Determination of trace and heavy metals in some commonly used medicinal herbs in Ayurveda.

Traditionally, the herbal drugs are well established for their therapeutic benefits. Depending upon their geographical sources sometimes the trace and heavy metals' content may differ, which may lead to severe toxicity. So, the toxicological and safety assessment of these herbal drugs are one of the major issues in recent days. Eight different plant species including Aloe vera, Centella asiatica, Calendula officinalis, Cucumis sativus, Camellia sinensis, Clitoria ternatea, Piper betel and Tagetes erecta were selected to determine their heavy and trace metals content and thereby to assure their safer therapeutic application. The trace and heavy metals were detected through atomic absorption spectrometry analysis. The selected medicinal plant materials were collected from the local cultivated regions of West Bengal, India, and were digested with nitric acid and hydrochloric acid as specified. Absorbance was measured through atomic absorption spectrometer (AA 303) and the concentration of different trace and heavy metals in the plant samples were calculated. The quantitative determinations were carried out using standard calibration curve obtained by the standard solutions of different metals. The contents of heavy metals were found to be within the prescribed limit. Other trace metals were found to be present in significant amount. Thus, on the basis of experimental outcome, it can be concluded that the plant materials collected from the specific region are safe and may not produce any harmful effect of metal toxicity during their therapeutic application. The investigated medicinal plants contain trace metals such as copper (Cu), chromium (Cr), manganese (Mn), iron (Fe) and nickel (Ni) as well as heavy metals such as arsenic (As), lead (Pb) and mercury (Hg), which were present within the permissible limit.

Quantification of glycyrrhizin in Glycyrrhiza glabra extract by validated HPTLC densitometry.

Glycyrrhiza glabra Linn (Family-Fabaceae) is active as an anti-allergic, anti-inflammatory, spasmolytic, mild laxative, antistress, antidepressive, antiulcer, liver protective, estrogenic, emmenagogue, and antidiabetic substance, and is widely used in the Indian system of medicine. The major bioactive constituent is glycyrrhizin. A simple HPTLC method has been developed to control the quality of raw as well as finished glycyrrhiza using glycyrrhizin as the bioactive marker. The solvent system was optimized to chloroform-methanol-water (65 + 36 + 7.5, v/v/v). Extract and standard were dissolved in 70% methanol and applied on a precoated TLC plate. After development, the plate was scanned at 254 nm to create a chromatogram, then the quantity of glycyrrhizin was determined in the extract. The method was validated in terms of specificity, linearity, precision, LOD, and LOQ. Linearity range was found to be 0.96-4.80 microg per spot. The linearity relationship was described by the equation: Y = 612.706 + 1.091X (with r = 0.99904 and SD = 2.52%), where Y is the area under curve and X is the amount of glycyrrhizin (ng). The amount of glycyrrhizin found in the extract was 9.1%. Thus, the method provides a rapid and cost-effective quality measure for Glycyrrhiza glabra hydroalcoholic extract.

Anti-biofilm activity of Marula - a study with the standardized bark extract.

ETHNOPHARMACOLOGICAL RELEVANCE: Marula (Sclerocarya birrea; family - Anacardiaceae) is an African plant, which enjoys wide socio-economic importance particularly in southern part of Africa. The fruits are consumed as food and also as alcoholic beverage (cream liquor). In different parts of Africa, the decoction of the bark is traditionally used for the treatment of dysentery, diarrhoea, and various other infectious conditions. The aim of the study was to investigate the anti-biofilm properties of the methanol extract of Marula bark (stem bark of Sclerocarya birrea), with a view towards combating the emergence of antimicrobial resistance often associated with bacterial biofilms. MATERIALS AND METHODS: The standardized methanol extract was initially tested for its antimicrobial property. The crystal violet assay was used for evaluating anti-biofilm (biofilm formation by Pseudomonas aeuginosa) activity. Further in order to study the mechanism of anti-biofilm activity, the same was evaluated for understanding its role on various quorums sensing mediated phenomenon (swarming motility assay, protease and pyoverdin assay) that are known to be associated with the formation of biofilms and pathogenicity. RESULTS: The methanol extract showed no inhibition of bacterial growth up to a concentration of 200 µg/ml. Interestingly, the sample produced anti-biofilm activity (around 75% decrease; 100 µg/ml) at sub-lethal concentration. Further it also significantly reduced the QS mediated swarming motility. The release of various virulent factors (protease and pyoverdin) was found to be lowered when pre-treated with the extract. CONCLUSION: The present study illustrates the anti-biofilm property Sclerocarya birrea. The standardized extract significantly disrupted the quorum sensing mediated production of biofilm formation and also inhibited swarming ability of the cells. The extract displayed a regulatory role on the secretion of protease and pyoverdin, two QS dependent pathogenic factors found in Pseudomonas aeruginosa. This study also validates the ethnobotanical use of Marula.

