TLC Method Research Articles

TLC with densitometric and image analysis in the control of anthocyanin content in fruits of Rubus occidentalis and Rubus idaeus cultivars and hybrids

A TLC method was developed for the determination of anthocyanins in the fruits of some cultivated varieties of Rubus occidentalis (black raspberries) and Rubus idaeus (red raspberries) and their hybrids. Anthocyanin sets in fruits’ extracts were determined on TLC silica gel 60 F254 plates using mobile phase consisted of ethyl acetate-water-formic-acid (100:30:30, V/V), at 4°C (±1°C), at a distance of 70 mm with the use of densitometry and videoscanning. The obtained results were comparable to the results of quantitative analysis performed using by the HPLC-UV/VIS method performed on a Kinetex C-18 column using gradient elution comprising an increase concentration of acetonitrile-water-trifluoroacetic acid (50:50:0.1, V/V) in water-trifluoroacetic acid (99.9:0.1, V/V) from 12 % to 40 % within 40 minutes at 520 nm. The TLC method with videoscanning provides immediate capture of data for unstable compounds like anthocyanins before their partial degradation.

Green and Sensitive Analysis of the Antihistaminic Drug Pheniramine Maleate and Its Main Toxic Impurity Using UPLC and TLC Methods, Blueness Assessment, and Greenness Assessments

For the first time, two direct and eco-friendly chromatographic approaches were adapted for the simultaneous estimation of pheniramine maleate (PAM) and its major toxic impurity, 2-benzyl pyridine (BNZ). Method A used reversed-phase ultra-performance liquid chromatography; separation was achieved within 4 min using a C18 column with a developing system of methanol/water (60:40 v/v) with a 0.1 mL/min flow rate. Photodiode array detection was adjusted at 215 nm. The method was linear in the ranges of 5.0–70.0 and 0.05–10.0 µg/mL for PAM and BNZ, correspondingly. Method B used thin-layer chromatography; separation was applied on silica gel TLC F254 using ethanol/ethyl acetate/liquid ammonia (8:2:0.1, in volumes) at room temperature, at 265 nm. Linearity was assured at concentration ranges 0.5–8.0 and 0.1–3.0 µg/band for the two components, respectively. Generally, the new UPLC and TLC methods outperform the old ones in terms of quickness, greenness, and sensitivity. Concisely, the greenness features were partially achieved using the Green Analytical Procedure Index (GAPI) and the Analytical Greenness (AGREE) pictograms. In contrast, the usefulness of the novel approaches was assured via the Blue Applicability Grade Index (BAGI) tool.

Ivermectin-based products in the context of green pharmaceutical analysis

Abstract Background Ivermectin (IVE), a broad-spectrum antiparasitic, is used in human and animal health. Analytical methods for evaluating IVE in pharmaceutical products are found in the literature and in official compendiums. However, the vast majority of them do not have an eco-friendly approach. Objective and Method The aim of this review is to present an overview of existing analytical methods for evaluating IVE in pharmaceutical matrices in the context of Green Analytical Chemistry (GAC) and show possibilities for increasing their greenness. Results GAC is a current alternative to promote sustainable development in laboratories and chemical-pharmaceutical industries, therefore, through its principles, such as reducing the use of aggressive solvents, it is possible to make processes more ecological. However, the vast majority of analytical methods available in the literature and official compendiums do not present an eco-friendly approach. 70% of the methods are by HPLC. Among the various pharmaceutical matrices, the most evaluated are tablets (37%). Of all the solvents used in HPLC, UPLC, HPLC-MS/MS, UV and TLC methods, the combination of methanol and acetonitrile is the most chosen, accounting for more than 50% of occurrences. Conclusions Analytical methods for evaluating IVE-based products can be leveraged within the scope of GAC, bringing sustainable work opportunities to analytical development laboratories around the world. Highlights This review shows an overview of the analytical methods present in the literature and official compendiums to evaluate pharmaceutical IVE matrices, in the context of green analytical chemistry.

