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The species Alpinia purpurata is scarcely cited as to ethnopharmacology and phytochemistry. This study aimed to analyze bioactive compounds through high-performance liquid chromatography (HPLC). Hydroalcoholic crude extract was obtained from A. purpurata dried leaves. Folin-Ciocalteau method was used to quantify total phenols, using gallic acid as standard. The obtained result was 15.6 mg GAE g-1. The crude extract was partitioned with the solvents ethyl acetate and butanol, followed by thin-layer chromatography (TLC) and HPLC. The flavonoids kaempferol-3-O-glucuronide and rutin were detected at a higher concentration in ethyl acetate and butanolic extracts. The butanolic extract contains the highest flavonoid percentage (94.3%). A. purpurata presents important flavonoids of therapeutic use, already verified for A. zerumbet. This is the first study verifying the presence of flavonoids in A. purpurata extracts.
Recebido para publicação em 29/02/2008
Aceito para publicação em 06/10/2008
Detection of flavonoids in Alpinia purpurata (Vieill.) K. Schum. leaves using high-
performance liquid chromatography
1 Laboratório de Fisiologia Vegetal, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de
Janeiro, Av. Carlos Chagas Filho, s/n, CCS, Bloco G, sala G2-050. Rio de Janeiro - RJ, 21941-902. Brazil.
* 2 Laboratório de Fitoquímica, Núcleo de Pesquisas de Produtos Naturais (NPPN), Universidade
Federal do Rio de Janeiro, Av. Carlos Chagas Filho, s/n, CCS, Bloco H, Rio de Janeiro - RJ, 21941-902. Brazil.
ABSTRACT: The species Alpinia purpurata is scarcely cited as to ethnopharmacology and
phytochemistry. This study aimed to analyze bioactive compounds through high-performance
liquid chromatography (HPLC). Hydroalcoholic crude extract was obtained from A. purpurata
dried leaves. Folin-Ciocalteau method was used to quantify total phenols, using gallic acid as
standard. The obtained result was 15.6 mg GAE g-1. The crude extract was partitioned with the
solvents ethyl acetate and butanol, followed by thin-layer chromatography (TLC) and HPLC. The
flavonoids kaempferol-3-O-glucuronide and rutin were detected at a higher concentration in ethyl
acetate and butanolic extracts. The butanolic extract contains the highest flavonoid percentage
(94.3%). A. purpurata presents important flavonoids of therapeutic use, already verified for A.
zerumbet. This is the first study verifying the presence of flavonoids in A. purpurata extracts.
Key words: TLC, polyphenols, medicinal plants, rutin, Zingiberaceae
RESUMO: Detecção de flavonóides em folhas de Alpinia purpurata (Vieill.) K. Schum.
por cromatografia líquida de alta eficiência. A espécie Alpinia purpurata apresenta poucas
citações referentes a etnofarmacologia e fitoquímica. Este estudo propõe a análise de substâncias
bioativas através da técnica de cromatografia líquida de alta eficiência (CLAE). O extrato bruto
hidroalcóolico foi obtido a partir de folhas secas de A. purpurata. A quantificação de fenóis totais
foi realizada pelo método de Folin-Ciocalteau, usando ácido gálico como padrão. Como resultado,
foi verificado 15,6 mg EAG g-1. O extrato bruto foi particionado com os solventes acetato de etila
e butanol e depois analisado por cromatografia em camada delgada e CLAE. Nos extratos acetato
de etila e butanólico foi detectada a presença dos flavonóides kaempferol-3-O-glicuronídeo e
rutina, em maior concentração. O extrato butanólico contém a maior porcentagem de flavonóides
(94,3%). Esta espécie possui flavonóides importantes no uso terapêutico, já antes verificados
para a espécie A. zerumbet. Este é o primeiro trabalho que verifica a presença de flavonóides em
extratos de A. purpurata.
Palavras-chave: CCF, polifenóis, plantas medicinais, rutina, Zingiberaceae
The genus Alpinia (Zingiberaceae family,
Alpinioideae subfamily, Alpinieae tribe) is native to
tropical and subtropical Asia (Kress et al., 2002).
Nowadays, it is cultivated in several places around
the world due to the attractive beauty of its
inflorescences and its therapeutic potential (Soares
de Moura et al., 2005; Victório, 2008). In addition,
these plants are important sources of raw material for
many useful products: foods, spices, medicines,
perfumes, dyes and fiber paper (Tomlinson, 1969).
A. purpurata (Vieill.) K. Schum (red
inflorescence) is an herbaceous perennial plant,
internationally known in the ornamental plant market
as potted plant, landscape accent and cut flower
(Morón, 1987; Kress et al., 2002). An ethnobotanical
study developed in a community from Trujillo State,
Venezuela, investigated the use of its flowers as
decoction for cough (Bermúdez & Velásquez, 2002).
