Design and methodological evaluation of rate method for determination of serum total cholesterol con-centration

LI Ying; LI Xiaoyan; LI Yuanhong

Abstract：Objective:To establish rate method for the detection of serum total cholesterol concentration,take meth-odological evaluation,then explore the practical feasibility of the method.Methods:A series of cholesterol standards with clinically available concentration range were prepared,and the process curve was drawn by observing the absor-bance change curve of chromogenic substances at a certain wavelength in the process of cholesterol oxidase-peroxidase method(COD-PAP)reaction.Determined reagent and sample size,incubation time,linear reaction time,K factor and other parameters of the reaction system and a series of parameters for the detection of serum total cholesterol by auto-matic equipment rate method werµe established.Finally,took methodological evaluation of the precision,linear charac-teristics,and reliability of this method.Results:The sample size was determined to be 10 pL and reagent was 1 mL.Af-ter incubating at 37 ℃ for 5 s,the absorbance change of chromogenic substances at 510 nm entered a linear phase,and the linear reaction phase basically ended at 18 s.The measurement time was set as 5 s and 15 s to determine.The K fac-tor was 23.28.The methodological evaluation showed that the reaction was linear in the concentration range of 1.32～10.00 mmol/L,with R2＞0.99.The method has good accuracy and CV value was 2.62％.The end-point method and rate method were used to determine the total cholesterol concentration in 20 batches of human serum,and paired T-test showed that there was no significant difference between the two methods(P＞0.05)and the rate method was ac-ceptable for the determination of serum total cholesterol concentration.Conclusion:The rate method with good lineari-ty,precision,and reliability could be used for total cholesterol concentration detection.