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GC/MS¿¡ ÀÇÇÑ ´ã¼®°ú ´ãÁó³»ÀÇ Cholesterol, Bile Acids ¹× SterolsÀÇ µ¿½Ã Profiling

Simultaneous Determination of Cholesterol, Bile Acids and Sterols in Human Bile Juices and Gallstones Using GC/MS

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¾çÀ±Á¤, À̼±È­, ±èÇöÁÖ, Á¤ºÀö,
¼Ò¼Ó »ó¼¼Á¤º¸
¾çÀ±Á¤ ( Yang Yoon-Jung ) - Korea Institute of Science and Technology Bioanalysis and Biotransformation Research Center
À̼±È­ ( Lee Seon-Hwa ) - Korea Institute of Science and Technology Bioanalysis and Biotransformation Research Center
±èÇöÁÖ ( Kim Hyun-Joo ) - Chungju National University Division of General Education
Á¤ºÀö ( Chung Bong-Chul ) - Korea Institute of Science and Technology Bioanalysis and Biotransformation Research Center

Abstract

´ãÁó»ê ºÐºñ ÀúÇÏ ¹× ÄÝ·¹½ºÅ×·ÑÀÇ °úµµÇÑ ºÐºñ´Â ´ã¼® Çü¼ºÀÇ ÁÖµÈ ¿øÀÎÀ¸·Î ´ãÁó³»ÀÇ ´ãÁó»ê°ú ÄÝ·¹½ºÅ×·Ñ ¹× ±× Àü±¸ ¹°ÁúÀÎ ½ºÅ×·ÑÀÇ »ó´ëÀûÀÎ ³óµµ´Â ´ãÁó ¼ººÐÀÇ ÀÌ»óÀ» ¾Ë¾Æº¸±â À§ÇÑ Áß¿äÇÑ ÁöÇ¥·Î ¾Ë·ÁÁ® ÀÖ´Ù. ÀÌ¿¡ º» ¿¬±¸¿¡¼­´Â ¿°±â¸¦ ÀÌ¿ëÇÑ °¡¼ö ºÐÇØ¿Í pH 14¿Í 1¿¡¼­ µÎ ¹øÀÇ ¾×ü-¾×ü ÃßÃâÀ» °ÅÄ£ ÈÄ, MSTFA/NH4I È¥ÇÕ¾×À¸·Î À¯µµÃ¼È­ ½ÃŰ´Â Àüó¸® ¹æ¹ý°ú GC/MS¸¦ ÀÌ¿ëÇÑ »õ·Î¿î ºÐ¼® ¹æ¹ýÀ» ¼³Á¤, ´ãÁó»ê°ú ½ºÅ×·Ñ ±×¸®°í ÄÝ·¹½ºÅ×·ÑÀÇ µ¿½Ã ºÐ¼®À» ½ÃµµÇÏ¿´´Ù. ±× °á°í ȸ¼öÀ²Àº 73.56-96.95% À̾ú°í within-a-day ¹× day-to-day ºÐ¼®ÀÇ RSD °ªÀº °¢°¢ 1.72-13.79%, 0.68-14.10% À̾úÀ¸¸ç, ÀÌ ¹æ¹ýÀ» °£³» ´ã¼®Áõ ȯÀÚ¿¡ Àû¿ëÇÏ¿© ´ãÁó°ú ´ã¼®³»¿¡ Á¸ÀçÇÏ´Â 7Á¾ÀÇ ½ºÅ׷Ѱú 5Á¾ÀÇ ´ãÁó»ê, ±×¸®°í ÄÝ·¹½ºÅ×·ÑÀÇ ³óµµ ¹× ±×µé°£ÀÇ »ó´ëÀûÀÎ ³óµµ ºÐÆ÷¸¦ ÃøÁ¤ÇÑ °á°ú ´ãÁó°í kekatjr¿¡¼­ ÇöÀúÈ÷ ´Ù¸¥ Â÷À̸¦ º¸¿´´Ù.

Cholesterol supersaturation in bile, which causes gallstone formation, is the result of low bile acid secretion or high cholesterol secretion. The quantitative analysis of cholesterol, bile acids and sterols which are precursors of cholesterol have been used to examine the changes in bile component. We described a simple, sensitive and reproducible method for simultaneous determination of cholesterol, five bile acids and seven sterols in human bile juices and gallstones by capillary column gas chromatography-mass spectrometry (GC/MS). Clinical samples were hydrolyzed by alcoholic KOH, extracted twice (pH 14 and 1) and derivatized to trimethylsilyl (TMS) ether with MSTFA/NH4I (N-methyl-N-trimethylsilyltrifluoroacetamide/ammonium iodide) mixture in order to be detected on the GC/MS. The good quality control data were obtained through within-a-day and day-to-day test (RSD values were 1.72-13.79%, 0.68-14.10%, respectively) and the recovery range of them was 73.56-96. 95%. Using this method, biliary and gallstone compositions in the patients with intrahepatic stones were analyzed. The amounts and its relative distribution of cholesterol, bile acids and sterols showed different pattern in bile juices and gallstones.

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