Rev. Bras. Pl. Med., Botucatu, v.11, n.2, p.147-153, 2009.
Zoghbi et al. (1999) analyzed A. purpurata essential
oil composition, which showed notable antibacterial
activity. However, studies about A. purpurata
phytotherapeutic potential are scarce; most scientific
works with this species are directed to the
improvement of its production as an ornamental plant,
including the evaluation of postharvest treatment as
an alternative to chemical insecticides, indications of
harvest and postharvest procedures, tolerance to
fumigation, biological control of pests, in vitro storage
of multiple shoots and micropropagation (Morón, 1987;
Dekkers et al., 1991; Illg & Faria, 1995; Hara et al.,
1997; Chen & Paull, 1998; Anderson & Gardner, 1999;
Gonzalez & Mogollon, 2001; Sangwanangkul et al.,
2008). The presence of A. purpurata in several parts
of Brazil allows easy access to it. Thus, this species
is an available resource to phytotherapeutic treatment.
Several species of the Zingiberaceae family present
antioxidant property mainly due to the considerable
presence of flavonoids such as rutin, quercetin,
alpinetin and different types of kaempferol in the genus
Alpinia (Table 1) (Williams & Harborne, 1977;
Mpalantinos et al., 1998; Vankar et al., 2006). Pugialli
et al. (1993) studied Zingiberaceae chemotaxonomy
and considered that flavonoids and their structural
variety are taxonomic markers. Zingiberaceae family
is at a higher level within the superorder Zingiberiflorae
due to the use of protection mechanisms of phenolic
hydroxyl groups: glycosylation and methylation.
Investigations of phytochemical compounds have been
important tools to study plant classification and
evolution (Kaplan & Gottlieb, 1982). This study
proposes a phytochemical approach to A. purpurata
based on the chemotaxonomy of the genus Alpinia,
which presents high therapeutic potential (Bleier &
Chirikdjian, 1972; Mendonça et al., 1998; Mpalantinos,
2001; Kim et al., 2006).
The aim of the present study was to evaluate
the phytochemistry of hydroalcoholic extracts from
A. purpurata leaves for the presence of the flavonoids
rutin, kaempferol-3-O-rutinoside and kaempferol-3-O-
glucuronide, reported in the scientific literature for their
therapeutic action.
Plant material
A. purpurata leaf samples were collected from
plants growing in the city of Rio de Janeiro in the
Federal University of Rio de Janeiro (Rio de Janeiro
State, Brazil). The voucher specimen was identified
and deposited at the Herbarium of Rio de Janeiro
Botanical Garden under the accession number RB
Preparation of extracts and Fractions
A. purpurata leaves were collected from adult
plants in the morning; then, the plant material was
dried and ground in 70% ethanol for a week. After the
first extraction, leaves were kept in ethanol (100%)
until exhaustive extraction. Crude extracts were filtered
and dried through evaporation at 60ºC in a rotary
evaporator and through freeze-drying. From 1009 g
dried leaves, 112.5 g dried crude extract was obtained.
The yield was calculated as percentage, according
to the formula: (crude extract weight/plant material
weight) x 100. A 59.4g crude extract fraction was
resuspended in methanol:water (9:1, v/v) and
partitioned in different solvents of increasing polarity
range: hexane, dichloromethane, ethyl acetate and
n-butanol. Hexane partition was separated and
evaporated to dryness (4.1 g). The residue was dried
TABLE 1. Flavonoids reported for the genus Alpinia.
**The abstract did not cite.
Rev. Bras. Pl. Med., Botucatu, v.11, n.2, p.147-153, 2009.
using a rotary evaporator until methanol elimination;
then, the aqueous layer was further partitioned using
solvents and evaporated to dryness in order to obtain
dichloromethane (0.29 g), ethyl acetate (0.31 g) and
n-butanol (4.8 g) partitions. Each solvent extractor
(50 to 60 mL) was used five times.
Flavonoid standards
Flavonoids of the kaempferol class were
isolated from Alpinia zerumbet Roxb. and identified
using Nuclear Magnetic Resonance (NMR)
(Mpalantinos et al., 1998). Rutin was purchased from
Merck. Kaempferol-3-O-glucuronide had 82% purity,
kaempferol-3-O-rutinoside, 91%, and rutin, 98%.
Purity was verified using three replicate injections of
standards into HPLC.
Evaluation of phenolic compound content
Total phenolic compounds were determined
using the Folin-Ciocalteau method. Hydroalcoholic
extracts were dissolved in ethanol (70%) at 1mg mL-1.
A 0.5 mL aliquot of diluted extract and 2 mL Folin-
Ciocalteau reagent (10%) were added after 3 min,
together with 2 mL of 7.5% sodium carbonate, and
mixed. The mixture was homogenized and incubated
at 50oC for 30 min. Absorbance was measured at 740
nm in a spectrophotometer using gallic acid as
standard. Two controls were used: (1) Folin-Ciocalteau
+ sodium carbonate and (2) crude extract solution.
