Determination of 25 characteristic components in Danhe Granules by LC-MS and consistency analysis of preparations / 中草药

ABSTRACT

Objective: A high performance liquid chromatography coupled to triple quadrupole mass spectrometry method (HPLC- MS/MS) was established to simultaneously determine the content of 25 characteristic components (gallic acid, tanshinol, catechin, chlorogenic acid, caffeic acid, epicatechin, rutin, polydatin, hyperin, astragalin, naringin, hesperidin, rosmarinic acid, resveratrol, salvianolic acid B, quercetin, emodin-8-O-β-D-glucoside, isorhamnetin, emodin, coclaurine, nuciferine, cryptotanshinone, tanshinone I, dehydronuciferine, tanshinone IIA) in Danhe Granules (DG), and the consistency between different batches was investigated. Methods The analysis was conducted on Agilent Rapid Resolution HD C18 column (50 mm × 2.1 mm, 1.8 μm) with the mobile phase of 0.1% formic acid water-0.1% formic acid acetonitrile. The dynamic multi-response detection (dMRM) scanning mode was used in the mass spectrometry. Results: Based on the established HPLC-MS/MS method, the simultaneous quantitative analysis of 25 characteristic components could be completed within 10 min, with the quantitative limits of isorhamnetin, coclaurine, and dehydronuciferine of 0.025 ng/mL; emodin and tanshinone I of 0.050 ng/mL; emodin-8-O-β-D-glucoside of 0.200 ng/ml; gallic acid, chlorogenic acid, caffeic acid, epicatechin, rutin, hyperin, astragalin, resveratrol, quercetin, nuciferine, tanshinone IIA of 0.250 ng/ml; catechin, rosmarinic acid, and cryptotanshinone of 0.500 ng/mL; tanshinol, hesperidin, and salvianolic acid B of 1.000 ng/mL; polydatin of 2.000 ng/mL; naringin of 5.000 ng/mL, respectively. The linear relationships of the 25 constituents within their respective mass concentrations were good, with the average recovery of 85.16%-113.46% and the RSD of 2.01%-8.80%. Furthermore, this method also included the main components named monarch, minister, assistant and guide of herbs in a relatively comprehensive way. The total content of the 25 components was 31.49 mg/g, among which the content of salvianolic acid B (9.44 mg/g) and hesperidin (7.60 mg/g) was the highest, and the content of isorhamnetin (0.79 μg/g) was the lowest. According to boxplot analysis, the content of 25 components in 10 different batches of DG fluctuated (P value) within 75% < P < 125%; and the RSD value of 25 components ranged from 2.58% to 13.10% by statistical analysis. The above results showed that the consistency of the component content among 10 batches of DG was acceptable. Conclusion: The analytical method established in this study is fast and sensitive. Furthermore, the results of this study are reliable and can provide scientific methods and basis for quality control and consistency analysis of DG.