RESUMO
The first phytochemical investigation of the twig extract of Uvaria leptopoda resulted in the isolation and identification of three new tetrahydroxanthene-1,3(2H)-diones, uvarialeptones A-C, two new oxidized hexadiene derivatives, uvarialeptols A and B, together with ten known compounds. Their structures were elucidated by spectroscopic techniques and mass spectrometry. Uvarialeptones A and B were unprecedented tetrahydroxanthene-1,3(2H)-dione dimers which exhibited a cyclobutane ring via [2 + 2] cycloaddition from uvarialeptone C and 9a-O-methyloxymitrone, respectively. The structure of uvarialeptone A was confirmed by X-ray diffraction analysis using Mo Kα radiation. Compound 3 inhibited NO production at an IC50 value of 6.7 ± 0.1 µM.
Assuntos
Uvaria , Uvaria/química , Estrutura Molecular , Animais , Óxido Nítrico/biossíntese , Camundongos , Xantenos/farmacologia , Xantenos/química , Cristalografia por Raios X , Oxirredução , Células RAW 264.7RESUMO
Piper species is one of the most widely consumed spices for culinary purposes. Piperine (PIP) present in Piper species has a wide range of therapeutic activity including hepatoprotection. However, the major biological limitation of PIP is its low bioavailability after oral administration. Purpose of the study was to prepare an optimized and adequately characterized PIP-phospholipid complex (PPC) as a delivery system to overcome these limitations and to investigate the pharmacokinetics and hepato-protectivity of the formulation in the animal model. Response surface methodology was adopted to optimize the process parameters for PPC preparation. FT-IR, DTA, PXRD, SEM, molecular docking etc. were used for characterization. Solubility, log P, dissolution efficiency and in vivo pharmacokinetics were also investigated. PPC showed enhanced hepatoprotective potential as compared to pure PIP at the same dose level (25 and 50 mg/kg). PPC restored the levels of serum marker and antioxidant enzymes. PPC also increased the bioavailability of PIP in rat serum by 10.40-fold in comparison with pure PIP at the same dose level and enhanced the elimination half-life (t1/2 el) from 0.477 ± 1.76 to 9.80 ± 1.98 h. Results concluded that PPC enhanced the hepatoprotection of PIP which may be due to the improved bioavailability and pharmacokinetics of PIP in rats.
Assuntos
Alcaloides/administração & dosagem , Alcaloides/metabolismo , Benzodioxóis/administração & dosagem , Benzodioxóis/metabolismo , Fígado/metabolismo , Fosfolipídeos/administração & dosagem , Fosfolipídeos/metabolismo , Piperidinas/administração & dosagem , Piperidinas/metabolismo , Alcamidas Poli-Insaturadas/administração & dosagem , Alcamidas Poli-Insaturadas/metabolismo , Alcaloides/síntese química , Animais , Benzodioxóis/síntese química , Disponibilidade Biológica , Fígado/efeitos dos fármacos , Masculino , Simulação de Acoplamento Molecular/métodos , Fosfolipídeos/síntese química , Piperidinas/síntese química , Alcamidas Poli-Insaturadas/síntese química , Substâncias Protetoras/administração & dosagem , Substâncias Protetoras/síntese química , Substâncias Protetoras/metabolismo , Ratos , Ratos WistarRESUMO
The first phytochemical investigation of Polyalthia cinnamomea led to the isolation and identification of two new oxoprotoberberine alkaloids, (-)-(13aS)-polyalthiacinnamines A and B, together with eleven known compounds. The structures of the new compounds were elucidated by extensive spectroscopic methods. The absolute configuration of miliusacunine E and consanguine B was established by X-ray diffraction analysis using Cu Kα radiation and ECD spectra, whereas the absolute configurations of polyalthiacinnamines A and B were established by comparison of their ECD spectra and specific rotations with those of miliusacunine E and consanguine B. Compounds 1-4, 6, and 8 exhibited α-glucosidase inhibitory activities (IC50 values ranging from 11.3 to 57.9 µM) better than a positive control (acarbose, IC50 83.5 µM). Compound 2 also exhibited NO production inhibitory activity with an IC50 value of 24.4 µM (indomethacin, a positive control, IC50 = 32.2 µM).
