Planta Med 2015; 81 - PW_224
DOI: 10.1055/s-0035-1565848

Development and validation of a method for standardization of infusions of Herniaria hirsuta

I van Dooren 1, K Foubert 1, A Vlietinck 1, L Pieters 1, S Apers 1
  • 1NatuRA, Department of Pharmaceutical Sciences, University of Antwerp, Wilrijk, Belgium

Herniaria hirsuta L. is traditionally used in Morocco to treat kidney stones, and shows cholesterol lowering effects and activity against gallstones [1, 2]. An HPLC-DAD method for the quantification of flavonoids and saponins, the main compounds, was developed and validated according to the ICH guidelines in order to standardize the extract. Several analytical columns and mobile phases were tested. For each solvent system, the gradient was optimized. The extraction solvent composition and the sonication time were evaluated. Sonication for 20 minutes with 50% (v/v) methanol gave the best extraction results. A gradient, starting with 5% acetonitrile (ACN) + 0.05% formic acid (FA) going to 100% ACN + 0.05% FA, using the Apollo C18 column, gave the best resolution. The flavonoïd content was expressed as rutin and the saponin content as hederacoside C. The calibration model appeared to be linear. Though for most of the flavonoids and for the saponins, there is a significant effect of the factor day and/or factor concentration, the precision of the method is acceptable taking into account the complexity of the analysis. The RSDI(Tc) for the determination of flavonoids is smaller than 3% and values for saponins ranged from 3.13% to 14.54%. Saponins are detected at a wavelength of 210nm causing higher variation in results, especially when the concentration is low, which is the case for one of them. Therefore only the total content of saponins can be determined with an acceptable RSDI(Tc) (3.13%). The method can be considered as accurate for flavonoids (mean recovery 99.18%). For the saponins the mean recovery is 109.20%, and because this falls out of the generally accepted limits (97% to 103%), it should be mentioned with every result obtained with this method.

References:

[1] Settaf A et al. Herniaria hirsuta dissout les calculs biliaires cholestéroliques. Espérance Médicale 1999; 6: 79 – 82

[2] Foubert K et al. Cholesterol lowering effect in the gall bladder of dogs. Abstract.