Phenolic compounds in crude extracts were quantified
through the regression equation of calibration curves:
y = 0.0229x + 0.0968 (R2 = 0.9993), and expressed
as mg gallic acid equivalents (GAE) per 1 g dried
leaves. All measurements were done in triplicate. The
analyses included A. purpurata and A. zerumbet
samples collected in June (2006) and February (2007).
Thin-layer chromatography (TLC)
Aliquots of standards, and crude, ethyl
acetate and butanolic extracts were spotted on TLC
plate (silica gel 60 F254 nm, Merck) and developed in
ethyl acetate, formic acid and distilled water (65/20/
15, v/v/v) mobile phase. TLC was observed under UV
spectrum at 254 and 360 nm before and after spraying
with NP/PEG reagent. The flavonoid standards of rutin
(Rf = 0.69), kaempferol-3-O-rutinoside (Rf = 0.76) and
kaempferol-3-O-glucuronide (Rf = 0.83) were verified
in the extracts after concomitant running with
standards (Victório et al., 2007).
High-performance liquid chromatography
(HPLC) Crude, ethyl acetate and butanolic extracts
were dissolved in methanol (70%) at 20 mg mL-1 and
filtered under vacuum; HPLC-UV analyses were
performed in a Shimadzu apparatus equipped with
SPD-M10A diode array detector, LC-10AD pump and
CBM-10 interface. Data were obtained and processed
in a reversed phase column (Lichrosorb RP-18, 25
cm x 5 mm), at room temperature. Separation was
done in the following mobile phase: MilliQ water +
0.1% phosphoric acid (A) and methanol (B): 1-10 min
(30% B); 20 min (40% B); 60 min (100% B). The
prepared mobile phase was degassed using ultrasonic
agitation. After 61 min, the gradient was recycled to
the initial conditions and held for 10 min before a new
injection. The flow rate was kept constant at 1 mL
min-1 and peaks were detected at 254 nm and 360
nm. All chemicals used in the analysis, such as
methanol and phosphoric acid, were of HPLC grades
and were purchased from Merck. MilliQ water was
used in HPLC mobile phase and sample preparation.
Standards were dissolved in 70% methanol at 1 mg.mL-
1 and analyzed in the same elution. Injections were
done in triplicate. Linearity was observed in the
concentration range 0.0078 - 0.0625 mg mL-1 rutin and
0.01325 - 0.25 mg mL-1 kaempferol-3-O-glucuronide.
Flavonoids in the extracts were quantified against
calibration curves of standards, where y is the peak
area and x the concentration in mg mL-1 (Figure 1).
FIGURE 1. Calibration curves of rutin and kaempferol-3-O-glucuronide standards.
Rev. Bras. Pl. Med., Botucatu, v.11, n.2, p.147-153, 2009.
Flavonoid detection
Flavonoids were detected through retention
times (RT), ultraviolet spectrum compared with
flavonoid standard spectrum (Figure 2) and coinjection
with authentic samples analyzed under the same
conditions. For coinjection, a mixture (1:1, v/v) of
extracts at 20 mg mL-1 and standard at 1 mg mL-1
was prepared. The purity of each flavonoid peak in
Figures 4 and 5 was assessed by comparing the UV
spectra at upslope and downslope inflexion points for
both wavelengths (254 and 360 nm).
Phenolic compounds present antioxidant
activity; thus, they are considered important therapeutic
agents. As already known, antioxidants reduce the
effects of excessive free radical production in critically
ill patients affected by diseases like cancer,
cardiovascular disturbances, and brain dysfunction
(Atoui et al., 2005). Folin-Ciocalteau method can quantify
the presence of flavonoids and other phenolic
compounds in plant material. There were significant
dif ferences in total phenolic content (p<0.03)
between A. purpurata and A. zerumbet in both
evaluated months (Figure 3). A. zerumbet was used
for comparison since it is greatly employed in folk
medicine and has been extensively reported
concerning phytochemistry and phytotherapy.
The chromatographic profile of A. purpurata
shows rutin and kaempferol-3-O-glucuronide peaks,
similarly to those for A. zerumbet (Figure 4) in opposite
proportions. These flavonoids were verified in ethyl
FIGURE 2. UV spectra of (A) rutin (RT= 32.5 min) and (B) kaempferol-3-O-glucuronide (RT= 35.2 min).
FIGURE 3. Comparison of total phenolic content extracted from Alpinia zerumbet and A. purpurata collected in two
different periods of the year. A (total phenols in mg GAE g-1 dried leaves), B (June, 2006) and C (February, 2007).
Rev. Bras. Pl. Med., Botucatu, v.11, n.2, p.147-153, 2009.
acetate and butanolic extracts between 20 and 40
min (Figure 5). The main difference was detected
between 40 and 60 min, when A. zerumbet crude
extract showed more compounds in the
chromatographic profile than A. purpurata (Figure 4).