Assuntos
Alcaloides/farmacologia , Inibidores de Glicosídeo Hidrolases/farmacologia , Óxido Nítrico/antagonistas & inibidores , Extratos Vegetais/farmacologia , Polyalthia/química , alfa-Glucosidases/metabolismo , Alcaloides/química , Alcaloides/isolamento & purificação , Animais , Sobrevivência Celular/efeitos dos fármacos , Cristalografia por Raios X , Relação Dose-Resposta a Droga , Inibidores de Glicosídeo Hidrolases/química , Inibidores de Glicosídeo Hidrolases/isolamento & purificação , Camundongos , Modelos Moleculares , Estrutura Molecular , Óxido Nítrico/biossíntese , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Células RAW 264.7 , Relação Estrutura-Atividade , Árvores/químicaRESUMO
Three previously undescribed aporphine alkaloids, phaeanthuslucidines E-G, one previously undescribed naphthoquinone derivative, phaeanthusnaphthoquinone, and three known compounds were isolated from an EtOAc extract of the leaves of Phaeanthus lucidus Oliv. The structures of all previously undescribed compounds were established through extensive spectroscopic investigations and high-resolution mass spectroscopy. The 6aR configuration of phaeanthuslucidines E-G was assigned by comparing their ECD spectra and specific rotation values with the reported known compounds. Some isolated compounds were evaluated for their α-glucosidase inhibitory activity. Among these compounds, phaeanthuslucidine E showed the highest α-glucosidase inhibitory activity with an IC50 value of 17.9 ± 0.4 µM. The molecular docking of phaeanthuslucidine E was further studied.
Assuntos
Alcaloides , Aporfinas , alfa-Glucosidases , Simulação de Acoplamento Molecular , Estrutura Molecular , Alcaloides/farmacologia , Alcaloides/química , Aporfinas/farmacologia , Aporfinas/química , Inibidores de Glicosídeo Hidrolases/farmacologiaRESUMO
Phytochemical investigations of the twig and leaf extracts of Uvaria dac Pierre ex Finet & Gagnep. resulted in the isolation and identification of five new highly oxygenated cyclohexenes, uvaridacols M - Q (1-3, 5, and 6), and six known compounds (4 and 7-11). All new structures were elucidated by spectroscopic methods and HRESITOFMS data. The absolute configuration of 1, 5, and 6 was confirmed by single X-ray diffraction analysis with Cu Kα radiation. In contrast, other compounds were established by comparing their specific rotation and ECD spectra with those of known compounds. Some of the isolated compounds with sufficient quantity were evaluated for their α-glucosidase inhibitory activity. Of these, (-)-1,6-desoxypipoxide (10) showed α-glucosidase inhibitory activity with an IC50 value of 28.6 µM. The in silico molecular docking of active compounds was also studied.
Assuntos
Cicloexenos , Inibidores de Glicosídeo Hidrolases , Simulação de Acoplamento Molecular , Compostos Fitoquímicos , Folhas de Planta , Uvaria , Inibidores de Glicosídeo Hidrolases/farmacologia , Inibidores de Glicosídeo Hidrolases/isolamento & purificação , Inibidores de Glicosídeo Hidrolases/química , Estrutura Molecular , Uvaria/química , Folhas de Planta/química , Compostos Fitoquímicos/farmacologia , Compostos Fitoquímicos/isolamento & purificação , Cicloexenos/isolamento & purificação , Cicloexenos/farmacologia , Cicloexenos/química , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Caules de Planta/química , ChinaRESUMO
Two new styryl lactone derivatives, goniothapic acids A (1) and B (2), and 18 known compounds, were isolated from the twig and leaf extracts of Goniothalamus tapis Miq. The structures of new compounds were characterised by spectroscopic methods and HRESITOFMS. Their absolute configuration was established by comparing the experimental and calculated ECD spectra. Eleven compounds were evaluated for their α-glucosidase inhibitory activity. Of these, (-)-goniothalamin (5) and oldhamactam (16) showed the best α-glucosidase inhibitory activity with IC50 values of 54.8 and 57.9 µM, respectively.
