High-performance Thin-layer Chromatography Profiling Research Articles

Chemical profiling of endangered Citrus macroptera leaf extracts and evaluation of its cytotoxic activity

The main component of drug production and formulation, with minimal to no adverse effects, is plant-based medicine. Citrus macroptera is an endangered species of North-Eastern India and has a diverse array of pharmaceutical applications owing to the presence of several bioactive compounds. The study aimed to evaluate the presence of bioactive metabolites, antioxidants, and cytotoxic mechanisms of the in vitro and ex-vitro leaf extracts of C.macroptera. Gas chromatography–mass spectrometry (GC-MS) profiling was performed using the ethanolic leaf extracts. Methanolic and aqueous extracts were profiled using High-performance thin-layer chromatography (HPTLC). Moreover, the cytotoxicity of the methanolic leaf extracts was assessed using the 3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide (MTT) test against SH-SY5Y, human neuroblastoma cell lines. GC-MS analysis revealed the existence of numerous pharmacologically significant components such as esters, hydrocarbons, fatty alcohols, and ester compounds. The presence of flavonoids and stigmasterol was exhibited by the HPTLC profile. The in-situ HPTLC analysis revealed white fluorescence after derivatization under white light that explicitly confirms the antioxidant potential of the extracts and can serve as a lead source of natural antioxidants. The cytotoxic efficacy of the extracts was observed in a concentration-dependent manner. However, the in vitro leaf extract displayed better suppressive effects against the SH-SY5Y cells with a 24-h IC50 of 167.71?g/mL compared to the ex-vitro leaf extract. Further, a comprehensive study is necessary to identify the cytotoxic compounds and their mode of action. The outcome of our investigation provides empirical evidence that phytoconstituents present in the leaf extracts of C. macroptera can be used as an effective pharmaceutical agent to combat several diseases.

Pharmacognostical (Ingredient Analysis) and Pharmaceutical Evaluation of Yashtimadhu Taila (Glycyrrhiza glabra Linn.)

Abstract Context: In Ayurveda, Yashtimadhu is being used as one of the healing drugs in the management of different kinds of wounds. The preparations made from Yashtimadhu such as Churna, Ghrita, Taila, and Malahar are being practiced in wound care unit to treat different kinds of wounds as local application. Multiple treatment modalities are mentioned in Sushruta Samhita for the management of various types of Vrana. Aim and Objective: This study was to standardize the finished product Yashtimadhu Taila to confirm its identity, quality, and purity. Yashtimadhu Taila is standardized using different parameters like pharmacognostical study including organoleptic characters such as color, odor, and taste. Subjects and Methods: Yashtimadhu Taila is prepared as per Sneh Kalpana mentioned in Sharangdhara Samhita. The pharmacognostical study includes organoleptic characters such as color, odor, and taste. The pharmaceutical study of Yashtimadhu Taila includes specific gravity, acid value, saponification, iodine value, and refractive index by the following standard procedures as per the Ayurvedic Pharmacopoeia of India, and chromatographic examination includes high-performance thin-layer chromatography (HPTLC) study. Results: All the physicochemical parameters show acid value (0.4883), saponification (178.7), iodine value (82.91), and refractive index (1.4820). HPTLC profile of the methanolic extract of the drug showed 14 spots at 254 nm and 8 spots at 366 nm. Conclusions: This study will be helpful to establish the safety profile, efficacy, and acceptance of classical parameters for further research studies.

Standardisation of different extracts of detoxified Nux-vomica seeds with its comparative study by TLC and HPTLC

