Evaluation of Unani Anti-Inflammatory Polyherbal Formulation using Validated HPLC Methods

Unani system of medicine (USM) is commonly practiced in India and many other countries as an alternative system of medicine, which mainly believes in holistic approaches. In this system, mainly plant based drugs are used, although drugs of mineral, animal and marine origins are practised to a lesser extent [1]. Habb-e-Khardal is one of the important polyherbal formulations, which is commonly prescribed by the Unani physicians as an anti inflammatory drugs, which contain Commiphora mukul, Boswellia serrata, Piper nigrum and Brassica nigrum [2]. The drug was investigated for its bioactive constituents using HPLC. Analytical methods were developed and validated as per the ICH guidelines for the quantification of piperine, boswellic acids (BBA, ABBA, KBBA and AKBBA) and guggul sterones (E & Z), separately in formulation and crude drug. All the chromatographic experiments were conducted on YL9100 HPLC system with UV detector using reverse phase C18 column (250mm x 4.6µm). Piperine and guggul sterones were eluted using acetonitrile: water (50:50, v/v) and methanol: water (45: 55, v/v) in 1.0 mL/min at 340 and 240nm, respectively. Boswellic acids were analyzed in gradient elution method using 0.005% acetic acid and acetonitrile at 1.0 mL/min at 210nm (BBA and ABBA) and 254nm (KBBA and AKBBA). Methods were validated as per the ICH guidelines for linearity, accuracy, precision, robustness, LOD and LOQ and found satisfactory. The herbal formulation was found rich in constituents like piperine, boswellic acid and guggul sterones (E & Z), which have already been reported for its anti inflammatory properties [3, 4].

Acknowledgement: Authors are grateful to CCRUM-AYUSH, Ministry of Health and Family Welfare, Government of India, for providing financial assistance.

References: [1] Ahmad S, et al. (2010)J Pharm Bioalli Sci, 2: 93–99. [2] Anonymous, (2006) National Formulary of Unani Medicine, Ministry of Health and Family Welfare, Department of AYUSH, Govt. of India, New Delhi, 30–31. [3] Bang JS, Oh da H (2009) Arthritis Res Ther, 11: 5–24. [4] Bernstein S (1992) Steroids, 57: 392–402.