STUDY ON APPLICATION OF KINETIC CHROMOGENIC METHOD FOR THE DETERMINATION OF ENDOTOXIN IN VACCINES AND MEDICAL

Abstract

Endotoxin residue in vaccines and medical biologicals is an ingredient that needs to be determined by its allowable limits. Currently, the Institute of Vaccines and Medical Biologicals (IVAC) is using the Gel Clot, a semi-quantitative method, it may not be suitable for products that need to determine the amount of endotoxin accurately. Therefore, the research team applied the Kinetic Chromogenic method to determine the endotoxin content in

our vaccines to enhance the level of endotoxin control for these products. The study uses the Kinetic-QCL kit (Lonza) in combination with the BioTek Elx808LBS plate reader/incubator (Lonza) and WinKQCLTM software (version 5.3.3) to perform the reaction according to the manufacturer's instructions. We are performing the test with 3 samples of each product type, repeating 3 times for each technician to confirm the calibration criteria, precision and recovery, repeatability, and robustness of the test method. Recovery (PPC%) and coefficient of variation (CV%) values were calculated by each criterion. The Kinetic Chromogenic method has been validated and the criteria required by the manufacturer, in which the absolute value of the r coefficient of the standard curve reaches 0.999, precision and recovery range from 103 to 125%, repeatability and robustness have CV% < 10%. The study has also discovered the sample preparation suitable for each product, which is heat treatment for influenza vaccine and combining both heat treatment and dilution in MgCl 2 solution for the serum sample. Good preparation helps eliminate inhibition and enhancement factors to LAL reaction.