Quality Control Of Herbal Materials Research Articles

Application of the herbal chemical marker ranking system (Herb MaRS) to the standardization of herbal raw materials: a case study

IntroductionPhytochemical standardization of herbal materials involves establishing consistent levels of one or more active ingredients or markers. It ensures the authenticity and quality of herbal materials, extracts, and their products. This research aimed to apply the herbal chemical marker ranking system (Herb MaRS) originally proposed for quality assurance of complex herbal products to establish markers for controlling the quality of herbal raw materials.MethodsThe assessment of compounds for suitability as markers was based on the Herb MaRS, with minor modifications as follows: for more objective scoring, evidence of biological activity of the potential marker compound(s) was determined at three levels based on the number of symptoms of the disease condition a compound can treat or alleviate: (i) one symptom (1 point), two symptoms (2 points), and 3 or more symptoms (3 points). The reported concentrations of the compounds were also scored as follows: concentration not determined (0 points), concentration ≥ 5 ppm (1 point), concentration ≥ 50 ppm (2 points) and availability of analytical standards (1 point). Finally, the compounds were scored for the availability of an analytical method (1 point). The compounds were scored from 0 to 8, where 8 indicated the most suitable chemical marker.ResultsThe selected markers were as follows: aromadendrine, α-terpineol, globulol, and 1,8-cineol (in Eucalyptus globulus Labill. ); aloin, aloe emodin, acemannan (in Aloe barbadensis (L.) Burm.f. ), lupeol, lupenone, betulinic acid, betulin, and catechin (in Albizia coriaria Oliv.); mangiferin, catechin, quercetin, and gallic acid (in Mangifera indica L.); polygodial (in Warburgia ugandensis Sprague); azadirachtin, nimbin, nimbidin (in Azadirachta indica A. Juss. ); and 6,8,10-gingerols, and 6-shogaol (in Zingiber officinalis Roscoe).ConclusionsHerb MaRS can be efficiently applied to select marker compounds for quality control of herbal materials. However, for herbs whose phytochemicals have not been sufficiently researched, it is difficult to establish evidence of activity, and there are no analytical standards and/or methods; this is the case for plants exclusively used in Africa. The markers identified should be incorporated into chromatographic fingerprints, their quantitative methods developed, and evaluated for applicability at the various stages of the production chain of herbal medicines; then, they can be included in future local plant monographs. There is also a need to build local capacity to isolate marker compounds, particularly those that are not sold by current vendors.

Antidiabetic activity of Ayurvedic polyherbal formulations Avipattikara Churna and Triphla Churna in streptozotocin and nicotinamide induced diabetic wistar rats

Objective : The aim of the present study is to formulate a polyherbal formulation whose formula is given in AFI and evaluate its antidiabetic potential in animals. Method: The polyherbal formulation was prepared using the methanol extracts and aqueous extracts of the given parts of herbs in AFI. The quality of the finished product was evaluated as per the World Health Organization’s guidelines for the quality control of herbal materials. The quality testing parameters of the polyherbal formulation were within the limits. Results : The acute toxicity studies of the polyherbal formulation did not show any toxic symptoms in doses up to 2000 mg/kg over 14 days. The extracts of both the formulations were evaluated for In-vitro and In-vivo anti-diabetic study. The oral anti-diabetic activity of the polyherbal formulation (200 and 400 mg/kg, p.o) was screened against intraperitoneal (i.p.) STZ (35 mg/kg) 15 min after i.p. administration of nicotinamide (200 mg/kg) continuously for 2 days, to induced diabetes mellitus in Wistar rats. The investigational drug was administered for 28 consecutive days, and the effect of the polyherbal formulation on blood glucose levels was studied at regular intervals.

