Establishment and verification of headspace gas chromatography method for organic solvent residues in PEGylated recombinant Candida uricase for injection
Objective To establish and verify a headspace gas chromatography method for the determination of residual solvents,isopropanol(IPA),dichloromethane(DCM)and toluene(TOL),in PEGylated recombinant Candida uricase for injection(SSS111).Methods A headspace gas chromatography was established using a capillary column DB-1301(30m× 0.53 mm,1.0 μm),with the detector temperature of 250 ℃,the injector temperature of 200 ℃,the carrier gas of nitrogen,the split ratio of three,the column flow rate of 3.0 mL/min,the hydrogen flow rate of 60 mL/min,and the air flow rate of 400 mL/min.The column temperature was programmed raising:the initial temperature was 50℃,maintaining for 12 min,and rose to 200 ℃ at the rate of 40 ℃/min,maintaining for 5 min.The equilibrium temperature of the headspace bottle was 120 ℃,and the equilibrium time was 20 min.The linear range,precision and accuracy of the method were verified,and the limit of detection(LOD)and limit of quantitation(LOQ)were determined.IPA,DCM and TOL residues in three batches of SSS111 samples were detected by using the established method.Results IPA,DCM and TOL showed a good linear relationship with the peak area in the range of 0.001 982-7.928,0.002 118-8.472 and 0.041 960-8.392 μg/mL respectively,with each r of more than 0.99.The RSDs of IPA,DCM and TOL residues in the control solution in six repeated detections were all lower than 5％.The recoveries of IPA,DCM and TOL were all in the range of 80％-120％after control solution was added to SSS111 sample.The LODs of IPA,DCM and TOL were 1.982,2.118 and 41.96 ng/mL,and the LOQs were 6.941,6.236 and 126.92 ng/mL,respectively.The residues of IPA in the three batches of samples were all lower than the LOD,the residues of DCM were about 1 ppm,and the residues of TOL were all less than 1 ppm.Conclusion The estab-lished headspace gas chromatography method has good precision and accuracy,simple operation and high sensitivity,and can be used for the determination of organic solvent residues in SSS111.
PEGylated recombinant Candida uricase for injectionHeadspace gas chromatographyOrganic solventIsopropanol(IPA)Dichloromethane(DCM)Toluene(TOL)
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