Determination of Eight Bufadienolides in the Skin of Bufo Bufo Gargarizans Cantor and Bufo Melanostictus Schneider Using HPLC Coupled With Triple Quadrupole Mass Spectrometry

Chan Pi (the toad skin) is an effective Chinese medicine in the treatment of boils, pyogenic infections, and cancer in clinical settings. However, the inhomogeneity of its quality is often observed due to confusing sources, namely Bufo bufo gargarizans Cantor and Bufo melanostictus Schneider. In the present paper, a simultaneous determination method of eight bufadienolides in two species of Chan Pi was established using HPLC coupled with triple quadrupole mass spectrometry (HPLC-QqQ MS). Eight bufadienolides in 7 batches of B. bufo gargarizans skin and 8 batches of B. melanostictus skin were analyzed with an acceptable linear regression relationship (r ≥ 0.9990), precisions (RSD, 1.97–4.38%), repeatability (RSD, 2.65–4.70%), stability (RSD, 2.39–4.51%), and recovery (96.39–103.82%). Additionally, cluster analysis was used for the successful discrimination of these two species of toad skin by the ratio of these eight bufadienolides. The present study greatly facilitates clarification of the difference between these two species of Chan Pi, which improves the quality control of this traditional Chinese medicine.