Simultaneous Determination of Pharmacological Active Phytocompound Gallic Acid and Quercetin in Aerial Parts of Solanun indicum and Solanum xanthocarpum by Validated High Performance Thin‑layer Chromatography (HPTLC) Method

Objective: Quantitative standardization of plant‑based products is challenging albeit essential to maintain their quality. This study aims to develop and validate high‑performance thin‑layer chromatography (HPTLC) method for the simultaneous determination of gallic acid and quercetin from aerial parts of Solanum indicum and Solanum xanthocarpum. Materials and Methods: The stock solution (1-mg/mL) of standard, gallic acid and quercetin in methanol: Water (1:1) was serially diluted and spotted (5 µL) on silica gel 60 F254 thin‑layer chromatography plates. Toluene: Ethyl acetate: Formic acid: Methanol (3.5:4.8:1.0.:0.7, v/v/v/v) was selected as mobile phase for analysis at 280 nm. Results: The developed method was robust and resolved gallic acid and quercetin at Rf0.34 ± 0.03 and 0.55 ± 0.01, respectively. The limit of detection (26 and 47 ng band-1 ) limit of quantification (78 and 141 ng.band-1 ), recovery (99.6–99.8 and 98.5–99.7%), and precision (≤1.98 and 1.97) were satisfactory for gallic acid and quercetin respectively. Linearity range for gallic acid and quercetin were 100–1000 (r 2 = 0.9993) and 150–900 ng band-1 (r 2 = 0.9956) and the contents estimated as 0.63 ± 0.01% and 0.57 ± 0.01% w/w in SI and 0.51 ± 0.01% and 0.69 ± 0.01% w/w for gallic acid and quercetin respectively. Conclusion: The developed HPTLC method was rapid, accurate, precise, reproducible, and specific for the concurrent estimation of gallic acid and quercetin. The method has been successfully applied in the analysis and routine quality control of herbal material and formulations containing Solanum Species.

Validated High-Performance Thin-Layer Chromatography-Mass Spectrometry Method and Stability Study of Linalool in the Volatile Oil of the Rhizomes of Homalomena aromatica Schott.

Homalomena aromatica is a herb of tremendous ethnomedicinal importance to various communities residing in northeast India. In this study, a high-performance thin-layer chromatography-based densitometric method was developed for identification, quantification and stability study of linalool. Mass spectrometry was hyphenated to HPTLC for streamlining the method. The stability of linalool was studied by analyzing the effect of acid, base, UV, sunlight, thermal stress and H2O2 on linalool. The chromatographic plates were developed to a height of 70mm in toluene:ethyl acetate solvent system at a ratio of 9.5:0.5 and visualized with p-anisaldehyde reagent. The developed method was found to be precise, accurate and reproducible according to International Conference on Harmonization guidelines, and compact bands of linalool were observed at Rf of 0.351 ± 0.001. The content of linalool in the volatile oil of H. aromatica was found to be 58% v/v. By application of the hyphenated MS technique, linalool was identified at m/z 137, (M + H)+. It was observed that acidic pH has the highest effect on linalool with a percentage degradation of 65. The developed method can be used in the analysis and quality control of herbal materials and volatile oils containing linalool and quality control of rhizomes of H. aromatica.

DEVELOPMENT AND EVALUATION OF ANTIDIABETIC POTENTIAL OF POLYHERBAL FORMULATION IN STREPTOZOTOCIN INDUCED ANIMAL MODEL

Vitex trifolia L, Vitex parviflora and Terminalia chebula are well-known plants available throughout India and are commonly used for the treatment of various diseases including diabetes mellitus. The antidiabetic activity of the individual plant parts is well known, but the synergistic or combined effects are unclear. The concept of polyherbalism has been highlighted in Sharangdhar Samhita, an Ayurvedic literature dating back to 1300 AD. Polyherbal formulations enhance the therapeutic action and reduce the concentrations of single herbs, thereby reducing adverse events. The aim of the present study is to formulate a polyherbal formulation and evaluate its antidiabetic potential in animals. The polyherbal formulation was formulated using the hydroalcoholic extracts of the leaves of V. trifolia, L. parviflora and T. chebula in the ratio of 1:1:1. The quality of the finished product was evaluated as per the World Health Organization's guidelines for the quality control of herbal materials. The quality testing parameters of the polyherbal formulation were within the limits. The acute toxicity studies of the polyherbal formulation did not show any toxic symptoms in doses up to 2000 mg/kg over 14 days. The oral antidiabetic activity of the all extract and PHF (100 and 200mg/kg) was screened against streptozotocin (STZ) induced diabetes in rats and glibenclamide was used (500 mcg/kg body weight) as standard drug. The investigational drug was administered for 21 consecutive days, and the effect of the extract and polyherbal formulation on blood glucose levels was studied at regular intervals. At the end of the study, the blood samples were collected from all the animals for biochemical estimation. Polyherbal formulation showed significant antidiabetic activity at 100 and 200mg/kg, respectively and this effect was comparable with that of glibenclamide. The antidiabetic activity of polyherbal formulation is supported by biochemical analysis. The formulation has emerged as potential combination which can challenge the synthetic drug.

