contributes to ketum’s pharmacological activities. This study describes the usage of a green extraction method to yield botanical leaf extracts of ketum. The leaf extracts were assessed for mitragynine content, total phenolic and flavonoid content, and cytotoxicities. The Ultrasound Assisted Extraction (UAE) method showed a varying dry yield of the extracts (0.22–1.92 g) that were obtained with solvents of variable polarities. However, the mitragynine content was consistent among the organic solvent extracts (7.22–9.40%). This method calls for a minimal extraction solvent volume (solid to solvent ratio, 1:30) and a shorter extraction period (20 min). Of the solvents tested, the methanolic extract showed the highest content of total phenolic (419.50 ± 2.50 GAE mg/g) and flavonoids (177.33 ± 3.00 QE mg/g). The extract was nontoxic towards kidney (HEK-293) and Chang liver (HeLa) cell lines. Analysis via UHPLC-ESI-QTOF-MS/MS made it possible to identify mitragynine congeners, such as mitragynine, paynantheine, and speciociliatine, in the leaves extract. In conclusion, the UAE method using methanol as the extraction solvent provides a noncytotoxic ketum botanical extract for future preclinical and clinical studies.
... Moreover, these alkaloid active ingredients play a very important role with antibacterial, anti-inflammatory, anti-viral, antioxidant, hypolipidaemic, central nervous system inhibition and anticancer activities. The alkaloids currently used in clinical medicine include morphine, ephedrine, vinblastine, paclitaxel, camptothecin and berberine for treatments such as analgesia, hypnosis, anaesthesia, cough and asthma, cancer cell inhibition, antibacterial and anti-inflammatory effects, and blood lipid lowering (Orio et al., 2012;Cushnie et al., 2014;Wang et al., 2014). Due to the rich content of alkaloid components coupled with the availability of raw materials and good biological activity, a large amount of scientific research has focused on the extraction of alkaloid components and their medical applications (Freire et al., 2010;Quijia and Chorilli, 2020). ...
... Alkaloids mainly exist in the form of free bases, salts, glycosides, amides and nitrogen oxides in plants, and the solvents used for their extraction are very different due to the structural differences of the various alkaloid forms (Orio et al., 2012;Cláudio et al., 2013;Hossain et al., 2014;Jiang et al., 2019). For example, alkaloids in their salt form are easily soluble in water and insoluble in organic solvents, but free alkalis have good solubility in organic solvents and are insoluble in water (Jiang et al., 2019;Quijia and Chorilli, 2020). ...
... To solve the above problems, it is necessary to design an appropriate solvent and combine it with the appropriate physical methods for enhanced extraction. Compared to conventional extraction methods, ultrasound-assisted extraction (UAE) has been continuously applied for the extraction of active plant compounds, and auxiliary methods have emerged that effectively reduce the extraction time, energy consumption and environmental pollution to greatly improve the extraction yield of active substances (Orio et al., 2012;Passos et al., 2014;Belwal et al., 2018). The UAE process is a scientific design based on the existence, polarity, solubility, etc. of the active ingredients and active ingredient groups in the plant substance. ...
Solvent application in industrial processes is inevitable due to its required role in dissolving solutes, promoting mass and heat transfer, and facilitating separation and purification processes. Notably, woody plants have a high degree of fibrosis and lignification with a mass transfer barrier effect from their multi-scale structure and complex component effects, which makes it difficult for solvents to seep into these plant cells. An ultrasound-assisted complete extraction in ionic liquid non-aqueous solvent system (UACE-ILNAS) method was successfully designed and applied to achieve excellent extraction of five alkaloid components (berberine, palmatine, jatrorrhizine, magnoflorine and phellodendrine) from the bark and leaves of Phellodendron amurense Rupr. From single-factor design and response surface methodology (RSM), the significant factors affecting the average yield of the alkaloids, the ionic liquid concentration, liquid-solid ratio and ultrasonic irradiation temperature, were optimized. Under the optimal conditions, the average yields of the alkaloids from the bark and leaves of P. amurense were 4.22 ± 0.20 and 2.47 ± 0.12 mg/g, respectively, which were highly consistent with the response surface optimization and kinetics analysis results. The non-aqueous aprotic solvent dimethyl sulfoxide (DMSO) was innovatively used as a co-solvent for the ionic liquid [C4mim][OAc]. The DMSO plays a positive role in promoting penetration, helping cell wall dissolve, reducing viscosity, etc. The proposed method overcomes the poor water solubility of isoquinoline alkaloids and the negative effects from the high viscosity of ionic liquids to effectively guarantee the alkaloid quality and yield. In addition, ultrasound plays a reinforcing role in achieving full extraction. Therefore, the developed method shows good application prospects for the separation of low-polarity constituents, especially those present in woody plants.
... McCurdy and co-workers obtained 34-39% of MG from ethanolic extraction combined with the acid-base treatment of Kratom leaves [39]. Applying ultrasonic irradiation (21.4 kHz, 70 W, 1 h, 25 • C) with methanol produced five times more MG than was obtained from common methanolic extraction (50 • C, 3 h) [50]. Ultrasonic irradiation with methanol was 1.3-1.7 more effective in extracting MG as well as 7-hydroxymitragynine (7-OHMG) compared with ethanol and acetonitrile [44]. ...
... acid-base treatment of Kratom leaves [39]. Applying ultrasonic irradiation (21.4 kHz, 70 W, 1 h, 25 °C) with methanol produced five times more MG than was obtained from common methanolic extraction (50 °C, 3 h) [50]. Ultrasonic irradiation with methanol was 1.3-1.7 more effective in extracting MG as well as 7-hydroxymitragynine (7-OHMG) compared with ethanol and acetonitrile [44]. ...
Doxorubicin is an effective chemotherapeutic agent in the treatment of solid hematological and non-hematological carcinoma. However, its long-term usage could result in side effects, such as cardiomyopathy, chronic heart failure, neurotoxicity and cancer cell resistance. In this study, we reported the sensitivity enhancement of A549 human lung cancer cells on doxorubicin at a low dose (0.1 ppm) in combination with 10–60 ppm of crude and alkaloid extracts derived from the leaves of Kratom (Mitragyna speciosa (Korth.) Havil. Rubiaceae). A549 cancer cell lines were insensitive to the crude extract containing low mitragynine (MG) (4–5%), while these cells were moderately inhibited by the alkaloid extract containing 40–45% MG (IC50 of 48–55 ppm). The alkaloid extract was found to inhibit A549 cancer cells via apoptosis as suggested by the higher relative fluorescence intensity with Annexin compared to that in propidium iodide (PI), i.e., a positive Annexin and a negative PI. The combination of crude extract and doxorubicin sensitized A549 cancer cells to doxorubicin by 1.3 to 2.4 times, while the combination with the alkaloid induced a 2.6- to 3.4-fold increase in sensitivity. The calculated combination index (CI) for doxorubicin with the crude and alkaloid extracts was 0.6 and 0.3, respectively, showing potential synergistic combinations to reduce the level of dosage of doxorubicin used in chemotherapy. In addition, the synergistic enhancement effect of crude extract on the cytotoxic activity of doxorubicin provides insights into the plausibility of non-alkaloids to influence the biological activities of Kratom.
... Although results were inconclusive, shaking seemed to improve precision [111]. Other assays [112] compared ultrasound assisted extraction (UAE), microwave-assisted extraction (MAE), and supercritical carbon dioxide extraction (SFE-CO 2 ) (using methanol, ethanol, water, and binary mixtures). Using LC/ESI-MS analysis, MAE (methanol:water, 1:1) gave the highest alkaloid fraction amount, while UAE showed the best yield for mitragynine. ...
... Using LC/ESI-MS analysis, MAE (methanol:water, 1:1) gave the highest alkaloid fraction amount, while UAE showed the best yield for mitragynine. The authors concluded that UAE (methanol:water) seemed to be the most effective method to obtain a large quantity of the alkaloid [112]. ...
The abuse of psychotropic substances is a well-known phenomenon, and many of them are usually associated with ancestral traditions and home remedies. This is the case of Mitragyna speciosa (kratom), a tropical tree used to improve work performance and to withstand great heat. According to several published studies, the main reasons for kratom consumption involve improving sexual performance and endurance, but also social and recreational uses for the feeling of happiness and euphoria; it is also used for medical purposes as a pain reliever, and in the treatment of diarrhea, fever, diabetes, and hypertension. However, this plant has gained more popularity amongst young people over the last years. Since it is available on the internet for purchase, its use is now widely as a drug of abuse, namely as a new psychoactive substance, being a cheaper alternative to opioids that does not require medical prescription in most countries. According to internet surveys by the European Monitoring Centre for Drugs and Drug Addiction in 2008 and 2011, kratom was one of the most widely supplied new psychoactive substances. The composition of kratom is complex; in fact, more than 40 different alkaloids have been identified in Mitragyna speciosa so far, the major constituent being mitragynine, which is exclusive to this plant. Besides mitragynine, alkaloids such as corynantheidine and 7-hydroxamitragynine also present pharmacological effects, a feature that may be attributed to the remaining constituents as well. The main goal of this review is not only to understand the origin, chemistry, consumption, and analytical methodologies for analysis and mechanism of action, but also the use of secondary metabolites of kratom as therapeutic drugs and the assessment of potential risks associated with its consumption, in order to aid health professionals, toxicologists, and police authorities in cases where this plant is present.
... Unfortunately, even though these fractions were very rich in alkaloids, their yields were very low and available only in very small quantities. According to the literature, SFE-CO 2 methods typically result in yields between 0.3% and 0.9%, and the mitragynine content is less than 2 mg/g in the extract by applying the following parameters: 243 g of leaf powder was extracted using 28.8% EtOH as a cosolvent at 65 • C, 300 bar, 12 kg/h of CO 2 flow, for 45 min [21]. Tohar et al. stated that the highest extraction yield, 1.56%, was achieved at 5000 psi (34.5 MPa) and 40 • C using a 60% ethanol modifier. ...
Mitragynine is an “atypic opioid” analgesic with an alternative mechanism of action and a favorable side-effect profile. Our aim was to optimize the alkaloid extraction procedure from kratom leaves and to determine and isolate the most relevant compounds capable of penetrating the central nervous system. The PAMPA-BBB study revealed that mitragynine and its coalkaloids, speciociliatine, speciogynine, and paynantheine, possess excellent in vitro BBB permeability. An optimized sequence of CPC, flash chromatography, and preparative HPLC methods was used to isolate the four identified BBB+ alkaloids. To improve the bioavailability of the isolated alkaloids, their cyclodextrin (CD) complexation behavior was investigated via affinity capillary electrophoresis using almost 40 CD derivatives. The apparent alkaloid–CD complex stability constants were determined and compared, and the most relevant CDs phase-solubility studies were also performed. Both the neutral and negatively charged derivatives were able to form complexes with all four kratom alkaloids. It was found that cavity size, substituent type, and degree of substitution also influenced complex formation. The negatively charged Sugammadex, Subetadex, and the sufoalkylated-beta-CD analogs were able to form the most stable complexes, exceeding 1000 M−1. These results serve as a good basis for further solubility and stability enhancement studies of kratom alkaloids.
... Alternative methods such as microwave-assisted extraction, ultrasound-assisted extraction, and supercritical fluid extraction (SFE) are efficient, fast and sustainable. However, their application has been limited due to the higher capital investment for complex equipment (Bligh and Dyer, 1959;Chang et al., 2017;Desgrouas et al., 2014;Zeković et al., 2017;Orio et al., 2012). ...