Quantitative Thin Layer Chromatography Analysis, Antioxidant, and Anti-Inflammatory Activities of Polyherbal Formulation (Ammarit-Osot) Extracts

This study was carried out to develop a TLC method for quantifying one of bioactive, piperine, from polyherbal formulation (Ammarit-Osot) extracts. The extracts were prepared by different solvents such as methanol (ME) and water (WE). The mobile phase of toluene:acetone:formic acid (7:2:1, v/v/v) was used for separation. The calibration curve showed a good linearity (r2 = 0.9972) in the range of 50 – 500 ng/spot. This assay was assessed by intra-and interday precision (RSD 0.25 – 2.49%), and accuracy (101.05 – 101.56 %). The piperine content was found to be 23.06 ± 0.06 and 0.44 ± 0.03% w/w for ME and WE, respectively. Both ME and WE demonstrated significant activity in DPPH and FRAP assays, indicating their antioxidant potential. The TPC of ME and WE were 53.40 ± 2.21 and 101.18 ± 0.46 (mg GAE/g extract), respectively. The TFC of ME and WE were 43.35 ± 3.94 and 25.31 ± 1.74 (mg catechin/g extract), respectively. In addition, the cytotoxicity of extracts on RAW264.7 was investigated by MTT assay. The ME and piperine showed IC50 with 162.40 ± 7.84 and 115.68 ± 11.96 µg/mL, respectively. While the WE exhibited low cytotoxicity (IC50 > 1,000 µg/mL), the anti-inflammatory activity was assessed by LPS-stimulated NO overproduction. The ME and piperine displayed the ability to inhibit NO production, but WE not inhibited. The results indicated that the different solvent extracts of Ammarit-Osot have the potential to be antioxidant, anti-inflammatory and immune-stimulate agent.

High Performance Thin Layer Chromatography (HPTLC) Analysis of Anti-Asthmatic Combination Therapy in Pharmaceutical Formulation: Assessment of the Method's Greenness and Blueness.

A cost-effective, selective, sensitive, and operational TLC-densitometric approach has been adapted for the concurrent assay of Hydroxyzine Hydrochloride (HYX), Ephedrine Hydrochloride (EPH), and Theophylline (THP) in their pure powder and pharmaceutical forms. In the innovative TLC-densitometric approach, HYX, EPH, and THP were efficaciously separated and quantified on a 60F254 silica gel stationary phase with chloroform-ammonium acetate buffer (9.5:0.5, v/v) adjusted to pH 6.5 using ammonia solution as a mobile liquid system and UV detection at 220 nm. The novel TLC method validation has been performed in line with the international conference for harmonization (ICH) standards and has been effectively used for the estimation of the researched medicines in their pharmaceutical formulations without intervention from excipients. Additionally, parameters affecting the chromatographic analysis have been investigated. The new TLC approach's functionality and greenness were appraised using three modern and automated tools, namely the Blue Applicability Grade Index (BAGI), the Analytical Greenness metric (AGREE), and the Green Analytical Procedure Index (GAPI) tools. In short, the greenness characteristics were not achieved as a result of using mandatory, non-ecofriendly solvents such as ammonia and chloroform. On the contrary, the applicability and usefulness of the novel TLC approach were attained via concurrent estimation for the three drugs using simple and straightforward procedures. Moreover, the novel TLC method outperforms previously published HPLC ones in terms of the short run time per sample and moderate pH value for the liquid system. According to the conclusions of comparisons with previously recorded TLC methods, our novel HPTLC method has the highest AGREE score, so it is the greenest HPTLC strategy. Moreover, its functionality and applicability are very appropriate because of the simultaneous assessment of three drugs in one TLC run. Furthermore, no tedious and complicated extraction and evaporation processes are prerequisites.