The flavonoid kaempferol-3-O-rutinoside was not
detected through TLC or HPLC in the extracts. HPLC
and TLC methods were reproducible. UV spectrum of
peaks 1 and 3 of the chromatogram (Figure 5) are
characteristic of flavonoids; however, for an accurate
identification, they should be isolated in SEPHADEX
column using polar solvents from ethyl acetate and
butanolic fractions, followed by structural elucidation
using the spectroscopy technique. Rutin and
kaempferol-3-O-glucuronide were isolated from A.
zerumbet and also detected in A. purpurata at the
following concentrations, respectively: 17.8 and 8.7
mg g-1 dried leaves (ethyl acetate) and 356 and 85.5
FIGURE 4. Comparison between chromatographic
profiles of crude extracts from Alpinia purpurata (A)
and A. zerumbet (B): rutin (1) and kaempferol-3-O-
glucuronide (2). Values obtained at 254 nm.
FIGURE 5. Chromatographic profile of Alpinia
purpurata: crude extract (A), butanolic extract (B) and
ethyl acetate extract (C). Rutin (2) and kaempferol-3-
O-glucuronide (4). Values obtained at 254 nm.
TABLE 2. Quantitative analysis of flavonoids rutin and kaempferol-3-O-glucuronide in extracts of dried leaves of
Alpinia purpurata. Values obtained at 254 nm.
Data represent mean of triplicates.
Rev. Bras. Pl. Med., Botucatu, v.11, n.2, p.147-153, 2009.
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... The~13 times TPC difference was similar to previous studies on TPC and AA in various herbs and spices that C. longa had the highest TPC [23,27]. Other high TPC herbs such as C. xanthorrhiza [27,28], A. purpurata [29], B. rotunda [30], and Z. cassoumounar [31] were also reported. ...
... Among 12 selected Indonesian herbs samples, the TFC (as mg QE/g) was 279.87, i.e., the highest in C. longa and was 1.31, i.e., the lowest in Z. officinale ( Figure 3B) similar to reported high curcuminoid content. Previous studies have found lesser TFC in Z. officinale, Z. zerumbet, and C. heynana [27,29,30]. The~214 times TFC difference might be attributed to the type of flavonoids present in the herbs. ...
... Typical TPC contribute more to plant AA are mainly phenolic acids and flavonoids. The matrix difference among plant species from different geographical origin and variance in genetics and cultivation conditions make the correlation analysis between AA with TPC and TFC a challenging work [21,23,28,29,31,35,36]. We adopted the Pearson correlation coefficient (PCC), also referred to as Pearson's r, to express the strength and direction of the linear relationship of correlation. ...
Full-text available
The antioxidant activity (AA), total phenolic content (TPC), and total flavonoid content (TFC) of selected Indonesian Zingiberaceae herbs were determined. An optimization extraction procedure was conducted by using Taguchi L16 orthogonal array. Four chemical assays were applied, including 2,2-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity assay, H2O2 scavenging activity assay, Folin–Ciocalteau (F–C) assay, and NaNO2-AlCl3-NaOH assay, which revealed remarkable differences in AA, TPC, and TFC. The result indicated the diversity of AA composition among the herbs, and C. longa exhibited the highest AA. HPLC-PAD analysis revealed that curcumin was present in five high antioxidant herbs, and the highest amount was in C. longa. Pearson correlation analysis indicated that the identified TPC and TFC were significant contributors to AA, and curcumin was likely the main contributing antioxidant compound. Our approach concluded that C. longa is the greatest source of natural antioxidants among 12 Indonesian indigenous Zingiberaceae herbs. The use of a mixed-method approach to augment the findings of solitary methods might facilitate future researchers to uncover deeper and hidden meanings.
... Micropropag., Lavras, v.4, n.2, p. 92-98, 2008 used for cough in Venezuela and presents antibacterial activity (BERMÚDEZ & VELÁSQUEZ, 2002;ZOGHBI et al., 1999). Recent studies have showed the phytochemical potential of this species (VICTÓRIO et al., 2007(VICTÓRIO et al., , 2008. ...
... Leaves of A. purpurata from field and in vitro systems were dried and macerated in 70% ethanol for 45 min using ultrasonic bath according to Victório (2008). Crude extracts were filtered and dried by evaporation at 60ºC in rotary evaporator and by lyophilization. ...