Phytochemical and therapeutic potential of cucumber.

Cucumber (Cucumis sativus L.) is a member of the Cucurbitaceae family like melon, squash and pumpkins. It is a popular vegetable crop used in Indian traditional medicine since ancient times. This vegetable is very high in water content and very low in calories. It has potential antidiabetic, lipid lowering and antioxidant activity. Cucumber has a cleansing action within the body by removing accumulated pockets of old waste materials and chemical toxins. Fresh fruit juice is used for nourishing the skin. It gives a soothing effect against skin irritations and reduces swelling. Cucumber also has the power to relax and alleviate the sunburn's pain. The fruit is refrigerant, haemostatic, tonic and useful in hyperdipsia, thermoplegia etc. The seeds also have a cooling effect on the body and they are used to prevent constipation. Several bioactive compounds have been isolated from cucumber including cucurbitacins, cucumegastigmanes I and II, cucumerin A and B, vitexin, orientin, isoscoparin 2″-O-(6‴-(E)-p-coumaroyl) glucoside, apigenin 7-O-(6″-O-p-coumaroylglucoside) etc. Despite huge exploration of cucumber in agricultural field, comparatively very few studies have been published about its chemical profile and its therapeutic potential. This article reviews the therapeutic application, pharmacological and phytochemical profile of different parts of C. sativus. In this review we have explored the current phytochemical and pharmacological knowledge available with this well known plant and several promising aspects for research on cucumber.

Simultaneous estimation of hydroxychavicol and chlorogenic acid from Piper betel L. through RP-HPLC.

A RP-HPLC method was developed (λ (max) =280) to quantify hydroxychavicol and chlorogenic acid in Piper betel Linn. The method was validated for linearity, limit of detection (LOD=3:1σ/S), limit of quantification (LOQ=10:1σ/S), precision, accuracy and ruggedness. The response was linear with good correlation between concentration and mean peak area through a coefficient of determinants (r (2)) of 0.9940, y=1.98e +004x +5.19e +004 and 0.9945, y=2.76e+004x+1.40e+005 with LOD 1.6 µg mL(-1), 1.0 µg mL(-1) and LOQ 5.0 µg mL(-1) and 3.0 µg mL(-1), respectively, for hydroxychavicol (28.56% w/w) and chlorogenic acid (0.40% w/w). The %RSD of precision and recovery of hydroxychavicol and chlorogenic acid were <2.0%. The proposed method was simple, accurate, specific, precise and reproducible.

Standardized Clitoria ternatea leaf extract as hyaluronidase, elastase and matrix-metalloproteinase-1 inhibitor.