Quantitative Determination of Aloin in Aloe vera and Its Antioxidant Activity

Aloe vera is a popular medicinal plant used widely by the cosmetic, pharmaceutical and food industries. The Aloe vera gel, which is used mostly for its positive effects on human health,contains over 75 different bioactive compounds, including aloin. A sensitive and reliable densitometric High Performance Thin Layer Chromatography method has been developed for the quantification of aloin, an anthraquinone present in Aloe vera leaves. Chromatographic analysis was performed using methanol extract of leaves of Aloe vera using solvent system ethyl acetate: Methanol: water (100:13.5:10). Detection and quantification of aloin was done by densitometric scanning at 350nm. The results of linearity range and correlation coefficient show that there was a good correlation between peak area and corresponding concentration of aloin. The method developed here in can be implemented in the analysis and routine quality control of herbal materials. Antioxidant activity of Aloe vera was also done by using DPPH method and found that Aloe vera is also a natural antioxidant.

Ethnopharmacological analysis from Thai traditional medicine called prasachandaeng remedy as a potential antipyretic drug

Ethnopharmacological relevancePrasachandaeng (PSD) remedy is a famous antipyretic drug for adults and children in Thai traditional medicine used and is described in Thailand's National List of Essential Medicine. Relationship between the taste of this herbal medicine, ethnopharmacological used and its pharmacological properties was reviewed. Aims of studySince there has been no scientific report on the antipyretic activity of PSD, aim of this study was to investigate the efficacy related antipyretic drug of the remedy and its 12 herbal ingredients. It involved quality evaluation of raw materials, extraction of PSD and its ingredients, in vitro evaluation of their inhibitory activities on fever mediators, i.e. NO and PGE2 production in murine macrophage (RAW 264.7) cell line stimulated by lipopolysaccharide, and its stability study of the 95% ethanolic extract of PSD remedy. Materials and methodsPSD remedy was extracted by maceration with 50% and 95% ethanol (PSD50 and PSD95), by decoction with distilled water (PSDW), and hydrolysis of PSDW with 0.1 N HCl (PSDH). The 12 plant ingredients were extracted with 95% ethanol. Quality evaluation of PSD ingredients was performed according to the standard procedures for the quality control of herbal materials. The inhibitory activity on nitric oxide production was determined by the Griess reaction and the inhibition of prostaglandin E2 production was determined using the ELISA test kit. ResultsPSD ingredients passed the quality standard stipulated for herbal materials. PSD95 exhibited the highest inhibitory activities on the production of NO and PGE2 with the IC50 values of 42.40 ± 0.72 and 4.65 ± 0.76 μg/mL, respectively. A standard drug acetaminophen (ACP) exhibited inhibition of NO and PGE2 production with the IC50 values of 99.50 ± 0.43 and 6.110 ± 0.661 μg/mL, respectively. The stability study was suggested two years shelf-life of PSD95. This is the first report on the activity related antipyretic activity of PSD remedy and its ingredients against two fever mediators, NO and PGE2. ConclusionThe results suggested that the 95% ethanolic extracts of PSD remedy and some of its ingredients, were better than ACP in reducing fever. PSD should be further studied using in vivo models and clinical trials to support its use as an antipyretic drug in Thai traditional medicine.

