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Kamini Vidrawan Ras is a classical Ayurvedic medicine, referred to in Bhaisjya Ratnavali, a book recognized by the Drugs and Cosmetics Act of India. Its usefulness has been stated in erectile dysfunction, impotence, and premature ejaculation. Opium is the major ingredient of formulation which is highly addictive as it contains narcotic alkaloids (morphine, codeine, and thebaine). Opium is a natural product; hence, the morphine content varies from 4–21%. The aim of this study was to develop a simple, precise, rapid, and reliable high-performance thin-layer chromatography— densitometry method for the quantitative estimation of morphine in the tablets of the said Ayurvedic medicines. Aluminum-backed silica gel F254 (20 cm × 10 cm) was used as the stationary phase and ethyl acetate, methanol, and ammonia (85:10:5, v/v) as the mobile phase. The R F value for morphine was 0.36, and the quantitative evaluation of the bands over plates was performed in the reflectance—absorbance mode at 280 nm. The regression analysis of the calibration plot showed good linear relationship between peak area vs. morphine concentration. Linearity was found in the range of 400 to 1200 ng per band, and the amount of morphine was estimated by comparing the peak area of the standard morphine.

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The technological improvement in the structural elucidation of natural compounds has made it probable to generate appropriate strategies for the analysis and standardization of plant-based medicines. An appliance of highly oriented hyphenated techniques provides a definite tool for herbal investigations. Therefore, the present study was directed towards the standardization of biomarkers gallic acid and berberine in polyherbal formulation Entoban capsules to ensure the quality of the herbal drugs. A rapid, simple, accurate, and specific high-performance thin-layer chromatography (HPTLC) method for the quantitative estimation of biomarkers berberine and gallic acid has been developed. HPTLC was performed to evaluate the presence of gallic acid and berberine applying toulene—ethyl acetate—formic acid—methanol (12:9:4:0.5 v/v) and ethanol—water—formic acid (90:9:1 v/v), as the mobile phase, respectively. The R F values (0.58 for gallic acid and 0.76 for berberine) in both sample and reference standard were found comparable under ultraviolet (UV) light at 273 nm and 366 nm, respectively. The method developed resulted in good-quality peak shape and enabled high-quality resolution of biomarkers. The present standardization undertaken reveals compliance with the analytical procedure; therefore, it is concluded that Entoban capsule is a well-standardized product. Standardization falls under the specific guidelines of quality herbal medicine following the prerequisite for global harmonization.

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Biomphalaria glabrata snails were infected with Schistosoma mansoni and maintained at different dilutions of artificial ocean water for up to 4 weeks. Glucose and maltose concentration of the digestive gland-gonad complex were analyzed by high-performance thin-layer chromatography at different stages of the infection. B. glabrata snails were divided into three experimental groups: Group A, snails with early prepatent infection (10 days post-infection); Group B, snails with late prepatent infection (22 days post-infection); and Group C, snails with patent infection (45 days post-infection). Infected snails in A were maintained at different salinities for 2 weeks and then necropsied, and their two main simple sugars, i.e., glucose and maltose, were analyzed. Groups B and C contained two subgroups: the first subgoups were analyzed after 2 weeks, and the second after 4 weeks. Controls for these experiments were maintained identically in either deionized water or artificial spring water. Maltose and glucose were extracted from the digestive gland-gonad complex in ethanol-water (70:30). 1-Butanol-glacial acetic acid-diethyl ether-deionized water (27:18:5:3) mobile phase was used to separate sugars on EMD Millipore silica gel preadsorbent plates. Sugars were detected using α-naphthol-sulfuric acid reagent and quantified with a CAMAG TLC Scanner 3 at 515 nm. The obtained data were compared using analysis of variance (ANOVA) single factor statistical analysis. Statistical differences were not found in any sugars in Group A snails. For glucose, a significant difference was found after 4 weeks in both B and C snails. For maltose, a significant difference was found after 4 weeks in B snails and after 2 weeks in C snails. Different salinity levels affect the maltose and glucose concentrations of adult B. glabrata snails infected with S. mansoni.

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Context

Bacoside A, a triterpenoid saponin, is a major constituent isolated from Bacopa monnieri (L.) Wettst. (Scrophulariaceae), used as a memory enhancer. Bacoside A and B are active ingredients in Bacopa herb and have antioxidant and hepatoprotective activities

Objective

A new rapid, simple, and economical high-performance thin-layer chromatographic (HPTLC) method was developed and validated for densitometric quantitative analysis of bacoside A in powdered leaves from different geographical regions of India.

Materials and methods

An amount of 10 mg mL−1 methanol extract of powdered leaves from different geographic regions was used for sample application on precoated silica gel 60 F254 aluminum sheets. Standard bacoside A (1 mg mL−1) was used for calibration curve. HPTLC separation was performed on percolated silica gel aluminum plate 60 F254 (20 cm × 10 cm with 0.2 mm thickness) as a stationary phase using ethyl acetate–methanol–water (4:1:1) as the mobile phase. Quantification was achieved by densitometric analysis at 598 nm over the concentration range of 500–4000 ng band−1.

