Beljean-Leymarie M, Bruna E
U.F.R. des Sciences Pharmaceutiques, Caen, France.
Anal Biochem. 1988 Aug 15;173(1):174-84. doi: 10.1016/0003-2697(88)90175-3.
A method for the measurement of malondialdehyde (MDA) using GLC with a nitrogen phosphorus detector (GLC/NPD) is described and evaluated. The method uses 2-hydrazinobenzothiazole (HBT) which forms condensation derivatives with MDA, acetoacetaldehyde, and acetylacetone (AA). GC/MS and 13C NMR studies of the three derivatives obtained showed that they are 2-(pyrazol-1'-yl)benzothiazoles and that they can be separated by GLC/NPD. Any one of these derivatives can be used as an internal standard for the measurement of the other two. The optimal conditions for the measurement of MDA were studied. At pH 2.5 and 70 degrees C, the condensation derivative is quantitatively formed in 30 min. Its extraction is obtained by a mixture of n-hexane/isoamyl alcohol 98/2 (v/v) containing HBT-AA as internal standard. The GLC detection limit is 0.04 pmol. Inter- and intrassay coefficients of variation were 2.9 and an average of 4.0%, respectively. The method is specific, and there was no interference from other carbonyl compounds. The artifactual formation of MDA from carbohydrates during the derivatization reaction is negligible. The method is proposed as a reference method for the standardization of working solutions of MDA or MDA-generating solutions.
描述并评估了一种使用带氮磷检测器的气相色谱法（GLC/NPD）测量丙二醛（MDA）的方法。该方法使用2-肼基苯并噻唑（HBT），它与MDA、乙酰乙醛和乙酰丙酮（AA）形成缩合衍生物。对得到的三种衍生物进行的气相色谱/质谱联用（GC/MS）和碳-13核磁共振（13C NMR）研究表明，它们是2-（吡唑-1'-基）苯并噻唑，并且可以通过GLC/NPD分离。这些衍生物中的任何一种都可以用作测量其他两种的内标。研究了测量MDA的最佳条件。在pH 2.5和70℃下，缩合衍生物在30分钟内定量形成。通过含有HBT-AA作为内标的正己烷/异戊醇98/2（v/v）混合物进行萃取。GLC检测限为0.04皮摩尔。批间和批内变异系数分别为2.9%和平均4.0%。该方法具有特异性，不受其他羰基化合物的干扰。衍生化反应过程中由碳水化合物人为形成的MDA可忽略不计。该方法被提议作为MDA工作溶液或产生MDA的溶液标准化的参考方法。