A comparative study of aged and contemporary Chinese herbal materials by using delayed luminescence technique

BackgroundInvestigation of aged Chinese herbal materials will help us to understand their use and sources in ancient time and broaden the historical perspective of Chinese material medica. To reach this aim, the basic understanding of aged herbal materials, including physical and chemical characters, is of great importance. Delayed luminescence (DL) technique was developed as a rapid, direct, systemic, objective and sample loss-free tool to characterize the properties of Chinese herbal materials. In this study, we measured DL values in aged Chinese herbal materials that were transported from Asia to Europe during the 20th century and stored in Naturalis Biodiversity Center and the Utrecht University museum, and compared these with modern material of the same species.MethodsA hyperbolic function was used to extract four properties from the DL curves of Chinese herbal material from 1900, the 1950s and recently harvested products. Statistical tools, including the Student’s t test, One-way analysis of variance and Principal Component Analysis, were used to differentiate the DL properties of aged and contemporary collections of Glycyrrhiza spp. Curcuma aromatica Salisb., Zingiber officinale Roscoe, Alpinia officinarum Hance and Acorus calamus L.ResultsOur results showed that DL properties were significantly different between historical and contemporary Chinese herbal materials. Changes in DL values were species-dependent: the effects of storage time of DL properties were specific for each species. These outcomes help us not only in the identification of historical Chinese medicine products but also provides valuable data of the effect of storage time on herbal materials.ConclusionThe simple, direct, rapid, and inexpensive measurements offered by DL provide a novel tool to assess the taxonomic identity of Chinese and other herbal materials and assess the differences in chemical properties with increasing storage time. Our results contribute to the further development of novel digital tools for the quality control of herbal materials.

Application of delayed luminescence measurements for the identification of herbal materials: a step toward rapid quality control

BackgroundHerbal materials are widely used as medicinal products, dietary supplements, food, and spices. With increased consumption, the safety, quality, and efficacy of herbal materials are becoming more relevant. The authenticity of herbal materials plays an important role in herbal quality control, and there is an urgent need to develop a simple, direct, objective, rapid, and inexpensive measurement tool for the identification of herbal materials for the purpose of quality control.MethodsDelayed luminescence (DL) was used to measure authentic and counterfeit herbal materials. A hyperbolic function was used to extract four properties from the DL curves of the herbal materials. Statistical tools, including Student’s t test and Principal Component Analysis, were used to differentiate authentic and counterfeit herbal materials based on the DL properties.ResultsOur results showed that authentic and counterfeit herbal materials could be identified based on the DL properties as follows: (a) authentic versus counterfeit materials; (b) authentic versus adulterated materials; (c) authentic versus sulfur-fumigated materials; as well as (d) authentic versus dyed materials.ConclusionThe simple, direct, rapid, and inexpensive measurements offered by DL potentially offer a novel technique for the identification of Chinese herbal materials. However, the establishment of a valid database will be the next step toward the possible application of this technique, which would contribute significantly to the development of a novel digital tool for the quality control of herbal materials.

Quantitative Determination of thymoquinone in Nigella Sativa and its nano formulation using validated stability indicating HPTLC densiometric method

An accurate and stability indicating high-performance thin layer chromatographic method (HPTLC) was developed and validated for the quantification of thymoquinone (TQ) as per the ICH guidelines. The analysis was carried out on the aluminum plate using n-hexane: ethyl acetate: methanol (7:2:1 v/v/v) as the mobile phase and the densitometric determination was carried out by TLC scanner (CAMAG) at 254 nm. The developed method was validated for different parameters like linearity, precision, recovery, robustness, and stressed stability study. The developed analytical method was found to be linear in the concentration range of 75-500 ng band-1 with regression value closer to unity (r2 = 0.997). The developed system was found to give compact spots for thymoquinone (Rf 0.77) with the limit of detection and limit of quantification (18 and 54 ng band-1) respectively. Further, the study showed accuracy, precision and repeatability were all within the required limits. The stress degradation study of TQ showed well separated degraded peak from the pure thymoquinone. The mean recoveries measured at three concentrations were more than 95% with RSD ≤ 3%. The method has been successfully applied in the analysis and routine quality control of herbal material and formulations containing TQ.Taleuzzaman et al., International Current Pharmaceutical Journal, September 2017, 6(10): 53-60http://www.icpjonline.com/documents/Vol6Issue10/01.pdf

Determination of carnosic acid in Rosmarinus officinalis L. using square wave voltammetry and electrochemical behavior

