一种用于测定硫代巴比妥酸反应性物质（TBARS）的新型高效液相色谱法。与市售试剂盒的比较。

A novel HPLC method for the measurement of thiobarbituric acid reactive substances (TBARS). A comparison with a commercially available kit.

作者信息

Seljeskog Esben, Hervig Tor, Mansoor Mohammad Azam

机构信息

Division of Medical Biochemistry, PO Box 8100, N-4068 Stavanger, Norway.

出版信息

Clin Biochem. 2006 Sep;39(9):947-54. doi: 10.1016/j.clinbiochem.2006.03.012. Epub 2006 Apr 4.

DOI:10.1016/j.clinbiochem.2006.03.012

PMID:16781699

Abstract

OBJECTIVES

Malondialdehyde (MDA) as a part of thiobarbituric acid reacting substances (TBARS) is frequently used as an indicator of lipid peroxidation. Most methods for the measurement of TBARS require long derivatization time and addition of antioxidants in the samples. Furthermore, comparison of these methods with commercially available HPLC kits is lacking.

DESIGN AND METHODS

We investigated column performance of five different columns, tested eight different acids for the hydrolysis of the samples, and estimated stability of derivatized plasma samples with different anticoagulants. The samples were derivatized with TBA. The peak for the TBA(2)-MDA adduct was separated and detected by HPLC.

RESULTS

Performance of the Phenomenex Gemini column was best. PCA at the concentration of 0.1125 N was used in this method. Coefficient of variation (CV %) within the run and between the run was 4.1% and 6.7%, and analytical recovery was 90-94%. The retention time of the TBA(2)-MDA peak was 1.8 min. Reference intervals for TBARS in serum from 250 individuals were 0.53 and 2.1 micromol/l using our HPLC method and 0.07 and 0.24 micromol/l using the Chromsystems assay. Linear regression with log converted values revealed weak relationship between the two methods (r(2) = 0.064).

CONCLUSIONS

Our HPLC method for the analysis of TBARS in serum and plasma is fast and accurate and therefore can be used in clinical studies.

摘要

目的

丙二醛（MDA）作为硫代巴比妥酸反应物质（TBARS）的一部分，常被用作脂质过氧化的指标。大多数测定TBARS的方法需要较长的衍生化时间，且需在样品中添加抗氧化剂。此外，缺乏这些方法与市售高效液相色谱（HPLC）试剂盒的比较。

设计与方法

我们研究了五种不同色谱柱的柱性能，测试了八种不同的酸用于样品水解，并评估了不同抗凝剂处理的衍生化血浆样品的稳定性。样品用硫代巴比妥酸（TBA）进行衍生化。通过HPLC分离并检测TBA(2)-MDA加合物的峰。

结果

菲罗门Gemini色谱柱性能最佳。该方法使用浓度为0.1125 N的PCA。批内和批间变异系数（CV%）分别为4. l%和6.7%，分析回收率为90 - 94%。TBA(2)-MDA峰的保留时间为1.8分钟。使用我们的HPLC方法，250名个体血清中TBARS的参考区间为0.53和2.1微摩尔/升，使用Chromsystems检测法的参考区间为0.07和0.24微摩尔/升。对数转换值的线性回归显示两种方法之间的关系较弱（r(2)=0.064）。