HPLC-DAD-ELSD Combined Pharmacodynamics and Serum Medicinal Chemistry for Quality Assessment of Huangqi Granule.
OBJECTIVE
To more scientifically and reasonably control the quality of Huangqi Granules, preliminary studies on the pharmacodynamics and serum pharmacochemistry of this medicine were performed. DPPH and MTT experiments showed that water extracts of Huangqi Granules had good antioxidant activity and increased immunity. Timed blood samples collected 5 min, 15 min, and 30 min after oral administration of a set amount of Huangqi Granules were collected and tested using UPLC-ESI-MS/MS. As a result, calycosin-7-O-β-D-glucoside, ononin, calycosin, astragaloside IV, and formononetin were found to exist in rat blood after dosing, indicating that the five chemical compounds might have pharmacological activity, and based on this result, they were designated biomarkers for quality control of Huangqi Granules. Consequently, a simple, rapid and efficient method was developed in the present study for the simultaneous determination of the five characteristic compounds in Huangqi Granules using HPLC-DAD-ELSD.


METHODS
The separation was performed using an Agilent Hypersil ODS column (4.6 × 250 mm, 5 μm) at 30 ℃. The mobile phase was composed of water (solvent A) and acetonitrile (solvent B) with a flow rate of 1 mL/min. The drift tube temperature of the ELSD system was set to 85 ℃, and the nitrogen pressure was 3.5 bar.


RESULTS
All five characteristic compounds had good linear behavior with r2 values greater than 0.9972. The recoveries varied from 96.31% to 101.22%. Subsequently, the developed method was applied to evaluate the quality of Huangqi Granules from different batches, and hierarchical clustering analysis (HCA) was used to analyze the classification of the samples based on the values of the five compounds.


CONCLUSIONS
The established HPLC method combined with HCA proved to be effective to evaluate the quality of Huangqi Granules.
Guizhou Engineering Laboratory for Quality Control & Evaluation Technology of Medicine, Guiyang, Guizhou, P. R. China; The Research Center for Quality Control of Natural Medicine, Guizhou Normal University, Guiyang, Guizhou, P. R. China; Key laboratory for Information System of Mountainous Areas and Protection of Ecological Environment, Guizhou Normal University, Guiyang, Guizhou, P. R. China.
Huaguo
H
Chen
Guizhou Engineering Laboratory for Quality Control & Evaluation Technology of Medicine, Guiyang, Guizhou, P. R. China; The Research Center for Quality Control of Natural Medicine, Guizhou Normal University, Guiyang, Guizhou, P. R. China; Key laboratory for Information System of Mountainous Areas and Protection of Ecological Environment, Guizhou Normal University, Guiyang, Guizhou, P. R. China.
Xin
X
Zhou
Guizhou Engineering Laboratory for Quality Control & Evaluation Technology of Medicine, Guiyang, Guizhou, P. R. China; The Research Center for Quality Control of Natural Medicine, Guizhou Normal University, Guiyang, Guizhou, P. R. China; Key laboratory for Information System of Mountainous Areas and Protection of Ecological Environment, Guizhou Normal University, Guiyang, Guizhou, P. R. China.
Yang
Y
Zhao
Guizhou Engineering Laboratory for Quality Control & Evaluation Technology of Medicine, Guiyang, Guizhou, P. R. China; The Research Center for Quality Control of Natural Medicine, Guizhou Normal University, Guiyang, Guizhou, P. R. China; Key laboratory for Information System of Mountainous Areas and Protection of Ecological Environment, Guizhou Normal University, Guiyang, Guizhou, P. R. China.
Xiao-Jian
XJ
Gong
Guizhou Engineering Laboratory for Quality Control & Evaluation Technology of Medicine, Guiyang, Guizhou, P. R. China; The Research Center for Quality Control of Natural Medicine, Guizhou Normal University, Guiyang, Guizhou, P. R. China; Key laboratory for Information System of Mountainous Areas and Protection of Ecological Environment, Guizhou Normal University, Guiyang, Guizhou, P. R. China.
Yan
Y
He
Guizhou Engineering Laboratory for Quality Control & Evaluation Technology of Medicine, Guiyang, Guizhou, P. R. China; The Research Center for Quality Control of Natural Medicine, Guizhou Normal University, Guiyang, Guizhou, P. R. China; Key laboratory for Information System of Mountainous Areas and Protection of Ecological Environment, Guizhou Normal University, Guiyang, Guizhou, P. R. China.
Feng-Wei
FW
Ma
Guizhou Engineering Laboratory for Quality Control & Evaluation Technology of Medicine, Guiyang, Guizhou, P. R. China; The Research Center for Quality Control of Natural Medicine, Guizhou Normal University, Guiyang, Guizhou, P. R. China; Key laboratory for Information System of Mountainous Areas and Protection of Ecological Environment, Guizhou Normal University, Guiyang, Guizhou, P. R. China.
Mei
M
Zhou
Guizhou Engineering Laboratory for Quality Control & Evaluation Technology of Medicine, Guiyang, Guizhou, P. R. China; The Research Center for Quality Control of Natural Medicine, Guizhou Normal University, Guiyang, Guizhou, P. R. China; Key laboratory for Information System of Mountainous Areas and Protection of Ecological Environment, Guizhou Normal University, Guiyang, Guizhou, P. R. China.
Chao
C
Zhao
Han Fang Pharmaceutical co., LTD of Guizhou province, Guiyang, Guizhou, P. R. China.
Yi
Y
Niu
Han Fang Pharmaceutical co., LTD of Guizhou province, Guiyang, Guizhou, P. R. China.
Jie
J
Deng
D000818:Animals; D000975:Antioxidants / Q000737:chemistry* / Q000493:pharmacokinetics / Q000494:pharmacology; D027885:Astragalus propinquus; D002851:Chromatography, High Pressure Liquid; D004365:Drugs, Chinese Herbal / Q000737:chemistry* / Q000493:pharmacokinetics / Q000494:pharmacology; D005960:Glucosides / Q000032:analysis; D007529:Isoflavones / Q000032:analysis; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley
D000975:Antioxidants; D004365:Drugs, Chinese Herbal; D005960:Glucosides; D007529:Isoflavones; C027492:Huang Qi