Citation: ZHOU Lang, LI Qiao-Jing, LI Yong-Sheng, GAO Xiu-Feng. Rapid Determination of Pyruvic Acid in Erythrocytes by Lysing-heating Fluorescence Capillary Method[J]. Chinese Journal of Analytical Chemistry, ;2012, 40(11): 1725-1729. doi: 10.3724/SP.J.1096.2012.20271 shu

Rapid Determination of Pyruvic Acid in Erythrocytes by Lysing-heating Fluorescence Capillary Method

1.
¹ School of Chemical Engineering, Sichuan University, Chengdu 610065, China;

Corresponding author: LI Yong-Sheng,
Received Date: 9 March 2012
Available Online: 25 May 2012

In the conventional method of erythrocytes treatment, there was the efflux of pyruvic acid in erythrocytes (PAE) during washing and separation of erythrocytes/leukocytes using the physiological saline. This phenomenon caused the deviation of the measured value of PAE from its real concentration. This research designed a method comprising the steps of employing the erythrocyte lysis buffer to specifically lyse the red blood cells, separating leukocytes and platelets by centrifugation, removing the protein by heating the hemolysate, and detecting PAE using the pyruvate-enzyme fluorescence capillary analysis. The optimized procedures for the blood pretreatment were as follows: at the centrifugal speed of 16000 r/min, the blood sample was centrifuged for 1 min to obtain erythrocytes, after the erythrocytes were lysed by the lysis buffer, the hemolysate was heated for 5 min at 100℃. Through the contrast experiment, it was found that this method was superior to other erythrocyte treatment method. The pyruvate determination method in this research had a good linearity in the range of 10-120 mmol/L, a detection limit of 0.98 mmol/L, and a sensitivity of 5.64 F L/mmol, it was 60 times higher than that of references method. This method offered a relative standard deviation of PAE less than 2.8% (n=11), and a recovery range of 98.3%-104.1%.
- Enzyme fluorescence capillary analysis,
- Pyruvic acid,
- Erythrocytes,
- Lysing-heating method
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沈阳化工大学材料科学与工程学院沈阳 110142