In this study a Machine Learning model was employed to predict the lipid profile from supercritical fluid extraction (SFE) of microalgae Galdieria sp. USBA-GBX-832 under different temperature (40, 50, 60°C), pressure (150, 250 bar), and ethanol flow (0.6, 0.9 mL min⁻¹) conditions. Six machine learning regression models were trained using 33 independent variables: 29 from RD-Kit molecular descriptors, three from the extraction conditions, and the infinite dilution activity coefficient (IDAC). The lipidomic characterization analysis identified 139 features, annotating 89 lipids used as the entries of the model, primarily glycerophospholipids and glycerolipids. It was proposed a methodology for selecting the representative lipids from the lipidomic analysis using an unsupervised learning method, these results were compared with Tanimoto scores and IDAC calculations using COSMO-SAC-HB2 model. The models based on decision trees, particularly XGBoost, outperformed others (RMSE: 0.035, 0.095, 0.065 and coefficient of determination (R²): 0.971, 0.933, 0.946 for train, test and experimental validation, respectively), accurately predicting lipid profiles for unseen conditions. Machine Learning methods provide a cost-effective way to optimize SFE conditions and are applicable to other biological samples.
... The leaves were pulverized in a multipurpose grinder (Thaigrinder model WF-04, Nonthaburi, Thailand) into a fine powder form (with a particle size of 1.0 mm). The resulting fine powder (100 g) was mixed with an extraction solvent at a concentration of 10% (w/v) [4]. The extraction solvent consisted of 1% of acetic acid in 80% ethanol. ...
This work aimed to test the efficacy of an ultrasound (US) and pulsed electric field (PEF) apparatus to extract mitragynine from dried Mitragyna speciosa cv. Karn Dang leaves. Four modes of the device were tested: PEF, US, US + PEF, and PEF + US, and the modes were compared using a conventional technique (maceration, as the control). The liquid chromatography/mass spectrometry (LC-MS/MS) analysis revealed that the mitragynine contents from M. speciosa leaves using the four different modes were significantly different (p < 0.05). The highest extraction (106.63 ± 0.85 mg/L) of mitragynine was obtained by the mode using a combination of PEF + US, followed by US + PEF (97.27 ± 1.33 mg/L), with increased extraction efficiencies of 45.81 ± 0.59% and 33.00 ± 1.85%, respectively. Moreover, the total energy consumption under the combination technique was 25.0% lower than that with PEF assistance. Scanning electron microscopy (SEM) and Fourier-transform infrared spectroscopy (FTIR) were used to analyze the structural and functional features of the alterations in M. speciosa leaves. This study demonstrated that a combination of PEF and US devices may be regarded as a green alternative technique and can assist in streamlining the implementation of agricultural products.
... MAE was more effective than UAE for the extraction of olive leaves [27] and gave higher extraction yields of TPC, TFC and tannins from Pistacia lentiscus leaves [28]. Comparative study of UAE, MAE, SFE-CO2 and classical methods for extraction of alkaloids from Mitragyna speciosa leaves showed that MAE gave higher alkaloid yields than other techniques [29]. For UAE, the principal mechanism of extraction is based on the collapse of bubbles which can produce physical, chemical, and mechanical effects and results in the disruption of biological membranes. ...
Supercritical fluid extraction (SFE) is an innovative green technology for the extraction of phytochemicals from plants. Therefore, this study aimed to evaluate the application of SFE and to optimize the extraction condition of the Thai herbal formula, Kleeb Bua Daeng (KBD). A Box-Behnken design (BBD) with response surface methodology was used to determine the effect of extraction time, temperature, and pressure on response variables including extraction yield, total phenolic content (TPC), total flavonoid content (TFC), total carotenoid content (TCC) and total anthocyanin content (TAC) of KBD formula. The highest percentage extraction yield, TPC, TFC, and TCC of the extracts were 3.81%, 464.56 mg gallic acid equivalents/g extract, 217.19 mg quercetin equivalents/g extract, and 22.26 mg β-carotene equivalents/g extract, respectively. The results indicated that SFE is a suitable method of extraction for green recovery of some phytochemicals from the KBD formula.
... A comparative study of UAE, MAE, SFE-CO2, and classical methods for extraction of alkaloids from Mitragyna speciosa leaves also showed that MAE gave higher alkaloid yield than other techniques [28]. ...
Supercritical fluid extraction (SFE) is an innovative green technology for the extraction of phytochemicals from plants. Therefore, this study aimed to evaluate the application of SFE and to optimize the extraction conditions of the Thai herbal formula, Kleeb Bua Daeng (KBD). A Box–Behnken design (BBD) with response surface methodology (RMS) was used to determine the effect of the extraction time (30–90 min), temperature (30–60 °C), and pressure (200–300 bar) on response variables including the extraction yield, total phenolic content (TPC), total flavonoid content (TFC), total carotenoid content (TCC), and total anthocyanin content (TAC) of the KBD formula. The highest percentage extraction yield (3.81%) was achieved at 60 °C, 300 bar, and 60 min of the extraction time. The highest TPC (464.56 mg gallic acid equivalents/g extract), TFC (217.19 mg quercetin equivalents/g extract), and TCC (22.26 mg β-carotene equivalents/g extract) were all achieved at 60 °C, 250 bar, and 90 min of the extraction time. On the contrary, it was not possible to quantify the total anthocyanin content as anthocyanins were not extracted by this method. The results indicated that SFE-CO2 is a suitable method of extraction for a green recovery of phytochemicals with low and moderate polarity from the KBD formula.
... The dried MS leaves extract of 9.96 g was obtained with a 19.92% percentage yield. Based on previous studies, the percentage yields of MS leaves extract were 15.0% (Sabetghadam et al., 2010), 18.0% (Yuniarti et al., 2020), 19.0% (Harizal et al., 2010) and 20.50% (Orio et al., 2012) respectively. The different results percentage may be prior to the extraction techniques and conditions of the extraction such as different temperatures and solvents which play a crucial role in the process efficiency and chemical composition of the extract. ...
Nanostructured lipid carriers (NLC) have evolved as effective vehicles for improving the bioavailability of poorly water-soluble drugs as well as protecting them from degradation and metabolism. The present study investigated the preformulation and its impact on different liquid lipids and solid lipids with diverse chemical structures and hydrophilic-lipophilic balance (HLB) for the critical quality attributes of Mitragyna speciosa leaves extract (MS)-based nanostructured lipid carriers suitable for drug delivery system. The presence of bioactive alkaloids (mitragynine, 7-hydroxymitragynine, speciogynine), flavonoids (quercetin, apigenin, kaempferol), saponins (daucosterol, quinovic acid 3-o-beta-d-quinovopyranoside, 1-o-feruloyl-beta-d-glucose), and other bioactive phytochemicals (chlorogenic acid, umbelliferone, ursolic acid) in MS leaves extract was detected using LCMS/MS analysis to support its medicinal properties. The total phenolic content, total flavonoid content, and DPPH assay of MS were 90.88 ± 0.30 GAE mg/g, 50.43 ± 0.47 QE mg/g, and IC50 at 0.0397 ± 0.0035 mg/mL, respectively. The screening preparation of NLC requires the selection of liquid lipid (oil), solid lipid, and surfactant. Based on the solubility test, the maximum solubility of MS leaves extract in oleic acid (liquid lipid) and compritol ATO 888 (solid lipid) were 2.859 ± 0.010% and 53.02 ± 0.39% respectively at ratio 70:30. Tween 80 was selected as the main surfactant for the preparation of NLC due to its highest emulsification efficiency (51.53%). The antimicrobial study showed that MS leaves extract gives antimicrobial effect towards Streptococcus aureus bacteria. The preformulation information collected offers a better, efficient, and cost-effective choice for further formulation development of MS leaves extract in drug delivery systems.
... Mitragynine is the most abundant indole alkaloid (11) consisting of 66% of the alkaloids content (11). 7-Hydroxymitragynine (7-HMG), an Ivyspring International Publisher active metabolite of mitragynine, is made up 0.04% of the alkaloid content of kratom leaves extracts (11)(12)(13). 7-HMG is formed as the break down product of mitragynine by the activity of CYP3A4 (14). In addition, 7-HMG may be formed post-harvest in kratom products that possibly contributing to increased reports of kratom toxicity in the Western world (14). ...
Background: Mitragyna speciosa Korth or Kratom is widely used traditionally for its medicinal values. The major alkaloid content of kratom leaves is mitragynine, which binds to opioid receptors to give opioid-like effects. This study aimed to analyse the brain proteome of animals that displayed addictive behaviors.
Design and Methods: Six groups (n=6-8) of rats made up of negative control, positive control using morphine (10 mg/kg), and treatment groups at low (1mg/kg) and high doses of mitragynine (30 mg/kg) for 1 and 4 days. The rats' behaviors were evaluated and subsequently the rats' brains were harvested for proteomic analysis that was performed by using 2D gel electrophoresis and LC/MS/MS.
Results: The rats developed physical dependence only on day 4 following morphine and mitragynine (1 and 30mg/kg) treatments. Among the proteins that were up-regulated in treatment groups were four calcium-binding proteins, namely calretinin, F-actin, annexin A3 and beta-centractin.
Conclusions: Upregulation of calretinin acted as low Ca²⁺ buffering upon the blockage of Ca²⁺ ion channel by mitragynine in the brain, which subsequently caused a reduction of GABA released and inversely increased the dopamine secretions that contributed to dependence indicators.
... This plant has been utilised for the treatment of musculoskeletal pain, diarrhoea, cough, and intestinal infection for centuries in this region, and due to its exceptional pharmacological properties, it has recently gained popularity in the Western world [12][13][14][15][16]. In addition, it is common knowledge that this plant possesses distinctive qualities, such as pigments that are rich in alkaloids and a shade of dark green that is brought about by the presence of chlorophyll pigments [17]. The chlorophyll content was reported to be proportional to the greenness of the leaves, which is the primary pigment involved in photosynthesis, with three other pigments playing supporting roles [18][19][20]. ...
This study investigates the MgSO4 salt compound as the dye additive for dye treatment in dye-sensitized solar cells (DSSCs). The properties of the chlorophyll-based dye molecule sensitizer extracted from Mitragyna speciosa (MS) under the treatment of MgSO4 additive that contributed to the highest power conversion efficiency, PCE were examined. Field emission scanning electron micrograph, FESEM, X-ray diffraction, XRD, Fourier transform-
IR, FTIR, UV-Vis spectroscopy, current to voltage characteristics, I-V and incidence photon to current efficiency,
IPCE were used to investigate the structural, optical, chemical and also the electrical properties of the DSSC cells and their components. It was discovered that the extracted MS dye contained chlorophyll pigment, a powerful light-harvesting pigment required for the production of charge transfer and electricity. The MgSO4-treated dye provides stable pH conditions, faster electron injection due to higher band energy, Eg position, higher dye molecules adsorption, and reduced back recombination of electrons in the fabricated DSSC. As a result, all of the treated MgSO4 dye cell DSSCs outperformed the untreated dye cell, and at their optimal dye additive of 0.3 g (3-TM), the DSSC’s current density, JSC, and associated PCE were higher at 2.08 mA/cm2 and 0.39%, respectively. Thus, using MS as a dye sensitizer in conjunction with an optimized MgSO4 compound as an additive resulted in improved photovoltaic effects, increased solar light absorption, and improved photon energy utilization.
... For example, it is employed in various processes including sterilization, preservation, extraction, cleaning, cutting, filtering and many others where ultrasound is shorten processing times and increase productivity [22][23][24]. Among numerous applications is the ultrasound-assisted extraction (UAE) technique which reduces the time required to extract edible oils [25][26][27]. ...
In engineering problems associated with acoustic wave propagation in a liquid, cavitation onset could be an adverse phenomenon, or, conversely, a required process. In both cases, knowledge of the ultrasonic parameters that lead to cavitation onset under given external conditions is relevant and necessary for solving both fundamental and practical problems. The present work proposes experimental results of studying the threshold of acoustic cavitation, which was implemented at different ultrasound frequencies with a change in external pressure, power of transducer and temperature of the liquid. The experiments were carried out for sunflower oil. The test findings demonstrated how the cavitation threshold changes with varying the power of ultrasound exposure in time. In addition, the effect of external pressure fluctuations on cavitation onset was investigated. The obtained results contribute to the understanding of cavitation processes and could be necessary for verification of theoretical models.
