Identification and Quantitation of Bile Acids in Bear Bile by HPLC
DONG-LIANG LIN1,2*, HSIEN-CHEH CHANG3 AND CHAU-YANG CHEN4
1.Scientific and Technical Research Center, Ministry of Justice Investigation Bureau, P.O. Box 3562, Taipei, Taiwan, R.O.C.
2.Graduate Institute of Pharmaceutical Sciences, Taipei Medical College, Taipei, Taiwan, R.O.C.
3. China Medical College, Taichung, Taiwan, R.O.C.
4. Tajen Institute of Technology, Pingtung, Taiwan, R.O.C.
(Received: November 6, 1999; Accepted: March 17, 2000)
ABSTRACT
A method for simultaneous quantitation of tauroursodeoxycholic acid (TUDC), taurocholic acid (TC) and taurochenodeoxycholic acid (TCDC) using high performance liquid chromatography with a Vydac reversed phase C18 column is described. Peak area ratios versus concentration calibration curve for TUDC, TC and TCDC showed linearity over the range of 50-2000 mg/mL with correlation coefficient (r) of 0.9999, 0.9989 and 1.0000, respectively. Intraday and interday precisions range from 0.08 to 5.63% for TUDC, 0.28 to 6.70% for TC and 0.14 to 4.82% for TCDC. Quantitative HPLC analysis was performed on 93 Ursidae bile salts. Average concentrations were 3087 mg/mL for TUDC, 212.6 mg/mL for TC and 1968 mg/mL for TCDC. Our data suggests that North American and polar bears contain a higher concentration of TC (relative to TUDC and TCDC), while the relative concentration of TC in Asiatic bears (wild or farmed) is much lower. Analytical results of 217 gall bladder (11 cases) items submitted as criminal evidence from customs officials indicate: 147 (67.74%) were of domestic pig origin; 42 (19.36%) were from Asiatic bears, 14 (6.45%) were from goat gall bladders and 14 (6.45%) were absent of animal bile.
Key words: bear gall bladder, identification, quantitation, HPLC