IntroductionDetoxified Nux-vomica (detoxified Strychnos nux-vomica Linn.) is a very popular crude herb used in Unani and Ayurvedic pathy after detoxification for many years. The first person to identify Nux-vomica with the Indian drug kuchla was Haji Zainuddin. Detoxified Nux-vomica has significant effects on various disorders like rheumatic arthritis, inflammation, and gout (Anonyms, 2007; Bentley and Trimen, 1983). It has proved the best remedy for paralysis, nervous disorders, nervous debility, tremor, diarrhoea, and mental emotions (Nadkarni, 2000). The present study is designed to develop the qualitative as well as quantitative parameters of crudely detoxified Nux-vomica seeds and their comparative study by Thin layer chromatography (TLC) and High-performance thin layer chromatography (HPTLC). MethodsOrganoleptic characters, ash values, extractive values, phytochemical analysis, fluorescence analysis, TLC and HPTLC profiles, etc. were the parameters used for the standardisation of the test drug. ResultsTotal ash values, water, alcohol, ether-soluble extractive values, and volatile oil percentages were found to be 1.925%, 6.54%, 3.25%, 3.20%, and 2.00%, respectively. The TLC profile of Detoxified Nux-vomica shows three spots for aqueous extract and five spots for hydroalcoholic extract in Toluene: Ethyl Acetate: Triethylamine (7:2:1) and four spots in Methanol: Ethyl Acetate: Triethylamine (0.5:8.5:1) with exposure in Dragendorff’s Reagent. The percentage yield of strychnine in the hydro-alcoholic and aqueous extracts of Detoxified Nux-vomica is 7.48 and 2.10, while it is 6.17 and 1.85 for brucine, respectively. ConclusionThe present study used secondary data for the identification and quantification of active constituents in detoxified Nux vomica seeds.

Analytical Profiling of Saffron (Crocus sativus) using 1H-NMR and FTIR based Metabolomics approach and UV-Vis, HPTLC and TLC Chromatography Fingerprinting

Crocus sativus L. commonly known as saffron or Kesar in India, is an important medicinal herb in Ayurveda and has been traditionally used for treatment of neurological disorders, for depression, anxiety and sleep disorders. It is uses as a coloring and flavouring agent in the preparation of various foods. Modern research has high lighted its beneficial effects in treatment of cardiovascular action, diabetic cataract, and as a potent antiinflammatory herb. Due to its high cost its quality control is of utmost importance to ensure its authenticity, purity and its medicinal properties. In the preset study the we have used Thin Layer Chromatography (TLC) analysis, High performance thin layer chromatography (HPTLC) fingerprint, UV-Vis spectrophotometric analysis, 1H nuclear magnetic resonance (1H NMR) and Fourier transform infrared spectroscopy (FTIR) based metabolomic study for quality control and characterization of saffron. The antibacterial and antifungal activity were evaluated using agar well-diffusion method in two pathogenic bacterial strains, Escherichia coli, Staphylococcus aureus and two pathogenic fungal strains, Candida albicans, and Aspergillus brasiliensis. The 1H NMR spectroscopy couples with FTIR analysis leads to identifcation of the secondary metabolites of saffron like crocetin, picrocrocin and safranal on basis of reported diagonostic signals and peaks. The antimicrobial activity showed moderate antibacterial and antifungal activity. The TLC and HPTLC profile reveals the characteristic fingerprint. Overall the present study showed that the 1H-NMR, FTIR based metabolomics approach and TLC and HPTLC metabolite profiling can be powerful strategy for maintaining the holistic quality of the saffron.

HPTLC PROFILING AND FTIR FINGERPRINTING COUPLED WITH CHEMOMETRIC ANALYSIS OF MALAYSIAN STINGLESS BEE PROPOLIS

Propolis is a natural substance produced by the bees to protect their hive. The present work reports the utilization of chemical profiling and fingerprinting combined with multivariate analysis for discrimination of the geographical origin of Malaysian stingless bee propolis. High Performance Thin Layer Chromatography (HPTLC) has been performed to profile the chemical composition of Geniotrigona thoracica propolis from different localities in East Coast of Peninsular Malaysia namely Besut, Terengganu (BST), Dungun, Terengganu (DGN), Lundang, Kelantan (LDG), Tanah Merah, Kelantan (TM) and Gua Musang, Kelantan (GM). The obtained HPTLC profiles showed the presence of flavonoids, phenolics and terpenoids in propolis. The chemical fingerprinting was obtained through Fourier Transform Infrared (FTIR) spectroscopy. Chemometric analysis on FTIR dataset using principal component analysis (PCA) and hierarchical cluster analysis (HCA) have classified the propolis into three major groups according to their sampling locations. Chemical fingerprinting analysis on the functional group via FTIR and chemometric revealed that the locations of propolis have direct correlation with the chemical composition, thus affecting the biological activities of propolis. Both chemical marker and chemical fingerprinting analysis are important tools in propolis standardization.