... This plant has been utilised for the treatment of musculoskeletal pain, diarrhoea, cough, and intestinal infection for centuries in this region, and due to its exceptional pharmacological properties, it has recently gained popularity in the Western world [12][13][14][15][16]. In addition, it is common knowledge that this plant possesses distinctive qualities, such as pigments that are rich in alkaloids and a shade of dark green that is brought about by the presence of chlorophyll pigments [17]. The chlorophyll content was reported to be proportional to the greenness of the leaves, which is the primary pigment involved in photosynthesis, with three other pigments playing supporting roles [18][19][20]. ...
the extracted MS dye contained chlorophyll pigment, a powerful light-harvesting pigment required for the production of charge transfer and electricity. The MgSO4-treated dye provides stable pH conditions, faster electron injection due to higher band energy, Eg position, higher dye molecules adsorption, and reduced back recombination of electrons in the fabricated DSSC. As a result, all of the treated MgSO4 dye cell DSSCs outperformed the untreated dye cell, and at their optimal dye additive of 0.3 g (3-TM), the DSSC's current density, JSC, and associated PCE were higher at 2.08 mA/cm2 and 0.39%, respectively. Thus, using MS as a dye sensitizer in conjunction with an optimized MgSO4 compound as an additive resulted in improved photovoltaic effects, increased solar light absorption, and improved photon energy utilization.
... In this research, chlorophyll based natural dye sensitizer was simulated. Green leaves are known to have unique characteristics that contain alkaloid pigments and a dark green colour [11], which is generally associated with chlorophyll (green) pigments [12]. An experiment done by L. Leenawaty et al. [13] reported that the higher the chlorophyll content in the leaves, the higher the green colour intensity of the leaves. ...
Dye-sensitized solar cell (DSSC) has been actively researched due to its lower fabrication cost. In DSSC, sensitizer imitates photosynthesis process in plants by absorbing the photon energy from the sunlight. This research investigated methanolic extracted chlorophyll dye sensitizer in comparison to water extracted and bare chlorophyll. Density Functional Theory (DFT) and Time-Dependent Density Functional Theory (TD-DFT) methods in Gaussian 03/Gaussview 03 software were used to investigate its optical and electronic properties. Each of the molecular structure (methanol and water) was individually attached to chlorophyll,
and their performances were determined. Results demonstrated that the absorbance spectrum of chlorophyll shows a broad peak in the visible range from 400 to 700 nm with its highest peak at 430 nm. Energy gap of chll-m between the E(LUMO) and the conduction band of TiO2 semiconductor, EC(TiO2) or so called as VOC is slightly smaller compared to water (chll-w). Meanwhile, the highest occupied molecular orbital (HOMO) and lowest unoccupied molecular orbital (LUMO) bandgap of chll-m is slightly higher at 2.13 eV. Finally, position of LUMO to redox electrolyte in chll-m is the lowest, which is favourable for faster electron transition. From observation, chll-m demonstrated better optical and electronic properties compared to its counterparts and hence good for chlorophyll sensitizer extraction.
... Compared with extraction without ultrasonication, ultrasonication can increase mitragynine yield by 16.88% with an optimum time of 10 min. The findings agreed with previous reports that ultrasound can improve extraction efficiency by increasing mitragynine content [4,15]. Ultrasonication produces a cavitation phenomenon, which means the production, growth, and collapse of bubbles that can determine mass transfer and accelerate the entry of solvents into plant cell materials. ...
Kratom (Mitragyna speciosa Korth) leaves contain a major alkaloid, mitragynine, which has opioid pain-relieving activity. Kratom and its derivatives are abused as alternatives to other narcotics. Therefore, this preliminary study was aimed to optimize the extraction of mitragynine from kratom investigating the effect of extraction time and ultrasonication on yield and quality of extract (the content of mitragynine). The extraction was carried out sequential extraction at room temperature and treatment of ultrasonication before extraction which was followed the purification process. Ultrasound-assisted extraction increased mitragynine yield to16.88%, compared to extraction without ultrasonication. The extract was purified by column chromatography, eluted with a mixture of n-hexane - ethyl acetate - 25% ammonia with a ratio of 30:15:1 (v/v). Likewise, the highest mitragynine purity obtained was 86.46%.
... 7-hydroxymitragynine (2) isospeciofoleine (3) speciofoline (6) corynoxine A (7) corynoxeine (8) rhynchophylline (9) ajmalicine (10) paynantheine (11) mitra-gynine (12) speciogynine (13) isopay-nantheine (14) specio-ciliatine (15) Other literature has established sonication as a rapid and useful method to extract alkaloids from M. speciosa [24,34]. This method allows a higher throughput of plant samples, which will prove useful in future analyses of many kratom plants and products. ...
Many consumers are turning to kratom (Mitragyna speciosa) to self-manage pain and opioid addiction. In the United States, an array of capsules, powders, and loose-leaf kratom products are readily available. Additionally, several online sites supply live kratom plants. A prerequisite to establishing quality control and quality assurance standards for the kratom industry, or understanding how alkaloid levels effect clinical outcomes, is the identification and quantitation of major and minor alkaloid constituents within available products and preparations. To this end, an ultra-high performance liquid chromatography-high resolution mass spectrometry method was developed for the analysis of 8 indole alkaloids (7-hydroxymitragynine, ajmalicine, paynantheine, mitragynine, speciogynine, isopaynantheine, speciociliatine, and mitraciliatine) and 6 oxindole alkaloids (isomitraphylline, isospeciofoleine, speciofoline, corynoxine A, corynoxeine, and rhynchophylline) in US-grown kratom plants and commercial products. These commercial products shared a qualitatively similar alkaloid profile, with 12 – 13 detected alkaloids and high levels of the indole alkaloid mitragynine (13.9 ± 1.1 – 270 ± 24 mg/g). The levels of the other major alkaloids (paynantheine, speciociliatine, speciogynine, mitraciliatine, and isopaynantheine) and the minor alkaloids varied in concentration from product to product. The alkaloid profile of US-grown M. speciosa “Rifat” showed high levels of the indole alkaloid speciogynine (7.94 ± 0.83 – 11.55 ± 0.18 mg/g) and quantifiable levels of isomitraphylline (0.943 ± 0.033 – 1.47 ± 0.18 mg/g). Notably, the alkaloid profile of a US-grown M. speciosa seedling was comparable to the commercial products with a high level of mitragynine (15.01 ± 0.20 mg/g). This work suggests that there are several M. speciosa chemotypes.
... CO 2 -SFE is a green technology that has evolved as a cost-effective, ecologically friendly, and scalable technique for the synthesis of essential oils and VOC-enriched extracts. SFE has a lower solvent recovery, heat degradation of molecules, and extraction time than other standard methods [23]. Side fractions characterized by VOCs combined with polyphenolic compounds and lipophilic compounds are formed during the extraction of VOCs from aromatic plant matrices [24]. ...
The antimicrobial and immunostimulant proprieties of aromatic plant extracts have been widely described, but their effects on serum blood biochemistry in fish have not. For this study, we assessed the changes in serum blood biochemical parameters in rainbow trout fed with a fish diet supplemented with a basil supercritical extract (F1-BEO). Our hypothesis was that treatment and time would be associated with changes in 10 serum blood biochemical parameters. F1-BEO was added to a commercial feed (0.5, 1, 2, 3% w/w). The fish were fed for 30 days, and the blood samples were collected at 2 time points (15 and 30 days). A two-way ANOVA showed a significant effect of treatment, time, and interaction treatment × time on creatinine, urea, total protein, albumin, magnesium, and phosphorus (p < 0.05), a significant effect of both time and interaction treatment × time on cholesterol (CHOL), glutamic oxaloacetic transaminase (GOT), and glutamic pyruvic transaminase (GPT) levels, and a significant effect of time on triglycerides (p < 0.05). While changes in several of the parameters were observed, the levels remained within the normal range for rainbow trout. Notably, after 30 days there was a significant decrease in CHOL in fish treated with 0.5% and 1% w/w F1-BEO (p < 0.05). The statistically non-significant increase in GOT and GPT in the fish fed with F1-BEO up to 3% w/w indicated a positive effect of basil on liver health. Our findings suggest a potential use for basil extracts (for example., F1-BEO) in fish feed to reduce antibiotic use and improve fish welfare.
... CO 2 -SFE is one of the green technologies that has emerged as an environmentally friendly, efficient and scalable process for the production of both oils and VOC-enriched extracts. Compared to other conventional methods, SFE has a lower solvent recovery, thermal degradation of molecules and extraction time [25]. However, during the extraction of VOCs from aromatic plant matrices, a series of fractions characterized by VOCs mixed with polyphenolic compounds and lipophilic compounds are produced [26]. ...
Phytotherapy is based on the use of plants to prevent or treat human and animal diseases. Recently, the use of essential oils and polyphenol-enriched extracts is also rapidly increasing in the aquaculture sector as a means of greater industrial and environmental sustainability. Previous studies assessed the antibacterial and antiparasitic effects of these bioactive compounds on fish. However, studies on the modulation of oxidative stress biomarkers are still scant to date. Thus, in this study, the modulation of antioxidant defense against oxidative stress exerted by fish diets supplemented with a basil supercritical extract (F1-BEO) was assessed in rainbow trout Oncorhynchus mykiss. The F1-BEO extracted with supercritical fluid extraction was added to the commercial feed flour (0.5, 1, 2, 3% w/w) and mixed with fish oil to obtain a suitable compound for pellet preparation. Fish were fed for 30 days. The levels of stress biomarkers such as superoxide dismutase, catalase, glutathione peroxidase, glutathione S-transferase, glutathione reductase, glyoxalase I, glyoxalase II, lactate dehydrogenase, glutathione and malondialdehyde showed a boost in the antioxidant pathway in fish fed with a 0.5% F1-BEO-supplemented diet. Higher F1-BEO supplementation led to a failure of activity of several enzymes and the depletion of glutathione levels. Malondialdehyde concentration suggests a sufficient oxidative stress defense against lipid peroxidation in all experimental groups, except for a 3% F1-BEO-supplemented diet (liver 168.87 ± 38.79 nmol/mg prot; kidney 146.86 ± 23.28 nmol/mg prot), compared to control (liver 127.76 ± 18.15 nmol/mg prot; kidney 98.68 ± 15.65 nmol/mg prot). Our results suggest supplementing F1-BEO in fish diets up to 0.5% to avoid potential oxidative pressure in farmed trout.
... MAE gave higher extraction yields of TPC, TFC and tannins from Pistacia lentiscus leaves . A comparative study of UAE, MAE, SFE-CO 2 and classical methods for extraction of alkaloids from Mitragyna speciosa leaves showed that MAE gave higher alkaloid yields than other techniques (Orio et al., 2012). MAE returned higher triterpene saponin contents from Xanthoceras sorbifolia Bunge than UAE and HRE (Li et al., 2010). ...
Ultrasound-assisted extraction (UAE) and microwave-assisted extraction (MAE) of the Thai herbal formula Kleeb Bua Daeng (KBD) were optimized separately and compared. A Box-Behnken design with response surface methodology was used to determine the effect of extraction time, temperature, solvent concentration, and material-to-solvent ratio for UAE and microwave power, extraction time, solvent concentration, and material-to-solvent ratio for MAE on response variables including extraction yield, total phenolic content (TPC), total flavonoid content (TFC), total carotenoid content (TCC) and total anthocyanin content (TAC) of KBD extracts. For UAE, the highest percentage extraction yield, TPC, TFC, TCC, and TAC of the extracts were 13.51%, 77.73 mg GAE/g extract, 35.40 mg QE/g extract, 17.27 mg β-carotene/g extract, and 0.33 mg cyanidin-3-glucoside/g extract, respectively. For MAE, the highest percentage extraction yield, TPC, TFC, TCC, and TAC were 18.95%, 476.61 mg GAE/g extract, 88.24 mg QE/g extract, 17.80 mg β-carotene/g extract, 0.97 mg cyanidin-3-glucoside/g extract, respectively. MAE extracts had higher yield, TPC, TFC, TCC, and TAC than UAE extracts and MAE appeared to be an efficient extraction method for this Thai herbal formula. This environmentally friendly extraction method could potentially be used to develop new KBD formulations with higher content of active ingredients.
