Applicability of monocyte activation test for pyrogen detection in succinylated gelatin injection

Background: Pyrogens are fever-inducing substances and pyrogen detection is mandatory in parenteral pharmaceuticals. Succinylated Gelatin Injection (SGI) is a biopharmaceutical product, containing multi-component, and it is administered parenterally. Objective: The study aimed to assess pyrogen in SGI and to evaluate the feasibility of the Monocyte Activation Test (MAT) for pyrogen detection in a multi-component pharmaceutical product. Method: In the present study, the Bacterial Endotoxin Test (BET) and the Monocyte Activation Test (MAT) were employed to assess pyrogen in SGI. The MAT method was developed on the basis of the HL-60/IL-6 assay. HL-60 cells were incubated with lipopolysaccharide (LPS) standards and sample solutions. The endotoxin produced by the incubation, interleukin-6 (IL-6), was measured by ELISA. The MAT method was validated and main parameters were investigated. Finally, the pyrogenicity of SGIs from two different enterprises was determined by the developed MAT method. Results: The BET failed in the test for interfering factors and the MAT was proved suitable for the pyrogen detection of SGI. All the products examined showed negative results in the pyrogen detection test. Conclusion: The MAT method is feasible in pyrogen detection of SGI. It can be applied in pyrogen detection for quality and safety control of multi-component biological products.