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AlpiniapurpurataK.Schumislargelyusedinornamentation and scarcely present itself in folk medicine. Studiesusing plant tissue culture technique under standardized conditionpermittoevaluateplantphysiological,morphologicalandphytochemicalresponsestoexogenousplantgrowthregulator.This work consisted in the establishment of in vitro A. purpurataplants using different concentrations of auxin and cytokinins inmg L-1:IAA 2, TDZ 2 and 4, IAA 2 + TDZ 2, IAA 2 + BAP 2.Additionally,itwasdeterminedthecontentofphenoliccompounds in the leaves of field and in vitro plants using FolinCiocalteau method. Buds from inflorescence A. purpurata weredisinfested and introduced in solid MS medium to initiate cultures.Thedirectorganogenesisprocesswasfastandsatisfactoryproducingelongatedplantswithadevelopedradicularsystem.The rooting percentage reached 100% for all treatments after 2months. Cytokinins and auxins were important for the increasingof the leaves quantity (4.2 to 5.1) and shoot elongation (4.0 cm).LiquidMS0presentedthefavorableresultsforfastmicropropagationandhigherproductionofshoots,providingplants ready to be acclimatized. In vitro plants cultivated underMS0 produced 93% of phenolic compounds in comparison withfieldplants(100%).ThroughHighPerformanceLiquidChromatographic, it was verified similar chromatographic profilesbetween field and in vitro plant extracts
... K. Schum and Alpinia galanga (L) Willd. In pharmacological tests, galangal extract exhibits promising potentials, which has a reduced blood pressure effect [2], antioxidant activity, anticancer [3], antibacterial, antiviral, antifungal, antiparasitic [4], antiinflammatory [5], antitumor, analgesics, and antiflatulence [6]. Phytochemical test results from red galangal ethanol extract contain alkaloid compounds, phenols, flavonoids, and tannins [5]. ...
... Tao (2006) evaluated the quality of A. officinarum Hance for the assessment of two main bioactive flavonoids: galangin and 3-O-methyl galangin [14]. Research conducted by Victorio showed that ethyl acetate and n-butanol fractions of Alpinia purpurata (red galangal) leaves detected kaempferol-3-O-glucoronide and rutin flavonoids using HPLC [2]. In this study, the identification and quantification of flavonoid compounds (quercetin and rutin) will be carried out in red galangal and white galangal rhizomes using HPLC method. ...
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Research of flavonoid determination (quercetin and rutin) in Alpinia purpurata and Alpinia galanga rhizomes using HPLC method has been conducted. This study aims to determine the qualitative and quantitative presence of rutin and quercetin compounds in Alpinia purpurata and Alpinia galanga rhizomes. Fractions of ethyl acetate and n-butanol have been obtained by TLC, followed by total flavonoid analysis with UV - Vis spectrophotometer method and quercetin and rutin determination with HPLC. The results obtained in this study are as follows: Total flavonoid from ethyl acetate fraction obtained is 46,48 mg EQ/gram in Alpinia purpurata and 70,60 mg EQ/gram in Alpinia galanga , and total flavonoid from nbutanol fraction obtained is 68,50 mg EQ/gram for Alpinia purpurata and 103,80 mg EQ/gram for Alpinia galanga . The ethyl acetate fraction of Alpinia purpurata detected quercetin content at 5469.64 mg / kg and rutin at 4955.59 mg / kg, while Alpinia galanga detected quercetin at 5764.10 mg / kg and rutin at 5327.93 mg / kg. From n-butanol fraction, quercetin are only detected at 6737.14 mg / kg in Alpini purpurata and 9098.74 mg / kg in Alpinia galanga . From these results, it can be concluded that HPLC method can be used to detect the presence of flavonoid quercetin and rutin in Alpinia purpurata and Alpinia galanga rhizomes.
... According to Kochuthressia et al. [27] , [20,21,28] , which present antimicrobial properties [29] . albicans, but a marked inhibition of other fungi, such as C. neoformans, F. pedrosoi, T. rubrum, M. canis and M. gypseum. ...
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Revista Fitos, Rio de Janeiro. 2021; Ahead of Print | e-ISSN: 2446-4775 | | CC-BY 4.0 ARTIGO DE PESQUISA In vitro antimicrobial activity of Alpinia zerumbet and A. purpurata nonpolar fraction of leaf extract Abstract This study aimed to evaluate the in vitro antimicrobial activity of fractions of leaf hydroalcoholic crude extracts: hexane, dichloromethane, ethyl acetate and butanolic of Alpinia zerumbet and A. purpurata by the agar drop diffusion method in order to screen the main compounds involved in antimicrobial activity. Leaves of A. zerumbet (Pers.) B.L. Burtt et R.M. Sm. and A. purpurata (Vieill) K. Schum adult plants were collected and then dried and macerated in 70% ethanol. Leaf extracts were further partitioned using solvents in increasing polarity. Dichloromethane fractions were analyzed by gas chromatography/mass spectrometry (GC/MS). The major compound in the dichloromethane fraction of both species was the hexadecanoic acid (palmitic acid). By drop diffusion assay, the antimicrobial activity of crude extract, as well as fractions of hexane, dichloromethane, ethyl acetate and butanolic, was evaluated against pathogenic bacteria and fungi. No bacteria were inhibited. However, the dichloromethane fraction exhibited promising antifungal activity against the following fungi tested: Cryptococcus neoformans, Fonsecaea pedrosoi, Trichophytoon rubrum, Microsporium canis and M. gypseum.
