High-performance Thin-layer Chromatography Fingerprinting Research Articles

Investigating Flavonoids by HPTLC Analysis Using Aluminium Chloride as Derivatization Reagent

This is the first study to report on high performance thin layer chromatography (HPTLC) generated spectrophotometric data to systematically capture flavonoid compounds using optimized derivatization with either AlCl3 or NaNO2-AlCl3-NaOH as visualisation reagents. While the traditional AlCl3 colorimetric method using UV–Vis analysis provides valuable insights into the presence of flavonoids and allows derivation of the total flavonoid content (TFC) of a sample, HPTLC fingerprints obtained after spraying with AlCl3 or NaNO2-AlCl3-NaOH enable the visualization of the various flavonoids present in a sample based on their respective absorption shifts, thus complementing the traditional TFC assay. In this study, 40 different flavonoids representing different classes (flavonols, flavanolols, flavan-3-ol, flavones, flavanones, and isoflavonoids) were analysed. Upon derivatization with AlCl3 most of the investigated flavonoids recorded bathochromic shifts, yielding characteristic λmax values between 370 and 420 nm, while spraying with NaNO2-AlCl3-NaOH triggered hyperchromic shifts, and thus an increase in absorbance intensity in flavonoids with particular substitution patterns. A few non-flavonoid components with structural similarities to flavonoids (e.g., rosmarinic acid, gallic acid, aspirin, salicylic acid) served as the negative control in this study to determine whether the derivatization reagents allowed exclusive detection of flavonoids. The method was then applied to the analysis of flavonoid containing supplements as well as red clover honey to demonstrate the method’s application in the analysis of natural products.

Quality control and shelf-life study of Ayush CCT formulation

BACKGROUND: One of the most widely popular brews globally is green tea/black tea, prepared by steeping the leaves of Camellia sinensis (L.) Kuntze plant. The examination of shelf-life proves to be a crucial tool in Ayurvedic formulations, particularly for ensuring the prolonged durability of the finished products. METHODS: The formulation underwent shelf-life studies maintaining conditions at 30°C ± 2°C and 65% RH ± 5% RH. The shelf-life dynamics were scrutinized through withdrawals at 0 day and at 3-month intervals to maintain the efficacy over a period of 24 months. In the current research, notable alterations were not detected in diverse physical, chemical, and safety parameters studied for shelf-life assessment. These parameters include pH, loss on drying at 105°C, ash values, extractive values, microbial contamination, high-performance thin-layer chromatography (HPTLC) profile, thermal analysis, and aflatoxins, all evaluated at different intervals throughout the entire study period. RESULTS: The current research study comprises of HPTLC fingerprint profiling of C. sinensis leaf and formulation, employing the solvent system of toluene: ethyl acetate: formic acid (6.5:3.4:0.1) with caffeine as the reference standard and safety parameters along with total phenolic content (6.9379 %w/w) and total flavonoid content (3.31 %w/w). The quantitative estimation values of caffeine in Ayush CCT and in C. sinensis leaf were 0.83% and 2.44%, respectively. CONCLUSION: It can be inferred that the Ayush CCT formulation demonstrates a shelf-life of more than 2 years. The study encompassed the assessment of shelf-life, as well as the qualitative and quantitative estimation of caffeine as a bioactive constituent in Ayush CCT formulation and C. sinensis extract.

Chemotaxonomy of Southeast Asian Peperomia (Piperaceae) Using High-Performance Thin-Layer Chromatography Colour Scale Fingerprint Imaging and Gas Chromatography–Mass Spectrometry