A new, fast, sensitive and simple voltammetric method is established for the direct determination of carnosic acid (CA). And the electroreduction of carnosic acid (CA) was studied using electrochemical methods. The number of electrons transferred in electrode mechanisms were calculated for reversible and adsorption-controlled electrochemical reduction of CA at 17 mV versus Ag/AgCl at pH 7.0 in Britton–Robinson buffer (BR) on a hanging mercury drop electrode. Square-wave voltammetry was developed and validated for direct determination of CA. Square-wave parameters were optimized as accumulation potential = 0.0 mV, accumulation time = 5 s, frequency = 50 Hz, pulse amplitude = 50 mV, and staircase step potential = 5 mV. The developed method displays three linear responses from 2 to 9 μM, 10 to 30 and 40 to 90 μM for carnosic acid with a correlation coefficient of 0.996, 0.999 and 0.999. The detection limits were found to be 1.5 μM, 4.0 μM and 40.1 μM, respectively. The interference effect of most common organic and inorganic species was investigated. Proposed method was successfully applied for determination of CA in natural extract of rosemary and the average content was determined as 11.9 ± 1.0 (μg CA/1 g rosemary). The results were in agreement with that obtained by HPLC-UV comparison method. The developed method can be widely used in routine quality control of herbal materials as well as other in foods, medicinal, pharmaceutical and environmental analysis.

Is It Possible To Identify Constituent Herbs In Decoctions By Molecular Approach?

Genomic DNA provides a rich source of material for species identification [1]. However, degradation of DNA during the processing procedures makes people believe that molecular authentication is unsuitable for herbal decoctions, a common form of herbal products. Using ginseng decoctions prepared by boiling for 30, 60 and 120 min as examples, we show that DNA may be extracted from these preparations for polymerase chain reaction (PCR) and DNA sequencing. We also found: (a) sample pulverization before decoction boiling improved PCR detection, (b) prolonged boiling increased the DNA concentration but decreased the intactness of DNA. At 120 min, only DNA with size of less than 191 bp was detected. We have further shown that molecular authentication could be performed on a decoction consisting of four herbs and a commercial ready-to-serve soup [2]. We are investigating if molecular techniques can be used to quantify herbal components in decoctions. Our study has enhanced the value of molecular techniques in the quality control of herbal material.

Standardization of phaseolus vulgaris valves by macro- and microscopic characteristics

During the last years on the Ukrainian pharmaceutical market a tendency to increase the range of the national herbal remedies is observed. Native flora herbs, having a sufficient resource base, deserve special attention and are characterized by a number of economic benefits for manufacturers. The implementation of herbal remedies in medical practice is accompanied by the necessity to improve the quality control of herbal material and to develop normative documents. Macro- and microscopy analysis becomes important at standardization of herbal material especially cut and powdered.Aim. The aim of our work was to carry out macro- and microscopy research of Phaseolus vulgaris and to determine its diagnostic morphologic and anatomic features.Methods. The conventional research methods in pharmacognosy have been used; photomicrography has been taken by using microscope MC-10 and Samsung PL 50 camera.Results. As a result of research, the microscopic diagnostic characteristics for Phaseolus vulgaris valves have been proposed, e.g.: the valves are elongated, straight, navicular or grooved, twisted to some extent; the external surface is smooth or slightly undulating, mat, pale yellow or yellow; the internal surface is shiny, white or yellowish-white. Sometimes carpophore (stalk) may occur. The microscopic diagnostic features have been determined and offered, e.g.: cells of outer epidermis are pentagonal and hexagonal, with plicated cuticle; stomata are surrounded by 3–4 subsidiary narrow cells, additionally surrounded by 5–6 narrow cells; cover trichomes and glandular trichomes are elongated, pointed and unbranched; xylem vessels and tracheids; non-bundled structure stalk fragments with pericyclic sclerenchyma and secretory cells in the phloem.Conclusion. Research on determination of the macro- and microscopic characteristics of Phaseolus vulgaris valves has been carried out. The diagnostic morphological and anatomic elements in herbal material have been determined for its identification. The data obtained have been used for development of the State Pharmacopoeia of Ukraine draft Monograph «Phaseolus vulgaris valves»

Validation evaluation of the technological process of leonurus turkestanicus liquid extract production