... Phytochemical analysis for Rhizome Alpinia purpurata showed positive results for alkaloids (14.9%), phenols (9.5%), flavonoids (0.85%) and tannins (13.8%) [3]. Investigation of the flavonoids compound contained by kaempferol-3-owned glucoronide, kaempferol-3-uliocronide and rutin of ethanol extract has been done using HPLC [4]. Ethanolic extracts of Alpinia purpurata showed highest inhibitory activity against Enteobacter aerogens [5]. ...
... Tosse: Decocção das flores 23,24 . ...
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O livro faz parte da série "Extensão Universitária em Farmacobotânica" e foi desenvolvido através da integração do grupo PET-Farmácia com o projeto de extensão ¨Uso e cultivo racionais de plantas medicinais e plantas alimentícias não convencionais (PANC) pelos agricultores de Magé e Guapimirim, RJ¨. O tema do livro é a caracterização de espécies da ordem Zingiberales que são cultivadas em Magé e Guapimirim para uso medicinal, ornamental e alimentício. As características de cada espécie estudada são apresentadas em linguagem acessível para difusão e estímulo do uso racional de recursos botânicos.
... The samples were subsequently stored in separate sample bottles for further study. The modified method of Victório et al. (2009) was applied for the extraction of plant material. The samples (2 gm each) were extracted with 20 ml methanol at room temperature for 24-h with occasional shaking. ...
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In vitro culture provide a suitable condition for regeneration of arsenic tolerant plants from stem calli of Andrographis paniculata (Burm.f.) Nees. The regenerated plants could be valuable material applicable to soil remediation. In vitro culture of stem explants on MS basal salts, 3.0% sucrose, 0.8% agar medium fortified with various concentrations of As2O3 (0.0-9.0 µM) along with NAA (3.5 mg L⁻¹) and KIN (1.0 mg L⁻¹) influenced resistant callus formation. Growth of callus was slightly inhibited with increased resistance up to 22% (53±0.8 mg fwt and 27±1.5 mg dwt) on 7.0 µM As2O3 selective medium. The resistant callus line inoculated on MS medium supplemented with different concentrations of As2O3 along with BA (2.5 mg L⁻¹) and NAA (3.0 mg L⁻¹) induced the development of shoots. Shoot organogenesis was slightly inhibited by arsenic metal stress. However the growth tolerance has increased up to 14.5% on medium with 7.0 µM As2 O3. The rate of adventitious rooting of plantlets was gradually decreased with more tolerance (11.1±1.4 rootlets per plantlets) in 7.0 µM As2 O3 selected media. After acclimatization, about 40% of plants were survived as arsenic tolerance than control plants in pots containing soil treated with 7.0 µM As2 O3 solutions. The level of arsenic detectability was 0.96 ppb and 4.67 ppb in control and 7.0 µM As2 O3 treated plants, respectively by AAS analysis. Moreover, the production of Andrographolide was found quite high (4.41 mg/g) in tolerant plants grown at 7.0 µM As2 O3 treatment than control by HPLC analysis. 1D¹ H NMR profile revealed the metabolic changes significantly in control and 7.0 µM As2 O3 treated plant samples. This is the first report confirming the suitability of in vitro selection for obtaining of vigorous and proliferative clones of A. paniculata plants tolerant to elevated arsenic concentration. © 2017 J. Vijayakumar, P. Ponmanickam, P. Samuel, B. Pavithra, A. Manjula, S. Aswathi and G. Shobana Rathi.
The antimicrobial activity of three solvent extracts (ethanol, petroleum ether & chloroform) of three different plant parts of Alpinia purpurata (leaves, roots and rhizomes) was studied in vitro against six reference strains of bacteria and four pathogenic fungi using disc diffusion assay. Ethanolic extracts of rhizomes showed a wide spectrum of activity against all tested bacteria, but no notable activity against any fungi except Candida albicans. Highest inhibition zone of (14.4 ± 0.4 mm) was recorded for the ethanolic extracts of rhizomes against Enterobacter aerogens. Petroleum ether and chloroform extracts did not show any highlighting results. The antimicrobial potential of rhizomes of A. purpurata is well substantiated than any other parts tested.