RESUMO
BACKGROUND: Reducing the risk of falling by improving balance and leg strength may be a preventive strategy. This study evaluated the integrated effects of Thai essential oil and balance exercises on parameters associated with Falls in community-dwelling older adults at risk of falling. METHODS: Fifty-six participants were randomly allocated to either the intervention group (IG), which performed balance exercises while smelling Thai essential oil scents of Zanthoxylum limonella (Dennst.) Alston, or the control group (CG), which performed balance exercises while receiving a control patch. Balance exercises were practiced for 12, 30-minute sessions over 4 weeks. Static and dynamic balance with eyes open and eyes closed (EC), leg muscle strength, agility, and fear of falling were assessed at baseline, after the 4-week intervention, and at 1 month after the last intervention session. RESULTS: Both groups showed significant improvements in static and dynamic balance, ankle plantarflexor strength, and agility after the 4-week intervention (p<0.05), which persisted at the 1-month follow-up (p<0.05). Compared to the CG, the IG demonstrated significantly better static balance in terms of elliptical sway area (p=0.04) and center of pressure (CoP) velocity (p=0.001) during EC, as well as ankle plantarflexor strength (p=0.01). The IG also maintained a significantly greater improvement in CoP velocity during EC (p=0.01). CONCLUSION: Integrated Thai essential oil and balance exercises improved static balance and ankle plantarflexor strength compared to the balance exercise with a control patch in older adults at risk of falling.
RESUMO
A new geranylated xanthone, nigrolineaxanthone AA (1) together with 18 known compounds (2-19) were isolated from latex and twig extracts of Garcinia nigrolineata Planch. ex T. Anderson. Some of the isolated compounds were assessed for their antidiabetic activities and cytotoxicity against three cancer cell lines. Of these, compounds 12 (IC50 value of 25.8 ± 0.2 µM), 16 (IC50 value of 124.8 ± 0.7 µM), and 17 (IC50 value of 44.4 ± 1.1 µM) exhibited the highest α-glucosidase inhibitory, α-amylase inhibitory, and glycation inhibition activities, respectively. Compound 11 showed glucose consumption and glucose uptake with IC50 values of 14.2 ± 0.8 µM and 3.1-fold. Compound 10 displayed cytotoxic activity against colon cancer (SW480) with an IC50 value of 4.3 ± 0.1 µM), while compound 2 showed cytotoxicity against leukemic cancer (K562) with IC50 value of 4.4 ± 0.3 µM.
Assuntos
Antineoplásicos , Clusiaceae , Garcinia , Xantonas , Látex , Hipoglicemiantes/farmacologia , Estrutura Molecular , Extratos Vegetais/farmacologia , Xantonas/farmacologiaRESUMO
The first phytochemical investigation of the twigs of Phaeanthus lucidus Oliv. resulted in the isolation and identification of four undescribed alkaloids, including two aporphine dimers, phaeanthuslucidines A and B, a hybrid of aristolactam-aporphine, phaeanthuslucidine C, and a C-N linked aporphine dimer, phaeanthuslucidine D, together with two known compounds. Their structures were determined by extensive analysis of spectroscopic data, and by comparison of their spectroscopic and physical data with previous reports. Phaeanthuslucidines A-C and bidebiline E were analysed and resolved by chiral HPLC to yield the (Ra) and (Sa) atropisomers, whose absolute configurations were respectively determined by ECD calculations. Phaeanthuslucidines A and B, bidebiline E, and lanuginosine showed α-glucosidase inhibitory activities with IC50 values in the range of 6.7-29.2 µM. Moreover, molecular docking simulations of α-glucosidase inhibition of active compounds were studied.
