Development of a rapid and simple LC-MS/MS method for identification and quality control of natural Calculus bovis and Calculus bovis sativus

Abstract

Calculus bovis sativus (C. bovis sativus, CBS), with the same pharmacodynamics activities as those of natural C. bovis (CB), has been approved as an ideal substitute in China. However, in the latest edition of the Chinese Pharmacopoeia, only cholic acid and bilirubin are used as the markers for quality control. To overcome current limitations and to further improve its quality control method, a rapid and reliable method was herein developed for simultaneous qualitative and quantitative analyses of taurine and twelve bile acids (BAs) in natural CB and CBS by liquid chromatography coupled with tandem mass spectrometry. Taurine and BAs were separated using a Diamonsil C 18 column (150 mm  2.1 mm i.d., 5 mm) with acetonitrile:water containing 10 mM ammonium acetate (pH 3) as mobile phase in a gradient elution program. Mass spectra were obtained in the negative ion mode through multiple reaction monitoring of respective mass transitions. This method had satisfactory separation efficiency, high sensitivity, little potential interference, and short running time. The average recoveries ranged from 93.0% to 107.3%, with relative standard deviations of less than 9.0% for all thirteen analytes of interest. Taurine and characteristic BAs, especially conjugated BAs, may partially reflect the internal quality of CBS and natural CB. Combining the ratios of cholic acid/deoxycholic acid and unconjugated BAs/conjugated BAs with quantitative data of single BA and chromatographic profiles may be able to control the quality of CBS. In conclusion, this method can be used for explicit identification and quality control of CBS and natural CB.

Investigation of the synergistic effects of haloperidol combined with <i>Calculus Bovis Sativus</i> in treating MK-801-induced schizophrenia in rats

“…Calculus Bovis Sativus (Lot: 2016–05-16) was provided by Wuhan Jianmin Dapeng Pharmaceuticals Co., Ltd., (Wuhan, China), and dissolved in normal sterile saline. The effective components (taurine and bile acids) in CBS were quantified by a HPLC-MS/MS approach described in our previous study [ 10 ].…”

“…Bile acids, bilirubin, and taurine are considered the main effective components in CBS [ 38 , 41 ]. In our previous work, we simultaneously quantified taurine and 12 bile acids using high-performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS) to control the quality of CBS [ 10 ].…”

Hepatoprotective effect of calculus bovis sativus on nonalcoholic fatty liver disease in mice by inhibiting oxidative stress and apoptosis of hepatocytes

“…CBS and natural CB are similar in characteristics, ingredients, contents, and therapeutic effects. 10 Although the therapeutic effects of CB and CBS in the liver are well documented, the underlying mechanism of action has not yet been identified.…”

Upregulation of PDZK1 by Calculus Bovis Sativus May Play an Important Role in Restoring Biliary Transport Function in Intrahepatic Cholestasis

“…Given that Calculus Bovis is insufficient of nature sources, Calculus Bovis Sativus (Tiwai Peiyu Niuhuang, CBS) was invented as the substitute in the medicinal preparations. Many studies have shown that CBS shares basically the same property, structure, composition, content, and clinical efficacy with those of Calculus Bovis [6–8]. Moreover, because of its complex components including bilirubin, cholic acid, bile acids and cholesterol, and so forth, we have established HPLC fingerprints for the effective quality control of CBS [8].…”

“…Many studies have shown that CBS shares basically the same property, structure, composition, content, and clinical efficacy with those of Calculus Bovis [6–8]. Moreover, because of its complex components including bilirubin, cholic acid, bile acids and cholesterol, and so forth, we have established HPLC fingerprints for the effective quality control of CBS [8]. Our previous study indicated that CBS treatment had a beneficial effect on 17 α -ethynylestradiol- (EE-) induced intrahepatic cholestasis rats and significantly increased the mRNA and protein expressions of multidrug resistance-associated protein 2 (MRP2) and breast cancer resistance protein (BCRP), both important proteins on the apical membrane of hepatocytes involving in the transport of bile acids, bilirubin, drugs, and so on [9].…”