The morphological characters of Southeast Asia’s indigenous Peperomia species are very similar, especially in their flower structures. The flowers are simple, hermaphrodite and lack a perianth. Therefore, many species are hard to distinguish using morphological characters alone. Here, we apply chemometric data for species identification and classification, gathered using multiwavelength detection combined with the colour scale High-Performance Thin-Layer Chromatography (HPTLC) fingerprinting procedure and chemical compounds determined by Gas Chromatography–Mass Spectrometry (GC-MS). Fourteen taxa were investigated using hexane, ethyl acetate and ethanol solvent extractions. Principal component analysis (PCA) and hierarchical cluster analysis (HCA) were used with the colour scale fingerprints to classify the Peperomia species. The PCA and HCA using the chromatogram profile from hexane divided the taxa into six groups compared to the profile from ethyl acetate and ethanol, which each detected seven groups. The chromatogram from the combined dataset of all three solvents can differentiate all the species. The GC-MS data detected a total of 40 compounds from the hexane extract, and these differed among Peperomia species. This approach based on HPTLC fingerprinting and GC-MS analysis can therefore be used as a tool for authentication and identification studies of Peperomia species.

Rapid mitragynine quantification and fingerprinting of products from Mitragyna speciosa Korth. leaf (Kratom) using high-performance thin-layer chromatography.

Kratom (leaves from Mitragyna speciosa Korth.; Rubiaceae) is a herbal medicine known for its analgesic properties and psychoactive effects. Kratom in Thailand is currently legal; however, it is prohibited in some countries and considered a narcotic plant. Our aim was to establish a reliable, simple, and rapid method for quantifying mitragynine in Kratom leaves and related products through a combination of high-performance thin-layer chromatography (HPTLC) and densitometry. A densitometric HPTLC method was developed and validated in terms of specificity, linearity, limit of detection (LOD), limit of quantification (LOQ), accuracy, precision, and robustness. The fingerprints of kratom leaves, Mitragyna spp., and related products were constructed. For HPTLC, samples were applied to silica gel 60 F254 plates, and the mobile phase comprised n-hexane, ethyl acetate, and triethylamine (1:1:0.15, v/v/v). Densitometric detection was carried out under ultraviolet light at a wavelength of 226 nm. The validated method exhibited a range of 14.31-143.10 μg/mL, yielding a correlation coefficient of 0.9993. Spiked recovery rates were within a range of 98.3%-100.9%, and the LOD and LOQ were 3.80 and 11.53 μg/mL, respectively. Kratom samples were analyzed with the developed method, and the correlation coefficient was 0.9641, compared to the high-performance liquid chromatography-diode-array detection (HPLC-DAD) method. The HPTLC fingerprints displayed a distinctive pattern, facilitating discrimination among different plant parts and Mitragyna spp. The established method offers the advantages of simplicity, ease of use, and speed of analysis, serving as a practical alternative for mitragynine quantification in kratom leaf and its related products.

HPTLC FINGERPRINTS AND FTIR ANALYSIS FOR AUTHENTICATION AND QUALITY CONTROL OF Holostemma ada-kodien Schult.

Plant-derived pharmaceuticals are gaining increased acceptance in treating various ailments, but ensuring their quality is crucial. Both conventional and modern techniques are employed to validate the quality. High performance thin layer chromatography (HPTLC) and Fourier transform infra-red (FTIR) analysis were conducted to analyse and identify the various phytoconstituents in serial root extract of Holostemma ada-kodien. The peaks observed in HPTLC fingerprint profile corresponding to different phytoconstituents served as markers for standardizing drug; while FTIR analysis aids in identifying the major functional groups present. Thirteen bands were noted for hexane extract of H. ada-kodien in short UV in track I, while chloroform and methanol extracts showed eight bands each. In track II, the hexane extract exhibited eight bands, while chloroform and methanol extracts showed eleven and eight bands, respectively, under similar conditions. The hexane extract in long UV showed a maximum area of 78.4 and 71.59%, in track 1 and 2, respectively, with respective spanning from Rf 0.61 to 0.75 and 0.51 to 0.73. The methanol extract in visible light in track I showed a maximum area of 46.87%, spanning from Rf 0.73 to 0.98, and in track II, it displayed a maximum area of 36.59%, spanning from Rf 0.75 to 0.96 with 10 µL applied. Methanolic root extract of H. ada-kodien showed clear banding patterns in HPTLC fingerprints which seemed to be a dependable technique to detect adulteration while FTIR technique revealed functional groups of methanol root extracts.