Aim. The transfer of Leonurus turkestanicus liquid extract production technology and its validation at LLC «FitOleum» (GMP compliance report conclusion CT PK 1617-2006 «Good Manufacturing Practice. Remedies production. Basic provisions» № 18, Nov. 21, 2014.). Methods. Different known statistical methods have been used to evaluate and to interpret both technological parameters and indicators that were determined during quality control of herbal material, intermediate product, and final product. A Statistical process control (SPC) concept is applied as a basis for all accepted international instruments ICH Q8 «Pharmaceutical Development», ICH Q10 «Pharmaceutical Quality System», PAT Concept, and FDA Guidance for Process Validation. Results. As a result of research normative documents that regulate manufacturing process in the test conditions have been developed. A comparable data across technological parameters of three consistently manufactured test production series have been obtained, as well as validity of the extract production technological process has been proved. Conclusion. On the basis of research, the experimental-industrial procedure for Leonurus turkestanicus herb liquid extract production at the LLC «FitOleum» has been developed

Herbal Medicines for Acute Kidney Injury: Evidence, Gaps and Frontiers

Acute kidney injury (AKI) is a major health threat worldwide. The literature on herbal intervention in AKI was searched from English and Chinese databases and reports were critically analyzed in terms of preventing AKI, promoting repair and regeneration, enhancing extrarenal clearance of uremic toxins, and preventing progression to chronic kidney disease (CKD). Altogether, 16 herbal formulae and a few extracts derived from individual herbs were reported to prevent or mitigate AKI in animal models induced by renal ischemia/reperfusion, cisplastin, gentamicin, glycerol, adenine, sepsis or physical exhaustion. Four formulae and six individual herbs were reported to accelerate recovery and/ or to prevent CKD in established AKI animal models. Intrarectal herbal medicines, with or without simultaneous oral administration, were reported in six clinical trials and in an animal model to increase extrarenal clearance of uremic toxins. Additional 13 clinical trials reported oral or intravenous herbal interventions in AKI of different etiologies. Despite recurring problems, notably poor compliance with good practice guidelines for clinical trials and for authentication, naming and quality control of herbal materials, accumulating experimental data on the preventive effects of herbal medicines in AKI look encouraging and urge for better, definitive trials to guide clinical practice. Herbal enemas promoting extrarenal clearance of uremic toxins seem cost-effective, but better clinical evidence is certainly needed before any affirmative recommendation be made for AKI patients without access to dialysis. New frontiers, however, lie in those herbal remedies that promote repair/ regeneration and prevent chronicity after AKI. Recent experimental data suggest that this may be possible. Abbreviations: AKI: acute kidney injury; AKIN: Acute Kidney Injury Network; αSMA: α smooth muscle actin; ARF: acute renal failure; ATN: acute tubular necrosis; BUN: blood urea nitrogen; CAT: catalase; CKD: chronic kidney disease; CXCL12: chemokine C-X-C motif ligand 12; CXCR4: chemokine C-X-C motif receptor 4; CLP: cecal ligation and puncture; Cu-Zn-SOD: copper-zinc superoxide dismutase; ECM: extracellular matrix; eGFR: estimated glomerular filtration rate; EMT: epithelial-to-mesenchymal transition; ESWL: extracorporeal shock wave lithotripsy; FSP: fibroblast-specific protein 1; GSH: glutathione; GSH-Px: glutathione peroxidase; HMP: herbal medicinal product; HO-1: heme oxygenase 1; ICAM-1: intercellular adhesion molecule-1; ICU: intensive care unit; IFN-γ: interferonγ; IL: interleukin; iNOS: inducible nitric oxide synthase; i.p.: intraperitoneally or intraperitoneal; I/R: ischemia/reperfusion; i.v.: intravenous or intravenously; JNK: c-Jun N-terminal kinase; KDIGO: Kidney Disease Improving Global Outcomes; KIM-1: kidney injury molecule 1; LPS: lipopolysaccharides; MCP-1: monocyte chemotactic protein 1; MDA: malondialdehyde; MMP9: matrix metalloproteinase 9; Mn-SOD: manganese superoxide dismutase; NF-κB: nuclear factor κB; NGAL: neutrophil gelatinase-associated lipocalin; NO: nitric oxide; PCI: percutaneous coronary intervention; PCNA: proliferating cell nuclear antigen; p.o.: per os (oral administration); pRIFLE: modified RIFLE guideline for pediatric use; RCT: randomized controlled clinical trial; ROS/RNS: reactive oxygen/nitrogen species; RIFLE: Risk, Injury, Failure, Loss, and End-stage renal disease; RPTECs: renal proximal tubular epithelial cells; Scr: serum creatinine; TCM: traditional Chinese medicine; TGF-β: transforming growth factor β; TIMP1: tissue inhibitor of metalloproteinase 1; TNF-α: tumor necrosis factor-α; UUO: unilateral ureteral obstruction.

