Taurine as a Marker for the Identification of Natural Calculus Bovis and Its Substitutes
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Calculus Bovis (C. Bovis) is a commonly used animal-derived therapeutic preparation. To meet the increasing clinical demand for the preparation, two artificial substitutes for Bos Taurus have been introduced in China: artificial C. Bovis and in vitro cultured C. Bovis. However, information on their efficacy and safety is inadequate. Therefore, we investigated the biological differences between the commonly used natural preparation and its two substitutes, with the aim of not only identifying the differences but also providing a procedure to distinguish between the different preparations.
In the study, we prepared 9 natural C. Bovis, 2 artificial C. Bovis, and 2 in vitro cultured C. Bovis preparations for evaluation. Differences were noted between the three preparations relative to their effect on viability of cardiac fibroblasts from 1-day-old Wistar rats. Although natural C. Bovis had no effect on cell viability, 1-h treatment of the cells with 0.25 mg/ml of the substitutes significantly reduced cell viability, as detected by the MTS assay. Based on liquid chromatography and inductively coupled plasma mass spectrometry, the preparations also differed in composition. Indeed, the substitutes contained more taurine, cholic acid, iron, magnesium, and calcium than the natural preparations. They also differed spectroscopically.
The present results reveal significant biological differences between natural C. Bovis and two of its substitutes. Since the substitutes appear to contain more taurine, cholic acid, and elements, these constituents may serve as markers to distinguish between natural C. Bovis and its substitutes.
KeywordsInductively Couple Plasma Mass Spectrometry Cholic Acid Cardiac Fibroblast Taurocholic Acid Organic Composition
Inductively coupled plasma mass spectrometry
Kyoto University Research Reactor Institute
Principal components analysis
We thank Mr. Yoichi Yokota for the support of the materials, and Dr. Tadashi Saito for help with ICP-MS. This study was supported by a Grant-in-Aid for “Scientific Research (B),” No. 22300310, in 2010–2012 from the Japan Society for the Promotion of Science (JSPS). Partial support was also provided by Grant-in-Aid for JSPS Fellows, No. 23·2852, in 2011–2012 from JSPS.
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