Exploring the therapeutic potential of Cynanchum tunicatum (Retz.) Alston- assessment of phytochemicals and biological activities

Cynanchum tunicatum (Retz.) Alston is native to the Asian region and is distributed in the tropical areas of India and Sri Lanka. The aim of this study is to explore the phytochemical composition, antioxidant potential, and anti-inflammatory properties of C. tunicatum. Metabolic profiling was carried out using phytochemical screening to detect and quantify the secondary metabolites. To evaluate the potential secondary metabolites using the standard methods, Fourier transform infrared (FTIR), High-Performance Thin Layer Chromatography (HPTLC) and Gas Chromatography–Mass Spectrometry (GC–MS) from organic extracts of C. tunicatum and its biological activities. FTIR investigated peaks that represent alkane and aromatic compounds. GC–MS revealed the presence of 22 constituents such as 1-Hexacosene (0.145 %), l-(+)-Ascorbic acid 2,6-dihexadecanoate (8.129 %), Campesterol (5.243 %), Beta −Amyrin (10.614 %), Lupeol (13.061 %), Octadecanoic acid (0.751 %) are the major active compounds present. HPTLC fingerprinting confirms the bioactive compounds such as colchicine, strychnine, coumarin etc. which are represented with corresponding Rf values. Among all the extracts of C. tunicatum methanolic extract showed highest antioxidant activities. In 2,2-diphenylpicrylhydrazyl method exhibit the IC50 of (38.91 µg/mL), Ferric reducing antioxidant power assay (1.6 µg/mL), total antioxidant assay (IC50 = 32.91 µg/mL) and IC50 of anti-inflammatory activity (42.31 µg/mL) respectively. These findings enrich the knowledge of the species Cynanchum tunicatum for the possible application as a source of bioactive compounds in drug discovery.

Phytochemical Investigation of Alysicarpus vaginalis by Gas Chromatography-mass Spectrometry and High-performance Thin-layer Chromatography

Introduction: Alysicarpus vaginalis belongs to the Fabaceae family, commonly known as buffalo clover and one-leaf clover. Alysicarpus vaginalis is medicinally used in disease-related kidney, diuretics, leprosy, pulmonary problems, jaundice, skin problems, respiratory difficulties, and as a hepatoprotective. Alysicarpus vaginalis is a medicinally important plant according to our review of literature on pharmacological activities, modern pharmacological activities, ethnomedicinal surveys, and traditional systems. However, very limited research related to phytochemical investigation by chromatographic techniques was studied. Methods: So, to investigate the phytochemicals of Alysicarpus vaginalis methanol extract by High-performance thin-layer chromatography and gas chromatography-mass spectrometry. Result: Phytochemical screening revealed the presence of steroids, flavonoids, polyphenols, terpenoids, carbohydrates, alkaloids, and tannins. GC-MS revealed, the presence of six out of nine phytochemicals found medicinally important. 4-O-Methylmannose, Undecane, Neophytadiene, 2-amino-5-[(2-carboxy) vinyl]- Imidazole, Lup-20(29)-en-3-one and stigmasterol are some of the important phytochemical useful as anticancer or as anti-inflammatory. A solvent system was developed by thin-layer chromatography, followed by the HPTLC analysis using the mobile phase toluene: methanol: ethyl acetate (7:3:1). High-performance thin-layer chromatography fingerprinting detected total of 8 peaks with Rf value of 0.007, 0.089, 0.142, 0.211, 0.468, 0.585, 0.772, and 0885 at 254nm and total of 9 peaks with Rf value of 0.007, 0.089, 0.140, 0.208, 0.468, 0.661, 0.772, 0887, and 0.940 at 366nm, respectively. Conclusion: Chemoprofiling by sophisticated spectral techniques such as GC-MS and HPTLC fingerprinting can serve as an important tool for quality control of assuring purity, safety, quality, and efficacy of herbal formulation, and herbal extracts from Alysicarpus vaginalis.

