Abstract
Objective: The main objective of this research is to develop and validate a simple, specific, precise, sensitive, cost-effective, and rapid reversed-phase high-performance liquid chromatography method for simultaneous quantification of glucosamine (GLU), diacerein (DIA) and methyl sulfonyl methane in bulk and pharmaceutical dosage forms, and micro-sample of rat plasma using ultraviolet (UV) detection, to perform the studies of drug dissolution from tablets.
 Methods: Sprague-Dawley rats were used for pharmacokinetic study after intravenous administration of the drug samples at dose 5 mg/kg. The drug samples were extracted by liquid-liquid extraction technique using acetonitrile, which also acted as a deproteinization agent. The separation of the analyte was carried out on a phenomena C18 column with a mobile phase composed of 0.1 % orthophosphoric acid:acetonitrile (80:20 v/v) delivered at a flow rate of 1.0 ml/min, and separation has been monitored by a UV detector, at detection of the wavelength of 285 nm.
 Results: This method was proven to be linear over a concentration range of 30–450 μg/ml for GLU, 2–30 μg/ml for DIA, and 10–150 μg/ml for methyl sulfonyl methane with a correlation coefficient of 0.999. The retention time of GLU, DIA, and methyl sulfonyl methane were 2.89, 6.32, and 9.87 min, respectively. Recovery of the drugs was found to be in the range of 98.0–102.0%. Validation results were found to be satisfactory and the method applicable for bulk and formulation analysis. Hence, it was evident that the proposed method was said to be a suitable one for the regular analysis and quality control of pharmaceutical preparations which contain these active drugs either individually or in combination.
 Conclusion: The validation results were in good agreement with acceptable limits. Relative standard deviation values which are less than 2.0% are indicating the accuracy and precision of this method. The usefulness of the method is that the common chromatographic conditions have been adopted for assay, dissolution, and pharmacokinetic studies. This developed method showed reliable, precise, and accurate results under optimized conditions.
Highlights
Glucosamine (GLU) (Fig. 1) is chemically called (3R, 4R, 5S)-3-amino6-(hydroxymethyl) oxane-2, 4, 5-triol
The purpose of this study is to develop accurate, specific, cost-effective, and a single isocratic HPLC method for the simultaneous quantification a b c Fig. 9: Chromatograms for (a) accuracy 50%, (b) accuracy 100%, and (c) accuracy 150%
The validation results were in good agreement with acceptable limits
Summary
Glucosamine (GLU) (Fig. 1) is chemically called (3R, 4R, 5S)-3-amino6-(hydroxymethyl) oxane-2, 4, 5-triol It can act as an amino sugar and a prominent precursor in the biochemical synthesis of glycosylated proteins and lipids [1, 2]. It is found in the hard covering of shellfish [3] It is one of the most common non-vitamin, non-mineral, dietary supplements, and natural products. Diacerein (DIA) (Fig. 2) chemically called 4, 5-bis (acetyloxy)-9,10dioxo-2-anthracenecarboxylic acid [4,5] It is slow-acting drug and is used in the treatment of osteoarthritis, analgesic, and nonsteroidal antiinflammatory drugs [6,7,8]. It can inhibit interleukin-1 and retards all pathological processes initiating in osteoarthritis [12] and inhibits superoxide production, chemo taxis, and phagocytic activity of neutrophils
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