... The percentages of mitragynine are found in different levels depend on the age of the plants, the environment in where it is grown and how it is cultivated [3,6]. Kratom leaves originated from Indonesia contained the highest level of mitragynine and alkaloid fraction related to mitragynine than those from Southern Thailand and Northern Malaysia [7]. Different varieties, according to the color of kratom leaf veins e. g. red or green kratom and geography are associated with the diversity of alkaloid distribution in each plant and thus impact to its effect on users [8,9]. ...
Objective: This study details the determination of mitragynine in various kratom samples using the thin-layer chromatography (TLC) technique and validation of analytical methods for quantifying the concentration of mitragynine in various kratom samples using liquid chromatography with photodiode array detector (LC-PDA). Methods: TLC technique using n-hexane: ethyl acetate: ammonia 25% (30: 15: 1 v/v/v) was applied to isolate mitragynine from kratom samples. Several interesting spots obtained were visualized under UV light at 254 nm. Samples were also prepared with organic solvent extraction directly prior to LC analysis (non-isolation method of preparation) to quantify the concentration of mitragynine. Mobile phases used were acetonitrile (MP A) and 0.1% formic acid in water (MP B). Samples and standards were run by gradient elution with a flow rate of 0.3 ml/min, detection using PDA detector at 254 nm. Results: Mitragynine was successfully isolated from kratom samples in Rf 0.50 by TLC system applied. The validation of analytical methods for mitragynine passed the acceptance criteria as described by UNODC Guidance. The concentration of mitragynine in various kratom samples seized in Indonesia ranged from 0.37%-1.70% (%w/w). Conclusion: Both TLC and LC analytical methods could be applied to determine and quantify the concentration of mitragynine in each examined sample, respectively.
... The fraction containing the mitragynine was collected and the solvent was removed using rotary evaporator. The purity of the mitragynine obtained was estimated to be 94.17% which was analysed using GC-MS (Orio et al., 2012). ...
Mitragynine is one of the dominant alkaloids present in Mitragyna speciosa. The compound possesses several pharmacological properties such as antinociceptive, anti-inflammatory, and anti-cancer. Studies have reported various methods in extracting mitragynine, both conventional and renewable technology combined with acid-base techniques for the enrichment and purification of mitragynine from M. speciosa. Several chromatography and spectroscopy instruments such as HPLC, LC-MS, GC-MS, and NMR have been used to identify mitragynine and its content in both the extract and fraction mixtures. In this review, we aim to provide insight on how the methods of extraction, purification, and identification of mitragynine have been developed over the last few decades. This report shows comparison among the various approaches in extracting mitragynine and points out the facts that different methods gave different yields of the compound.
... The fraction containing the mitragynine was collected and the solvent was removed using rotary evaporator. The purity of the mitragynine obtained was estimated to be 94.17% which was analysed using GC-MS (Orio et al., 2012). ...
Mitragynine is one of the dominant alkaloids presents in the Mitragyna speciosa which possesses several pharmacological properties such as antinociceptive, anti-inflammatory, and anti-cancer. Studies have reported various methods in extracting mitragynine, both conventional and renewable technology combined with acid-base techniques for the enrichment and purification of mitragynine from the extract of M. speciosa. Several chromatography and spectroscopy instruments such as HPLC, LC-MS, GC-MS, and NMR have been used to identify mitragynine and its content in both the extract and fraction mixtures. In this review, we aim to provide insight on how the methods of extraction, purification, and identification of mitragynine have been developed over the last few decades. This report also shows comparison among the various approaches in extracting mitragynine and points out the facts that different methods gave different yields of the compound.
... Extraction techniques, time, and type of solvents play a vital role in extracting targeted and non-targeted phytochemicals from plants [16,17]. For the first time, the accelerated solvent extraction (ASE) technique was employed to extract the kratom leaves. ...
Mitragyna speciosa Korth (kratom) is known for its psychoactive and analgesic properties. Mitragynine is the primary constituent present in kratom leaves. This study highlights the utilisation of the green accelerated solvent extraction technique to produce a better, non-toxic and antinociceptive active botanical extract of kratom. ASE M. speciosa extract had a dry yield (0.53–2.91 g) and showed a constant mitragynine content (6.53–7.19%) when extracted with organic solvents of different polarities. It only requires a shorter extraction time (5 min) and a reduced amount of solvents (less than 100 mL). A substantial amount of total phenolic (407.83 ± 2.50 GAE mg/g and flavonoids (194.00 ± 5.00 QE mg/g) were found in ASE kratom ethanol extract. The MTT test indicated that the ASE kratom ethanolic leaf extract is non-cytotoxic towards HEK-293 and HeLa Chang liver cells. In mice, ASE kratom ethanolic extract (200 mg/kg) demonstrated a better antinociceptive effect compared to methanol and ethyl acetate leaf extracts. The presence of bioactive indole alkaloids and flavonols such as mitragynine, paynantheine, quercetin, and rutin in ASE kratom ethanolic leaf extract was detected using UHPLC-ESI-QTOF-MS/MS analysis supports its antinociceptive properties. ASE ethanolic leaf extract offers a better, safe, and cost-effective choice of test botanical extract for further preclinical studies.
... Telah dilaporkan pula bahwa daun Kadamba mengandung senyawa metabolit sekunder berupa flavonoid, terpenoid, polifenol, glikosida, dan alkaloid sebagai senyawa mayor (mengandung 40 jenis alkaloid terutama mitraginin) (Ikhwan et al., 2018). Sejauh ini, penggunaan metode ekstraksi non-konvensional untuk proses ekstraksi metabolit sekunder target dari daun Kadamba pernah dilaporkan oleh (Orio et al., 2012). Akan tetapi, optimasi metode ekstraksi berbantu microwave dengan pelarut hijau terhadap kadar polifenol total dari daun Kadamba belum pernah dilaporkan. ...
Kadamba (Mitragyna speciosa Korth Havil) merupakan salah satu tumbuhan endemik di Asia Tenggara, di Indonesia, tumbuhan ini banyak di jumpai di pulau Kalimantan. Tumbuhan ini dipercaya oleh masyarakat dapat mengobati berbagai macam penyakit. Namun, karena kandungan alkaloidnya, tumbuhan ini dilarang digunakan sebagai bahan baku obat herbal oleh Badan Pengawas Obat dan Makanan. Oleh karena itu, perlu dilakukan pengembangan metode ekstraksi untuk menarik senyawa metabolit sekunder target dan meminimalkan senyawa yang tidak diinginkan yaitu dengan menggunakan metode ekstraksi berbantu mikrowave dengan pelarut hijau (Natural Deep Eutectic Solvent) yang dioptimasi menggunakan response surface methodology (RSM). Simplisia kering daun Kadamba diekstraksi menggunakan metode ekstraksi berbantu mikrowave dengan pelarut hijau (asam sitrat-glukosa) dengan berbagai kondisi ekstraksi dengan empat faktor dan tiga level (Box Behnken Design) yang dioptimasi menggunakan RSM dengan aplikasi perangkat lunak Design Expert versi 12 berlisensi. Penetapan kadar polifenol total dilakukan menggunakan reagen Folin-Ciocalteau dan diukur absorbansinya pada spektrofotometer UV-Vis pada panjang gelombang 770 nm, serta asam gallat sebagai standar. Berdasarkan hasil penelitian, diperoleh kondisi optimum pada perbandingan rasio pelarut NADES (asam sitrat : glukosa) 5:1 g/g, rasio pelarut-sampel sebesar 1:20 g/mL, waktu ekstraksi selama 20 menit dan kekuatan gelombang mikro sebesar 30% dengan kadar polifenol total prediksi sebesar 314,924 ±35,95 mg GAE/g sampel, sedangkan dari hasil proses verifikasi (scale-up confirmation) yaitu dengan meningkatkan jumlah sampel yang digunakan sebanyak sepuluh kali lipat dengan Kadar polifenol total diperoleh sebesar 427,12 ±35,95 mg GAE/g. Penelitian ini merupakan langkah awal dalam pengembangan metode ekstraksi untuk memperoleh senyawa target secara cepat, mudah, efisien, dan ramah lingkungan.
... Telah dilaporkan pula bahwa daun Kadamba mengandung senyawa metabolit sekunder berupa flavonoid, terpenoid, polifenol, glikosida, dan alkaloid sebagai senyawa mayor (mengandung 40 jenis alkaloid terutama mitraginin) (Ikhwan et al., 2018). Sejauh ini, penggunaan metode ekstraksi non-konvensional untuk proses ekstraksi metabolit sekunder target dari daun Kadamba pernah dilaporkan oleh (Orio et al., 2012). Akan tetapi, optimasi metode ekstraksi berbantu microwave dengan pelarut hijau terhadap kadar polifenol total dari daun Kadamba belum pernah dilaporkan. ...
The present study aims to optimize the natural deep eutectic solvent (NADES) as a green solvent-based microwave-assisted extraction (MAE) of total polyphenols content (TPC) from Mitragyna speciosa Korth Havil leaves using response surface methodology (RSM). Preparation of natural deep eutectic solvent (NADES) as a green solvent was performed by melting the two-component of malic acid and glucose using a magnetic stirrer. The leaves sample was extracted using the NADES-MAE method with various extraction conditions with four factors and three levels (Box Behnken Design) and optimized using RSM with licensed software of Design Expert V12. TPC was determined using a Folin-Ciocalteau reagent and absorbance was measured on a UV-VIS spectrophotometer at 770 nm and gallic acid as a standard. This study was obtained the optimum extraction conditions include: NADES ratio (malic acid: glucose) of 1:2 g/g, 50% microwave power, a solvent-sampel ratio of 12:1 mL/g, and extraction time for 7 minutes. The optimum conditions were obtained according to the equation formula: TPC = 165.17 – 33.97X1 + 38.36X2 – 6.08X3 + 16.12X4 – 62.77X1X2 + 79.61X12X2, where a R2 value = 0.7102 with a TPC prediction of 402.087±43.81 mg GAE/g sample. These conditions can be used to extract TPC from this plant efficiently, quickly, easily, and environmentally friendly.
... There is no work reported on % yield crude methanolic extract. However, Orio et al. [31] revealed that % crude extract in methanol: water, 1:1 was 24.8% where the conditions were at 25 °C for 1 hour. In the present study, the TFC of crude methanolic extract was similar with the previous study done by Parthasarathy et al. [23] which was 90.3 mg RE/g and 91.1 mg RE/g respectively. ...
In this study, Response surface methodology (RSM) was applied to optimize the yield of crude methanolic extract of Mitragyna speciosa leaves using Ultrasound-assisted extraction (UAE). The crude methanolic extract and its fractions were quantified in terms of total phenolic content and total flavonoid content, along with characterized using Fourier-transform infrared and Gas chromatography–mass spectrometry. The results showed the maximum yield of 49.72% at the optimal conditions (temperature, 34 °C; time, 25 min; and volume of solvent, 166 mL). The recovery crude methanolic extract for TPC and TFC were 137.3 ± 15.7 mg GAE/g and 90.3 ± 15.3 mg RE/g, respectively.
... An alkaloid extract was obtained from Maeng Da Micro Powder (MoonKratom, Austin, TX, USA) as described previously byOrio, Alexandru, Cravotto, Mantegna and Barge (2012). In brief, as shown inFigure S1A, extraction was performed by treating kratom powder in hloromethane. ...