Pharmacognostic study and preliminary phytochemical investigation of Andrographis paniculata (Burm.f.) Wall. ex Nees

Andrographis paniculata (family –Acanthaceae) is an annual herbaceous plant with branches that grows up to a height of 50-150 cm in moist, shaded areas. Its stem is abruptly quadrangular, heavily branched, and has a delicate texture that is readily broken. It is found in Southeast Asia, tropical and subtropical Asia, and a few other nations like Vietnam, Cambodia, and the Caribbean islands, Indonesia, Laos, Malaysia, Myanmar, Sri Lanka and Thailand etc. It is widely used in different traditional medicinal systems or countries for medicinal purposes by the traditional practitioners, tribes or community as a folklore remedies. The present communication provides a detailed account of the pharmacognostic study carried out on various parts of the Andrographis paniculata. The study includes macroscopy, microscopy, powder microscopic studies, physicochemical tests, preliminary phytochemical screening, development of HPTLC (High Performance Thin Layer Chromatography) fingerprints profile and heavy metal tests. Physicochemical parameters were performed and found average values of three parts (root, stem and leaf) such as root loss on drying at 1050C 4.93% w/w, total ash value 10.16% w/w, acid insoluble ash value 0.21% w/w, alcohol soluble extractive value 13.98% w/w and water soluble extractive value 25.77%w/w. Stem loss on drying at 1050C 6.23% w/w, total ash value 6.88% w/w, acid insoluble ash value 0.25% w/w, alcohol soluble extractive value 18.46% w/w and water soluble extractive value 21.94%w/w and leaf loss on drying at 1050C was found 6.29% w/w, total ash value 7.30% w/w, acid insoluble ash value 0.13% w/w, alcohol soluble extractive value 21.93% w/w and water soluble extractive value 38.68%w/w. HPTLC (High Performance Thin Layer Chromatography) fingerprints profile of methanolic extract was done by using mobile phase toluene: ethyl acetate (7:3). Andrographolide and Ferullic acid standard markers were applied, major spots Rf values with colour were recorded before derivatization at 254nm. Rf values are 0.25 black, leaf, stem and root with Andrographolide standard marker, and 0.20 light black color of stem, and root with Ferullic acid standard marker. Andrographolide is higher present in leaf which was range from 1.51-162 than the stem range1.46-153 and root range 1.45-1.150, while Ferulic acid was higher present in stem range from 0.91-0.96 than root 0.76-0.82 but absent in leaf. Heavy metals i.e Pb, Cd, As, & Hg were tested and found under WHO limits Established parameters can be used as standards for quality control and identification of the plant in herbal compound formulations and also preparation of a monograph of the plant.

Phytochemical Profiling for Leaf, Stem, Roots, and Seeds of Withania somnifera (L.) Dunal

Background: Withania somnifera (L.) Dunal, belonging to the Solanaceae family and commonly known as Ashwagandha, is a traditional Ayurvedic herb renowned for its adaptogenic properties. It plays a significant role in various classical Ayurvedic formulations and proprietary medicines. Traditionally, the roots of the plant are utilized over the aerial parts, such as leaves and stems. In recent years, a global increase in the use of Ashwagandha roots has been spurred by several modern pharmacological studies supporting its benefits. Methods: This study examined different parts of the herb, including leaves, stems, roots, and seeds, using methods like phytochemical analysis, thin layer chromatography (TLC) analysis, high-performance thin layer chromatography (HPTLC) phytochemical profiling, UV-visible spectral analysis, and 1H nuclear magnetic resonance (NMR) profiling. Results: The findings indicated that phytosterols, triterpenoids, and alkaloids were predominantly found in the leaves and stems, with lower concentrations in the roots and seeds. Ethanol extracts were particularly rich in total flavonoids, whereas aqueous extracts contained higher levels of phenolic compounds. High-performance thin layer chromatography fingerprinting varied for each part of the herb, effectively separating pigment-based components such as chlorophyll at 366 nm in the aerial parts. The 1H NMR spectra showed distinct metabolite profiles for each part of the plant. Conclusions: The analytical methods proposed in this study offer enhanced quality control and evaluation for the medicinal use of W. somnifera roots and its aerial components.