Assuntos
Alcaloides , Annonaceae , Antineoplásicos , Aporfinas , Simulação de Acoplamento Molecular , alfa-Glucosidases , Estrutura Molecular , Alcaloides/química , Aporfinas/química , Annonaceae/químicaRESUMO
A phytochemical investigation of the twig extract of Trivalvaria costata (Hook.f. & Thomson) I.M.Turner has identified ten undescribed dimeric aporphine alkaloids, trivalcostatines A-J, one undescribed isoquinoline alkaloid, trivalcostaisoquinoline, and four known aporphine alkaloids. Their structures were elucidated by detailed analysis of NMR and HRESITOFMS data. Three of the dimeric aporphine structures were confirmed by single crystal X-ray diffraction analysis. All of the dimeric aporphine alkaloids were isolated as mixtures of atropisomers. Several of them were resolved by chiral-phase HPLC and the absolute configurations of the pure atropisomers were assigned by calculated and experimental ECD analysis. Bidebilines A and B, heteropsine, and urabaine showed α-glucosidase inhibitory activities with IC50 values in the range of 4.1-11 µM.
Assuntos
Alcaloides , Annonaceae , Aporfinas , Estrutura Molecular , Aporfinas/farmacologia , Aporfinas/química , Alcaloides/farmacologia , Alcaloides/química , Annonaceae/química , Espectroscopia de Ressonância MagnéticaRESUMO
A new C-benzylated flavone, uvariaruflavone (1), along with 13 known compounds (2-14) were isolated from the twig and leaf extracts of Uvaria rufa Blume. Their structures were established by extensive spectroscopic methods. Flavones (5-8) and cyclohexene (10) were isolated from U. rufa for the first time. Most of the isolated compounds were evaluated for their α-glucosidase and α-amylase inhibitory activities. Of these, uvariaruflavone (1) showed the highest α-glucosidase inhibitory activity with an IC50 value of 44.3 µM, while ferrudiol (12) displayed the highest α-amylase inhibitory activity with an IC50 value of 73.5 µM.
Assuntos
Uvaria , Uvaria/química , alfa-Glucosidases , Inibidores de Glicosídeo Hidrolases/farmacologia , Extratos Vegetais/farmacologia , Extratos Vegetais/química , alfa-AmilasesRESUMO
Special Thai oolong tea is oolong tea (Camellia sinensis (L.) Kuntze) steamed with selected Thai botanical drugs. Oolong tea steamed with ginger (Zingiber officinale), lemongrass (Cymbopogon citratus), and celery (Anathallis graveolens L.) is called eternity tea (EN), whereas peaceful rest (PR) tea is made of oolong tea leaves steamed with Indian gooseberry (Phyllanthus emblica), Turkey berry (Solanum torvum), and wild betel leaf bush leaves (Piper sarmentosum). Oolong tea is known for its numerous biological activities including antidiabetic properties. However, the effect of the additional botanical drugs on the biological activities of special oolong teas has not yet been explored. From the results, the PR extract exhibited the best activity in the in vitro assays relevant to antidiabetic properties such as chemical antioxidant, anti-inflammation, anti-adipogenesis, enzyme inhibition, and glucose uptake and consumption. The UHPLC-QTOF-MS/MS profiles of PR and EN extracts indicated chemical profiles different from oolong tea. For instance, gingerdiol and gingerol were detected in EN, whereas piperettine I was detected in PR. Therefore, it was inferred that among the three tea extracts, the additional compounds in PR contributed to good activities compared to oolong and EN. It is also important to highlight that the PR extract inhibited glucose uptake and consumption by adipocytes and skeletal muscles at concentrations of 500 and 100 µg/ml, respectively, as well as metformin activity (p < 0.05). Findings from this study support the antidiabetic potential of PR tea.
RESUMO
Phytochemical investigations of the leaf and pod extracts of Millettia brandisiana Kurz led to the isolation and identification of four previously undescribed rotenoids, (-)-(6aS,12aS)-millettiabrandisins A-C and (-)-(6aS,12aS)-6-deoxyclitoriacetal, two previously undescribed isoflavones, millettiabrandisins D and E, and 20 known compounds. The structures of previously undescribed compounds were determined on the basis of NMR and MS data. The absolute configurations of (-)-(6aS,12aS)-millettiabrandisins A-C were determined from the comparison of their experimental and calculated ECD spectra. (-)-(6aR,12aR)-12a-Hydroxy-α-toxicarol was also confirmed by X-ray crystallographic data. Some isolated compounds were evaluated for their cytotoxicity against three cancer cell lines, including lung cancer (A549), colorectal cancer (SW480), and leukemic cells (K562). Of these, α-toxicarol displayed the best cytotoxicity against lung cancer (A549) and leukemic cells (K562) with the IC50 values of 104.4 and 67.5 µM, respectively. 6â³,6â³-Dimethylchromene-[2â³,3â³:7,8]-flavone showed the highest cytotoxicity against colorectal cancer (SW480) with an IC50 value of 97.2 µM.