Antiarthritic activity of a polyherbal formulation against Freund's complete adjuvant induced arthritis in Female Wistar rats.

Objectives:To formulate a polyherbal formulation and evaluate its antiarthritic activity against Freund's complete adjuvant induced arthritis in Female Wistar rats.Materials and Methods:Glycosmis pentaphylla, Tridax procumbens, and Mangifera indica are well-known plants available throughout India and they are commonly used for the treatment of various diseases including arthritis. The polyherbal formulation was formulated using the ethanol extracts of the stem bark of G. pentaphylla, whole plant of T. procumbens, and leaves of M. indica. The polyherbal formulation contains the ethanol extracts of G. pentaphylla, T. procumbens, and M. indica in the ratio of 2:2:1. The quality of the finished product was evaluated as per the World Health Organization's guidelines for the quality control of herbal materials. Arthritis was induced in female Wistar rats using Freund's complete adjuvant (FCA), and the antiarthritic effect of polyherbal formulation was studied at doses of 250 and 500 mg/kg. The effects were compared with those of indomethacin (10 mg/kg). At the end of the study, blood samples were collected for biochemical and hematological analysis. The radiological examination was carried out before terminating the study.Results:Polyherbal formulation showed significant antiarthritic activity at 250 and 500 mg/kg, respectively, and this effect was comparable with that of indomethacin. The antiarthritic activity of polyherbal formulation is supported by biochemical and hematological analysis.Conclusion:The polyherbal formulation showed signinicant antiarthritic activity against FCA-induced arthritis in female Wistar rats.

HPTLC densitometry method for simultaneous determination of flavonoids in selected medicinal plants

The aim of the present work was to develop a high-performance thin-layer chromatography (HPTLC) densitometric analysis technique using the solvent system ethyl acetate:methanol:formic acid:water (20:3:1:2, v/v), which has been developed and validated for simultaneous quantification of four marker compounds (quercetin, rutin, luteolin and vitexin) in Asparagus racemosus, Withania somnifera, Vitex negundo, Plumbago zylenica, Butea monosperma, and Tephrosia purpurea plant extracts. The analysis was performed on HPTLC precoated silica gel 60F254 plates with as the mobile phase (distance of 6.5 cm). Densitometric detections of flavonoids were performed at 254 and 366 nm respectively. The amount of these marker compounds was calculated using the regression equations of the calibration curves, which were linear within a range of 0.40–0.90 µg/spot (r2 > 0.99). The system was found to provide compact spots for quercetin, rutin, luteolin and vitexin (Rf values of 0.96, 0.53, 0.96 and 0.78, respectively). The amounts of these marker compounds present was measured by the method developed and expressed in mg ml–1 of dry extracts of plants. This simple, precise and accurate method gave good resolution from other constituents present in the plant extract and has been successfully applied in the analysis and routine quality control of herbal materials and formulations containing these studied plants.

Comparative Mass Spectrometry & Nuclear Magnetic Resonance Metabolomic Approaches for Nutraceuticals Quality Control Analysis: A Brief Review

The number of botanical dietary supplements in the market has recently increased primarily due to increased health awareness. Standardization and quality control of the constituents of these plant extracts is an important topic, particularly when such ingredients are used long term as dietary supplements, or in cases where higher doses are marketed as drugs. The development of fast, comprehensive, and effective untargeted analytical methods for plant extracts is of high interest. Nuclear magnetic resonance spectroscopy and mass spectrometry are the most informative tools, each of which enables high-throughput and global analysis of hundreds of metabolites in a single step. Although only one of the two techniques is utilized in the majority of plant metabolomics applications, there is a growing interest in combining the data from both platforms to effectively unravel the complexity of plant samples. The application of combined MS and NMR in the quality control of nutraceuticals forms the major part of this review. Finally I will look at the future developments and perspectives of these two technologies for the quality control of herbal materials.