A rapid HPTLC fingerprinting technique for identifying the various geo-floral origins of honeys

Honey has several nutritional and therapeutic uses due to the presence of different bioactive compounds.These substances are derived from floral nectar. Therefore, it becomes essential to identify the distinct chemical profiles of honey samples. The aim of this research was to provide an accurate, straightforward, and sensitive HPTLC approach for different types of honey verification. Eight mono floral honeys (mustard, eucalyptus, litchi, orange, tea, Indian plum, black plum and pineapple) were collected from different origin of West Bengal (eastern India) and examined. Standard procedures were followed to check the quality of each honey.The flora was identified by microscopically examining the pollen found in honeys. High-performance thin layer chromatography (HPTLC) was used to analyze lipophilic fractions of each honey. Chromatographic results identified distinct patterns of bands with specific Rf values for each type of mono floral honey. HPTLC is a simple and effective method for routine analysis and verification. It can serve as authentication for different types of honey.

Comparative Evaluation of Rasa (Taste), Phytochemical Characterisation, HPTLC Fingerprinting and Antioxidant Activity of Taruni peya (Modified Rose Tea) Brewed using Five Methods: A Research Protocol

Introduction: A “rose” is usually a flowering shrub of the genus Rosa that has thorny stems and fragrant blooms. Humans have been using roses and their preparations since ages for health benefits. Rose tea is an infusion prepared from rose buds and petals that is popular for its mild flavour and possible health benefits, including its antioxidant content. It can be drunk as a hot or cold infusion. Need of the Study: No research has been done to date on the effects of brewing procedures on Rasa, phytochemical characterisation, High-performance Thin-layer Chromatography (HPTLC) fingerprinting, and the antioxidant properties of modified rose tea. A new field for standardising the brewing process for beverages like modified rose tea will be made possible by the study of these factors. Aim: To evaluate of Rasa (taste), phytochemical characterisation, HPTLC fingerprinting, and antioxidant activity of Taruni peya {Modified Rose (Rosa centifolia Linn.) tea} prepared by five different brewing methods. Materials and Methods: An experimental study will be conducted in the Department of Dravyaguna Vigyan, Mahatma Gandhi Ayurved College and Hospital and Research Centre (MGACH and RC), Salod, Wardha, Maharashtra, India, from April 2024 to June 2025. Roses (Rosa centifolia Linn.) will be collected from the natural habitat of the plant. The plant material will be authenticated and identified from Foundation for Revitalisation of Local Health Traditions (FRLHT), Bengaluru, Botanical Survey of India (BSI), or the Botany Department or by an authorised person of the Dravyaguna (Pharmacology) Department. Rasa Nirdharana (Taste) of the samples will be analysed using parameters mentioned in Samhitas. Charaka, Sushruta, and other acharyas, have described the method of the perception and determination of taste by putting a substance on the tongue in which the Rasa perceived soon after the substance comes in contact with the tongue is called Rasa. Standard Pharmacognostical character and phytochemical characterisation will be performed on all samples using standard methods like macroscopy-microscopy and HPTLC fingerprinting will be studied as given in Ayurvedic Pharmacopoeia of India. Antioxidant properties will be studied by using 2, 2-diphenyl-1- picrylhydrazyl (DPPH), Ferric ion Reducing Antioxidant Potential (FRAP), and phosphomolybdenum assay. Analysis of Variance (ANOVA) test will be used for statistical analysis with a level of significance at p-value<0.05.

Chromatographical fingerprint analysis, in silico investigation, and identification of hepatoprotective active compounds from Capparis decidua Edgwe (Forssk.)