Pharmacognostical and phytochemical studies of Crotalaria retusa L. (Shanapuspi) dried roots

Abstract BACKGROUND: Crotalaria retusa L. belongs to the Fabaceae family and is found in tropical and subtropical regions of the globe. In Ayurveda, its roots are used against hemoptysis. The roots of C. retusa are used in folklore medicine in many countries for the treatment of numerous diseases. Roots are used against fever, cough, dyspepsia, cardiac disorders, stomatitis, diarrhea, scabies, impetigo, and several other disorders. But so far, the pharmacognostical and phytochemical standardization of root has not been reported for proper identification. Hence, the present study was carried out for the establishment of quality parameters of the root of this plant. METHODS: Macroscopy, anatomical studies, powder analysis, physiochemical properties like ash values, extractive values, and high-performance thin layer chromatography profile of roots were done as per Ayurvedic Pharmacopoeia of India guidelines. RESULTS: In powder microscopy, acicular crystals, elongated tracheids were observed. Water-soluble and alcohol-soluble extractive values were found to be 4.78% and 7.63%, respectively. CONCLUSIONS: The qualitative and quantitative standards established in this study can serve as a valuable authentication source of C. retusa L. roots.

Combining DNA and HPTLC profiles to differentiate a pain relief herb, Mallotus repandus, from plants sharing the same common name, "Kho-Khlan".

The pain relief formula "Ya Pa Som Kho-Khlan (YPSKK)" or "ยาผสมโคคลาน" in Thai is officially recorded in the Natural List of Essential Medicines (NLEM) of Thailand. The main component is Mallotus repandus (Willd.) Müll. Arg.; however, Anamirta cocculus (L.) Wight & Arn and Croton caudatus Gleiseler share the same common name: "Kho-Khlan". Confused usage of A. cocculus or C. caudatus can have effects via toxicity or unsuccessful treatment. This study aimed to combine a high-performance thin-layer chromatography (HPTLC) technique and DNA barcoding coupled with high-resolution melting (Bar-HRM) to differentiate M. repandus from the other two species. The M. repandus extract exhibited a distinct HPTLC profile that could be used to differentiate it from the others. DNA barcodes of the rbcL, matK, ITS and psbA-trnH intergenic spacer regions of all the plants were established to assist HPTLC analysis. The rbcL region was selected for Bar-HRM analysis. PCR amplification was performed to obtain 102 bp amplicons encompassing nine polymorphic nucleotides. The amplicons were subjected to HRM analysis to obtain melting curve profiles. The melting temperatures (Tm) of authentic A. cocculus (A), C. caudatus (C) and M. repandus (M) were separated at 82.03±0.09°C, 80.93±0.04°C and 80.05±0.07°C, respectively. The protocol was applied to test crude drugs (CD1-6). The HPTLC profiles of CD2-6 showed distinct bands of M. repandus, while CD1 showed unclear band results. The Bar-HRM method was applied to assist the HPTLC and indicated that CD1 was C. caudatus. While ambiguous melting curves from the laboratory-made formulae were obtained, HPTLC analysis helped reveal distinct patterns for the identification of the plant species. The combination of HPTLC and Bar-HRM analysis could be a tool for confirming the identities of plant species sharing the same name, especially for those whose sources are multiple and difficult to identify by either chemical or DNA techniques.

Cyclooxygenase (COX) and 15-Lipoxygenase (15-LOX) Inhibitory Activity and HPTLC Profile of Asplenium Nidus, Diplazium Esculentum, and Drynaria Quercifolia in Bukidnon, Philippines

