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Acta Chromatographica
Authors: Y.T. Kamal, Sayeed Ahmad, Nanjaian Mahadevan, Prawez Alam, Shahana Salam, Yahya I Asiri, Abdullatif Bin Muhsinah and Abdulrhman Alsayari

order to improve the recovery of all nine marker constituents from polyherbal Unani formulations. Experimental Samples, chemicals, solvents and standards Three batches of Itrifal-e-Aftiomoon and Itrifal-e-Badiyan were chosen for analysis, batch I was

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Acta Chromatographica
Authors: Y.T. Kamal, Sayeed Ahmad, Nanjaian Mahadevan, Prawez Alam, Shahana Salam, Yahya I Asiri, Abdullatif Bin Muhsinah and Abdulrhman Alsayari

extraction procedure in order to improve the recovery of all nine marker constituents from polyherbal Unani formulations. Experimental Samples, chemicals, solvents and standards Three batches of Itrifal-e-Aftiomoon and Itrifal-e-Badiyan were

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Summary

A validated reversed-phase HPLC method has been developed for quantitative analysis of berberine in Berberis aristata fruits and in a polyherbal formulation. Separation of berberine was achieved on a C18 column with a mobile phase consisting of a 10–80% acetonitrile gradient in 0.05% aqueous orthophosphoric acid. The flow rate was 1 mL min−1. Detection was at 266 nm. A sharp, well defined peak was obtained at a retention time of 10.0 ± 0.4 min. The method was validated in accordance with ICH guidelines for accuracy, precision, robustness, and the limits of detection (LOD) and quantification (LOQ). Results from linear regression analysis were indicative of a good linear relationship (r 2 = 0.998 ± 0.0011) in a wide concentration range (5–500 μ g mL−1). LOD and LOQ were 1.5 and 5.3 μg mL−1, respectively. Satisfactory recovery results (94.6–103.1%) were obtained by the method of standard addition. Intra-day, inter-day, and intersystem precision was satisfactory, with relative standard deviation in the range 0.7–1.8%. The berberine content of fruit of Berberis aristata and the herbal formulation were 0.033% and 0.0089% (w/w), respectively. This HPLC method for quantification of berberine can be used for quality control and standardization of several crude drugs and different herbal formulations in which berberine is present as a phyto constituent.

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Marmelosin is one of the major phytochemical constituents of Aegle marmelos (L.) Corr. (Rutaceae). Till date, there are no methods reported for the estimation of marmelosin in plasma matrix using high-performance thin-layer chromatography (HPTLC) which is a versatile analytical technique promoting shorter analysis time and less consumption of solvents with efficient data acquisition and processing. This study is an attempt to develop a rapid and sensitive HPTLC method for the estimation of marmelosin from rat plasma and its application to evaluate pharmacokinetics of A. marmelos fruit pulp extract and a traditional Unani medicine Sharabat-e-Belgiri. Plasma processing involved liquid-liquid extraction with diethyl ether followed by HPTLC. Separation and determination of marmelosin were performed on silica gel 60 F254 TLC plate using toluene-ethyl acetate-glacial acetic acid as mobile phase. United States Food and Drug Administration (USFDA) guidelines for bioanalytical validation were followed to validate the HPTLC method. R F of marmelosin was found to be 0.57. The calibration curve was linear over the range of 1.0–150.0 μg mL−1 (r 2 = 0.995) in rat plasma. Limit of detection and limit of quantitation were found to be 0.5 μg mL−1 and 1.0 μg mL−1, respectively. The extraction recovery from plasma was in the range of 90.07–98.22%. The intra-day and inter-day precisions were found to be 90.89–104.12%, and 92.82–104.86%, respectively. The absorption of marmelosin from extract was found significantly higher than the formulation. The results indicated that HPTLC is a suitable technique for the estimation and pharmacokinetic evaluation of marmelosin from plant extract and Unani formulation in rat plasma.

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A novel HPTLC method has been developed for the estimation of glabridin in Licorice rhizome and its Unani polyherbal formulation (Qurs-e-Gul). Separation was achieved on silica using toluene, dichloromethane, and ethyl acetate in equal ratios. A compact, well resolved peak of glabridin with RF value 0.56 ± 0.02 was observed. Calibration curve revealed a good linear relationship with r2 value of 0.993 between the peak area and concentration in the range of 25–500 ng spot−1. The proposed method was validated as per the International Conference on Harmonization (ICH) guidelines. The stability assessment was carried out by studying degradation of glabridin stressed by acid, base, oxidation, thermal, and humidity. Photodegradation was also carried out after keeping the drug in sunlight, dark, and in UV lights. The method proposed can be used for routine determination of glabridin in crude drugs and in herbal formulations containing Licorice as one of the ingredients, for quality control as well as for stability testing with high precision, accuracy and a wide range of linearity.

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