Quality control of Andrographis herba and its preparations by HPLC-DAD-MS

Andrographis paniculata (Burm.f.) Nees, (Acanthaceae) is used in Asian traditional medicine for the treatment of upper respiratory infections, lower urinary tract infections, diarrhoea, as hepatoprotective agent [1]. In Europe it is used for the treatment of common cold [2 – 3] and standardised extracts have been used in preclinical and clinical trials [4]. In the framework of the preparation of TCM herbal drug monographs for the European Pharmacopoeia the analytical method for the quality control of andrographidis herba was undertaken. A simple and effective method for the quality control of andrographidis herba and its preparations was developed and accordingly validated. Three extraction protocols (soxhlet apparatus, ultrasonication and decoction) were tested. Among them ultrasonication was exhaustive, repeatable, accurate and ensured the stability of the active principles. For the first time decoctions were prepared and analysed. An RP-HPLC method was developed and optimised to quantify the amounts of andrographolides in the herbal drug and its preparations. A long isocratic step separated minor constituents, degradation products from the main constituents and gave the complete fingerprint of the herbal drug. The method was validated for linearity, LOD and LOQ, precision (intra and inter-day variations between 0.01 and 0.63 expressed as % RSD) and accuracy (recovery was 95 – 99%). The overall assay was finally validated for precision (% RSD ranged from 0.7 to 1.1) and accuracy (recovery ranged from 95 to 97%) in three different concentration levels. All validation criteria were fulfilled. The proposed protocol is proven suitable for the quality control of andrographidis herba and preparations thereof.

Keywords: Andrographis paniculata, quality control, HPLC-DAD-MS, validation of analytical method

References:

[1] WHO monographs on selected medicinal plants Vol 2, Geneva, 2004, 12 – 24.

[2] Kligler B, Ulbricht C, Basch E, Kirkwood CD, Abrams TR, Miranda M, Singh Khalsa KP, Giles M, Boon H, Woods J. Andrographis paniculata for the treatment of upper respiratory infection: a systematic review by the natural standard research collaboration. Explore, 2006; 2: 25 – 9.

[3] Saxena RC, Singh R, Kumar P, Yadav SC, Negi MPS, Saxena VS, Joshua AJ, Vijayabalaji V, Goudar KS, Venkateshwarlu K, Amit A. A randomized double blind placebo controlled clinical evaluation of extract of Andrographis paniculata (KalmCold (TM)) in patients with uncomplicated upper respiratory tract infection. Phytomedicine 2010; 17: 178 – 185.

[4] European Medicines Agency. Committee on Herbal Medicinal Products (HMPC). Assessment report on Andrographis paniculata Nees, folium. EMA/HMPC/320433/2012.