Diplazium esculentum, Drynaria quercifolia, and Asplenium nidus are among the fern species found in Bukidnon, Philippines which are used as traditional herbal medicines. The HPTLC profile and the anti-inflammatory properties against cyclooxygenase (COX) and 15-lipoxygenase (15-LOX) of the frond ethanolic extracts of D. esculentum, D. quercifolia, and A. nidus were determined. The High-Performance Thin Layer Chromatography (HPTLC) profile was obtained using ethyl acetate: formic acid: water (16:2:2) as mobile phase and Natural Products (NP) as derivatizing reagent. The HPTLC profile of the D. esculentum, D. quercifolia, and A. nidus extracts showed 10 (Rf = 0.02-0.97), 13 (Rf = 0.03-0.90), and 14 (Rf = 0.02-0.99) bands, respectively. The profiles for each fern species may be used as marker for quality evaluation and standardization of herbal formulations containing these plants. For the anti-inflammatory properties, D. esculentum and D. quercifolia extracts which inhibited more than 50% of the COX enzymes showed significantly higher activity than A. nidus and were considered active against COX-2 and COX-1. D. esculentum, however, gave a selectivity ratio (COX-2/COX-1) of 1.03 making its inhibitory activity selective against COX-2. The percent 15-LOX inhibitory activity of D. quercifolia (58.62%) is significantly higher than that of A. nidus (38.70%) but statistically comparable to D. esculentum (51.19%). Among the extracts, D. quercifolia and D. esculentum which inhibited more than 50% of the 15-LOX were considered active. D. esculentum and D. quercifolia can therefore be potential sources of anti-inflammatory lead compounds for future drug development.

Rapid Screening of Lipase Inhibitors from Ophiopogonis Radix Using High-Performance Thin Layer Chromatography by Two Step Gradient Elution Combined with Bioautographic Method.

In this study, a high-performance thin layer chromatography (HPTLC) method by two step gradient elution with two mobile phases was developed for the simultaneous analysis of seven constituents in Ophiopogonis Radix. The chromatography was performed on silica gel 60 F254 plate with dichloromethane-methanol-ethyl acetate-water (70:25:12:3, v/v/v/v) and dichloromethane-methanol (300:1, v/v) as the mobile phase for two step gradient elution. Then, the HPTLC profiles were observed after derivatization with 10% sulfuric acid in ethanol solution. The obtained HPTLC images were further analyzed by chemometric approaches and the samples could be clustered based on regions and/or growth years, which were two important factors affecting the constituents in Ophiopogonis Radix. Furthermore, five compounds including ophiopogonin D, ophiopojaponin C, ophiopogonin D’, ophiopogonin C’ and methylophiopogonanone B were screened as potential lipase inhibitors from Ophiopogonis Radix by the HPTLC-bioautographic method. The binding modes and interactions between the five compounds and lipase were further explored by molecular docking analysis. The developed HPTLC method could be used for quality control of Ophiopogonis Radix and screening of the potential lipase inhibitors.

Comparative chemical profiling of purified and unpurified Strychnos nux-vomica Linn seeds: An attempt to reduce toxic Brucine content

Vishamusthi (Strychnos nux-vomica Linn.), a medicinal plant described as Upavisha (semi-poisonous) group of Ayurvedic Pharmacopoeia of India. Vishamusthi has widely been used and being practiced in several illness namely nervous debility, paralysis, weakness of limbs, sexual weakness, dyspepsia and etc. Ayurveda practices strictly recommend the use of Vishamusthi in therapeutics only after proper shodhana (purificatory procedure) through specific medias such as Gomutra (cow’s urine), Godugdha (cow’s milk), Goghrita (cow’s ghee), and etc. Although various shodhana procedures are recommended in Ayurvedic treatise, but updated scientific researches regarding the shodhana methods are lacking. The present study was undertaken to investigate the physicochemical and phytochemical parameters, quantitative estimation of brucine using cutting edge research tools such as high-performance thin layer chromatography (HPTLC), liquid chromatography-mass spectrometry (LC-MS) and gas chromatography-mass spectrometry (GC-MS) analyses of Vishamusthi seeds before and after purification. The remarkable changes have been observed in different physicochemical parameters, HPTLC, HPLC, GC-MS and LC-MS chromatographic profiling before and after shodhana process of Vishamusthi seeds. Quantitative HPLC studies revealed that the process of shodhana resulted in depletion of toxic brucine (chief poisonous constituent of Vishamusthi seeds) reduced to 79.66% in chloroform extract and 64.54% in ethanol extract after shodhana process.

Pharmacognostical studies on leaves of Mussaenda frondosa Linn.