Liver disorders are globally distributed with a high burden of disease costs. The methanolic extract of Capparis decidua stem part and its dichloromethane (DCM) and ethylacetate fractions showed significant hepatoprotective activity, so this study aimed to develop gas chromatography/mass spectrometry (GC/MS), high-performance thin layer chromatography (HPTLC) fingerprint profile and in silico docking of identified compounds from both fractions responsible for hepatoprotective activity. Hexadecanoic acid, 9-octadecenoic acid, 9,12,15-octadecatrienoic acid, and dihydrolinalool are known hepatoprotective and antioxidant agents that were identified from both fractions by GC/MS, they have various degrees of hepatoprotective and antioxidant properties, this is the first time to identify hexadecanoic acid in C. decidua stem. Both fractions were chromatogramed with known hepatoprotective standard compounds, they were rutin, quercetin, kaempferol, ferulic acid, oleanolic acid, and β-sitosterol; β-sitosterol and oleanolic were isolated and identified in the DCM fraction by HPTLC, but nothing of these standard compounds were identified in the ethylacetate fraction. According to International Council for Harmonization guidelines, the HPTLC fingerprint profile method was validated to confirm our isolation and identification process of β-sitosterol and oleanolic acid in the DCM fraction, also their contents were quantified and they were 9.21 and 7.73 μg/mg, respectively, in the total dry fraction weight, this was the first time to isolate and identify oleanolic acid from C. decidua stem part. In silico docking was carried and was found that hexadecanoic acid showed significant predicted binding properties on Nrf2, p38 mitogen-activated protein kinase, and IL-1 receptor proteins. 9-octadecenoic acid (elaidic acid) showed significant predicted interactions with IL-1 receptor, Nrf2, and peroxisome proliferator-activated receptor-α (PPAR-α) proteins. Oleanolic acid showed significant predicted bindings with p65, JNK, and NF-kb. β-sitosterol showed significant predicted interaction with IL-6α protein, while 9,12,15-octadecatrienoic acid revealed significant binding with PPAR-α protein. Therefore, we recommend considering our active compounds in the field of drug development (structure-activity relationship) and the possibility of the presence of synergism if they are combined and administered together. Moreover, we recommend liquid chromatography-mass spectrometry and nuclear magnetic resonance on both fractions to search for the possibility of the presence of other hepatoprotective compounds.

COMPARATIVE EVALUATION OF IN-HOUSE AND MARKETED SAMPLES OF VAISHWANARA CHURNA

The present study aims to evaluate the pharmacognostic, physicochemical, spectroscopic analysis which includes total phenolic content, total flavonoid content, high performance thin layer chromatography (HPTLC), antioxidant activity, antiurolithiatic activity, total microbial load of the in-house and marketed samples of Vaishwanara Churna. The study intends to highlight the variations observed between the in-house and two marketed samples of Vaishwanara Churna. The comparative standardization of IHVC, TVC, and SVC was performed based on pharmacognostic evaluation, physicochemical evaluation and phytochemical screening. The total phenolic content was found highest in TVC viz.161.57±5.426 mg GAE/g. The highest flavonoid content was found to be 82.67±6.549 mg QUE/g in SVC. The sample also shows good antioxidant activity which was indicated by three antioxidant methods. Antiurolithiatic activity was found to be dose dependent. HPTLC fingerprint technique showed that SVC has highest concentration of gallic acid.

Pharmacognostic standardization and HPTLC fingerprinting analysis of Sahacharadi Kashayam: a classical ayurvedic polyherbal formulation

Multiherbal compositions are widely practiced in ayurvedic medical system, and ka??yam (decoction) is one such dose. Sahacharadi Kashayam is a traditional ayurvedic polyherbal medicine recommended by ayurvedic doctors for chronic and acute back pains, lower limb disorders, and conditions that mainly affect the nutrient level in the blood and the bone. The aim of this study was to develop pharmacognostic parameters for the standardization and quality evaluation of the Sahacharadi Kashayam. In-house polyherbal formulation was prepared as per the previously prescribed method, and physicochemical parameters were evaluated as per standard protocol and showed loss on drying at 99.31%, total ash content at 4.911 ± 0.120%, water-soluble ash at 3.022, and acid-insoluble ash at 0.489. Total phenolic content was estimated with FC reagent and expressed as gallic acid equivalent (9.95 + 0.004 mg/g). High-performance thin-layer chromatography (HPTLC) fingerprint profiling was obtained, and the quantity of total tannins was estimated as tannic acid equivalent using HPLC (0.47%). The results obtained from the study could be utilized as a reference for the quality control and quality assurance of Sahacharadi Kashayam.

