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A high-performance liquid chromatographic (HPLC) method for the simultaneous determination of resibufogenin and cinobufagin in traditional Chinese medicine (heart-protecting musk pill, shexiang baoxin wan in Chinese) was developed. A reversed-phase system with a Hypersil (ODS2) C(18) column and tetrahydrofuran: methanol: water (8:31:61) mobile phase was employed for the separation of resibufogenin and cinobufagin. The detection was set at 299 nm and ethinyl estradiol was chosen as the internal standard. The limit of detection was 1.5 ng for resibufogenin and 2.0 ng for cinobufagin at a signal-to-noise ratio of 4:1. It is a rapid, simple and accurate method for quantitative analysis of resibufogenin and cinobufagin in heart-protecting musk pill.
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Hong-tao et al. (2000) studied this question.
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