Analysis of Inborm Errors by High-Performance Liquid Chromatography

高效液相色谱法分析 Inborm 误差

• 批准号: 60460221
• 负责人: TSUJI Akio
• 金额: $ 2.5万
• 依托单位: School of Pharmaceutical Sciences, Showa University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1985
• 资助国家: 日本
• 起止时间: 1985 至 1987
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-60460221/
• 关键词: Fluorescence high-performance liquid chromatography; chemiluminescence high-performance liquid chromatography; congenital hypothyroidism; congenital adrenal hyperplasia; inborn error; 3 -hydroxysteroid dehydrogenase; corticosteroid; bile acid; fat

In this project, we have developed highly sensitive HPLC methods for biological substances related to inborn errors; congenital hypothyroidism and congenital adrenal hyperplasia etc.. The results are summarized as follows :1. Fluorescence HPLC method using an immobilized 3 -hydroxysteroid dehydrogenase column as a post-column enzyme reactor has been developed for the determination of corticosteroids and their metabolites in the urine of subjects with congenital adrenal hyperplasia. Each steroid was measured at the 2.5 ug/dl at the highest sensitivity of the detector. The mean recoveries and reproducibilities were 91.5 - 108.2% with 0.9 - 6.5% (CV%).2. Chemiluminescence HPLC method based on the chemiluminescence reaction of lucigenin has been developed for the determination of corticosteroids and their metabolites, and phenacyl esters of bile acids. The detection limits of steroids were about 50 pmol - 1 nmol. The CV% was 4.0 - 9.7%.3. Chemiluminescence HPLC method using N-(4-aminobutyl)-N-ethylisolumionl as a pre-column reagent has been developed for the determination of bile acids. The chemiluminescent derivative of cholic acid was 20 fmol. This value was higher than that of chemiluminescence HPLC using 3 -hydroxysteroid dehydrogenase reactor.4. Chemiluminescence determination of NADH using flow injection system and its application to HPLC detection system have been developed. Bile acids and related 3 -hydroxysteroids could be assayed by this method.5. The HPLC method for the determination of free fatty acids in plasma by using phenacyl bromide as a pre-column reagent has also developed.

本项目开发了与先天性缺陷、先天性甲状腺功能减退症、先天性肾上腺皮质增生症等相关的生物物质的高灵敏度HPLC检测方法。主要研究结果如下：1.采用固定化3 -羟基类固醇脱氢酶柱作为柱后酶反应器，建立了测定先天性肾上腺皮质增生症患者尿液中皮质类固醇及其代谢产物的荧光高效液相色谱法。每种类固醇在检测器的最高灵敏度下以2.5 ug/dl测量。方法的平均回收率为91.5 - 108.2%，变异系数为0.9 - 6.5%（CV%）.基于光泽精的化学发光反应，建立了化学发光高效液相色谱法测定皮质类固醇及其代谢物和苯甲酰甲基胆汁酸酯。甾体化合物的检测限约为50 pmol - 1 nmol。CV%为4.0 - 9.7%。本文建立了以N-（4-氨基丁基）-N-乙基异柳胺为柱前试剂测定胆汁酸的化学发光高效液相色谱法。胆酸的荧光衍生物为20 fmol。该值高于3 -羟基类固醇脱氢酶反应器的化学发光HPLC法.本文研究了流动注射化学发光法测定NADH及其在高效液相色谱检测系统中的应用。该方法可用于胆汁酸及相关3 -羟基甾体的测定.建立了以苯甲酰甲基溴为柱前试剂测定血浆中游离脂肪酸的高效液相色谱法。

项目成果

Akio Tsuju: "Bioluminescence and Chemiluminescence:Instruments and Applications." CRC Press, Inc., Boca Roton, Florida, USA, 241 (1985)

Akio Tsuju：“生物发光和化学发光：仪器和应用。”

Takao Kawasaki, Maeda and Akio Tsuji: "Fluorescence HPLC of 3<alpha>-Hydroxysteroids in Urine of 21-Hydroxylase Deficiency" Biomedical Chromatography. 1. 1-6 (1986)

Takao Kawasaki、Maeda 和 Akio Tsuji：“21-羟化酶缺乏症尿液中 3α-羟基类固醇的荧光 HPLC”生物医学色谱法。

Takao Kawasaki: J.Biomedical Chromatography. 1. 1-6 (1986)

Takao Kawasaki：J.Biomedical Chromatography。

Takao Kawasaki: Biomedica Chromatography. 1. 1-6 (1986)

Takao Kawasaki：生物医学色谱法。

ohinori Uji: Biomedica Chromatography. 2. 110-114 (1987)

ohinori Uji：生物医学色谱法。