Background and Purpose
Mitragyna speciosa, more commonly known as kratom, is a plant that contains opioidergic alkaloids but is unregulated in most countries. Kratom is used in the self‐medication of chronic pain and to reduce illicit and prescription opioid dependence. Kratom may be less dangerous than typical opioids because of the stronger preference of kratom alkaloids to induce receptor interaction with G proteins over β‐arrestin proteins. We hypothesized that kratom (alkaloids) can also reduce alcohol intake.
Experimental Approach
We pharmacologically characterized kratom extracts, kratom alkaloids (mitragynine, 7‐hydroxymitragynine, paynantheine, and speciogynine) and synthetic carfentanil‐amide opioids for their ability to interact with G proteins and β‐arrestin at μ, δ, and κ opioid receptors in vitro. We used C57BL/6 mice to assess to which degree these opioids could reduce alcohol intake and whether they had rewarding properties.
Key Results
Kratom alkaloids were strongly G protein‐biased at all three opioid receptors and reduced alcohol intake, but kratom and 7‐hydroxymitragynine were rewarding. Several results indicated a key role for δ opioid receptors, including that the synthetic carfentanil‐amide opioid MP102—a G protein‐biased agonist with modest selectivity for δ opioid receptors—reduced alcohol intake, whereas the G protein‐biased μ opioid agonist TRV130 did not.
Conclusion and Implications
Our results suggest that kratom extracts can decrease alcohol intake but still carry significant risk upon prolonged use. Development of more δ opioid‐selective synthetic opioids may provide a safer option than kratom to treat alcohol use disorder with fewer side effects.
... Kratom's pharmacological properties are affected by where, when and how it is cultivated. Plants from Indonesia contain higher levels of mitragynine and related alkaloids than those from Malaysia and Thailand (Orio et al., 2012). Kratom varieties with red-veined leaves (only found in Thailand) are reported to be more sedating than those with green or white leaves, which are more stimulating, according to drug user fora (Domingo et al., 2017). ...
Background
Kratom ( Mitragyna speciosa Korth) use has increased in Western countries, with a rising number of associated deaths. There is growing debate about the involvement of kratom in these events.
Aims
This study details the characteristics of such fatalities and provides a ‘state-of-the-art’ review.
Methods
UK cases were identified from mortality registers by searching with the terms ‘kratom’, ‘mitragynine’, etc. Databases and online media were searched using these terms and ‘death’, ‘fatal*’, ‘overdose’, ‘poisoning’, etc. to identify additional cases; details were obtained from relevant officials. Case characteristics were extracted into an Excel spreadsheet, and analysed employing descriptive statistics and thematic analysis.
Results
Typical case characteristics ( n = 156): male (80%), mean age 32.3 years, White (100%), drug abuse history (95%); reasons for use included self-medication, recreation, relaxation, bodybuilding, and avoiding positive drug tests. Mitragynine alone was identified/implicated in 23% of cases. Poly substance use was common (87%), typically controlled/recreational drugs, therapeutic drugs, and alcohol. Death cause(s) included toxic effects of kratom ± other substances; underlying health issues.
Conclusions
These findings add substantially to the knowledge base on kratom-associated deaths; these need systematic, accurate recording. Kratom’s safety profile remains only partially understood; toxic and fatal levels require quantification.
... M. speciosa preparations have been used by Malay and Thai native people for their opium-and coca-like effects in order to enhance the tolerance of physical labor in high-temperature conditions (Grewal, 1932a,b;Suwanlert, 1975;Tanguay, 2011), as well as an opium substitute in the treatment of opium addiction (Ahmad and Aziz, 2012;Beckett et al., 1965;Burkill, 1935;Burkill and Haniff, 1930;Tanguay, 2011;Vicknasingam et al., 2010;Wray, 1907a,b). The main indole alkaloids present in the young leaves of M. speciosa are mitragynine and its analogues, speciogynine, paynantheine, and speciociliatine (Leon et al., 2009;Orio et al., 2012;Ponglux et al., 1994;Shellard et al., 1978a,b). Mitragynine constitutes 66% of the total alkaloid mixture, making it the most abundant active alkaloid derived from the leaves of M. speciosa (Chittrakarn et al., 2008;Jansen and Prast, 1988;Shellard, 1974Shellard, , 1989Shellard et al., 1978a,b;Takayama et al., 1998). ...
Background:
Mitragynine is the major alkaloid of Mitragyna speciosa (Korth.) or Kratom, a psychoactive plant widely abused in Southeast Asia. While addictive effects of the substance are emerging, adverse cognitive effects of this drug and neuropharmacological actions are insufficiently understood.
Aims:
In the present study, we investigated the effects of mitragynine on spatial learning and synaptic transmission in the CA1 region of the hippocampus.
Methods:
Male Sprague Dawley rats received daily (for 12 days) training sessions in the Morris water maze, with each session followed by treatment either with mitragynine (1, 5, or 10 mg/kg; intraperitoneally), morphine (5 mg/kg; intraperitoneally) or a vehicle. In the second experiment, we recorded field excitatory postsynaptic potentials in the hippocampal CA1 area in anesthetized rats and assessed the effects of mitragynine on baseline synaptic transmission, paired-pulse facilitation, and long-term potentiation. Gene expression of major memory- and addiction-related genes was investigated and the effects of mitragynine on Ca2+ influx was also examined in cultured primary neurons from E16-E18 rats.
Results/outcomes:
Escape latency results indicate that animals treated with mitragynine displayed a slower rate of acquisition as compared to their control counterparts. Further, mitragynine treatment significantly reduced the amplitude of baseline (i.e. non-potentiated) field excitatory postsynaptic potentials and resulted in a minor suppression of long-term potentiation in CA1. Bdnf and αCaMKII mRNA expressions in the brain were not affected and Ca2+ influx elicited by glutamate application was inhibited in neurons pre-treated with mitragynine.
Conclusions/interpretation:
These data suggest that high doses of mitragynine (5 and 10 mg/kg) cause memory deficits, possibly via inhibition of Ca2+ influx and disruption of hippocampal synaptic transmission and long-term potentiation induction.
Mitragynine is a natural compound found in the leaves of the mitragyna speciosa tree, commonly known as kratom, which is primarily sourced from Southeast Asia. Mitragynine and its derivatives possess various pharmacological properties, including pain relief, anticancer effects, anti-inflammatory properties, anxiety reduction, stimulant properties, antidepressant effects, anti-addictive properties, antioxidant properties, and potential benefits for diabetes management. This review article highlights the methodologies of extraction techniques for isolating mitragynine, purification, characterization, biosynthesis, including the complete synthesis of mitragynine and its derivatives, and briefly summarizes their biological activities and toxicology of mitragynine. It is important to acknowledge that ongoing research is being conducted to explore the pharmacological effects of mitragynine, and further investigation is necessary to fully comprehend its impacts and potential medical applications. Moreover, the utilization of mitragynine and kratom remains contentious, with legal limitations imposed in numerous countries.
Mitragyna speciosa is a perennial plant native to Asia, well known for its psychoactive properties. Its major alkaloid mitragynine is known to have sedative and euphoric effects. Hence, the plant has been a subject of abuse, leading to addiction, necessitating efficient analytical methods to detect its psychoactive constituents. However, current chromatography‐based methods for detecting the alkaloids are time consuming and costly. Quantitative nuclear magnetic resonance (qNMR) spectroscopy emerges as a promising alternative due to its nondestructive nature, structural insights, and short analysis time. Hence, a rapid and precise qNMR method was developed to quantify selected major psychoactive alkaloids in various parts of M. speciosa . Mitragynine, specioliatine, and speciogynine were quantified in relation to the integral value of the ‐OCH 3 groups of the alkaloids and the internal standard 1,4‐dinitrobenzene. The precision and reproducibility of the method gave a relative standard deviation (RSD) of 2%, demonstrating the reliability of the method. In addition, the method showed excellent specificity, sensitivity, high linearity range ( R ² = 0.999), and limits of detection (LOD) and quantification (LOQ) values. The analysis revealed that the red‐veined M. speciosa leaves contained higher levels of mitragynine (32.34 mg/g), specioliatine (16.84 mg/g) and speciogynine (7.69 mg/g) compared to the green‐veined leaves, stem bark, or fruits.
This study aimed to develop an antioxidative compound extraction method for Kratom (Mitragyna speciosa) leaves using cellulase and pectinase, in combination and alone. The enzymatic hydrolysis-assisted process (EA) successfully enhanced antioxidant extraction from kratom leaves. The single enzymatic hydrolysis-assisted process (SEA) with 6% cellulase (C6) provided the highest extraction yield. The mixed enzymatic hydrolysis-assisted process (MEA) was then developed using C6, mixed with different concentrations of pectinase. Based on the high extraction yield, the MEA using 6% cellulase and 2% pectinase (C6+P2) was selected for further studies in comparison with that from relevant SEA and the control (without EA). Different extraction processes altered the characteristics and antioxidative activities of resulting extracts. The antioxidative activity of C6+P2 was dramatically enhanced in the gastrointestinal tract model system. Fourier transform infrared (FT-IR) spectrometer results confirmed significant content of phenolic compounds, proteins, and polysaccharides in C6+P2. From in vitro cytotoxicity study, C6+P2 showed an IC50 value of 22.86 and 4.76 µg/mL in RAW264.7 and Caco-2 cells, respectively. The bioactive compounds from C6+P2 should be identified in further studies to facilitate their application in the food and pharmaceutical industries.
This study aimed to investigate the efficiency of extraction procedures for Mitragyna speciosa Korth (M. speciosa) as well as the anti-inflammatory, cytotoxicity, and transdermal delivery potential of M. speciosa ethanolic extract. Extraction of M. speciosa leaves was conducted using three different methods: maceration, ultrasonic-assisted extraction (UAE), and UAE with maceration. The extraction yields obtained were 9.3–23.1% with a mitragynine content of 7.9–10.2%. Mitragynine remaining of M. speciosa extracts was greater than 97% after 30-d storage at various conditions. Cytotoxicity study towards various skin cells reveals that cell growth half inhibitory concentrations (IC50) of M. speciosa extract ranged from 1.4 ± 0.0 to 2.5 ± 0.2 mg/mL. M. speciosa extract exhibited dose-dependent inhibitory effects on nitric oxide production (IC50 = 147.8 ± 9.0 µg/mL) and suppressed pro-inflammatory cytokine expression levels. Ex vivo assessment using porcine ear skin reveals that skin permeation of mitragynine from M. speciosa extract was restricted with permeation flux ranging from 0.4 ± 0.1 to 11.6 ± 1.1 µg·cm−2·h−1 and skin deposition at 24 h ranged from 8.7 ± 2.6 to 60.5 ± 16.2 µg·cm−2. Vehicle composition and lipophilicity were found to be determinants of mitragynine permeation. The mixture of hydrophilic chemical enhancers—propylene glycol and diethylene glycol monoethyl ether—as M. speciosa extract vehicle yielded the highest skin permeability coefficient of mitragynine. These findings offer valuable information that can be utilized for the transdermal and/or topical product development of mitragynine and M. speciosa extract.
In this research, response surface methodology was employed in order to find the most efficient conditions to produce peanut extracts with the highest antioxidative potential (by DPPH, ABTS, FRAP and ORAC tests) from ultrasound-assisted (UAE) and microwave-assisted (MAE) extractions. The optimal conditions for UAE were 75 mL of 30%v/v ethanol as extraction solvent, extraction temperature of 65°C, and 180 min extraction time. Optimal conditions for MAE were 15 mL of 30%v/v ethanol as extraction solvent, extraction temperature of 80°C and 90 s extraction time. The generated models presented a reliable antioxidative response by showing non-difference between experimental and predicted values within a 95% confidence level. Characteristics such as antioxidant activities, TPC, resveratrol, and TFC, were used to compare the effectiveness of the obtained extracts and the SLE extract from our previous work. The UAE extract displayed higher extraction efficiency than that of MAE in terms of higher levels of all responses (p<0.05). Slightly lower antioxidant values of MAE might be due to the limited solvent volume used, which was 5 times less than that of UAE. Moreover, the high efficacy of MAE was confirmed by its extraction time which was 120 times shorter than that of UAE. Ultimately, the correlation analysis implied that the antioxidant activity of peanut extracts was contributed by TPC rather than resveratrol or TFC, and the revealed TPC in this study was higher than previous reports.