Antidiabetic Activity of Polyherbal Formulation in Streptozotocin – Nicotinamide Induced Diabetic Wistar Rats

Glycosmis pentaphylla, Tridax procumbens, and Mangifera indica are well-known plants available throughout India and they are commonly used for the treatment of various diseases including diabetes mellitus. The antidiabetic activity of the individual plant parts is well known, but the synergistic or combined effects are unclear. The concept of polyherbalism has been highlighted in Sharangdhar Samhita, an Ayurvedic literature dating back to 1300 AD. Polyherbal formulations enhance the therapeutic action and reduce the concentrations of single herbs, thereby reducing adverse events. The aim of the present study is to formulate a polyherbal formulation and evaluate its antidiabetic potential in animals. The polyherbal formulation was formulated using the ethanol extracts of the stem bark of G. pentaphylla, whole plant of T. procumbens, and leaves of M. indica. The polyherbal formulation contains the ethanol extracts of G. pentaphylla, T. procumbens, and M. indica in the ratio of 2:2:1. The quality of the finished product was evaluated as per the World Health Organization's guidelines for the quality control of herbal materials. The quality testing parameters of the polyherbal formulation were within the limits. Fingerprint analysis of the polyherbal formulation showed effective separation at 366 nm, and it revealed that the active compound present in the polyherbal formulation and the active compounds present in all the three extracts were the same. The acute toxicity studies of the polyherbal formulation did not show any toxic symptoms in doses up to 2000 mg/kg over 14 days. The oral antidiabetic activity of the polyherbal formulation (250 and 500 mg/kg) was screened against streptozotocin (50 mg/kg; i.p.) + nicotinamide (120 mg/kg; i.p.) induced diabetes mellitus in rats. The investigational drug was administered for 21 consecutive days, and the effect of the polyherbal formulation on blood glucose levels was studied at regular intervals. At the end of the study, the blood samples were collected from all the animals for biochemical estimation, and the animals were sacrificed and the liver and pancreatic tissues were collected for histopathologic analysis. Polyherbal formulation showed significant antidiabetic activity at 250 and 500 mg/kg, respectively, and this effect was comparable with that of glibenclamide. The antidiabetic activity of polyherbal formulation is supported by biochemical and histopathologic analysis.

Rapid simultaneous determination of marmelosin, umbelliferone and scopoletin from Aegle marmelos fruit by RP-HPLC.

The surge of interest in naturally occurring phytochemicals with high therapeutic potential has led to the discovery of many molecules, out of which naturally occuring coumarins such as marmelosin, umbelliferone and scopoletin present in Aegle marmelos (Bael) fruit shows good therapeutic potential. The aim of the present work is to develop and validate Reverse Phase-High Performance Liquid Chromatography (RP-HPLC) method for simultaneous determination of marmelosin, umbelliferone and scopoletin in A. marmelos fruit extracts. The chromatographic separation was performed with isocratic elution of 55:45 (%, v/v) methanol-water containing 0.1% acetic acid as mobile phase. The method used to analyse the extract of A. marmelos showed good resolution with retention time within 12min. The relative concentrations of above phytoconstituent were determined in A. marmelos fruits. The method was found to give compact peaks for scopoletin, umbelliferone and marmelosin (Rt of 4.6, 6.5 and 11.3min respectively) and were linear over the range 5-30μgml(-1) (R(2) = 0.9655), 2-10μgml(-1) (R(2) = 0.9964) and 2-10μgml(-1) (R(2) = 0.9862) respectively. The mean recoveries for marmelosin, umbelliferone and scopoletin at three concentrations were in the range of 98.8-102.9, 98.8-101.1 and 94.2-98.3% respectively. The relative standard deviation of accuracy, precision and repeatability were within 2%, indicating the method produced highly reproducible results. Therefore this simple, precise and accurate method enables simultaneous separation of this phytoconstituent and hence can be successfully applied in analysis and routine quality control of herbal material and formulation containing A. marmelos.