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Essential oils from spices such as red galangal and cinnamon are known to have antimicrobial activity that is strong enough against pathogenic and food destroying bacteria, so it has the potential to be developed as a preservative, especially in food. High antimicrobial activity is generally obtained at high concentrations, which can affect sensory acceptance when applied to food. One way to overcome this problem is to combine the use of essential oils. This study aims to determine the effect of the ratio of the combination of red galangal and cinnamon essential oils to its antimicrobial activity on Escherichia coli and Staphylococcus aureus bacteria. The research was carried out with an experimental method which was analyzed descriptively. Antimicrobial activity testing was carried out by the agar diffusion method and contact method to determine the inhibition growth of E. coli and S. aureus bacteria. The treatment consists of 5 ratios combination of red galangal and cinnamon essential oils, which were 1:0, 0:1, 1:1, 1:2, and 2:1 (v/v). The results showed that the ratio of the combination of red galangal and cinnamon essential oils showed a strong effect on antimicrobial activity towards E. coli and S. aureus. The combination of essential oil at 1:1 (v/v) ratio showed the highest antimicrobial activity against E.coli which was 20.5 mm of inhibition zone, while toward S. aureus was 21.25 mm inhibition zone (strong activity); and could reduce the total test bacteria by reducing 16.85% of E. coli and 21.69% of S. aureus bacteria after 24-hour length incubation.
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Some of the secondary metabolites present in red algae are terpenoids, quinones, flavonoids, alkaloids, essential oils, diarylheptanoids, steroids, cardioglycosides, oils and fats, tannins, carbohydrates. Activity of rhizomes, leaves and flowers red leaf is as antimicrobial, anti-fungal, anti-oxidants, anti-tumor, anti-cancer and vasodilator. One way to improve the physical, chemical and bioactivity properties of natural compounds was to make them into nanoparticles. In this study, the isolation of bioactive compounds contained in red laos rhizome by maceration method using ethanol solvent was done, then partitioned with n-hexane and ethyl acetate. The extracts thus obtained are fabricated into nanoparticles. Extracts in bulk and nanoparticles were then tested for cytotoxic activity using BSLT method. Results of analysis with PSA showed that ethanol extract had size 410,8 nm, n-hexane extract 220,7 nm and ethyl acetate extract 208,3 nm. The results of cytotoxic tests showed that nanoparticle size increased cytotoxic activity. Ethyl acetate extract was most active compared to ethanol and n-hexane extracts with LC50values of 17.919; 84,956; 166,526 ppm. Whereas the nanoparticle size was respectively 10,491; 74,072 and 84,197 ppm. Cytotoxic activity increases with nanoparticle fabrication.
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The vascular relaxant effects of Alpinia officinarum (Zingiberaceae) were evaluated on isolated thoracic aorta from rats. The methanolic extract of the rhizome of Alpinia officinarum caused a concentration-dependent relaxation of rat aortic preparations precontracted with 0.3 μM phenylephrine. The intensive investigation of the extract by way of activity-guided fractionation led us to yield three kind of active components, galangin (1), kaempferide (2) and kaempferol (3), which were responsible for the vasodilating property of the extract. Structures of the isolated active components were established by chemical and spectroscopic means.
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A análise das características flavonoídicas da ordem Zingiberales (superordem Zingiberiflorae, Monocotyledonae) reforçou a separação das suas famílias em dois grupos: grupo I (Strelitziaceae, Heliconiaceae, Musaceae e Lowiaceae) e grupo II (Zingiberaceae, Costaceae, Marantaceae e Cannaceae). Baseado nos dados químicos e morfológicos sugeriu-se um esquema que caracteriza os diferentes estádios de avanço evolutivo dessas famílias.The analysis of flavonoids in the order Zingiberales (superorder Zingiberiflorae, Monocotyledonae) reinforced the separation of its families into two groups: group I (Strelitziaceae, Heliconiaceae, Musaceae and Lowiaceae) and group II (Zingiberaceae, Costaceae, Marantaceae and Cannaceae). Chemical and morphological data suggested the relative stages of evolutionary advancement of these families.
The green leaves tea of Alpinia zerumbet was extracted with ethyl acetate. The resultant material upon fractionation on a silica gel chromatographic column, furnished three substances identified by means of spectroscopic analysis (IV, RMN1H, RMN13C) as (+)-catechin, (-)-epicatechin and alpinetin. These flavonoids were rested in relation to the arterial pressure in rats and showed a significant antihypertensive activity.
OBJECTIVE: To establish the analysis method for alpinetin and cardamomin and evaluate the quality of Caodoukou (Alpinia katsumadai). METHOD: A HPLC method was deueloped by using a Shim-Pack CLC-ODS column (150 mm x 6 mm). A mixture of methanol-water (80:20) was used as the mobile phase with a flow rate of 1 ml per minute and the detection wavelength was set at 300 nm. The contents were calculated by external-standard method. RESULTS: The standard curves for alpinetin and cardamomin were linear over the range of injected amount of 0.05 μg ~ 0.26 μg (r = 0.9996 and 0.9998, respectively). The average recoveries of alpinetin and cardamomin were 101.9% (n = 5, RSD = 0.18%) and 100.0% (RSD = 3.6%), respectively. Results of analysis of 5 samples show the contents of alpinetin and cardamomin in the semens of Alpinia katsumadai were 0.50% ~ 1.45% and 0.07% ~ 0.72% respectively and their contents in the pericarp, stem leaves and rhizome were less than 0.2%. CONCLUSION: The method is convenient, reliable and can be used for analysis of crude drugs.