This article presents comprehensive information on applications and developments of supercritical fluid extraction (SFE) technique in plant oils and herbal volatile components analysis. Supercritical carbon dioxide (SC‐CO 2 ) is widely used for extraction and isolation of volatile and essential oils of plants. SFE possesses valuable advantages in the extraction of various analytes, especially nonpolar compounds, from plant matrices. Meanwhile, some modifications such as the usage of modifiers make this technique applicable for the extraction of polar targets as well. SFE in herbal field is used for the extraction of a wide range of analytes including food, agricultural agents, pharmaceuticals, cosmetics, and pesticides. This article aims to review, from plant oil analysis standpoint, the recent advances in the use of SC‐CO 2 for the extraction and separation of herbal analytes. The use of experimental designs, including screening and optimizing, in the SFE process for the extraction of plant oils is described. In addition, it presents a critical appraisal about the advantages and disadvantages of SFE technique in comparison with traditional extraction methods, that is Soxhlet, hydrodistillation (HD), and head space extraction. The abilities, advantages, shortcomings, and prospects of SFE for isolation and extraction of volatile components and essential oil of plant materials are also discussed.
Ethnopharmacological relevance:
The use of herbal tea infusions is widespread in ethnomedicine throughout the world. One such ethnobotanical is kratom (Mitragyna speciosa Korth., Rubiaceae) which has gained considerable interest as an herbal supplement in recent years in the West beyond its native Southeast Asia. Traditional, kratom leaves are either chewed fresh or made into a tea infusion to treat fatigue, pain, or diarrhea. However, dried kratom leaf powder and hydroalcoholic extracts are more commonly used in Western countries, raising the question of exposure to kratom alkaloids and related effects.
Aim of the study:
A specific kratom tea bag product was analyzed for mitragynine content using tea infusion preparation and methanolic extraction. Consumers of both the tea bag product and other kratom products completed an online anonymous survey to determine demographics, kratom use patterns, and self-reported beneficial and detrimental effects.
Materials and methods:
Kratom tea bag samples were extracted using pH-adjusted water or methanol and analyzed using an established LC-QTOF method. A modified kratom survey was distributed to consumers of the kratom tea bag products and other kratom products over a 14-month period.
Results:
Tea infusion extraction of tea bag samples resulted in lower mitragynine levels (0.062-0.131% (w/w)) compared to methanolic extraction (0.485-0.616% (w/w)). Kratom tea bag consumers did report similar, although often milder beneficial effects compared to consumers using other kratom products. Overall self-reported health was better among kratom tea bag consumers whereas improvement of a diagnosed medical condition was less in tea bag consumers compared to those using other kratom products.
Conclusions:
Traditional tea infusions of Mitragyna speciosa dried leaves provide benefits to consumers despite substantially lower mitragynine content. These effects may be less pronounced but indicate that tea infusions provide a potentially safer formulation compared to more concentrated products.
Mitragyna speciosa or kratom is emerging worldwide as a "legal" herbal drug of abuse. An increasing number of papers is appearing in the scientific literature regarding its pharmacological profile and the analysis of its chemical constituents, mainly represented by alkaloids. However, its detection and identification are not straightforward as the plant material is not particularly distinctive. Hyphenated techniques are generally preferred for the identification and quantification of these compounds, especially the main purported psychoactive substances, mitragynine (MG) and 7-hydroxymitragynine (7-OH-MG), in raw and commercial products. Considering the vast popularity of this recreational drug and the growing concern about its safety, the analysis of alkaloids in biological specimens is also of great importance for forensic and toxicological laboratories. The review addresses the analytical aspects of kratom spanning the extraction techniques used to isolate the alkaloids, the qualitative and quantitative analytical methods and the strategies for the distinction of the naturally occurring isomers.
Introduction:
Aconitum spp. are prime medicinal plants rich in alkaloids and have been used as the main constituents of traditional medicine in India and China. The whole plant can be toxic and creates pathophysiological conditions inside the human body. Therefore, simultaneous quantification of alkaloids within plant parts and herbal medicines associated with this genus is essential for quality control.
Objective:
We aimed to develop and validate methods using ultra-high-performance liquid chromatography-diode array detector-quadrupole time-of-flight ion mobility mass spectrometry (UHPLC-DAD-QTOF-IMS) and to develop an analytical strategy for the identification and quantification of alkaloid compounds (aconitine, hypaconitine, mesaconitine, aconine, benzoylmesaconitine, benzoylaconine, bulleyaconitine A, and deoxyaconitine) from Aconitum heterophyllum.
Methodology:
We developed a simultaneous identification and quantification method for eight alkaloids using UHPLC-DAD-QTOF-IMS. The method was validated as per International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use (ICH) guidelines and also in IMS mode.
Results:
The developed method has good linearity (r2 = 0.997-0.999), LOD (0.63-8.31 μg/mL), LOQ (0.63-2.80 μg/mL), recovery (86.01-104.33%), reproducibility, intra- and inter-day variability (<3.25%), and stability. Significant qualitative and quantitative variations were found among different plant parts (flower, leaf, stem, root, and tuber) and five market products of A. heterophyllum. Furthermore, a total of 21 metabolites were also profiled based on the fragmentation pattern of MS2 using the validated method.
Conclusion:
An appropriate mobile phase using acetonitrile and water in a gradient elution gave a satisfactory chromatographic separation of eight Aconitum alkaloids with their adjacent peaks. Therefore, this method could provide a scientific and technical platform for quality control assurance.
Natural product-based drug discovery necessitates highly interdisciplinary approaches. The benefits of phytochemicals for the human health are countless. Modern green extraction procedures—such as the use of cavitational reactors or the green applications of supercritical CO2 and subcritical water—have revolutionized production methods improving the quality of the products and reducing or eliminating the use of nonbenign solvents, thus making the large-scale obtainment of these compounds more sustainable.
The possibility of computationally studying natural products and analysing the relationship between structure and activity provides a means for a preliminary virtual screening to identify promising lead compounds in early stages. Metabolomics screening and high throughput screening in silico further facilitate this identification. Besides the advances achieved in academic investigation, several companies have invested in new technologies and in preliminary computational predictions of structure-activity relationships.
The Mediterranean natural bio- and chemo-diversity offer an inexhaustible library of natural products that can act as sources of new lead bioactive compounds. The success of the Mediterranean diet was a major driving force to thoroughly investigate the pharmacological potential of enriched and purified fractions from Mediterranean plants. This chapter presents an overview of the medicinal properties of selected Mediterranean plants and highlights the role of computational studies in the understanding of the mechanisms of action of their active compounds and the role of green extraction techniques to guarantee high-quality products. The combination of these strategies is expected to pave the way for large commercial scale-up and significant payback on capital investment in designing Mediterranean plant-based products.
Mitragyna speciosa (K.) H. (Kratom) is a tree that possesses stimulant and opioid-like analgesic effects, and is indigenous to Southeast Asia and Indochina, where it has seen widespread use for hundreds of years. The principal pharmacologically active alkaloids in kratom leaves include mitragynine (MG), 7-hydroxymitragynine (HMG), speciociliatine (SC), speciogynine (SG) and paynantheine (P). The pharmacological effects induced and their potency can vary dramatically according to variations in the proportions of alkaloid compounds present, which are related to geographic origin, stage of maturity and ecotype. Much of the analgesic and opiate-like psychoactive effect of kratom has been associated with the MG and HMG detected in M. speciosa (K.). H. Five different strains of M. speciosa (K.) H., which present differing vein colours and geographic origin, have been studied herein; red vein strains from Thailand, Malaysia and Bali, named Red Thai, Red Malay and Red Bali, a white vein strain from Borneo (White Borneo) and a green vein strain from Malaysia (Green Malay) were included in the study. Plant leaves were extracted under magnetic stirring at room temperature in a MeOH/H2O 1:1 mixture. Purified alkaloids were isolated in a number of organic extraction steps, from either aqueous basic or acidic phases, that culminated in precipitation (yields between 0.94 and 1.43%). These samples have been analysed using HPLC-DAD, HPLC-MS, HPLC-MS/MS and GC-MS to optimize the identification and quantification of the principal alkaloids present in the different strains. 24 alkaloids were detected in Red Bali whereas 11 compounds were found in the other varieties. Red Thai, Red Bali, Green Malay and White Borneo strains had a higher w/w percentage for MG than for P, while P was more abundant in Red Malay. The Green Malay variety (GMK) showed the highest w/w percentages for MG and total alkaloids in its extracts (59.7 and 94.9% respectively). The Green Malay variety was therefore chosen for in vivo pharmacological studies. The Green Malay extract has shown remarkable and significant antinociceptive and anti-inflammatory activity in mouse hot plate and carrageenan-induced paw edema tests.
To study the application value of sweet cherry leaves before abscission, a supercritical carbon dioxide (SFE-CO2) extraction method was established for sweet cherry (Prunus avium L.) leaves. The extraction temperature, pressure and time were optimized with a Box-Behnken design, and the optimal conditions were 43 °C, 30 MPa, and 120 min, resulting in a yield of 2.52 ± 0.261% (w/w), which agrees well with the predicted value. The yield of the sweet cherry leaf extracts obtained by methanol solvent extraction is 2.03% (w/w); the SFE-CO2 extraction method was more efficient and had a higher yield than the methanol solvent extraction method, and the adverse effects of organic solvent residues and high temperatures on the product during traditional solvent extraction were avoided, resulting in a higher quality product. UPLC-MS/MS in negative ion mode provided 56 identifiable chromatographic peaks, including those of 31 acids (13.25%), 5 sugars (15.94%), 2 alcohols (6.19%), and 18 other compounds (3.86%). The fragmentation pathways of the 7 main components in the sweet cherry leaf extract were identified. The carbohydrates and bioactive substances in the extracts obtained from sweet cherry leaves suggested the potential use of sweet cherry leaves in the food and medical industries.
This review presents innovative extraction techniques and their role in promoting sustainable ingredients for the food, cosmetic and pharmaceutical industries. These techniques (such as microwave, ultrasound, pulse electric field, instant controlled pressure drop, sub- and super-critical fluid processing, extrusion, mechanochemistry, high pressure, and ohmic, UV and IR heating) use or produce less solvent, energy, and hazards. This review will provide the necessary theoretical background and some details about green extraction techniques, their mechanisms, some applications, and environmental impacts. We will pay special attention to the strategies and present them as success stories for research and education and at the industrial scale.
Plants represent a huge reservoir of bioactive molecules that are still little explored. Modern science is now focused on identifying the beneficial compounds from these sources to be used in pharmaceutical drugs. The search for new bioactive natural molecules is based on the choice of extraction, separation and structural identification techniques. This chapter provides a brief review of the specificity and usefulness of the most popular extraction methods in the phytochemistry field. In addition, this study describes the development of chromatographic separation and structural elucidation techniques of secondary metabolites, including analytical and preparative chromatography, mass spectrometry (MS), one- (1D) and two-dimensional (2D) Nuclear magnetic resonance (NMR) experiences. Today, the immense progress of these techniques has enabled chemists, biologists and pharmacists to discover bioactive molecules that have found applications in drug and food industries.
Ethnopharmacological relevance:
Ethnopharmacological knowledge is important for the identification of active compounds from natural products. Pain may have different aetiologies with complex mechanisms. Tabernaemontana catharinensis A. DC. is well known for indole alkaloids, being used empirically in folk medicine, with antimicrobial and anti-inflammatory as well as antiofidic actions among others.
Aim of the study:
This work aims to evaluate the antinociceptive and antioxidant effect in mice of the alkaloids extract from leaves of Tabernaemontana catharinensis A. DC. (AITc).