РАЗРАБОТКА МЕТОДА ИДЕНТИФИКАЦИИ И ОЦЕНКА КОЛИЧЕСТВЕННОГО СОДЕРЖАНИЯ ЭФИРНОГО МАСЛА В ХВОЕ МОЖЖЕВЕЛЬНИКА ОБЫКНОВЕННОГО

The aim of the study is to develop methods for the identification of terpenoids and to assess the quantitative content of essential oil in the conifers of juniper, fresh and dried by air-shade drying. The object of the study were samples of needles of juniper. The raw materials were used fresh and dried. To carry out qualitative reactions, well-known methods were selected, which are most often used in the analysis of essential oil raw materials. The identification of essential oil components was carried out by TLC method on Silufol plates. A-pinene solution was used as a standard. The quantitative content of essential oil in the raw material was determined by steam distillation in accordance with the OFS method.1.5.3.00010.15. Determination of the essential oil content in medicinal plant raw materials and medicinal herbal preparations. The authors proposed qualitative reactions for the identification of terpenoids in raw materials, which can be used to form the "Authenticity" section for a new type of raw material-needles of juniper. A-pinene was identified by the TLC method. A quantitative assessment of the essential oil content in fresh and dried raw materials was carried out. A decrease in the content of essential oil during drying of raw materials was revealed. Qualitative reactions were selected experimentally to confirm the presence of terpenoids in alcoholic extracts from juniper needles, as well as unsaturated compounds in essential oil obtained by steam distillation. The TLC method identified a-pinene in the analyzed samples. The presence of additional 11 adsorption zones in the samples of essential oil obtained from fresh raw materials and 9 in samples from dried raw materials was also revealed. The results obtained indicate changes in the composition of the terpenoid fraction occurring during the drying process and actualize further research in this direction. Assessment of the quantitative content of essential oil in the raw material, carried out according to the method 2 of the OFS.1.5.3.00010.15 "Determination of the essential oil content in medicinal plant raw materials and medicinal plant preparations", showed a decrease in the essential oil content in all dried samples compared with fresh raw materials. The data obtained during the experiment allow us to recommend the inclusion of an essential oil content of at least 1% in the quality indicators of juniper needles, as well as optimize the drying mode to better ensure the preservation of essential oil in raw materials.

Stability-Indicating TLC-Densitometric and HPLC Methods for Simultaneous Determination of Teneligliptin and Pioglitazone in Pharmaceutical Dosage Forms with Eco-Friendly Assessment.

The combination of teneligliptin hydrobromide hydrate and pioglitazone hydrochloride in pharmaceutical formulations has improved type 2 diabetes management. Two chromatographic methods TLC-densitometry and RP-HPLC were developed for simultaneous quantification of teneligliptin hydrobromide hydrate and pioglitazone hydrochloride in pharmaceutical formulations, ensuring accuracy and stability assessment. The TLC method uses a mobile phase of methanol, toluene, ethyl acetate and triethylamine (1:7:2:0.1, v/v/v/v) on TLC silica gel plates, scanned at 268nm. The RP-HPLC method employs isocratic elution with acetonitrile and sodium acetate buffer (adjust pH3.6 with glacial acetic acid, 60:40v/v) on a shimpack C18 column (250 × 4.6mm i.d., 5μm), detected at 235nm. Both methods offer high accuracy and reliability, making them valuable for pharmaceutical quality control. Additionally, an environmental impact assessment was conducted using eco-scale, Analytical Greenness Metric Approach, Green Analytical Procedure Index, and national environmental method index to evaluate solvent consumption, waste generation and energy usage. Statistical comparisons (t-tests and F-tests) validate the outcomes of both methods, ensuring their effectiveness in drug formulation analysis. These methods can enhance pharmaceutical quality control while fulfilling environmental responsibilities.

Thin layer chromatography of methylated derivatives of sodium alkylbenzenesulfonates in water analysis by GC-MS

Sodium alkylbenzene sulfonates or linear alkyl benzene sulfonates (LAS) are the most common synthetic anionic surfactants and water pollutants. They can cause both acute and chronic toxic effects on water organisms. Selective determination of sodium alkylbenzene sulfonates as a separate class of anionic surfactants is possible using gas chromatography coupled with mass spectrometry (GC-MS) in the form of linear alkylbenzenesulfonic acid methyl esters (LABSA ME). In order to purify the extracts and concentrate the analytes, we studied the behaviour of these compounds by ascending high-performance thin-layer chromatography using Kieselgel 60 F254 and Sorbfil plates. A mixture of n-hexane and methanol solvents in a ratio of 23:1 (by volume) was used as the mobile phase. Under these conditions, sodium alkylbenzenesulphonates remain on the start line, while their derivatives (ME ABSC), obtained by methylation with trimethyl orthoformate in the presence of trifluoroacetic acid (with a yield of η=98%), form zones characterised by retention coefficient values of Rf = 0.62 and Rf = 0.71 on Kieselgel 60 F254 and Sorbfil plates, respectively. The repeatability of the Rf values was characterised by a standard deviation of 6.1 and 5.9 %, respectively (n=16). The completeness of extraction (95.0–100.0 %) of analytes from the plates by descending TLC with acetonitrile was noted. The applicability of the TLC method for the concentration of analytes and pretreatment of extracts on the example of real water samples was shown. Using GC-MS with electron impact ionisation, we determined the concentrations of sodium alkylbenzenesulphonates in water sampled in the southern basin of Lake Baikal from a depth of 400 m (0.24±0.02 µg/dm3) and in snowmelt water from the ice of the Krestovka River where it flows into Lake Baikal near the village of Listvyanka (31.1 ± 1.0 µg/dm3).

