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Identification and Determination of Related Substances in Diosmin Bulk Drug and Pharmaceutical Formulations by HPLC and HPLC–MS

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Abstract

Six related substances were detected in diosmin bulk drug substances and products by a newly developed gradient reverse-phase high performance liquid chromatography (RP-HPLC) with UV detection. The chromatographic system consisted of an Intersil Wondasil TM ODS (C 18) column (250 × 4.6 mm; 5 μm). The mobile phase consisted of water/acetic acid 66:6 v/v (solvent A) and methanol (solvent B) using a gradient program at a flow rate of 0.8 mL min−1 with 345 nm detection and an injection volume of 20 μL. In addition, the linearity, quantitation limit (QL), accuracy, selectivity, robustness and precision were determined. Good linearity was obtained over the concentration range 0.5–200 μg mL−1 with the coefficient of determination (r 2) of 0.999. The QL was 0.125 μg mL−1 (relative standard deviation <2.0 %). The major impurities have been resolved and identified using two analytical systems, HPLC and HPLC/electrospray ionization-mass spectrometry operated in a negative ion mode. One of these impurities marked as 7-hexopyranosidal diosmetin was unknown and has not been reported previously. Based on mass spectrometry data the structure of the new impurity was proposed as 5-hydroxy-2-(3-hydroxy-4-methoxyphenyl)-4-oxo-4H-chromen-7-yl hexopyranoside. The newly developed RP–LC method for quantitative determination of diosmin-related substances was found to be precise, accurate, robust and specific. It has been successfully employed for the quality evaluation of different sources of raw material and generic formulations of diosmin.

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Acknowledgments

The authors would like to thank Chengdu Runde Pharmaceutical Co., Ltd (China) for the supply of raw material of diosmin and Chiatai Tianqing Co., Ltd (Nanjing, China) for the supply of drug tablets. All colleagues are also acknowledged for support and constant encouragement.

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  1. Department of Pharmaceutical Analysis, China Pharmaceutical University, Nanjing, 210009, People’s Republic of China

    Imene Adouani, Mingluo Du & Tai-Jun Hang

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  1. Imene Adouani
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Correspondence to Imene Adouani.

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Adouani, I., Du, M. & Hang, TJ. Identification and Determination of Related Substances in Diosmin Bulk Drug and Pharmaceutical Formulations by HPLC and HPLC–MS. Chromatographia 76, 499–508 (2013). https://doi.org/10.1007/s10337-013-2404-z

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