Materials and methods:
The AITc was produced by ultrasound and acid-base extraction, and the chemical composition was evaluated by high resolution mass spectrometry. Male mice (Mus musculus), Swiss, were used for in vivo tests. The AITc was administrated at doses of 1.0, 5.0, and 10.0 mg/kg in acetic acid model, formalin, tail-immersion, hot plate, and open field tests, and compared to saline, morphine, or diazepam controls, depending on the test. The toxicological, biochemical, haemogram and antioxidant effect were evaluated in mouse organs such as liver, brain, kidneys, spleen and stomach.
Results:
In total, 10 compounds were identified in the AITc, being from the indole alkaloids from the ibogan and corynanthean classes. The extract in doses ranging from 5.0 to 10.0 mg/kg showed an antinociceptive effect for acetic acid, inhibiting by 47.7% and 61.6%. In the same line, reductions of 47.1% (first phase) and 43.6% (second phase) were observed for the 5.0 mg/kg dose in the formalin test. However, tail-immersion and hot plate tests did not show considerable modifications in the latency period, while in the open field test there was an inhibition of only 5.1%. It was observed no differences in NO levels and total antioxidant status of the mice in any of the studie tissues.
Conclusions:
The results justify the use of this plant in traditional medicine. in vivo tests indicate that these compounds possess central and peripheral mechanisms of action. This is study that reports the nociceptive action of these alkaloids, also including toxicity tests, which are intended to guarantee the safety of use of extracts of this plant.
The leaves of Mitragyna speciosa were chewed as a substitute to opium in Thailand and Malaysia. A study was therefore undertaken to compare the antinociceptive activity of morphine and paracetamol to that of mitragynine, a major constituent of fresh leaves of M. speciosa. The tests employed were acetic acid induced writhings, hot tail-flick and cold tail flick. All test drugs were administered orally to mice. Results indicated that mitragynine (200 mg/kg) and morphine (5 mg/kg) reduced writhings from 17.5 ± 2.8 per 5 min to 9.6 ± 0.6 and 7.3 ± 0.6, respectively. Paracetamol (100 mg/kg) did not significantly reduce writhings in mice. All three drugs produced significant analgesia when tested by the hot tail-flick producing peak maximum possible analgesia (MPA) of 35.1 ± 4.8% (morphine), 27.8 ± 2.1% (paracetamol) and 42.8 ± 7.2% (mitragynine). Morphine produced significant and marked analgesia when tested by the cold tail-flick technique achieving a peak MPA of 66.2 ± 2.4% at 45 min following oral administration. Mitragynine produced a peak MPA of 49.0 ± 5.9% at 30 min whilst paracetamol did not appear to be active. Mitragynine may be a potential new analgesic and requires further study.
The leaves of Mitragyna speciosa, a tropical plant known as “kratom,” have been traditionally used as a substitute for opium in Thailand and Malaysia. Mitragynine,
a major constituent of M. speciosa, has an opioid agonistic activity, and its derivative 7-hydroxymitragynine (7-OH-mitragynine) (a minor constituent) is much
more potent than mitragynine or morphine. Recently, many products containing this plant have been distributed as “incense”
on the drug market in Japan for their expected narcotic effects. Despite their potency and their wide distribution for abuse,
there are no reports on the quantitative analysis of mitragynine and 7-OH-mitragynine in the raw materials or in the commercial
products of kratom. In this study, a method for simultaneous analysis of mitragynine, 7-OH-mitragynine, and other indole alkaloids
(speciogynine, speciociliatine, and paynantheine), present in the raw materials and commercial products of kratom, was developed
using liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS). By this method, mitragynine, 7-OH-mitragynine,
and the other alkaloids were detected in 11 of the 13 products. The content of mitragynine in the products ranged from 1%
to 6%, and that of 7-OH-mitragynine from 0.01% to 0.04%. Because 7-OH-mitragynine is much more potent than morphine, M. speciosa abuse is a matter of major concern. The present analytical method is considered useful for the screening of M. speciosa products in the drug market.
In Africa, Mitragyna inermis (Willd.) O. Kuntze (Rubiaceae) is commonly used in traditional medicine to treat malaria. Antimalarial activity is mostly due to the hydromethanolic extract of M. inermis leaves and especially to the main alkaloids, uncarine D and isorhynchophilline. In the present study, we describe for the first time an HPLC method for the simultaneous quantification of uncarine D and isorhynchophylline in biological matrices. SPE was used to extract the components and the internal standard naphthalene from human and pig plasma samples. Chromatographic separation was performed on a C-18 reversed column at a flow rate of 1 mL/min, using methanol-phosphate buffer (10:90, pH 7), as a mobile phase. Good linearity was observed over the concentration ranges of 0.0662-3.31 microg/mL for uncarine D and 0.0476-2.38 microg/mL for isorynchophylline. The precision was less than 12% and the accuracy was from 86 to 107% without any discrepancy between the two species. Uncarine D and isorhynchophylline recoveries were over 80%. These results allowed the quantification of both uncarine D and isorhynchophylline in pig plasma after intravenous administration of M. inermis extract.
Studies on the antioxidant and antimicrobial activities of Mitragyna speciosa leaf extracts are lacking. In this study the antioxidant properties of water, methanolic and alkaloid M. speciosa leaf extracts were evaluated using the DPPH (2,2-diphenyl-1- picrylhydrazyl) radical scavenging method. The amount of total phenolics and flavanoid contents were also estimated. The DPPH IC(50) values of the aqueous, alkaloid and methanolic extracts were 213.4, 104.81 and 37.08 microg/mL, respectively. The total phenolic content of the aqueous, alkaloid and methanolic extracts were 66.0 mg, 88.4, 105.6 mg GAE/g, respectively, while the total flavanoid were 28.2, 20.0 and 91.1 mg CAE/g respectively. The antioxidant activities were correlated with the total phenolic content. This result suggests that the relatively high antioxidant activity of the methanolic extract compared to aqueous and alkaloid extract could be possibly be due to its high phenolic content. The aqueous, alkaloid and methanolic extracts were screened for antimicrobial activity. The extracts showed antimicrobial activity against Salmonella typhi and Bacillus subtilis. The minimum inhibitory concentrations (MICs) of extracts determined by the broth dilution method ranged from 3.12 to 6.25 mg/mL. The alkaloid extract was found to be most effective against all of the tested organisms.
Ultrasound and microwave techniques were used to extract tobacco alkaloids, and response surface methodology was used to optimize extraction conditions. Ultrasonic technique factors were temperature, 30-85 degrees C; time, 3-45 min; solvent volume, 8-80 mL. Microwave extraction factors were pressure, 15-75 psi; time, 3-40 min; power, 30-90% of the maximum magnetron power of 650 W. Soxhlet and solvent AOAC-modified extraction methods were also applied after some improvements. Nicotine, nornicotine, anabasine, and anatabine were quantified by gas chromatography. A steam distillation International Standards Organization method for total alkaloid evaluation was used as reference. The results obtained by the different methods were compared using a least squares deviation test. The ultrasonic and the proposed modified-AOAC extraction method were the more convenient with regard to practicability and precision. The relative deviations (n = 5) were as follows: For the ultrasonic method in low-level alkaloid tobaccos, 0.7% nicotine and 1.4-14% minor alkaloids; in high-level alkaloid tobaccos, 2.4% nicotine and 4.5-5.1% minor alkaloids. For the modified AOAC method in low-level alkaloid tobaccos, 0.9% nicotine and 2.4-11.6% minor alkaloids; and in high-level alkaloid tobaccos, 1.7% nicotine and 2.0-2.4% minor alkaloids.
Acute administration of Mitragyna speciosa (MS) extract (45 and 50 mg/kg) significantly resulted in dose-dependent decreases in food and water intakes (P<0.05) in rats. Prolonged suppressing effects were observed following administration of the MS extract (40 mg/kg) for 60 consecutive days. Moreover, the long-term administration also significantly suppressed weight gaining.
Ultrasound-assisted extraction (UAE) and microwave-assisted extraction (MAE) techniques have been employed as complementary techniques to extract oils from vegetable sources, viz, soybean germ and a cultivated marine microalga rich in docosahexaenoic acid (DHA). Ultrasound (US) devices developed by ourselves, working at several frequencies (19, 25, 40 and 300 kHz), were used for US-based protocols, while a multimode microwave (MW) oven (operating with both open and closed vessels) was used for MAE. Combined treatments were also studied, such as simultaneous double sonication (at 19 and 25 kHz) and simultaneous US/MW irradiation, achieved by inserting a non-metallic horn in a MW oven. Extraction times and yields were compared with those resulting from conventional procedures. With soybean germ the best yield was obtained with a 'cavitating tube' prototype (19 kHz, 80 W), featuring a thin titanium cylinder instead of a conventional horn. Double sonication, carried out by inserting an immersion horn (25 kHz) in the same tube, improved the yield only slightly but halved the extraction time. Almost comparable yields were achieved by closed-vessel MAE and simultaneous US/MW irradiation. Compared with conventional methods, extraction times were reduced by up to 10-fold and yields increased by 50-500%. In the case of marine microalgae, UAE worked best, as the disruption by US of the tough algal cell wall considerably improved the extraction yield from 4.8% in soxhlet to 25.9%. Our results indicate that US and MW, either alone or combined, can greatly improve the extraction of bioactive substances, achieving higher efficiency and shorter reaction times at low or moderate costs, with minimal added toxicity.
This study used for the first time LC–MS/MS for the analysis of mitragynine (MIT), a mu-opioid agonist with antinociceptive and antitussive properties, in rat plasma. Mitragynine and the internal standard (amitriptyline)were extracted from plasma with hexane:isoamylalcohol and resolved on a Lichrospher®
RPSelectB column (9.80 and 12.90 min, respectively). The quantification limit was 0.2 ng/mL within a
linear range of 0.2–1000 ng/mL. The method was applied to quantify mitragynine in plasma samples of
rats (n = 8 per sampling time) treated with a single oral dose of 20mg/kg. The following pharmacokinetic
parameters were obtained (mean): maximum plasma concentration: 424 ng/mL; time to reach maximum
plasma concentration: 1.26 h; elimination halflife: 3.85 h, apparent total clearance: 6.35 L/h/kg, and apparent volume of distribution: 37.90 L/kg.
Three new monoterpenoid indole alkaloids, i.e., 3,4,5,6-tetradehydromitragynine, mitralactonal, and mitrasulgynine carrying a sulfonate function, were isolated, together with seven known compounds, from the leaves of Mitragyna speciosa native to Malaysia.
Over the centuries mankind has benefited from the natural materials that occur in plants. In earlier times the whole plant or an extract was used in cooking or as a medicine but nowadays the active constituents of plant extracts provide targets for the synthetic chemist. In this review we will explore the advantages that accrue from the incorporation of either ultrasound or microwaves in the extraction process. The two techniques offer different approaches in that ultrasound is generally used to improve conventional solvent extraction whereas microwaves are known for their ability to remove constituents via heating without solvents.
A new Corynanthe-type indole alkaloid, (−)-9-methoxymitralactonine (1), having a highly conjugated system was isolated from the young leaves of Mitragyna speciosa in Malaysia, and its structure was first deduced by spectroscopic analysis and then confirmed by chiral-total synthesis starting from optically pure epoxy-ketone and 5-methoxy-3,4-dihydro-β-carboline. The chiral HPLC analysis demonstrated that the natural 9-methoxymitralactonine contained predominantly the (−)-enantiomer over the (+)-enantiomer in the ratio of 62:38.
Mitragyna speciosa Korth (ketum) is widely used in Malaysia as a medicinal agent for treating diarrhea, worm infestations and also acts as an analgesic and antipyretic.
The aim of the study is to determine the acute toxicity of Mitragyna speciosa Korth standardized methanol extract in vivo in 4-weeks-old Sprague-Dawley rats.