Algorithm for the development of a multicomponent pharmaceutical substance of plant origin with antimicrobial action: from science search to dosage form

Introduction. Despite the proven clinical efficacy of antifungal and anti-inflammatory drugs for the prevention and treatment of seborrheic dermatitis, the search for new targets and the development of new substances with a beneficial effect on the scalp microflora, with a low risk of antimicrobial resistance and adverse effects, are relevant.Aim. Development of the antimicrobial multicomponent pharmaceutical substance of plant origin in stages: from a literature search for promising substances, analysis of their composition by GC-MS, in silico evaluation of the affinity of individual components to pathogenetic targets, selection of the optimal composition of a multicomponent substance based on the results of in vitro research of antimicrobial action and the making of a medical dosage form based on it – a medicinal shampoo for the treatment of seborrheic dermatitis.Materials and methods. Objects of research: tea tree essential oil, 1,8-cineole, α-(-)-bisabolol and the multicomponent substance based on them. Methods: molecular docking (AutoDock version 4.2), prediction of pharmacological activity (Phyto4Health), TLC, GC-MS, study of antimicrobial activity in vitro.Results and discussion. Based on the results of a literature search, 3 promising substances were selected for the development of a multicomponent plant-based substance: tea tree essential oil, 1,8-cineole and α-(-)-bisabolol. Molecular docking predicted the targeted activity of the phytochemicals of tea tree essential oil, 1,8-cineole and α-(-)-bisabolol on the domains of ABC-transporters of microorganisms involved in the pathogenesis of seborrheic dermatitis and justified the possibility of use for therapy. The multicomponent substance has been developed based on tea tree essential oil, 1,8-cineole and α-(-)-bisabolol in a mass ratio of 1 : 1 : 1. The qualitative composition of the substance was assessed by TLC and GC-MS methods, and 15 terpenes were quantitatively identified in its composition with a predominance of terpinen-4-ol (16.98 %), 1,8-cineole (25.63 %) and α-(-)-bisabolol (27.67 %). The synergistic antimicrobial activity of the substance has been established against S. epidermidis, S. aureus, C. albicans and M. furfur in comparison with benzalkonium chloride, ketoconazole and climbazole. The composition of a new medical shampoo based on the investigated substance has been developed, which has a pronounced antifungal effect (more than 99.0 %) against M. furfur without visible suppression of normal microflora. For the novel substance of plant origin and medical shampoo, quality parameters were assessed in accordance with the Russian Pharmacopoeia of XIV edition.Conclusion. A substance of plant origin with synergistic and targeted antimicrobial activity has been developed. It has an interest for further study as a drug and API for new products for the treatment of seborrheic dermatitis.

Two-Dimensional Thin-Layer Chromatography as an Accessible, Low-Cost Tool for Lipid-Class Profile Screening

The interest in lipid composition profiling is significantly increasing as research reveals the immense importance of lipids in medicine, plant science, food and agriculture. However, lipidomic analysis requires high-end specialty equipment. We used two-dimensional thin-layer chromatography (2D-TLC) as a readily available, low-cost tool for basic lipidomic profiling of lipid classes in algal samples in the models Chlamydomonas reinhardtii, Auxenochlorella protothecoides, and Euglena gracilis, validating lipid class identification using an LC-MS/MS analysis. Algal lipid extracts were separated on a 2D-TLC plate, and TLC analysis was followed by scraping individual TLC spots off the plate, and a subsequent liquid chromatography separation and tandem mass spectrometry (LC-MS/MS) analysis. For comparison, crude lipid extracts were also injected directly to the LC-MS/MS system. Lipid class annotation was achieved by a combination of accurate mass, retention time information, neutral loss and fragment ion analysis by MS2Analyzer, and by matching spectra to LipidBlast MS/MS library. Overall, we were able to identify 15 lipid classes, and to adequately profile the lipid classes in all three organisms. This TLC method is thus suggested as an accessible tool for lipid class profiling of algal, plant, and food lipids, alike, when a rapid and simple analysis is required, e.g., for screening purposes.