RESUMO
A phytochemical investigation of the root and leaf extracts of Millettia pachycarpa Benth resulted in the isolation and identification of 16 compounds, including six rotenoids (1-6) and 10 prenylated isoflavonoids (7-16). Compound 4 was isolated as a scalemic mixture, which was resolved by chiral HPLC to afford (-)-(6aS,12aS)-12a-hydroxy-α-toxicarol (4) and (+)-(6aR,12aR)-12a-hydroxy-α-toxicarol (4). (+)-(6aR,12aR)-Millettiapachycarpin (3) and (-)-(6aS,12aS)-12a-hydroxy-α-toxicarol (4) were isolated as new compounds. The absolute configuration of (-)-(6R)-pachycarotenoid (2), (+)-(6aR,12aR)-millettiapachycarpin (3), (-)-(6aS,12aS)-4 and (+)-(6aR,12aR)-12a-hydroxy-α-toxicarol (4), (+)-(6aS,12aS)-(5), and (-)-(6aS,12aS,2â³R)-sumatrol (6) were identified by electronic circular dichroism (ECD) data. (-)-(6aS,12aS,2â³R)-Sumatrol (6) was also confirmed by X-ray diffraction analysis using Cu-Kα radiation. Antidiabetic activities, including α-glucosidase and α-amylase inhibitory activities, and cytotoxicities against lung cancer A549, colorectal cancer SW480, and leukemic K562 cells of some isolated compounds were evaluated. Of these, isolupalbigenin (11) exhibited the highest α-glucosidase inhibitory activity, with an IC50 value of 11.3 ± 0.2 µM, whereas the scalemic mixture of 12a-hydroxy-α-toxicarol (4) displayed the best α-amylase inhibitory activity, with an IC50 value of 106.9 ± 0.2 µM. Euchrenone b10 (15) exhibited the highest cytotoxicity against lung cancer A549, colorectal cancer SW480, and leukemic K562 cells, with IC50 values of 40.3, 39.1, and 15.1 µM, respectively. In addition, molecular docking simulations of α-glucosidase inhibition of the active compounds were studied.
RESUMO
Red limestone is a mixture of turmeric (Curcuma longa L.) powder and limestone which is made from burning shells at high temperature. The yellow mixture turns to red color or deep orange because of the reaction between turmeric and calcium carbonate in limestone. Red limestone is traditionally used to treat many diseases such as abscess, cut wound and insect bite. The purpose of this study was to investigate anti-inflammatory activity and chemical constituents of red limestone. The chemical analysis of red limestone extract by liquid chromatography with tandem mass spectrometry revealed that red limestone consisted of alpha-turmerone and curcumanolide B as major components. These compounds were related with the chemical constituents in C. longa extract which is a main ingredient of red limestone. However, curcuminoids were not detected in red limestone extract. Cytotoxicity of red limestone extract was investigated. Macrophage cell lines (RAW 264.7) and human keratinocyte cell lines (HaCaT cells) were investigated cell viability using MTT assay. Red limestone extract was nontoxic to normal cells such as macrophage cells and human keratinocyte cells. Moreover, the inflammatory activity was detected nitric oxide (NO) secretion in RAW 264.7 cells. The result showed that the extracts inhibited NO in dose-dependent manner and IC50 was found to be 102.42 µg/ml. It suggested that red limestone extract had a potential for anti-inflammatory activity.
RESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Trimada is well-known polyherbal Ayurvedic formulation used in Indian Traditional medicine since ancient times. It consisted of three inebriant herbs including "Chitraka" (Plumbago zeylanica Linn. Family- Plumabaginaceae), "Musta" (Cyperus rotundus Linn. Family- Cyperaceae) and Vidanga (Embelia ribes Burm. F. Family- Myrsinaceae) in equal ratios as mentioned in Ayurveda. Trimada is traditionally used to increase the functioning of the digestive system and metabolism. Along with these, it also assists in the reduction of cholesterol as well as reduces stomach aches and chest pain. AIM OF THE STUDY: This study is aimed to identify the metabolites present in this polyherbal formulation. Further, the cytotoxicity and interaction potential of the formulation and individual herbs with Cytochrome P450 isozymes (CYP3A4, 2D6, 2C9, 1A2) was evaluated by MTT assay and CYP450 enzyme inhibition. The concentration of heavy metals was also determined. MATERIAL AND METHODS: Ultra-performance liquid chromatography-quadrupole/time-of-flight mass spectrometry (UPLC-QTOF-MS) analysis was performed to detect and identify the phytoconstituents in the formulation. Cytotoxicity of the formulation was evaluated by MTT assay. CYP450 enzyme interaction potential of the individual herbs and the Trimada formulation was carried out through CYP-CO assay and fluorometric high throughput screening (HTS) assay for individual isozymes. The content of heavy metal in the formulation was quantified by Atomic Absorption Spectroscopy. RESULTS: Trimada formulation exhibited lower cytotoxicity to human liver carcinoma cell line (HepG2). CYP-CO assay revealed that the interaction potential of individual herbs and Trimada on the liver microsomes was found to be lesser than the standard inhibitor ketoconazole. Individual herbs and Trimada formulation displayed higher IC50 values than the respective standard inhibitors in the fluorimetric assay. UPLC-QTOF-MS analysis showed the presence of a number of active phytoconstituents including sesquiterpenes, phenolic acids, benzoquinones, triterpenes and flavonoids. The heavy metal concentration in the traditional medicinal herbal formulation was found within the approved limit. CONCLUSIONS: This study suggested that the individual herbs and Trimada formulation exhibited low cytotoxicity and contributes insignificant interaction with CYP450 isozymes. So, the formulation is considered to be safe for its therapeutic management without any potential drug interaction involving CYP 450 isozymes.
Assuntos
Sistema Enzimático do Citocromo P-450/efeitos dos fármacos , Ayurveda , Microssomos Hepáticos/efeitos dos fármacos , Extratos Vegetais/farmacologia , Cromatografia Líquida de Alta Pressão , Cyperus/química , Sistema Enzimático do Citocromo P-450/metabolismo , Embelia/química , Células Hep G2 , Ensaios de Triagem em Larga Escala , Humanos , Concentração Inibidora 50 , Isoenzimas , Metais Pesados/análise , Metais Pesados/química , Metais Pesados/isolamento & purificação , Microssomos Hepáticos/metabolismo , Extratos Vegetais/química , Extratos Vegetais/metabolismo , Plumbaginaceae/químicaRESUMO
Garcinia cowa Roxb. ex Choisy (Clusiaceae) is a Thai local edible plant, which has been used for the treatment of diabetes. The aim of this study is to discover and identify bioactive compounds related to antidiabetic properties from the leaf extract of G. cowa. α-Glucosidase inhibitory bioassay-guided isolation of the ethyl acetate extract of the leaves of G.cowa resulted in the isolation and identification of 11 compounds. Of these, a decahydro-1H-xanthene derivative, garciniacowone K (1), was identified as a novel compound. Their structures were characterized by spectroscopic data and by comparison of their NMR spectroscopic data with those previously reported. All compounds were evaluated for their α-glucosidase inhibitory and glucose consumption activities. Compound 2 showed the highest efficacy in inhibiting α-glucosidase enzyme and promoting glucose consumption activity by 3T3-L1 cells, with IC50 values of 0.5 µM and 13.1 µM, respectively, without causing toxicity to cells.