Rats were orally administrated single dose of 100, 500 and 1000 mg/kg Mitragyna speciosa Korth standardized methanol extract and the control group received 430 mg/kg of morphine orally. There were 10 rats in each group. All animals were sacrificed after 14 days of treatment. Eight parameters were tested: cage side observation, body weight measurement, food and water consumption, blood pressure, absolute and relative organ weight, hematology, biochemical analysis and histopathology, to look for evidence of toxicity.
No mortality was noted after 14 days of treatment. In general, behavior, food and water consumption, hematological studies and organ weights showed no significant changes. The standardized methanol extraction of Mitragyna speciosa Korth increased rat blood pressure (systolic: 147.4+/-1.01, 131.64+/-4.94 and 137.8+/-4.46) after an hour of 100, 500 and 1000 mg/kg doses, respectively. Biochemical studies showed significant elevation of ALT, AST, albumin, triglycerides, cholesterol and albumin (p>0.05), at all levels of doses. But, nephrotoxicity evidenced by elevated creatinine was seen only at a dose of 1000 mg/kg. Histological examination showed congestion of sinusoids, hemorrhage hepatocytes, fatty change, centrilobular necrosis and increased number of Kuppfer cells in the liver of all Mitragyna speciosa Korth standardized methanol extract treated groups.
Oral administration of standardized methanolic extraction of Mitragyna speciosa Korth resulted in increasing rat blood pressure after an hour of drug administration. The highest dose of extract also induced acute severe hepatotoxicity and mild nephrotoxicity. However, Mitragyna speciosa Korth shows no effects on body weight, food and water consumption, absolute and relative organ weight and also hematology parameters.
The effects of pure alkaloid, mitragynine and a methanolic extract of kratom leaves were investigated on neuromuscular junction and compound nerve action potential.
Wistar rats were killed by cervical dislocation and decapitated. The phrenic nerve-hemidiaphragms, hemidiaphragms and sciatic nerve were isolated.
Kratom methanolic extract present at 0.1-1 mg/mL and mitragynine (0.0156 mg/mL) decreased the muscle twitch on the isolated phrenic nerve-hemidiaphragm and hemidiaphragm preparation. Muscle relaxation caused by kratom extract (1 mg/mL) was greater than the effect of mitragynine. Pancuronium and succinylcholine potentiated the effect of kratom extract. It also had a direct relaxation effect on the hemidiaphragm muscle. The muscle relaxation caused by kratom extract was not antagonized by neostigmine, tetraethylammonium and calcium chloride. High concentrations of kratom extract (10-40 mg/mL) and mitragynine (2 mg/mL) blocked the nerve conduction, amplitude and duration of compound nerve action potential.
The mechanism of action of kratom extract might not act as a competitive antagonist of acetylcholine yet its dominant effect was at the neuromuscular junction and not at the skeletal muscle or somatic nerve.
In recent years, chemistry in flowing systems has become more prominent as a method of carrying out chemical transformations, ranging in scale from microchemistry up to kilogram-scale processes. Compared to classic batch ultrasound reactors, flow reactors stand out for their greater efficiency and flexibility as well as lower energy consumption. This paper presents a new ultrasonic flow reactor developed in our laboratory, a pilot system well suited for reaction scale up. This was applied to the transesterification of soybean oil with methanol for biodiesel production. This reaction is mass-transfer-limited initially because the two reactants are immiscible with each other, then because the glycerol phase separates together with most of the catalyst (Na or K methoxide). In our reactor a mixture of oil (1.6 L), methanol and sodium methoxide 30% in methanol (wt/wt ratio 80:19.5:0.5, respectively) was fully transesterified at about 45 degrees C in 1h (21.5 kHz, 600 W, flow rate 55 mL/min). The same result could be achieved together with a considerable reduction in energy consumption, by a two-step procedure: first a conventional heating under mechanical stirring (30 min at 45 degrees C), followed by ultrasound irradiation at the same temperature (35 min, 600 W, flow rate 55 mL/min). Our studies confirmed that high-throughput ultrasound applications definitively require flow reactors.
Mitragynine is the primary active alkaloid extracted from the leaves of Mitragyna speciosa Korth, a plant that originates in South-East Asia and is commonly known as kratom in Thailand. Kratom has been used for many centuries for their medicinal and psychoactive qualities, which are comparable to that of opiate-based drugs. Kratom abuse can lead to a detectable content of mitragynine residue in urine. Ultra trace amount of mitragynine in human urine was determined by a high performance liquid chromatography coupled to an electrospray tandem mass spectrometry (HPLC-ESI/MS/MS). Mitragynine was extracted by methyl t-butyl ether (MTBE) and separated on a HILIC column. The ESI/MS/MS was accomplished using a triple quadrupole mass spectrometer in positive ion detection and multiple reactions monitoring (MRM) mode. Ajmalicine, a mitragynine's structure analog was selected as internal standard (IS) for method development. Quality control (QC) performed at three levels 0.1, 1 and 5 ng/ml of mitragynine in urine gave mean recoveries of 90, 109, and 98% with average relative standard deviation of 22, 12 and 16%, respectively. The regression linearity of mitragynine calibration ranged from 0.01 to 5.0 ng/ml was achieved with correlation coefficient greater than 0.995. A detection limit of 0.02 ng/ml and high precision data within-day and between days analysis were obtained.
Kratom is indigenous to Thailand. Market gardeners, peasants and labourers often become addicted to kratom leaf use. In certain respects, kratom addiction resembles addiction to a drug with narcotic properties, except that long term kratom addicts develop a dark skin, particularly on the cheeks. The age of onset is apparently later than in heroin addiction, and females are rare amongst those who use the substance. There were 5 cases of kratom addiction revealing psychotic symptoms; these had been seen by the author in the last yr (1974) in the outpatient department. Initially, 3 cases were suspected of having kratom psychosis of the basis of their history of addiction and their general appearance and on psychiatric examination. The measure chosen by lar to control kratom addiction by banning the cultivation of the tree has not been found to be effective, since it is a local law It is hoped that drug education for the rural youth in areas where kratom can be grown will be a more effective step towards its control.
An efficient system for the analysis of the total alkaloids extracted from leaves of Mitragyna inermis (Willd.) O. Kuntze (Rubiaceae) by HPLC using a reversed-phase column is described. The chromatographic conditions allowed the separation of indole and oxindole alkaloids in leaf extracts, and the quantification of uncarine D in samples collected in Burkina Faso and Mali. The HPLC method described was validated for its specificity, linearity and precision using an internal standard (naphthalene). The concentrations of uncarine D in various extracts were compared with their in vitro anti-plasmodial activity. The anti-proliferative activity on chloroquine-resistant strain (W2) of Plasmodium falciparum was not correlated with the concentration of uncarine D in leaves.
Mitragynine is an indole alkaloid isolated from the Thai medicinal plant Mitragyna speciosa. We previously reported the morphine-like action of mitragynine and its related compounds in the in vitro assays. In the present study, we investigated the opioid effects of 7-hydroxymitragynine, which is isolated as its novel constituent, on contraction of isolated ileum, binding of the specific ligands to opioid receptors and nociceptive stimuli in mice. In guinea-pig ileum, 7-hydroxymitragynine inhibited electrically induced contraction through the opioid receptors. Receptor-binding assays revealed that 7-hydroxymitragynine has a higher affinity for micro-opioid receptors relative to the other opioid receptors. Administration of 7-hydroxymitragynine (2.5-10 mg/kg, s.c.) induced dose-dependent antinociceptive effects in tail-flick and hot-plate tests in mice. Its effect was more potent than that of morphine in both tests. When orally administered, 7-hydroxymitragynine (5-10 mg/kg) showed potent antinociceptive activities in tail-flick and hot-plate tests. In contrast, only weak antinociception was observed in the case of oral administration of morphine at a dose of 20 mg/kg. It was found that 7-hydroxymitragynine is a novel opioid agonist that is structurally different from the other opioid agonists, and has potent analgesic activity when orally administered.
A simple HPLC technique for determining mitragynine levels in serum was developed. The separation system consisted of a C18 column heated to 35 degrees C, a methanol-water (80:20, v/v) mobile phase, a flow rate of 0.8 mL/min and detection in the ultraviolet at 225 nm. Mitragynine, with a retention time of 10.09 min, was well resolved from any interferences in human serum and the internal standard peak. The calibration curve was linear from 0.1 to 10 microg/mL (r = 0.9995). Extraction of mitragy-nine from alkalinized serum using diethyl ether gave a high recovery (>or=85%). The intra- and inter-day precisions of the method were 4.29-5.88%RSD and 7.06-8.45%RSD, respectively. The accuracy ranged from -9.54 to +0.67%DEV. The limit of detection was 0.03 microg/mL and the lower limit of quantification was 0.1 microg/mL. Mitragynine in the stock solution was stable during 30 days of storage at 4 degrees C. This method was successfully applied to determine the pharmacokinetic characteristics of mitragynine levels in the serum of rats after it was administered orally.
Administration of the aqueous extract of Mitragyna speciosa at a dose of 300 mg/kg significantly inhibited ethanol withdrawal-induced behaviors that included rearing, displacement and head weaving. The results also showed that at doses of 100, 300 and 500 mg/kg M. speciosa showed antidepressant activity without effect on the spontaneous motor activity.
Kratom (Mitragyna speciosa Korth.) is an indigenous plant of Thailand used traditionally in folk medicine although it is claimed to cause addiction. It is used to treat diarrhea, however, there is no scientific evidence to support the use. The aim of this study is to investigate the effect of methanolic extract of kratom leaves on the rat gastrointestinal tract. Kratom extract at 50, 100, 200 and 400 mg/kg (p.o.) caused a dose dependent protection against castor oil-induced diarrhea in rats and also inhibited intestinal transit. The antidiarrheal effect was not antagonized by naloxzone. The inhibition of intestinal transit by kratom extract was significantly different from the control when treated with a single dose for 1 day. For longer-term treatments of 15 and 30 days, kratom extract did not decrease the intestinal transit time indicating that adaptation had occurred. Kratom extract at a dose level of 200 and 400 mg/kg for 30 days and morphine at 3 mg/kg (i.p.) caused a decrease in the increment of body weight that was significantly different from the control and kratom extract at lower doses (50 and 100 mg/kg). However it had no effect on the level of plasma cholecystokinin. The results suggested that methanolic kratom extract exhibited its antidiarrheal effect on rat gastrointestinal tract. The effects may occur via pathways in addition to the action on opioid receptors. High does of kratom extract decreased the increment of body weight similar to the effect of morphine.
A new pilot flow-reactor for high-intensity ultrasound irradiation. Application to the synthesis of biodiesel 0 10 20 30 40 50 60 70 80 90 00 Malaysia Indonesia Thailand 1 2 3 4 5 Fig. 8. Alkaloid distribution within Thai, Malaysian and Indonesian M. speciosa leaves (USH, met/w 1:1). L. Orio et al
985-989
P Cintas
S Mantegna
E Calcio
G Gaudino
Cravotto
P. Cintas, S. Mantegna, E. Calcio Gaudino, G. Cravotto, A new pilot flow-reactor for high-intensity ultrasound irradiation. Application to the synthesis of biodiesel, Ultrason. Sonochem. 17 (2010) 985–989. 0 10 20 30 40 50 60 70 80 90 00 Malaysia Indonesia Thailand 1 2 3 4 5 Fig. 8. Alkaloid distribution within Thai, Malaysian and Indonesian M. speciosa leaves (USH, met/w 1:1). L. Orio et al. / Ultrasonics Sonochemistry 19 (2012) 591–595 595
V. Sinou, J. Fiot, N. Taudon, J. Mosnier, M. Martelloni, S.S. Bun, D. Parzy, E. Ollivier,High-performanceliquid quantificationof Mitragynainermis pharmacokinetic studies, J. Sep. Sci. 33 (2010) 1863–1869.
Improved extraction of natural matrices under high-intensity ultrasound and microwave, alone or combined