Objective: To establish a high performance liquid chromatography-charged aerosol detection (HPLC-CAD) method for the determination of cinobufagin, resibufogenin, and cholic acid in Niuhuang Xiaoyan tablets. Methods: The separation was conducted on a CAPCELL PAK MGII C18 column (250 mm×4.6 mm, 3 μm), employing the gradient elution with acetonitrile and 0.3% acetic acid aqueous solution as the mobile phase. The flow rate was 0.6 mL · min-1 and the column temperature was 30 ℃, and the injection volume was 10 μL. The CAD had a filter constant of 1 second, a frequency of 5 Hz, a evaporation temperature of 90 ℃, and a power function value of 1.1. Results: The linear ranges of cinobufagin, resibufogenin and cholic acid were 24.97-99.89 μg · mL-1 (r=0.999 2), 25.33-101.3 μg · mL-1 (r=0.999 5), and 100.5-602.9 μg · mL-1 (r=0.999 0), respectively. The RSDs of stability, precision and repeatability for the three components were all less than 3.0%. The average spike recovery rates (n=9) were 99.17%, 103.6%, and 102.0%, with RSD of 1.5%, 2.3%, and 3.0%, respectively. The contents of cinobufagin in six batches of samples were 0.122 1, 0.126 4, 0.122 6, 0.128 4, 0.122 5 and 0.125 6 mg per tablet respectively, the contents of resibufogenin were 0.051 50, 0.051 15, 0.051 79, 0.050 68, 0.054 43 and 0.054 62 mg per tablet respectively, and the contents of cholic acid were 0.235 3, 0.239 0, 0.236 0, 0.236 2, 0.233 8 and 0.236 8 mg per tablet respectively. Conclusion: This method is accurate, robust, and has good repeatability. It enables the simultaneous quantification of three components—cinobufagin, resibufogenin, and cholic acid in Niuhuang Xiaoyan tablets in single run, which is more convenient and offers higher analytical throughput.
LIU Hui-yi
,
ZUO Li-min
,
QIU Xiao-dan
,
ZHAO Ting
,
ZHAO Xue-jia
,
GUO Xin
,
LIAN Xiao-fang
,
JIA Qing-ying
,
GU Yong-sheng
,
LIU Xuan-lin
,
SHAN Guang-zhi
. Determination of three active components in Niuhuang Xiaoyan tablets by high performance liquid chromatography coupled with charged aerosol detection*[J]. Chinese Journal of Pharmaceutical Analysis, 2025
, 45(7)
: 1181
-1190
.
DOI: 10.16155/j.0254-1793.2024-1149
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