AIM: Plant Clitoria ternatea L. is claimed to possess a wide range of activities including antiinflammatory, local anesthetic and antidiabetic effect, etc. The aim of the present study was to evaluate the wound healing potential of standardized C. ternatea leaf extract in terms of different enzymatic models, which are mostly associated with skin wound. MATERIALS AND METHODS: The methanol extract and fractions were screened for its hyaluronidase, elastase, and matrix metalloproteinase-1 (MMP-1) inhibitory activity compared with standard oleanolic acid. The activity was rationalized through reverse phase high performance liquid chromatography (RP-HPLC) standardization of the extract and fractions with respect to its isolated biomarker taraxerol (yield 5.27% w/w). RESULTS: The extract showed significant (P < 0.001) hyaluronidase (IC(50) 18.08 ± 0.46 µg/ ml) and MMP-1 (P < 0.05) inhibition, but the elastase inhibition was insignificant (IC(50) 42.68 ± 0.46 µg/ml). Among the fractions, ethyl acetate fraction showed significant (P < 0.001) inhibition of hyaluronidase (IC(50) 28.01 ± 0.48 µg/ml) and MMP-1 (P < 0.01). The HPLC analysis revealed that the extract and the ethyl acetate fraction are enriched with taraxerol (5.32% w/w and 4.55% w/w, respectively). CONCLUSIONS: The experiment validated the traditional uses of C. ternatea and may be recommended for use in the treatment of different types of skin wounds, where taraxerol may be a responsible biomarker.

Changing scenario for promotion and development of Ayurveda--way forward.

ETHNOPHARMACOLOGICAL RELEVANCE: Ayurveda, derives from the Sanskrit words Ayus (life) and Veda (knowledge) is the most ancient system of traditional medicine of the world. It has been practiced in Indian peninsula since 5000 BC to offer natural ways to treat diseases and to promote healthcare. METHODOLOGY: We reviewed the literature on the history, principles and current status of Ayurveda. The data have been presented systematically including the initiatives from Government of India. Several aspects of administrative management, education, teaching and related aspects for promotion and development of Ayurveda and other Indian systems of traditional medicine have been discussed. RESULTS: This paper reviews on different aspects of development of Ayurveda. Presently, there are 2420 hospitals with about 42271 beds, 15017 dispensaries, 429246 registered practitioners, more than 320 educational institutions, 7699 drug-manufacturing units to promote Ayurveda into the health care delivery system in the country. Ayurvedic Pharmacopoeia of India is the official document for single Ayurvedic drugs (540 monographs) and different formulations (152 monographs). Several aspects in this regard for development of Ayurveda have been discussed. CONCLUSION: Considering the widespread use and popularity of Ayurveda worldwide, administrative management and infrastructure facilities, indigenous practices and standards for quality control and it's evaluation have been highlighted. In India, all such efforts for integration of Ayurveda provide potential role in the health care benefits.

Bioactive compounds from natural resources against skin aging.

Skin aging involves degradation of extracellular matrix (ECM) in both the epidermal and dermal layers, it leaves visible signs on the surface of skin and the physical properties of the skin are modified. Chronological aging is due to passage of time, whereas premature aging occurred due to some environmental factors on skin produces visible signs such as irregular dryness, dark/light pigmentation, sallowness, severe atrophy, telangiectases, premalignant lesions, laxity, leathery appearance and deep wrinkling. There are several synthetic skincare cosmetics existing in the market to treat premature aging and the most common adverse reactions of those include allergic contact dermatitis, irritant contact dermatitis, phototoxic and photo-allergic reactions. Recent trends in anti-aging research projected the use of natural products derived from ancient era after scientific validation. Ample varieties of phytomolecules such as aloin, ginsenoside, curcumin, epicatechin, asiaticoside, ziyuglycoside I, magnolol, gallic acid, hydroxychavicol, hydroxycinnamic acids, hydroxybenzoic acids, etc. scavenges free radicals from skin cells, prevent trans-epidermal water loss, include a sun protection factor (SPF) of 15 or higher contribute to protect skin from wrinkles, leading to glowing and healthy younger skin. Present era of treating aging skin has become technologically more invasive; but herbal products including botanicals are still relevant and combining them with molecular techniques outlined throughout this review will help to maximize the results and maintain the desired anti-skin aging benefits.

Cucumis sativus fruit-potential antioxidant, anti-hyaluronidase, and anti-elastase agent.

