Taurine as a Marker for the Identification of Natural Calculus Bovis and Its Substitutes - Advances in Experimental Medicine and Biology - page 134

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 distin-
guish between natural C . Bovis and its substitutes.

Abbreviations

ICP - MS Inductively coupled plasma mass spectrometry

THz Terahertz

LINAC Linear accelerator

KURRI Kyoto University Research Reactor Institute

PCA Principal components analysis

15.1 Introduction

Calculus Bovis ( C . Bovis : Bezoar Bovis ) is an important animal-derived therapeutic
agent that originates from gallstones formed in the gall sac of Bos Taurus Linné var.
domesticus Gmelin (Bovidae) in the Japanese pharmacopoeia 16th edition (Ministry
of Health, Labour and Welfare 2011 ). Only one out of every 1,000 cattle contains
gallstones for C . Bovis , making the preparation quite expensive. However, there is
increased therapeutic demand for it.
Many studies have been carried out to find substitutes for natural C . Bovis . There
are two kinds of substitutes of C . Bovis in Chinese pharmacopoeia that are available
for clinical practice: one is artificial C . Bovis (Bovis Calculus Artifactus), a mixture
of bile salts, bilirubin, taurine, and some undetermined ingredients. The other is
in vitro cultured C . Bovis (Bovis Calculus Sativus), produced in vitro under condi-
tions mimicking in vivo gallstone formation (Qin 2008 ; Chinese Pharmacopoeia
Committee of People's Republic of China 2010 ). Previously we showed that the
substitutes are distinguishable by multiple element analysis from natural C . Bovis
(Takahashi et al. 2010 ). Moreover, differences in organic composition (Yan et al.
2007 ; Kong et al. 2010 ) and morphology (Yamaguchi et al. 2008 ) between natural
C . Bovis and its substitutes have been observed. However, further information is
required to properly use the substitutes in clinical practice.
Therefore, the aim of the present study is to identify differences between natural
C . Bovis and its substitutes relative to biological activity and composition. The
intention of the report is to (1) establish the importance of these differences and (2)
introduce methodology to distinguish between natural and artificial preparations.