RESUMO
Phytochemical investigations from the flower and leaf extracts of D. cochinchinensis resulted in the isolation and structural elucidation of five new polyoxygenated seco-cyclohexene derivatives, desmoscochinchinenes A-E (1-5), together with 11 known compounds (6-16). The structures on the new compounds were elucidated from their spectroscopic data, including UV, IR, NMR, and HRESITOFMS. Some of the isolated compounds were evaluated for their α-glucosidase inhibitory activities. Chrysin (9), pinocembrin 7-O-benzoate (12), and (-)-(5R)-desmoscochinoxepinone B (16) inhibited α-glucosidase better than the standard control (acarbose, IC50 = 83.5 µM) with IC50 values of 5.7, 33.8, 53.3 µM, respectively.
RESUMO
BACKGROUND: Lawsonia inermis L. has been used as a traditional or folk medicine for the treatment of a wide range of skin infectious diseases. OBJECTIVE: The objective of this study was to determine the pharmacognostic specifications and lawsone contents of L. inermis leaves. MATERIALS AND METHODS: The pharmacognostic specifications of L. inermis leaves from 12 sources were evaluated according to the WHO guideline of quality control method for medicinal plant materials. The lawsone contents were analyzed by thin-layer chromatography (TLC) coupled with densitometry and image analysis. RESULTS: Microscopic evaluation of L. inermis powders showed the fragment of mesophyll, fragment of parenchyma, epidermis layer with stomata, and the rosette crystal of calcium oxalate. Physicochemical parameters revealed that total ash, acid-insoluble ash, loss on drying, and water content should be not <6.98, 1.12, 8.08, and 9.86% of dried weight, respectively, whereas ethanol and water extractive values should be not < 19.67 and 23.06% of dried weight, respectively. The content of lawsone in L. inermis leaves by TLC-densitometry was found to be 0.76 ± 0.05 g/100 g of dried crude drug, whereas the lawsone content evaluation by TLC image analysis was found to be 0.87 ± 0.11 g/100 g of dried crude drug. The validation of the methods revealed that both TLC-densitometry and TLC image analysis showed a good sensitivity and accuracy for lawsone quantitation in L. inermis. CONCLUSION: The pharmacognostic specifications could be used as the standardization data of L. inermis leaves, and the development of TLC method could be applied to determine lawsone content in this plant material. SUMMARY: The pharmacognostic specification of Lawsonia inermis leaves could be used as the standardization data of L. inermis leaves in Thailand.Both TLC-densitometry and TLC image analysis showed a good sensitivity and accuracy for lawsone quantitation. Abbreviations Used: LOD: Limit of detection; LOQ: Limit of quantitation; RSD: Relative standard deviation; TLC: Thin layer chromatography; UV: Ultraviolet; Rf value: Relative to front value.
RESUMO
Zanthoxylum limonella (Dennst.) Alston. (Rutaceae) or Ma-khwaen is one of the medicinal plants in Thai traditional medicine. To investigate the pharmacognostic specifications and chemical constituents of Z. limonella fruits and seeds. Fruits and seeds of Z. limonella were collected from 15 sources throughout Thailand; then examined the pharmacognostic specification following WHO guideline of quality control method for medicinal plant materials. Microscopic determination of Z. limonella powders demonstrated fragment of mesocarp, fragment of brown vitta, oil glands, fragment of endocarp, and endosperm containing oil globule, trichome and pale brown stone cells. Stomatal index and pellucid dots in mm2 were 19.87 and 4.2 respectively. Physico-chemical parameters unveiled that loss on drying, water content, total ash, and acid-insoluble ash should be not >17.90%, 9.18%, 4.50%, and 0.60% of dried weight respectively; while ethanol, water, and hexane extractive values and volatile oil content should be not <2.24%, 2.27%, 1.57% and 9.63% of dried weight respectively. Rf values of thin-layer chromatographic fingerprint of Z. limonella fruits and seeds ethanolic extract were 0.38, 0.45, 0.90, and 0.97 detected ultraviolet (UV) light 254 nm, 0.30, 0.44, 0.67, and 0.77 detected UV light 366 nm, and 0.24, 0.73, 0.78, and 0.93 detected 10% sulfuric acid. There are three main chemical compounds in Z. limonella oil including limonene (43.63%), (+)-sabinene (16.72%), and terpinen-4-ol (10.95%). The result gained from pharmacognostic specifications and chemical fingerprints could be used as standardization data of Z. limonella fruits and seeds to apply or provide for guarantee of quality.