Toxicological Studies on Grass Carp Bile Juices

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Project title

Code/計畫編號

DOH88-TD-1039

Translated Name/計畫中文名

草魚膽汁之毒性學研究

Project Coordinator/計畫主持人

Deng-Fwu Hwang

Funding Organization/主管機關

Environmental Protection Administration

Department/Unit

Department of Food Science

Website

https://www.grb.gov.tw/search/planDetail?id=479441

Year

1999

Start date/計畫起

01-07-1998

Expected Completion/計畫迄

01-06-1999

Bugetid/研究經費

900千元

ResearchField/研究領域

漁業

為知草魚膽汁中之膽酸、膽酸鹽和膽醇硫酸鹽之組成,乃首先從草魚膽汁中純化出5.alpha.-cyprinol,其過程為將草魚膽汁經鹼水解後,藉PHP-LH-20和Silica gel兩種管柱層析純化後,取其酒精溶出物,經減壓濃縮除去溶劑後,以氰甲烷做結晶,將此結晶物經紅外光譜儀(FT-IR)、氫原子(/sup 1/H)和碳-13 (/sup 13/C)核磁共振光譜儀(NMR)和質譜儀(MS)之化學分析,結果發現5.alpha.-cyprinol之分子式為C/sub 27/H/sub 48/O/sub 5/,分子量452,其在草魚膽汁係以5.alpha.-cyprinol sulfate存在。同時研發高效能液相層析法(HPLC)分析膽酸和膽鹽之方法,發現以ODS逆向層析管,溶煤系統0.3% ammonium carbonate/acetonitrile(73:27) 10 min.arrr.(68:32) 10 min.arrr.(50:50) 10 min,即可將13種bile acid分離。其中deoxycholic acid和glycodeocholic acid會在18分鐘時同時溶出,但另以溶出條件buffer 0.3% ammonium carbonate/acetonitrile(73:27)即可區分此兩種膽酸成分。經採樣14種魚類膽汁,經以HPLC方法分析bile acids,另外GC方法分析5.alpha.-cyprinol sulfate,得知草魚和鯉魚膽汁中之5.alpha.-cyprinol sulfate之含量為膽酸、膽酸鹽和膽醇鹽全量之94%以上,而鱒魚、鰻魚和鯖魚等12種魚類膽汁中膽酸主要成分為taurocholic acid和taurochenodeoxycholic acid,而香魚膽汁中膽酸主要成分為cholic acid和taurocholic acid。除草魚膽外,其他魚類膽汁中都未檢出5.alpha.-cyprinol sulfate。為知5.alpha.-cyprinol之毒性,乃以口胃管餵食老鼠生理食鹽水(0.6ml)、草魚膽汁(97.44mg/0.6ml)和5.alpha.-cyprinol(97.44mg/0.6ml), 6小時後,發現草魚膽汁和5.alpha.-cyprinol組之老鼠,其血漿中BUN和creatinine顯著上升(p<0.05),顯示兩組老鼠腎功能受損。但5.alpha.-cyprinol組之老鼠,胺基酸轉氨酵素AST和ALT未見變化,顯示5.alpha.-cyprinol之毒作用器官為腎臟,而草魚膽汁之毒作用器官為肝臟和腎臟。 To elucidate the responsible toxic components of grass carp in its bile juice. The bile alcohol 5.alpha.-cyprinol from grass carp bile juice was purified and identified. By IR, /sup 1/H-NMR, /sup 13/C-NMR and MS spectrometry analyses, a compound with molecular formula of C/sub 27/H/sub 48/O/sub 5/ and molecular weight of 452 was obtained from grass carp bile juice and identified as 5.alpha.-cyprinol. In the meantime, the analysis for bile acids in fish bile juices was developed. It was found that high performance liquid chromatography (HPLC), with a reversed ODS column and a buffer system of 0.3% ammonium carbonate/acetonitrite (73:27) 10 min.arrr.(68:32) 10 min.arrr.(50:50) 10 min, clearly separated 13 bile acids. Among them, deoxycholic acid and glycodeoxycholic acid appeared the same time. These two components were further separated by using other buffer 0.3% ammonium carbonate/acetonitrite (73:27). Based on the analyses of about HPLC method and GC method, 13 samples of bile juices obtained from 13 fish species. It was found that the major component of bile juices was 5.alpha.-cyprinol sulfate for grass carp and common carp. The rainbow trout, rock trout, japanese eel, spanish mackerel, right eyed flounder, rudder fish, black scraper, tongue sole, japanese jack and mackerel contained mainly taurocholic acid and taurochenodeoxycholic acid, but ayu fish contained mainly cholic acid and taurocholic acid. To compare the toxicity of 5.alpha.-cyprinol and grass carp, the rats were separately administered with saline (0.6ml), grass carp bile juice (97.44mg/0.6ml) and 5.alpha.-cyprinol (97.44mg/0.6ml). After 6hr, the plasma of rats were examined. The value of BUN and creatinine in the plasma of rats treated with grass carp bile juice and 5.alpha.-cyprinol were significantly higher than that of saline group. The function of liver was also affected by raw bile juice, but not by 5.alpha.-cyprinol.

Keyword(s)

草魚膽汁
毒性
膽醇
膽酸
腎衰竭
Grass carp bile juice
Toxicity
Bile alcohol
Bile acid
Renal failure

DSpace CRIS

Toxicological Studies on Grass Carp Bile Juices

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