The Mussaenda frondosa Linn. belonging to the family Rubiaceae, commonly known as Sriparnah in Sanskrit, is a scandent shrub traditionally used in the treatment of cough, bronchitis, fever, inflammation, wounds, ulcers, jaundice, leucoderma and pruritus. Though it is an important plant, till date, no pharmacognostical reports have been available on its leaf. Therefore, the present investigation was undertaken to ascertain the requisite pharmacognostical standards for the standardization of the Mussaenda frondosa leaves. Various investigations like Pharmacognostical studies, preliminary phytochemical screening and High-Performance Thin Layer Chromatography (HPTLC) analysis were carried out, and the salient qualitative parameters were reported. Microscopical evaluation of the leaf revealed the presence of paracytic stomata, microcrystal’s, Idioblast, collenchymas, sand crystals and unicellular unbranched covering Trichomes. The presence of flavonoids, steroids, glycosides, mucilage, saponins and proteins were confirmed through Preliminary phytochemical studies. The HPTLC profile of ethanol extract from M. frondosa L. revealed ten phytoconstituents of Rf value ranging from 0.11 to 0.88. The significant peaks are observed at Rf values of 0.11, 0.16, 0.23 and 0.81. These findings provide referential information for correct identification and standardization of the Musssaenda frondosa leaves, even in powder form. This information will also be useful to distinguish Mussaenda frondosa from the closely related other species of Mussaenda. The Pharmacognostic and phytochemical profiles reported in this research work for Mussaenda frondosa may play a major role in setting monograph of the plant, which might be helpful in proper identification of the plant.

Quality Analysis of Manjishta (Rubia cordifolia L.) with respect to pharmacognostical and high-performance thin-layer chromatography profile of the genuine sample: A cross-sectional market sample study

Background: The genuineness of Ayurvedic herbs causally determines the effectiveness of Ayurvedic treatment protocols. Research and subsequent evidence-based medicinal practices fundamentally dependant on appropriate identification and standardization of specific herbs are used for health-care purposes. Definite deficits that prevail in this regard in the form of adulteration, substitution, and compromised quality standards are the reasons why many scientific communities and health-related organizations question Ayurvedic sciences. This study aims at raising the very timely subject matter of genuine drug collection based on a model of Pharmacognostical and high-performance thin-layer chromatography (HPTLC) profiling of an Indian medicinal herb "Manjishta"' (Rubia cordifolia L.), a profoundly marketed Ayurvedic drug. Objectives: To compare the six market samples of Manjishta (Pharmacognostical and with HPTLC profiling) with the genuine root and stolon. Materials and Methods: The market samples of Manjishta from 6 districts of Kerala were collected, and these samples were compared with the genuine root and stolon (morphology, histology, and with HPTLC profiles). Results: Morphological and anatomical evaluation of the market samples was similar with that of the original stolon of Rubia cordifolia. HPTLC profiling yielded entirely different peaks in specific samples when compared with that of the genuine stolon. Conclusion: The collected market samples of Manjishta from different districts of Kerala were almost similar to the stolon of Rubia cordifolia L. A clear standard operative procedure should be prepared for medicinal plant part collection with respect to source plant maturity for Manjishta or as a matter of fact any other herb and thus the HPTLC profiles should be redefined.

Pharmacognostic characterization and antibacterial activity of Brugmansia suaveolens (Humb. & Bonpl. ex Willd.) Bercht. & J. Presl leaves

