Interference and blood sample preparation for a pyruvate enzymatic assay
Interference and blood sample preparation for a pyruvate enzymatic assay
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DOI：
10.1016/j.clinbiochem.2005.10.007
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发表时间：
2006-01-01
影响因子：
2.8
通讯作者：
Lin, SP
中科院分区：
文献类型：
--
作者：
Chuang, CK;Wang, TJ;Lin, SP
背景资料：为了评估循环衰竭的严重程度，使用乳酸脱氢酶对全血进行丙酮酸酶测定以催化丙酮酸转化为乳酸。我们调查了与血液样本采集和制备相关的可能影响结果的因素，包括血液脱蛋白的时间，样本储存的温度和溶血。将每个样品分成2份：一份储存在室温（RT）下，另一份保存在冰上。在采集后的不同时间，使用8%高氯酸（PCA）对样本进行脱蛋白;第一次脱蛋白是在抽血后立即进行（0 h），然后以1 h的间隔进行6 h，并在样本中过夜。在脱蛋白后不久，使用COBAS离心分析仪分析上清液样品。在另一组样品中，立即将血液脱蛋白，并将上清液储存在RT和4 ℃下，并如上所述在不同时间测定丙酮酸盐。最后，溶血对血液丙酮酸酶测定的影响也evaluated.Results：当样品储存在RT，丙酮酸水平保持恒定，直到第三个小时后，脱蛋白，当有一个约13.3%的丙酮酸浓度增加。当全血样品在脱蛋白前保持在4 ℃时，丙酮酸水平随时间显著降低，范围为37.8%至62.2%（配对t检验显示显著平均差异，P < 0.001）。在RT或4 ℃下储存的上清液中未观察到丙酮酸盐浓度的显著差异。溶血导致丙酮酸浓度增加33.7%，相当于0.18毫克丙酮酸每克每分升hemoglobin.Conclusions：对于丙酮酸酶法测定，保持在RT的全血样品不会导致丙酮酸水平的显着差异，只要样品立即脱蛋白。全血样本不应储存在冰浴中运输，溶血样本也不应用于血液丙酮酸酶测定。(c)2005年加拿大临床化学家协会。All rights reserved.
Background: To assess the severity of circulatory failure, a pyruvate enzymatic assay was performed on whole blood using lactate dehydrogenase to catalyze the conversion of pyruvate to lactate. We investigated factors related to blood sample collection and preparation that might influence the results, including the timing of blood deproteinization, temperature of sample storage, and hemolysis.Method: A total of 25 whole blood specimens were collected for this study. Each sample was divided into 2 parts: one stored at room temperature (RT) and another kept on ice. The samples were deproteinizied by using 8% perchloric acid (PCA) at varying times after collection; the first deproteinization was immediately after the blood was drawn (0 h), then at 1 h intervals for 6 h and also in samples kept overnight. The supernatant samples were analyzed soon after deproteinization using a COBAS Centrifugal Analyzer. In another set of samples, the blood was immediately deproteinized, and the supernatants were stored at RT and 4 degrees C and assayed for pyruvate at varying times, as above. Finally, the effect of hemolysis on the blood pyruvate enzymatic assay was also evaluated.Results: When samples were stored at RT, pyruvate levels remained constant until the third h after deproteinization, when there was an approximately 13.3% increase in pyruvate concentration. When whole blood samples were kept at 4 degrees C before deproteinization, pyruvate levels were significantly reduced over time, ranging from 37.8% to 62.2% (paired t test showed a significant mean difference, P < 0.001). No significant differences in pyruvate concentration were observed in supernatant stored at either RT or 4 degrees C. Hemolysis caused a 33.7% increase in the pyruvate concentration, equivalent to 0.18 mg pyruvate per gram per deciliter of hemoglobin.Conclusions: For a pyruvate enzymatic assay, keeping a whole blood sample at RT will not cause a significant difference in the pyruvate level as long as the sample is immediately deproteinized. Whole blood samples should not be stored in an ice bath for transport, nor should hemolyzed samples be used for a blood pyruvate enzymatic assay. (c) 2005 The Canadian Society of Clinical Chemists. All rights reserved.