Adulteration in dietary supplements: HPTLC profiling of Withania somnifera and Terminalia bellirica

In the realm of traditional medicine, (Withania somnifera), commonly known as Ashwagandha, has long been revered for its multifaceted benefits, earning it the moniker "Indian Ginseng" or winter cherry within Unani systems. Its reputation precedes it, boasting a spectrum of advantageous properties such as antiviral, antioxidant, and anti-inflammatory effects. Moreover, research suggests its potential efficacy against various cancers including breast, colon, prostate, ovarian, and lung cancer. Similarly, Terminalia bellirica, referred to as Behra, holds its own array of therapeutic potential, addressing maladies ranging from headaches to liver disorders, gastrointestinal discomforts, common colds, and constipation. However, the widespread popularity of these medicinal plants has led to concerns regarding their sustainability. Overexploitation poses a significant risk, potentially depleting these valuable resources. Compounded with this risk is the issue of adulteration or contamination, where other plant parts or species resembling the authentic herbs are utilized, compromising the safety and efficacy of the end the product. In a recent investigation, two samples each of authentic and commercial Withania somnifera and Terminalia bellirica were subjected to rigorous assessment through physiochemical, phytochemical, TLC, and HPLC chromatographic methods. Alarming findings surfaced, revealing instances of adulteration or contamination in both the Ashwagandha and Behra products. Phytochemical analyses and HPTLC profiles of various extracts unveiled the intentional or unintentional addition of undesired herbs or substitutions. These adulterants not only jeopardize the expected therapeutic outcomes but also dilute the potency and safety of the herbal medicines flooding the local markets. This study serves as a clarion call for stringent authentication measures in the procurement and utilization of herbal medicines. High-Performance Thin-Layer Chromatography (HPTLC) emerges as a reliable tool for confirming adulteration, urging stakeholders to adopt thorough authentication protocols to safeguard public health and preserve the integrity of traditional herbal remedies.

Pharmacognostic Evaluation and Antioxidant Activities of Tetracera indica (Christm. and Panz.) Merr

The community uses Tetracera indica (Christm. and Panz.) Merr extensively as a traditional medicine. This plant was the main source of raw materials for herbal medicinal products, so identification was needed to ensure its authenticity through pharmacognosy evaluation. The present study evaluated macros- and microscopic characteristics, performed phytochemical analysis, and performed total phenolic (TPC) and flavonoid content (TFC) analysis. The organoleptic and macroscopic studies were carried out through sensory organs. The microscopic characteristics of leaf powder and twigs identification using chloral hydrate and distilled water. Numerous chemical reagents, TLC methods and LC-MS/MS analysis were employed in the phytochemical study. Research employed DPPH and FRAP tests to assess the antioxidant activity. Furthermore, quercetin was used as a marker of antioxidant activity. The morphological marker of the leaf T. indica showed the surface of the leaves is rough bristles, shape elliptic-oblong, margin serratus and phyllotaxy alternate. The stem is lignosus, and the rough stem surface is slightly peeling. The lower leaf surface had paracytic stomata and uniseriate trichomes with tapering ends. Raphide crystals and cork cells were detected in the powder analysis of the twigs. The ethyl acetate extract's twig had the highest TPC and TFC values. Ethyl acetate extract from Tetracera indica twigs has potent antioxidant activity with IC50 = 99.84±0.011 µg/ml (DPPH) and 4296.67±0.024 mol/g (FRAP). The study showed that Tetracera indica contains possible active components that could be developed as novel antioxidants. The result of this study might provide early verification and identification to obtain quality medicinal raw materials.

Development of Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy Coupled with Multivariate Classification Chemometric Model for Routine Screening of Paracetamol, Ibuprofen, and Aspirin Adulteration in Herbal Products