BACKGROUND: Brugmansia suaveolens (Humb. & Bonpl. ex Willd.) Bercht. & J. Presl (synonym Brugmansia arborea; family Solanaceae) is one of the total seven species of Brugmansia which is commonly known as angel’s trumpet. It has analgesic, antinociceptive, anticancer, anti-inflammatory, antiasthmatic, and various other activities. Despite its numerous medicinal properties, it has never been explored for its potential against bacterial species; also, no publication has been made on pharmacognostical characterization and high-performance thin layer chromatography (HPTLC) investigation of its leaves. OBJECTIVE: The aim of this article is to evaluate the pharmacognostical, physicochemical, HPTLC profiles of B. suaveolens leaves along with the assessment of in-vitro antibacterial activity of its various fractions. MATERIALS AND METHODS: B. suaveolens collected from sub-Himalayan region of Himachal Pradesh, India was authenticated and then studied for various pharmacognostic and physicochemical parameters employing proper quality control methods for medicinal plant materials designed by the WHO. Extracts and fractions of air-dried coarse plant powder were prepared and were identified for the presence of various classes of compounds using preliminary phytochemical screening. HPTLC profile for quantitative evaluation of atropine in leaves of B. suaveolens was carried out and lastly, fractions were assessed for antibacterial activity by using disc diffusion method against chloramphenicol as standard. RESULTS: Transverse section, powder microscopy, leaf constants, and physicochemical parameters revealed valuable data to set up standards for the plant. Alkaloids, glycosides, flavonoids, steroids, triterpenoids, and carbohydrates were found to be present. HPTLC showed 7.79–13.20%w/w of atropine in the plant. Chloroform and ethyl acetate fraction showed encouraging results against four strains of Gram-negative bacteria with good zone of inhibitions. CONCLUSION: The present study provides referential information for identification, authentication, and standardization of this highly important crude drug.

Distribution of Quercetin in Different Organs of Male Sprague Dawley Rats Under Hypoxia: A High-Performance Thin Layer Chromatography Approach

Background: Quercetin (3’,3’,4,5,7-pentahydroxyflavonol), a natural flavonoid found in fruits, vegetables, beverages, and other phytoproducts, exerts multiple health benefits including a reduction in hypoxia-induced oxidative stress, inflammation, lipid peroxidation, allergic disorders, neurodegenerative disorders, and cardiovascular diseases. Objective: Despite knowledge of such therapeutic efficacy of quercetin to human health, there is limited literature available that sheds light on an organ-wise distribution of quercetin. Therefore, the current study was performed to accurately estimate the distribution of quercetin in its supplemented form in different tissues of a mammalian model, i.e., male Sprague Dawley (SD) rats. Method: The rats were exposed to different durations (1 h, 3 h, 6 h, and 12 h) of hypoxia in a simulated hypobaric hypoxia chamber, with parameters maintained at 8 % O2 and 282 mm Hg, following which they were sacrificed. Plasma and different tissue samples were duly collected. A high-performance thin layer chromatography (HPTLC) approach was employed for the first time, using our own reported method, along with an optimized sample preparation procedure for quercetin determination. Briefly, the samples were developed in a mobile phase constituted of ethyl acetate, dichloromethane, methanol, formic acid, and glacial acetic acid. Results: Distinct bands of quercetin in resultant HPTLC profiles verified that the amount of quercetin varied among different tissues, with varying durations to hypoxia exposure. Quercetin was substantially retained in vital organs namely, lungs, liver, and heart for relatively longer durations. Conclusion: The present study established HPTLC as an efficient and high throughput tool, leading to a satisfactory evaluation of the amount of quercetin present in various tissue samples under hypoxia.

Physico-chemical investigation of a polyherbal formulation - Vidangatandulaadi choorna

Vidangatandulaadi Choorna is a polyherbal formulation consisting of seven ingredients. Trivrith (Operculina turpethum (Linn.) Silva Manso) is the chief ingredient responsible for the purgative action of the formulation. This yoga is mentioned in Kalpasthana of Ashtangahridaya, intended for virechana (Purgation). It is useful in Kapha-vatha disorders. Even though many kinds of research have been done to identify the physicochemical constituents of individual drugs in the formulation, no studies were done to identify the physicochemical properties of the formulation. This analysis helps in understanding the mechanism for different pharmacological actions of the formulation. Hence, Physico-chemical study of Vidangatandulaadi Choorna along with high-performance thin-layer chromatography (HPTLC) fingerprinting is done to fix the standards. All the drugs included in the formulation is identified by the botanist and is prepared according to Standards mentioned for the preparation of Choorna mentioned in Ayurvedic Pharmacopoeia of India. The formulation is least encountered, but it has shown its significant action in Dyslipidemia in folklore practices. As there are no Standards mentioned for this formulation, the result observed in the present study may be considered suitable. The data obtained from Physicochemical investigation, high-performance thin-layer chromatography profile and ICP-MS (Inductively Coupled Plasma - Mass Spectrometry) could be used as the standards for the present formulation under study.

