Xây dựng quy trình định lượng acid asperulosidic trong cao chiết từ bài thuốc thoái hóa cột sống của Lương y Nguyễn Thiện Chung bằng phương pháp HPLC

A remedy of physician Nguyen Thien Chung which comprises 13 medicinal herbs (BT) has been using for spinal degeneration treatment due to it analgesic and anti-inflamatory activities. Quantification of asperulosidic acid, one major bioactive coumpound of BT, plays an important role in controlling the quality of the extract of BT. This study was conducted to develop  a method for asperulosidic acid quantification in extract of BT by high performance liquid chromatography (HPLC). Different contitions for asperulosidic acid extraction and HPLC determination from extract of BT was screened and the best condition was selected. The HPLC method was validated according to the ICH guidelines on system suitability, specificity, linearity, precision and accuracy. The method of sample preparation was optimized using methanol as a solvent for asperulosidic acid extraction and the ratio of BT extract and methanol was at 500:25 (mg/ml). The best HPLC condition for determination of asperulosidic acid employed a C18 Syncronis column (250×4.6 mm; 5 μm), a detection wavelength of 236 nm, a column temperature of 30 ^oC, a flow rate of 1 ml/min, and an injection volume of 10 μl. Mobile phase was mixture of acetonitrile and H₃PO₄ 0.1 %  at a ratio of 10:90 (v/v). There was good correlation between peak areas and asperulosidic acid concentrations (r² = 0.9998). The RSD values of inter-day precision were found to be at 0.41 %. The recovery percentages were of 95 – 105%. The HPLC method for asperulosidic acid quantification in the extract of BT was reported for the first time. The proposed method met the requirements of validation and could be useful for establishing the quality standards of the extract of BT and its related products.

Keywords: Asperulosidic acid, HPLC, the spinal degeneration remedy.

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