Bear bile powder (BBP) is a widely used traditional Chinese medicine (TCM), and bile acids (BAs) are the main active components in BBP. Due to the scarcity of BBP resources, adulterations often occur in the market. Conventional methods to distinguish them are usually complicated and time-consuming. To enhance effectiveness and accuracy, a rapid and rough analytical method is desperately needed.Methods: In this study, a rapid strategy using chip-based nano-electrospray ionization tandem mass spectrometry (nano-ESI-MS/MS) was established to distinguish BBP from other sources of bile powder (BP). In addition, the results were further verified by ultra-high-performance liquid chromatography combined with high-resolution mass spectrometry (UPLC/MS).
Results:The precision of the chip-based nano-ESI-MS/MS method was validated to be acceptable with relative standard deviation (RSD) <15%. The distinction between BBP and other sources of BP, including common adulterants of pig bile powder (PBP), cattle bile powder (CBP), sheep bile powder (SBP), and chicken bile powder (CkBP), can be observed in the spectra. By using orthogonal partial least-squares discriminant analysis (OPLS-DA), more potential m/z markers were investigated. A BAs-related m/z marker of 498.3 was discovered as a typical differential molecular ion peak and was identified as tauroursodeoxycholic acid (TUDCA) and taurochenodeoxycholic acid (TCDCA) in BBP.
Conclusions:The proposed strategy has simple sample pretreatment steps and significantly shortened analysis time. As an emerging technology, chip-based nano-ESI-MS not only provides a reference for the rapid distinction of adulterated Chinese medicines, but also provides some insights into the identification of other chemicals and foods.
| INTRODUCTIONAdulteration of traditional Chinese medicines (TCMs), dietary supplement materials, and herbal products is a global problem for regulators. [1][2][3] These adulterant or counterfeit drugs can seriously compromise the therapeutic effects of natural medicines. Bear bile powder (BBP) is dried gallbladder bile collected from the black bear (Selenarctos thibetanus), the brown bear (Ursus arctos), or other species of Ursidae, under the category of animal drugs in TCMs. BBP has been used as a TCM for over 1300 years since the Tang Dynasty. [4][5][6] According to the theory of TCM, BBP has efficacies of "cleansing heat symptoms, calming the liver, and brightening eyes", and has been used in the clinic to treat convulsion, conjunctivitis, and pharyngolaryngitis in China. 7 Bile acids (BAs) are the main components in BBP, and