Three medicinal Corydalis species of the Himalayas: Their ethnobotany, pharmacognosy, phytochemistry and pharmacology

Herbal medicines including the scholarly Bhutanese Sowa Rigpa medicine (BSM) contribute significantly to primary health care services and play a considerable part in modern drug discovery. BSM, which was integrated with modern health care system in 1967, currently uses more than 200 species of medicinal plants including three closely related members of the Corydalis species; Corydalis calliantha, C. crispa, and C. dubia. Maintaining the quality of these medicinal plants has been always challenging due to adulterants and also due to lack of traditional quality control parameters that could distinguish closely related species. Therefore there is an urgent need to develop reliable analytical methods and quality parameters. Given the significant medicinal properties attributed to the three Corydalis species which are extensively used in BSM, a thorough literature review was performed to investigate their existing quality control parameters including: 1) morphological assessment, 2) habitat and ecological evaluation, 3) macroscopical and microscopical examination, 4) physiochemical limit setting and comparative high-performance thin layer chromatography (HPTLC) profiling, and 5) phytochemical and biological activities. The study revealed that there are distinct species-specific features including ecological adaptation, micromorphology, anatomy, phytochemical values and HPTLC profiles presented by each Corydalis species, and that these quality parameters support traditional quality screening processes in BSM.

Development of a HPTLC method to profile the phytochemicals in Allanblackia parviflora (tallow tree) kernel and seed cakes

HPTLC is a useful and practical analytical tool to characterize plant compositions. This study was focused on exploring the results of high-performance thin-layer chromatography (HPTLC) analysis, particularly as a useful tool for the authentication of Allanblackia parviflora seed and kernel cakes. Bulked samples from sixteen different Ghanaian communities were analysed by HPTLC and their fingerprints were compared. The optimum experimental conditions were established: sample weight of 2.0 g, methanol:water (80:20 v/v) as extraction solvent, 30 min extraction time and twice extraction, ethyl acetate:methanol:water (100:16.5:13.5 v/v) as mobile phase, vanillic acid as derivatisation agent and 7 min of plate heating time after derivatisation. The HPTLC profile generated from extracts across 16 communities and 157 trees was very reproducible and demonstrates the robustness of the technique in characterising the profile.

Standard operating procedure of Purification of Chitraka (Plumbago zeylanica Linn.) along with pharmacognostical and analytical profiles of Plumbagin.

Introduction:Shodhana (purification) is the process by which one can remove the impurity or toxicity of the raw drug and make the drug suitable for therapeutic purpose. Chitraka (Plumbago zeylanica Linn.) is well known drug in Ayurveda and root of this plant is being used for therapeutic purpose and requires purification before used as a medicine.Aims and objective:There is no data available for pharmacognostical and analytical profile of processed Chitraka, hence it was planned to develop SOP of processed Chitraka for its identity, purity and strength through pharmacognostical and analytical profile.Materials and methods:Chitraka roots were procured from Pharmacy, Gujarat Ayurved University, Jamnagar. Purification was done in five batches with Churnodaka (lime water). Organoleptic characters, microscopic features, pH, loss on drying, ash value, water soluble extracts, methanol soluble extracts and plumbagin quantification through high-performance thin layer chromatography (HPTLC) were carried out, before and after the purification.Results:Average 98.07% yield of Chitraka was obtained after purification. Differences were found in the processed samples of Chitraka in organoleptic features, pharmacognostical characters and physicochemical parameters, which show the impact of purification procedure on Chitraka. In HPTLC profile, plumbagin content was 0.29% in unpurified Chitraka powder, where in it was noted 0.98% after purification.Conclusion:Increase in plumbagin content through pharmaceutical process of Chitraka purification with lime water indicates that, this operating procedure is simple, convenient and can be considered as standard procedure. The organoleptic features, pharmacognostical characters, values of physicochemical parameters and quantity of plumbagin of purified Chitraka powder may be utilized for quality assurance in future studies.