Objective: The objective of this study is to develop and validate a routine screening test for the determination of three common antipyretic-analgesic synthetic drugs (paracetamol, ibuprofen, and aspirin) adulteration in herbal products using Attenuated Total Reflectance Fourier Transform Infrared (ATR-FTIR) coupled with chemometric method. Methods: ATR-FTIR spectra of sixteen testing sets of herbal product samples for pain and fever indications were used for multivariate chemometrics model construction. Linear Discriminant Analysis (LDA) was selected as a method for model construction with IBM SPSS for statistical analysis. Model development employed feature selection, such as the stepwise method for variable selection. The model with a high %correct classification and cross-validation was selected and was then validated with an independent testing data set with an auto-prediction test, confusion matrix, and Receiver Operating Characteristic (ROC) curve. To validate the developed test for routine use, the result from ATR-FTIR method was compared with the standard HPLC and TLC analyses used for adulteration screening. Results: The selected model's overall %correct classification result was 97.7%, with a cross-validation of 93.8% rate in training set samples. External validation with an independent testing dataset gave an overall correct classification of 93.8%, with an area under the curve of ROC at 0.979. Comparative testing revealed that model performance was comparable with the HPLC and TLC methods, which routinely detect the presence of paracetamol, aspirin, and ibuprofen. The results of testing set samples classification were consistent with training set samples. Conclusion: Against the standard chromatographic methods, the multivariate chemometric model based on ATR-FTIR demonstrates comparable detection capability to determine adulteration of paracetamol, ibuprofen, and aspirin in herbal products.

Analysis of Hydroquinone Compounds in Whitening Body Lotion Preparations Circulating in The Community

Skin care has become an important part for today's women. Whitening body lotion is very necessary in maintaining skin when doing activities outside and indoors. Many Indonesian women desire to have white, clean and bright skin so they can appear attractive. An attractive appearance is very necessary in society. To achieve this, many women choose skin care products at affordable prices without paying attention to the dangers of the chemical content in them. One of the skin whitening ingredients that is often used in beauty products is the compound hydroquinone. The hydroquinone compound plays a role in preventing skin pigmentation. However, if the levels exceed 0.02% in the preparation, it will cause skin irritation, kidney function disorders and cancer. The type of research is a mixed type, namely qualitative and quantitative. Qualitatively, it will obtain data on the presence of hydroquinone and quantitatively in the form of hydroquinone levels. The samples were 5 whitening body lotion preparations circulating in Lubuk Pakam which were determined randomly and were coded A, B, C, D, and E. Qualitative analysis was carried out using the TLC method and quantitative analysis was carried out using the UV-spectrophotometric method. Visible. The results of qualitative analysis show that samples A, C, and D contain hydroquinone compounds which are characterized by the appearance of blackish brown spots with respective Rf values of 0.89; 0.96; and 0.89. Meanwhile, when qualitative analysis was carried out, the levels obtained for the three samples were 44.56% each; 5,604%; and 5,612%. Based on regulations issued by BPOM RI in 2022, it is stated that hydroquinone levels in beauty products must not exceed 0.02%. So it can be concluded that whitening body lotion preparations with sample codes A, C, and D are not recommended for use because the use limit for hydroquinone is classified as unsafe. It is recommended that people, especially women, be more selective in choosing and using beauty products.

Qualitative and quantitative analysis methods for quality control of rhubarb in Taiwan's markets.

Introduction: Rhubarb is a traditional Chinese medicine (TCM) used to release heat and has cathartic effects. Official rhubarb in Taiwan Herbal Pharmacopeias 4th edition (THP 4th) and China Pharmacopeia 2020 (CP 2020) are the roots and rhizomes of Rheum palmatum L., Rheum tanguticum Maxim. ex Balf., and Rheum officinale Baill. However, the Rheum genus is a large genus with many different species, and owing to the similarity in appearance and taste with official rhubarb, there needs to be more clarity in the distinction between the species of rhubarb and their applications. Given the time-consuming and complicated extraction and chromatography methods outlined in pharmacopeias, we improved the qualitative analysis and quantitative analysis methods for rhubarb in the market. Hence, we applied our method to identify the species and quality of official and unofficial rhubarb. Method: We analyzed 21 rhubarb samples from the Taiwanese market using a proposed HPLC-based extraction and qualitative analysis employing eight markers: aloe-emodin, rhein, emodin, chrysophanol, physcion, rhapontigenin, rhaponticin, and resveratrol. Additionally, we developed a TLC method for the analysis of rhubarb. KEGG pathway analysis was used to clarify the phytochemical and pharmacological knowledge of official and unofficial rhubarb. Results: Rhein and rhapontigenin emerged as key markers to differentiate official and unofficial rhubarb. Rhapontigenin is abundant in unofficial rhubarb; however, rhein content was low. In contrast, their contents in official rhubarb were opposite to their contents in unofficial rhubarb. The TLC analysis used rhein and rhapontigenin to identify rhubarb in Taiwan's markets, whereas the KEGG pathway analysis revealed that anthraquinones and stilbenes affected different pathways. Discussion: Eight reference standards were used in this study to propose a quality control method for rhubarb in Taiwanese markets. We propose a rapid extraction method and quantitative analysis of rhubarb to differentiate between official and unofficial rhubarb.

