以高效液相層析法鑑別及定量熊膽內膽酸成分   全文下載

Identification and Quantitation of Bile Acids in Bear Bile by HPLC

本研究利用高效液相層析法，鑑別各種動物膽汁成分並同時定量分析熊膽內之TUDC, TC及TCDC之含量，採用Vydac C18 4.6 × 250 mm逆相層析管柱，25 mM KH2PO4／K2HPO4甲醇溶液：水（85: 15, v／v）為移動相，流速為0.75 mL／min ，檢測波長為210 nm ，並以P-Cresol 為內標準品。三種成分之定量曲線於50-2000 μg／mL 濃度範圍內，均呈現良好的線性關係（r>0.998）。同日間及異日間試驗之相對標準偏差，TUDC為0.08 ?5.63%，TC為0.20 ?6.70%，TCDC為0.14 ?4.82%。應用上述高效液相層析法，定量分析93 個亞洲熊膽檢體，TUDC平均濃度為3087 μg／mL、TC為213.6 μg／mL、TCDC為1968 μg／mL，並發現TC含量的多寡是鑑別亞洲熊與北極熊、北美黑熊之關鍵，而非用來鑑別亞洲野生熊膽與飼養引流熊膽。分析市售183 個熊膽檢體， 56（30.6%）個為亞洲熊膽， 118（64.5%）個為豬膽；分析農委會查扣217 個熊膽檢體， 42（19.4%）為亞洲熊膽，147（67.7%）個為豬膽。"

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 μg/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 μg/mL for TUDC, 212.6 μg/mL for TC and 1968 μg/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. 本研究利用高效液相層析法，鑑別各種動物膽汁成分並同時定量分析熊膽內之TUDC, TC及TCDC之含量，採用Vydac C18 4.6 × 250 mm逆相層析管柱，25 mM KH2PO4／K2HPO4甲醇溶液：水（85: 15, v／v）為移動相，流速為0.75 mL／min ，檢測波長為210 nm ，並以P-Cresol 為內標準品。三種成分之定量曲線於50-2000 μg／mL 濃度範圍內，均呈現良好的線性關係（r>0.998）。同日間及異日間試驗之相對標準偏差，TUDC為0.08 ∼5.63%，TC為0.20 ∼6.70%，TCDC為0.14 ∼4.82%。應用上述高效液相層析法，定量分析93 個亞洲熊膽檢體，TUDC平均濃度為3087 μg／mL、TC為213.6 μg／mL、TCDC為1968 μg／mL，並發現TC含量的多寡是鑑別亞洲熊與北極熊、北美黑熊之關鍵，而非用來鑑別亞洲野生熊膽與飼養引流熊膽。分析市售183 個熊膽檢體， 56（30.6%）個為亞洲熊膽， 118（64.5%）個為豬膽；分析農委會查扣217 個熊膽檢體， 42（19.4%）為亞洲熊膽，147（67.7%）個為豬膽。