The inhibitory effects of an eight-herb formula (RCM-107) on pancreatic lipase: enzymatic, HPTLC profiling and in silico approaches

AimsObesity is a global, public health issue that causes or exacerbates serious medical disorders. Chinese herbal therapies have become one of the most popular alternatives due to intolerances of current anti-obesity treatments. The RCM-107 formula (granule) is modified from our previous studied RCM-104 formula, which has demonstrated significant effects on weight reduction in randomized clinical trials. Up to date, there is no published scientific evidence to evaluate the effect of this formula on the weight-loss target pancreatic lipase and therefore, the aim of this study is to investigate the inhibitory effect of RCM-107 and respective individual ingredient on the pancreatic lipase activities. Main methodsFluorometric based enzymatic assays, high-performance thin-layer chromatography (HPTLC) profiling and in silico molecular docking techniques were used to investigate the lipase inhibitory effects of the RCM-107 herbal formula and its respective individual herbs. Principle findingsThe results demonstrated the potent lipase suppressing effect of the RCM-107 formula. The majority of the ingredients from this formula also showed pancreatic lipase inhibitory activities. The presence of the known weight-loss compounds such as (-)-epigallocatechin-3-gallate (EGCG), epicatechin-3-gallate (ECG), (-)-epicatechin (EC), rutin, crocin and caffeine were identified in the RCM-107 and related single herbs using HPTLC profiling approaches. In addition, EGCG, EC and the known lipase antagonist orlistat acted on the same site. These compounds form hydrogen bonds with corresponding residues HIS152, ASP80 and GLY77, which can be considered as markers of important areas in the ligand-binding site. This may explain the details of their roles in inhibiting pancreatic lipase activities. ConclusionOur data has provided new knowledge to the mechanistic properties of the RCM-107 formula and its respective individual herbal ingredients for weight loss, in terms of reducing lipid absorption via the inhibition of pancreatic lipase.

Effect-directed screening of Bacillus lipopeptide extracts via hyphenated high-performance thin-layer chromatography

Bacillus species produce a wide array of biologically active metabolites, including nonribosomaly synthesized lipopeptides (LPs). The high-performance thin-layer chromatography (HPTLC) technique hyphenated with different bioassays and mass spectrometry was demonstrated as a valuable tool for effect-directed analysis of iturins, surfactins, fengycins and kurstakins homologues from complex mixtures of LPs. As proof of this straightforward strategy, the found surfactin and iturin A homologues were characterized and compared with reference substances. This study considered two different extraction methods for LPs produced by five Bacillus strains. The ethyl acetate extraction (Ex-1), and the acidic precipitation followed by methanol extraction (Ex-2) were investigated. Diverse enzyme inhibitions and antimicrobial potentials of LPs were analyzed, and in parallel, high-resolution mass spectra (HRMS) were online recorded from the HPTLC zones of interest. No antimicrobial effect against Gram-positive B. subtilis was evident for iturin, whereas a response was detected for surfactin. The nonpolar kurstakin compounds showed a pronounced B. subtilis antimicrobial activity in Ex-1 of almost all strains, whereas the fengycin homologues were detected in Ex-2 of SS-10.7 and SS-27.2. Iturin had also no activity against Gram-negative Aliivibrio fischeri, while again surfactin showed an enhancing luminescent activity. Contrary, kurstakin compounds caused a decrease in the luminescence in Ex-1 of all strains, particularly for SS-13.1. Both, iturin and surfactin showed a strong acetylcholinesterase (AChE) and α-glucosidase inhibition, but surfactin caused a much stronger inhibition. This was evident in all bacterial strains, except for SS-13.1 in Ex-1 and for SS-38.4 in Ex-2. Although, iturin and surfactin exhibited no DPPH˙ scavenging activity, Ex-1 of all strains contained more intense DPPH˙ scavenging compounds compared to Ex-2, and surfactin methyl esters showed a pronounced DPPH˙ activity, particularly in SS-12.6 in Ex-1. This study pointed to active metabolites of strains that can be used as therapeutics and biocontrol agents with beneficial effects on human health. The straightforward HPTLC profiling served as an excellent bioanalytical tool to control the formed bioactive metabolites. As the fermentation process is very sensitive to external influences, it could be a helpful control tool for standardization of the biotechnological processing.