QU Tong, HU Xiao-juan, LI Ning, LU Wen-jing, GENG Fei-fei, CHEN Yin-zi, CHEN Zhi-yong, REN Hui
Objective: To establish the fingerprint of Qingbai Tongbi capsules, and to screen out its indicative compounds for quality control combined with chemometrics methods and network pharmacology. Methods: Agilent 5 TC-C18(250 mm×4.6 mm, 5 μm) chromatographic column was used for separation. The mobile phase was acetonitrile-0.1% phosphoric acid solution for gradient elution at a flow rate of 1.0 mL · min-1, and the column temperature was 30 ℃. The injection volume was 10 μL, and the wavelength was 210 nm (0-45 min), 260 nm (45-70 min). The fingerprint was established and the common peaks were determined. By comparing with the relative retention time and UV spectra of the reference substances, the corresponding compounds of the chromatographic peak were identified. Then the common peaks were identified by single decoction. Chemometrics methods was used to evaluate quality of 10 batches of Qingbai Tongbi capsules, and OPLS-DA analysis was used to screen out the main marker components of Qingbai Tongbi capsules. Combined with network pharmacology, the core targets and key pathways were constructed a “component-target-pathway” network map through corresponding databases. Combined with the above results, indicative compounds for quality control of Qingbai Tongbi capsules were screened out, and an HPLC method was established to determine the content of the Q-Markers. Results: An HPLC fingerprint of Qingbai Tongbi capsules was established, identifying 24 common peaks, and assigning them to different peaks. Among them, peaks 1-3, 5-8, 10-11, 13 and 16 came from Qingfengteng. Peaks 4, 9, 12, 14, 16, 18-19 came from Baishao. Peaks 15, 17, 20-24 came from Zhigancao. Eight common peaks were identified, including catechin, sinomenine, gallic acid, magnoflorine, paeoniflorin, glycyrrhizin, 1, 2, 3, 4, 6-O-pentagalloylglucose, glycyrrhizic acid. The similarity evaluation showed that the similarity of 10 batches of Qingbai Tongbi capsules samples ranged from 0.934-1.000. Principal component analysis (PCA) showed that the cumulative variance contribution rate of the first four principal components was 93.998%, while orthogonal partial least squares-discriminant analysis (OPLS-DA) showed that 8 components had higher variable importance projection values. On this basis, the network pharmacology method was used to analyze and conclude that catechin, sinomenine, gallic acid, magnoflorine, paeoniflorin, glycyrrhizin, 1, 2, 3, 4, 6-O-pentagalloylglucose and glycyrrhizic acid may be the potential Q-Marker of Qingbai Tongbi capsules. The contents of the above eight components were determined simultaneously, and the methodological investigation results were good. The average sample recovery rate was 96.81%-103.44%, and the RSD was 0.6%-3.7%. The mass fractions of catechin, sinomenine, gallic acid, magnoflorine, paeoniflorin, glycyrrhizin, 1, 2, 3, 4, 6-O-pentagalloylglucose and glycyrrhizic acid in 10 batches of samples were 0.907 1-1.189 3 mg · g-1, 2.183 3-3.118 6 mg · g-1, 0.397 0-1.427 6 mg · g-1, 3.507 9-5.446 6 mg · g-1,14.207 7-19.570 1 mg · g-1, 1.412 8-3.577 5 mg · g-1, 0.442 0-1.697 7 mg · g-1, 2.738 8-4.761 2 mg · g-1. Conclusion: The established HPLC fingerprint method is simple and good repeatability. The quality control indicative compounds of Qingbai Tongbi capsules can provide a basis for its quality control.