A strategy for quality evaluation of complex herbal preparations based on multi-color scale and efficacy-oriented high-performance thin-layer chromatography characteristic fingerprint combined with chemometric method: Sanwujiao Pills as an example

To rapidly evaluate the quality of complex herbal preparations, a new strategy was proposed based on multi-color scale and efficacy-oriented high-performance thin-layer chromatography (HPTLC) characteristic fingerprint combined with chemometric method. Firstly, effective components were screened through high-performance liquid chromatography with ultraviolet detection and evaporative light-scattering (HPLC-UV-ELSD), using multi-wavelength fusion combined with network pharmacology and molecular docking techniques. Subsequently, guided by the effective components, the targeted HPTLC characteristic fingerprint was established by multi-color scale scanning. Finally, combined with the chemometric method, the consistency of the preparation quality was evaluated, the marker components leading to quality differences were screened, and the quality control limit was established. Sanwujiao Pills (SWJPs) is a herbal preparation composed of six herbs for treating rheumatoid arthritis (RA). Through this strategy, four HPTLC characteristic fingerprints were established, they were derived from five herbs and guided by eight effective components in SWJPs. Through similarity, clustering heatmap, principal component analysis (PCA), and orthogonal partial least squares discriminant analysis (OPLS-DA), the quality distinctions among the 12 batches of SWJPs were determined. These batches were categorized into two groups based on their production time, and eight components affecting the quality of the preparation were identified. Meanwhile, the quality control threshold for SWJPs was determined based on Hotelling's T2 and DModX methods. This strategy aims to rapidly evaluate the quality of complex herbal preparations by HPTLC and extends the application of HPTLC fingerprint chromatography for identifying herbal medicine species and activity-related quality detection. The proposed strategy is also helpful for the quality control of other complex herbal preparations.

Investigation on ethanolic stem extract of Ipomea sagittifolia to explore the presence of Saponins by High Performance Thin Layer Chromatography

Background: Saponins are a diverse group of naturally occurring plant compounds that play important roles in various aspects of plant physiology and ecology. An extract composed of several phytoconstituents may be easily analysed and interpreted using high performance thin layer chromatography (HPTLC) fingerprint analysis from a qualitative standpoint. Ipomea sagittifolia (Burn. f.) has a long history of use in traditional medical systems. Method: It was decided to interpret the phytoconstituents, namely the steroids in Ipomea sagittifolia (Burn. f.), in line with Current Good Manufacturing Practises, which highlight the significance of quality in relation to phytoconstituents, because HPTLC is a very trustworthy method of analysis. In this work, an HPTLC finger print profile for the saponins was made using an ethanolic extract of the stem of Ipomea sagittifolia (Burn. f.). This system includes the Scanner 4, TLC Visualizer, and Linomat 5 Applicator. Results: When phytoconstituents were evaluated using HPTLC densitometric screening at wavelengths of 366 nm and 540 nm, several peaks could be seen in the chromatogram. By analysing the peak regions, peak heights, and Rf values that were given in the proper tables, the phytochemicals are appraised. Conclusion: The study revealed the presence of saponins. This information is highly useful in studying chemical profiling and discovering bioactive components when the Rf values of these chemicals are compared with standards as a reference.

Phytochemical, Physicochemical and HPTLC Analysis of Siddha Herbal Formulation Muppirandai Chooranam

