Determination of hemocoagulase agkistrodon in a pharmaceutical preparation by high-performance liquid chromatography with pre-column derivatization and fluorescence detection.
Currently, there is no analytical method for the quantification of hemocoagulase agkistrodon (HCA) in pharmaceutical preparations. This study presents a pre-column derivatization method for the quantification of HCA, a compound extracted from the venom of Agkistrodon acutus, in a pharmaceutical preparation (trade name Suling). In the proposed method, 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate was used to tag the HCA substrate, and the derivatives were analyzed by high-performance liquid chromatography with fluorescence detection. Complete and homogeneous derivatization of HCA was confirmed by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry analysis. The specificity of the method was validated by forced degradation, and interference was assessed using a placebo. Under the optimum chromatographic conditions, the calibration curve was linear over a range of 10 to 500 ng/mL, featuring a correlation coefficient of 0.9999. The limits of detection and quantification of the method were 0.57 and 1.6 ng/mL, respectively. The percentage recovery of HCA in quality control samples ranged from 97.49 to 99.15%. Overall, this novel method can be applied to the quantitative determination of HCA in pharmaceutical preparations.
Institute of Biochemical and Biotechnological Drugs, School of Pharmaceutical Sciences, Shandong University, Jinan 250012, China.
Suyuan
S
Cheng
Chaozhong
C
Wang
Jing
J
Li
Chenggang
C
Liang
Fengshan
F
Wang
D000818:Animals; D012103:Batroxobin / Q000032:analysis* / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004304:Dosage Forms; D004355:Drug Stability; D016014:Linear Models; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013696:Temperature; D017819:Viperidae
D004304:Dosage Forms; D012103:Batroxobin