Result

Compact and well-resolved bands for bacoside A from powdered leaves of different geographic regions were found at retardation factor (R f) 0.53 ± 0.02. The linear regression analysis data for calibration curve showed good linear relationship with regression coefficient r 2 = 0.9996 and r 2 = 0.99810 with respect to peak area and peak height. The method was validated for precision, recovery, and robustness as per the International Conference on Harmonization (ICH) guidelines. Variation in quantitative analysis of bacoside A in powdered leaves sample from different geographic regions was found by HPTLC method.

Discussion and conclusion

The highest and lowest content of bacoside A in powdered leaves sample from Jammu and Kerala regions, respectively. The variety of B. monnieri in Jammu is superior to other regions of India. The proposed developed HPTLC method can be applied for the quantitative determination of bacoside A in powdered leaves of plant and its formulation.

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Hypoxis (Hypoxidaceae) consists of about 90 species of plants reported worldwide, of which 76 occur in Africa. As many as 41 species are indigenous to countries belonging to the Southern African Development Community (SADC), including South Africa. Of all the Hypoxis species, Hypoxis hemerocallidea has versatile application in traditional health care system of over 85% of South Africans and is regarded as one of the most ethnomedicinally important and most marketed species in South Africa. H. hemerocallidea corm’s water or alcoholic extract is widely used as traditional medicine for the treatment of benign hypertrophy and urinary tract infections as well as for boosting the immune system of people living with human immunodeficiency virus/acquired immune deficiency syndrome (HIV/AIDS) among others. However, the use of other parts of Hypoxis plant as medicine is vital for conservation purposes. The roots attached on the corm of H. hemerocallidea contain hypoxoside, but the roots are usually ripped off during the preparation of Hypoxis-containing traditional medicines and other herbal products. A developed and validated, affordable but reliable high-performance thin-layer chromatography (HPTLC) densitometry for the rapid and repeatable visualization and quantitative determination of hypoxoside from the roots of H. hemerocallidea was performed. After thin-layer chromatography analysis, the hypoxoside resolved and was visualized at R F of 0.30 in CHCl3‒MeOH‒H20 (70:30:2 v/v). The method was linear with R2 of 0.9876 over a calibration range of 0.20 × 10−4–1.80 × 10−3 mg mL−1. The limits of detection (LOD) and quantification (LOQ) were 5.08 × 10−4 and 1.65 × 10−3 mg mL−1, respectively, while the percentage recovery and the method repeatability (%RSD) were 84.10 and 4.98, respectively. The roots of H. hemerocallidea were found to contain 4.101 × 10−4 mg mL−1 of hypoxoside.

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Bottle gourd (Lagenaria siceraria (Molina) Standl.) is an important vegetable cucurbit cultivated in the warmer parts of the world. Forms bearing nonbitter fruits are used as vegetables, while those with bitter fruits are used in the Indian Traditional System of Medicine. The presence of cucurbitacins is probably the only distinguishing feature between the two forms. The present communication describes the development and validation of a high-performance thin-layer chromatographic (HPTLC)–densitometric method for the estimation of cucurbitacin B in bottle gourd using the International Conference on Harmonization (ICH) guidelines. The complete methanol extracts obtained by accelerated solvent extraction (ASE) were resolved in the mobile phase chloroform– methanol (9.5:0.5, v/v), wherein cucurbitacin B was resolved at R F 0.67 ± 0.02. Linear relationship was obtained between the values 200 and 1200 ng per spot (r = 0.9909). The limit of detection and limit of quantification were 40 and 200 ng per spot, respectively. The recovery of compound varied between 94 and 96%. Using the developed HPTLC method, the cucurbitacin B content in the vegetative parts (VP) and fruits (F) of the bitter form was found to be 0.017% and 0.062%, respectively, while it was not detected in the nonbitter form. The application of the method to study the accumulation of cucurbitacin B during progressive developmental stages of fruits of both forms revealed an increasing content of cucurbitacin B from 0.001 to 0.085% during the fruit development in the bitter form. In investigations of market samples, cucurbitacin B was detected in two out of six samples indicating admixture of bitter and nonbitter forms. To the best of our knowledge, this study is the first report of the evaluation of bitter and nonbitter forms of bottle gourd for cucurbitacin B using HPTLC. The developed method may be applicable in the routine quality control of marketed products and formulations of bottle gourd.

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A new rapid, simple, economical, and environment-friendly reversed- phase high-performance thin-layer chromatography (RPHPTLC) method has been established for the simultaneous determination of glycyrrhizin and glabridin in Glycyrrhiza glabra roots, rhizomes and selected herbal formulations. The method was carried out using RP-18 silica gel 60 F254S HPTLC glass plates and methanol–water (7:3 v/v) as the mobile phase. The developed plates were scanned and quantified densitometrically at 256 and 233 nm for glycyrrhizin and glabridin, respectively. Glycyrrhizin and glabridin peaks from G. glabra roots and rhizomes and herbal formulations were identified by comparing their single spots at R F = 0.63 ± 0.02 and R F = 0.28 ± 0.01, respectively. Linear regression analysis revealed a good linear relationship between the peak areas and the amounts of glycyrrhizin and glabridin in the ranges of 1000–7000 and 100–700 ng band−1, respectively. The method was validated, in accordance with the International Conference on Harmonization (ICH) guidelines for precision, accuracy, and robustness. The proposed method will be useful to determine the therapeutic doses of glycyrrhizin and glabridin in herbal formulations as well as in bulk drug.

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