Aim: The aim of the study was to evaluate the Phytochemical, Physicochemical, HPTLC analysis of Siddha herbal formulation Muppirandai Chooranam (MRC), indicated for various ailments including menorrhagia.
 Study design
 Place of study: The Phytochemical and Physicochemical analysis were conducted at the Tamilnadu Dr. MGR Medical University, located at no.69, Anna salai, Guindy, Chennai-32. The high-performance thin layer chromatography (HPTLC) was carried out at Noble research solutions, Kolathur, Chennai -99.
 Methodology: Siddha formulation Muppirandai Chooranam was prepared as per good manufacturing practices (GMP) guidelines and the phytochemical analysis, physicochemical analysis were carried out at the Tamilnadu Dr. MGR Medical University, located at no.69, Anna salai, Guindy, Chennai-32. As per PLIM guidelines (Pharmacopeia Laboratory Of Indian Medicine) in accordance with the guidelines established by AYUSH (Ayurveda, Yoga, Unani, Siddha, Homoeopathy), the governing body for traditional health systems in India. The HPTLC analysis was conducted at Noble Research Solutions, Kolathur, Chennai -99.
 Results: The Phytochemical screening of MRC shown the presence of Alkaloids, Carbohydrates, Saponins, Flavanoids, Diterpenes, Gum and Mucilage. The physicochemical analysis of MRC revealed that it had a loss on drying of 0.42% at 105°C, a total ash content of 23.3%, an acid-insoluble ash content of 3.02%, a water-soluble ash content of 2.30%, and a water-soluble extract content of 17.67%, and alcohol soluble extract content of 5.23%.HPTLC finger printing analysis of the sample reveals the presence of five prominent peaks corresponds to the presence of five versatile phytocomponents present within it. Rf value of the peaks ranges from 0.02 to 0.37.
 Conclusion: From the findings, it is concluded that it shown the compendious understanding of presence of Phytochemical components, Physicochemical characteristics, and HPTLC analysis of MRC and it is competent to assess the quality profile of Muppirandai Chooranam as a reference standard for the development of the standardised pharmaceutical product.

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.

High-performance thin-layer chromatography fingerprint profile analysis and spectro-densitometric evaluation of antiproliferative antioxidants such as ellagic acid and gallic acid from four widely used Terminalia species

High-performance thin-layer chromatography fingerprint profile analysis and spectro-densitometric evaluation of antiproliferative antioxidants such as ellagic acid and gallic acid from four widely used Terminalia species

Phytochemical Profiling and Pancreatic Lipase Inhibitory Activity of <i>Flacourtia inermis</i> Roxb. Fruits

Objectives: The present research work was carried out to explore the potential use Flacourtia inermis [FI] fruits for the prevention and treatment of obesity through pancreatic lipase inhibition in vitro. The study also aimed to investigate the chemical profiling of ethanol extract of FI using High-Performance Thin Layer Chromatography (HPTLC), High-Resolution Liquid Chromatography (HR-LC/MS), and Nuclear Magnetic Resonance Spectroscopy (NMR). Materials and Methods: Dried fruits of Flacourtia inermis were pulverised and subsequently extracted using various solvents in sequential steps of increasing polarity, such as hexane, ether, chloroform, ethyl acetate, ethanol, and water. After phytochemical analysis by preliminary chemical testing various extracts were evaluated for their ability to inhibit pancreatic lipase, and the ethanol extract was found to have an IC<sub>50</sub> close to that of reference drug orlistat. The most potent ethanol extract was analysed by HPTLC and separated through column chromatography, and further analysis was performed by HR-LC/MS and 1H-NMR techniques. Results: The presence of various phytoconstituents in this plant was detected using different types of analytical techniques. PL lipase inhibitory activity was observed in extracts in a dose dependent manner. Performing PL inhibition assay, it was found that the ethanol fruit extracts have lipase inhibitory activity with an IC<sub>50</sub> value of 377.15 μg/ml. HPTLC finger printing of the ethanol extract showed the presence of various bioactice compounds. HR-LC/MS study of the most active ethanol extract indicated the presence of different phytochemicals, such as phenolics and flavonoids. Column chromatographic separation of ethanol fruit extract of FI followed by structural elucidation using various spectral studies demonstrated the presence of two compounds namely myricetin and quinic acid. Conclusion: The study suggests that the edible fruits of Flacourtia inermis have the potential to inhibit pancreatic lipase enzyme and therefore, may be recommended for the management of obesity. Additionally, our research sheds light on the phytochemistry of flacourtia species and may lead to the development of novel chemical entities as potential pancreatic lipase inhibitors.