SIMPLE AND SENSITIVE ASSAY OF TOTAL SERUM BILE-ACIDS
SIMPLE AND SENSITIVE ASSAY OF TOTAL SERUM BILE-ACIDS
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DOI：
10.1016/0009-8981(76)90007-3
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发表时间：
1976-01-01
影响因子：
5
通讯作者：
OSUGA, T
中科院分区：
文献类型：
--
作者：
MASHIGE, F;IMAI, K;OSUGA, T
建立了一种简便、灵敏的测定血清总3-羟基胆汁酸的方法。血清(0.1ml)与三羟甲基氨基甲烷盐酸缓冲液混合，在67℃加热。C浸泡30分钟。在溶液中加入3α-羟基类固醇氧化还原酶(EC 1.1.1.50；3α-HSD)、NAD黄递酶(EC 1.6.4.3)和瑞珠灵。混合物在20度的温度下孵化。在560 nm激发下，在580 nm处测量间苯二酚的荧光强度。另取同一份血清0.1ml，不加3α-HSD，测得空白值，胆汁酸含量与荧光强度在1-150u.mol/L范围内呈线性关系，加样回收率为81.4.+-。胆酸盐、鹅去氧胆酸盐和脱氧胆酸盐为2.5(SD)%。血清胆汁酸含量为48.8u.mol/L，标准差为+-。0.42和.+-的变异系数。10次重复测定0.87%。正常空腹男性血清胆汁酸含量为8.0mu.mol/L(3.6~12.6mu.mol/L，n=12)，女性血清平均胆汁酸含量为6.8mu.mol/L(3.2~12.7mu.mol/L，n=13)。
A simple and sensitive method was developed for the quantification of serum total 3.alpha.-hydroxy bile acids. Serum (0.1 ml) was mixed with tris(hydroxymethyl) aminomethane hydrochloric acid buffer and heated at 67.degree. C for 30 min. To the solution were added 3.alpha.-hydroxysteroid:oxidoreductase (EC 1.1.1.50; 3.alpha.-HSD), NAD diaphorase (EC 1.6.4.3) and resazurin. The mixture was incubated at 20.degree. C for 1 h. The resultant fluorescence of resorfin was measured at 580 nm with the excitation at 560 nm. The blank value was obtained after the same treatment of another 0.1 ml of the same serum without 3.alpha.-HSD. A linear relationship was obtained between the amount of bile acids and the fluorescence intensities in the range of 1-150 .mu.mol/l. The recovery of bile acids added to the serum was 81.4 .+-. 2.5 (SD) % for cholate, chenodeoxycholate and deoxycholate. The bile acid content in the serum was 48.8 .mu.mol/l with a SD of .+-. 0.42 and a coefficient of variation of .+-. 0.87% in 10 replicate determinations. The mean bile acid content of normal fasting male sera was 8.0 .mu.mol/l (3.6-12.6 .mu.mol/l, n = 12) and female sera 6.8 .mu.mol/l (3.2-12.7 .mu.mol/l, n = 13).