Hashizume N, Ihara H, Ohtsuka M, Kadowaki H
Department of Laboratory Medicine, Ohashi Hospital, Toho University School of Medicine, Tokyo.
Rinsho Byori. 1997 Aug;45(8):785-9.
Since plasma is generally employed for amino acid analysis, we compared amino acid levels in plasma with those in serum for healthy individuals and examined the influence of separation and storage conditions on the stability of the samples. Then, we determined the amino acid levels of frozen serum samples obtained from sarin poisoned patients. A. Comparison of Amino Acid Levels in Plasma and Those in Serum Blood was collected from 5 healthy individuals. Then, heparinated plasma and serum were separated by centrifugation immediately after blood collection. Serum was also separated by centrifugation after standing whole blood at room temperature for 1 hour. Frozen plasma and serum were store at -40 degrees C for 5 months. All were subjected to analysis in an amino acid analyzer. It was found that the cystine (Cys) and 3-methyl-histidine (3-M-His) levels in serum and plasma were affected when stored in a frozen state, that the aspartate (Asp) level was changed according to the method of collecting serum, and that the taurine (Tau) and ornithine (Orn) levels were affected by standing blood. B. Analysis of Blood Taken from Sarin Poisoned Patients Twelve sarin poisoned patients were selected as subjects, and serum cholinesterase (Ch-E) and serum albumin (Alb) levels were determined. Amino acid analysis was conducted using an amino acid analyzer. Serum samples which had been obtained from the 6 patients and frozen and stored at -40 degrees C from 5 months were used for amino acid analysis. As a result, the serum Ch-E level decreased and the Alb level tended to rise. Since the Ch-E/Alb ratio was reduced in the sarin poisoned patients, it is considered useful for discrimination from liver cirrhosis in which both Ch-E and Alb levels decreased. Amino acid levels in the serum obtained from the sarin poisoned patients were compared with those of healthy individuals, both of which had been stored under the same conditions. There were significant differences in Asp, glutamate (Glu), phenylalanine (Phe), 3-M-His, glutamine (Gln), and Cys levels. The Glu, Phe, and Gln levels were not affected by storage of serum in a frozen state, while the Glu and Phe levels were elevated and the Gln level was reduced. Although Cys exhibited lower values in frozen serum samples, the Cys level was elevated with a rise in the serum Ch-E levels. Therefore, we deduced that Cys metabolism disorders also occur in sarin poisoning. As stated above, the Glu and Phe levels were elevated and the Gln and Cys levels were reduced, suggesting the presence of abnormal amino acid metabolism, in patients with sarin-poisoning.
由于血浆通常用于氨基酸分析,我们比较了健康个体血浆和血清中的氨基酸水平,并研究了分离和储存条件对样本稳定性的影响。然后,我们测定了从沙林中毒患者获得的冷冻血清样本中的氨基酸水平。A. 血浆和血清中氨基酸水平的比较 从5名健康个体采集血液。采血后立即通过离心分离肝素化血浆和血清。全血在室温下静置1小时后也通过离心分离血清。冷冻的血浆和血清在-40℃储存5个月。所有样本均在氨基酸分析仪中进行分析。结果发现,冷冻保存时血清和血浆中的胱氨酸(Cys)和3-甲基组氨酸(3-M-His)水平受到影响,天冬氨酸(Asp)水平根据血清采集方法而变化,牛磺酸(Tau)和鸟氨酸(Orn)水平受到血液静置的影响。B. 对沙林中毒患者血液的分析 选择12名沙林中毒患者作为研究对象,测定血清胆碱酯酶(Ch-E)和血清白蛋白(Alb)水平。使用氨基酸分析仪进行氨基酸分析。从6名患者获得并在-40℃冷冻保存5个月的血清样本用于氨基酸分析。结果,血清Ch-E水平降低,Alb水平有升高趋势。由于沙林中毒患者的Ch-E/Alb比值降低,因此认为这有助于与Ch-E和Alb水平均降低的肝硬化进行鉴别。将沙林中毒患者血清中的氨基酸水平与在相同条件下储存的健康个体的氨基酸水平进行比较。天冬氨酸、谷氨酸(Glu)、苯丙氨酸(Phe)、3-M-His、谷氨酰胺(Gln)和胱氨酸水平存在显著差异。Glu、Phe和Gln水平不受血清冷冻保存的影响,而Glu和Phe水平升高,Gln水平降低。虽然冷冻血清样本中的Cys值较低,但Cys水平随着血清Ch-E水平的升高而升高。因此,我们推断沙林中毒时也会发生Cys代谢紊乱。如上所述,沙林中毒患者的Glu和Phe水平升高,Gln和Cys水平降低,提示存在氨基酸代谢异常。