Chrysosplenium americanum (Saxifragaceae) accumulates a variety of partially O-methylated flavonol glucosides. Because of the semi-aquatic nature of this plant and its extensive contamination with endogenous organisms, the initiation of shoot and callus cultures could only be achieved after (a) using a special surface sterilization procedure, (b) production of new shoots from initial explants, and (c) selective elimination of organogenic structures during several subcultures. HPLC analysis of the cultured tissues established the presence of a number of flavonoids characteristic of the intact plant, though in lower and variable concentrations. However, shoot and callus cultures exhibited flavonoid profiles similar to those of the intact leaves and roots, respectively.
Kahili ginger (Hedychium gardnerianum) is an invasive weed in tropical forests in Hawaii and elsewhere. Bacterial wilt caused by the ginger strain of Ralstonia (=Pseudomonas) solanacearum systemically infects edible ginger (Zingiber officinale) and ornamental gingers (Hedychium spp.), causing wilt in infected plants. The suitability of R. solanacearum as a biological control agent for kahili ginger was investigated by inoculating seedlings and rooted cuttings of native forest plants, ornamental ginger, and solanaceous species to confirm host specificity. Inoculation via stem injection or root wounding with a bacterial–water suspension was followed by observation for 8 weeks. Inoculations on H. gardnerianum were then carried out in ohia-lehua (Metrosideros polymorpha) wet forests of Hawaii Volcanoes National Park to determine the bacterium's efficacy in the field. No native forest or solanaceous species developed wilt or other symptoms during the study. The bacterium caused limited infection near the inoculation site on H. coronarium, Z. zerumbet, Heliconia latispatha, and Musa sapientum. However, infection did not become systemic in any of these species, and normal growth resumed following appearance of initial symptoms. All inoculated H. gardnerianum plants developed irreversible chlorosis and severe wilting 3–4 weeks following inoculation. Systemic infection also caused death and decay of rhizomes. Most plants were completely dead 16–20 weeks following inoculation. The destructiveness of the ginger strain of R. solanacearum to edible ginger has raised questions regarding its use for biological control. However, because locations of kahili ginger infestations are often remote, the risk of contaminating edible ginger plantings is unlikely. The ability of this bacterium to cause severe disease in H. gardnerianum in the field, together with its lack of virulence in other ginger species, contributes to its potential as a biological control agent.
The sensitivity of cut flowers and foliage to irradiation used for insect disinfestation varies from species to species and to a lesser extent variety to variety. Flowers with low sensitivity to 250Gy of electron-beam irradiation were Red ginger (Alpinia purpurata) and Bird-of-Paradise (Strelitzia reginae) inflorescences, and Oncidium (Oncidium spp.–Gower Ramsey) sprays. Medium sensitive flowers were Dendrobium (Dendrobium spp.) ‘Royal Purples’, Protea hybrid ‘Pink Ice’ and heliconia ‘Red Stricta’ (Heliconia stricta). Flowers that showed high sensitivity to 250Gy were Heliconia ‘Keanae’, ‘Beehive’ ginger (Zingiber spectabilis), ‘Barbatus’ ginger (Costus barbatus), and ‘Indian head’ ginger (Costus woodsonii) inflorescences. White (UH 306) dendrobium was more sensitive than the Pink (UH 232) to irradiation (150Gy). Pretreatment of UH 306 with 1-methylcyclopropene (1-MCP: 50μgL−1, 3h) extended the vase life of dendrobium sprays and mitigated the effects of irradiation (250Gy) on dendrobium vase life. Hot water at 40°C for 30min or up to 47.5°C for 20–30min reduced the minimal irradiation injury to Bird-of-Paradise inflorescences. For the heliconia ‘Keanae’ inflorescences, a hot water treatment at 45–47.5°C for 40min effectively reduced irradiation injury. Red ginger infloresences were able to withstand 500Gy if pretreated with benzyladenine (BA) before irradiation. Green Ti (Cordyline terminalis) was not sensitive to irradiation doses from 0 to 500Gy. Leather-leaf fern (Dryopteris erythrosora) and baby eucalyptus (Eucalyptus pulverulenta) foliage showed little or no sensitivity to irradiation. Lycopodium (Lycopodium taxifolium) and Bracken fern (Pteridium aquilinum) were sensitive. Ruscus leaves (Ruscus aeulectus L.) showed chlorosis after irradiation and leaves treated with BA had less chlorosis compared to the non-irradiated foliage. ‘Pink Ice’ protea foliage withstood up to 500Gy, if first pulsed with 2% glucose solution for 12h.