The aim of this study was to screen the antioxidant, anti-hyaluronidase, and anti-elastase activity of the lyophilized juice of Cucumis sativus fruit (CSLJ). The CSLJ was subjected to DPPH and superoxide radical scavenging assay in reference to butylated hydroxytoluene. The hyaluronidase and elastase inhibitory assay was performed in reference to oleanolic acid. Furthermore, the activities have been rationalized with HPLC analysis of the CSLJ with standard reference compound of ascorbic acid. The CSLJ exhibited DPPH-free radical and superoxide radical scavenging activity, IC(50) at a concentration of 14.73 ± 1.42 and 35.29 ± 1.30 µg/mL, respectively. The CSLJ also showed strong anti-hyaluronidase ((c) P < 0.001) and anti-elastase ((c) P < 0.001) activity, IC(50) at a concentration of 20.98 ± 1.78 and 6.14 ± 1.74 µg/mL, respectively. The HPLC 'chromatogram' of standard and CSLJ showed specific peak at retention time 2.905 and 3.066 min, respectively. Content of ascorbic acid was calculated with respect to the standard compound and it was found to be 3.5 ± 0.23% w/w. CSLJ is the rich source of ascorbic acid and this study thereby rationalizes the use of C. sativus as potential anti-wrinkle agent in cosmetic products.

Exploring Tagetes erecta Linn flower for the elastase, hyaluronidase and MMP-1 inhibitory activity.

ETHNOPHARMACOLOGICAL RELEVANCE: Traditionally Tagetes erecta Linn flower is claimed to treat skin diseases like sores, burns, wounds, ulcers, eczema and several other skin ailments. The aim of the present experiment was to evaluate the anti-wrinkle potential of standardized flower extract of Tagetes erecta. MATERIALS AND METHODS: The Tagetes erecta extract and fractions were screened for hyaluronidase, elastase and matrix metalloproteinase (MMP-1) inhibitory activity compared with the activity of standard oleanolic acid. Syringic acid and ß-amyrin were obtained from the extract and quantified through RP-HPLC. Also the compounds were evaluated for anti-wrinkle activity. RESULTS: The methanol extract showed significant ((a)P < 0.05) hyaluronidase and elastase inhibition with IC50 of 11.70 ± 1.79 µg mL(-1) and 4.13 ± 0.93 µg mL(-1) respectively and better MMP-1 inhibition compared to standard oleanolic acid. The isolated compounds syringic acid and ß-amyrin found to inhibit enzymes comparable to oleanolic acid. The RP-HPLC analysis revealed that good amounts of syringic acid and ß-amyrin (2.30%, w/w and 0.06%) are present in Tagetes erecta. CONCLUSION: Tagetes erecta flower showed effective inhibition of hyaluronidase, elastase and MMP-1. Therefore, this experiment further rationalizes the traditional uses of this plant, which may be useful as an anti-wrinkle agent.

Mast cell stabilization and antihistaminic potentials of Curculigo orchioides rhizomes.

AIM OF THE STUDY: To investigate the mast cell stabilization and antihistaminic activities of the rhizomes of Curculigo orchioides (COR). Extract of Curculigo orchioides Gaertn. (Fam. Amaryllidaceae) has been reported to possess immunostimulant, and anti-inflammatory potentials. In Indian traditional system of medicine it is also used as anti-asthmatic and anti-inflammatory. MATERIALS AND METHODS: Estimation of histamine release is key parameter for evaluating any target for its anti-allergic potential. The stabilization potential of the alcoholic extract of COR (100-400mg/kg) against mast cell degranulation was studied on isolated mice peritoneal mast cells. The antihistaminic activity was performed by determining the mortality rate of mice upon exposure to compound 48/80 and effect on inhibition of histamine release upon degranulation. RESULTS: The raised number of intact mast cells intimates that the COR stabilized the mast cell degranulation (60.96+/-1.96%) and percentage antihistaminic potential of the extract (63.58+/-1.8 inhibition at dose of 400mg/kg) and it virtues further work towards the isolation of phytoconstituents from this plant. CONCLUSION: This finding provides evidence that COR inhibits mast cell-derived immediate-type allergic reactions and mast cell degranulation.