Increased Intracellular Insulin from Differentiated Stem Cells to Insulin- Producing Cells Incorporating Β-Carotene

Analyzes the effect of β-Carotene (BC) in the processes of differentiation of adipose tissue Human Mesenchymal Stem Cells (hADMSCs) and dental pulp Human Mesenchymal Stem Cells (hDPMSCs) for obtaining Insulin-Producing Cells (IPCs). The BC present in the methanolic extract was analyzed and quantified by UV-VIS, TLC and HPLC methods. The cell viability test was done by flow cytometry, a concentration of 1.5 μM of BC or AP extract no cytotoxic effect on stem cells, so this concentration was used for the Human Mesenchymal Stem Cells (hMSCs) differentiation protocols for obtaining IPCs. The differentiation tests were done with the AP extract or β-carotene standard addition which demonstrates the increased cell differentiation percentages by the presence of intracellular insulin. This was more outstanding in the hMSCs that were stimulated with the AP in stage 3 and applied at all stages mainly in hDPMSCs, reaching up to 74% of the insulin-positive population respect to 11% where the molecules were not added. RT-PCR was performed for the level of gene expression like MAFA, PDX1 and NKX6.1 and those are found well expressed on differentiated cells. This could be relevant in cell therapy in diabetes, where inclusion of AP extracts or BC may improve yield of IPCs.

An indicator color chart for quick detection of Pigment Red 53 in cosmetic products in Indonesia.

Pigment Red 53 is a dangerous synthetic dye that is often added to cosmetics, even though its use in cosmetic products has been prohibited because of possible impacts on health. Faster and more sensitive detection of Pigment Red 53 is needed for onsite analysis to protect the community from illegal cosmetics that contain the dye. Indicator color charts are a kind of analytical method that can be used to detect Pigment Red 53 in cosmetic products, including lipstick, rouge, and eyeshadow. Such charts are practical, fast, and can be used for onsite analysis. In this study, an indicator for Pigment Red 53 detection was obtained through a reagent reaction that caused a specific color change. An indicator color chart was then produced by setting out in paper form the series of colors which resulted from the reaction of specific chemical reagents and Pigment Red 53 solutions at concentrations of 10, 20, 40, 60, 80, and 100 ppm. The testing results showed that the indicator color chart may be used as an initial screening method for the detection of Pigment Red 53 in cosmetic products with a detectable minimum concentration of 10 ppm. Out of nine samples, only one (Eyeshadow 3) tested positive for Pigment Red 53. Further analysis was carried out on the indicator color chart and the results showed good agreement with TLC and UV-Vis spectrophotometry methods. The results reported in this paper demonstrate that the indicator color chart is a good prospective method for onsite analysis to detect Pigment Red 53 in cosmetic samples, with a lower detection limit compared to polymer-based indicators.

Flavonoid-Rich Trianthema decandra Ameliorates Cognitive Dysfunction in the Hyperglycemic Rats.

The present study was aimed at the evaluation of neuroprotective ability of methanolic extract of Trianthema decandra (METD) against hyperglycemia-related cognitive impairment in rats. The extract of T. decandra was standardized by TLC and HPTLC methods. To verify the identity and purity of isolated compounds, they were segregated and characterized using various techniques, including UV-visible spectrophotometry, FT-IR, H-NMR, and Mass spectroscopy. α-Amylase and α-glucosidase inhibition property of the extracts were assessed in-vitro. The screening of the neuroprotective effects of METD in hyperglycemic rats was done utilizing Morri's water (MWM) and elevated plus maze (EPM) model, as well as acetylcholinesterase (AChE) activity. The extracts of Trianthema decandra and its chemical constituents, namely quercetin and phytol, demonstrated a significant protective effect on enzymes like α-amylase and α-glucosidase. Methanol and hydroalcoholic extracts have shown the strongest inhibitory activity followed by chloroform extract. Quercetin and phytol were associated with the methanolic and chloroform extracts which were identified using TLC and HPTLC techniques. During the thirty days of the study, the induction of diabetes in the rats exhibited persistent hyperglycemia, hyperlipidemia, higher escape latency during training trials and reduced time spent in target quadrant in probe trial in Morris water maze test, and increased escape latency in EPM task. Regimen of METD (200 and 400 mg/kg) in the diabetic rats reduced the glucose levels in blood, lipid, and liver profile and showed positive results on Morri's water and elevated plus maze tasks. During the investigation, it was determined that Trianthema decandra extracts and the chemical constituent's quercetin and phytol in it had anti-diabetic and neuroprotective activities.

