黄芪桂枝五物汤基准样品 UPLC 指纹图谱建立及质量标志物（Q-Marker）定量测定

Objective To establish UPLC fingerprint and determination method for quality markers of Huangqi Guizhi Wuwu Decoction (HGWD, 黄芪桂枝五物汤) benchmark samples and evaluate the quality of 15 batches of HGWD, so as to provide quality control method and evaluation system for the subsequent development of compound preparation. Methods The benchmark samples of HGWD were prepared by Waters Acquity^TM T3 column (100 mm × 2.1 mm, 1.8 μm). Acetonitrile-0.15% phosphoric acid aqueous solution was used as mobile phase with gradient elution. Column temperature: 30 ℃; Detection wavelength: 0—18 min, 230 nm; 18.5—24.5 min, 260 nm; 24.5—40 min, 280 nm; Elution rate: 0.3 mL/min; Injection volume: 1 μL; The fingerprint methodology was investigated, 15 batches of UPLC fingerprints of HGWD benchmark samples were collected, and the data results were imported into the “TCM Chromatographic Fingerprint Similarity Evaluation System” (2012 edition). The similarity evaluation was carried out on the fingerprint of 15 batches of benchmark samples of HGWD, and the common peaks were attributed and cluster analyzed. At the same time, the contents of quality markers (calycosin-7-glucoside, paeoniflorin, cinnamic acid, 6-gingerol) in 15 batches of benchmark samples were determined. Results The methodological verification of the UPLC fingerprint of the HGWD benchmark samples was good. The UPLC fingerprints of 15 batches of the benchmark samples of HGWD were well verified. The similarity between the UPLC fingerprints and the control fingerprints was greater than 0.9. A total of 24 common peaks were identified, and eight chromatographic peaks were identified by comparing with mixed controls, namely peak 8 oxypaeoniflorin, peak 10 paeoniflorin, peak 12 calycosin-7-glucoside, peak 14 coumarin, peak 18 cinnamic acid, peak 19 cinnamaldehyde, peak 20 quercetin and peak 24 6-zingerol. The mass fractions of four quality markers, including calycosin-7-glucoside, cinnamic acid, paeoniflorin and 6-gingerol, in 15 batches of HGWD were determined to be 115.49—494.96, 254.68—760.79, 3 106.72—6 049.40, 232.61—704.10 μg/g, respectively. The paste extraction rate was 11.63%—14.14%, and the transfer rates of calycosin-7-glucoside, cinnamic acid, paeoniflorin and 6-gingerol in the decoction pieces-soup liquid were 8.23%—19.66%, 23.52%—41.21%, 29.15%—41.23% and 14.58%—30.07%, respectively. The transfer rates of calycosin-7-glucoside, cinnamic acid, paeoniflorin and 6-gingerol in the benchmark samples were 86.35%—97.67%, 90.34%—98.13%, 91.51%—98.88% and 93.53%—98.29%, respectively. Conclusion The established method of UPLC fingerprint and quality marker content determination of HGWD is accurate and reliable, which can be used for the quality evaluation of HGWD benchmark samples and could be used as the basis for the development and quality control of subsequent preparations.