Taurine dynamics in serum during the oestrous cycle in buffaloes.

Estrus identification is one of the common issues in buffaloes because of their short estrus duration and silent estrus problem. Hence, specific biomarkers facilitating in identifying the estrus stage would be helpful to buffalo farmers and researchers. In our previous studies, taurine, a non-protein amino acid that helps in the secretion of reproductive hormones such as GnRH, was found to be associated with postpartum anestrus in buffaloes. Therefore, the present study was conducted to explore the level of taurine in serum during different stages of the oestrous cycle in healthy cyclic buffaloes. Blood samples were collected from healthy cyclic buffaloes (n = 4), and taurine was estimated at the estrus (0th day), proestrus (-2nd day), metestrus (3rd day) and diestrus (+10th day) stages using TLC method. The days of the oestrous cycle were determined by ultrasonography and observation of behavioural signs by trained professionals. The results revealed that taurine was consistently present in the serum. However, the highest concentration of taurine was observed at the proestrus (0.20 ± 0.03 mg/mL) stage, which was greater (p < .05) than metestrus (0.10 ± 0.05 mg/mL) and diestrus (0.13 ± 0.03 mg/mL) stages, but comparable with the estrus stage. These results were also validated in the simulated population datasets of population size 6 to 10,000. Further, ROC curve analysis for the large simulated population indicated the efficiency of taurine to distinguish proestrus from metestrus and diestrus stages at a lower cutoff value of <0.1643 mg/mL with 60% sensitivity and specificity. Therefore, the present study concludes that serum taurine concentration could help in detecting proestrus stage of buffalo estrous cycle.

Studying the acids and amino acid composition of horse chestnut flowers

Introduction. Currently, there is an increasing trend towards assessing the full spectrum of chemical compounds in plant materials. One of the first steps in the formation of secondary metabolites of complex structure, participating in biochemical processes in the plant body, which have their own effect on the human and animal body, are organic acids (OA) and amino acids (AA). However, due attention is not always given to these compounds when studying the component composition of plant raw materials.Aim. The purpose of the work was to study the profile and quantitative assessment of the content of organic acids and amino acids of Aesculus hippocastanum L. flowers.Materials and methods. The object of the study were horse chestnut flowers, collected during flowering in the Voronezh region in 2021 and dried by air-shade method. Quantitative assessment of the amount of TC in terms of malic acid and ascorbic acid was carried out titrimetrically according to the methods presented in the State Pharmacopoeia of the Russian Federation, XIV edition. The composition of the OA and AA profiles of horse chestnut flowers was determined using the capillary electrophoresis method ("Kapel®-105/105M", Group of Companies "Lumex", Russia).Results and discussion. The method of capillary electrophoresis revealed the presence of 12 compounds of the OA group in a total amount of 4.7 % (in terms of absolutely dry raw materials). The rationality of recalculating the content of the sum of OA to the major component – citric acid, which amounted to 1.65 ± 0.04 %, was shown. The content of ascorbic acid was low (0.033 ± 0.007) %. Using the TLC method, the presence of 5 zones of compounds belonging to the AA class was established in the aqueous extract of horse chestnut flowers. The quantitative content of the sum of free AA in terms of glutamine, determined by the spectrophotometric method, was 2.25 ± 0.07 % (n = 5). The presence of 17 AA was determined by capillary electrophoresis, which amounted to 7.13 %. Glutamic acid is present in the greatest quantity in horse chestnut flowers (1.19 %). Essential amino acids amounted to 2.27 %; leucine was present in the predominant amount (0.58 %).Conclusion. A study was carried out of the qualitative composition and quantitative content of organic acids and amino acids of horse chestnut flowers. The composition of the profile and the quantitative content of organic acids and amino acids have been established. It was revealed that citric acid prevails in the total organic acids; glutamic, aspartic acid, arginine and proline are found in greater quantities in the total amino acids.