Development and verification of a gas chromatography method for determination of β-propiolactone residues in stock solution and semi-finished products of freeze-dried rabies vaccine for human use (Vero cells)
10.13200/j.cnki.cjb.004574
- VernacularTitle:冻干人用狂犬病疫苗（Vero细胞）原液、半成品中β-丙内酯残留量气相色谱检测方法的建立及验证
- Author:
WANG Wentong
- Publication Type:Journal Article
- Keywords:
Freeze-dried rabies vaccine for human use(Vero cells);
β-propiolactone(BPL);
Residues;
Gas chromatography
- From:
Chinese Journal of Biologicals
2025;38(10):1231-1234+1240
- CountryChina
- Language:Chinese
-
Abstract:
Objective To develop and verify a gas chromatography method for the determination of β-propiolactone(BPL)residues in stock solution and semi-finished products of freeze-dried rabies vaccine for human use(Vero cells), so as to provide experimental basis for the vaccine quality control.Methods The gas chromatography conditions were as follows:The chromatographic column was Agilent DB-624 capillary column(30 m × 0. 250 mm × 1. 40 μm) with the injection port temperature of 180 ℃, the detector temperature of 250 ℃, the column oven temperature of 120 ℃, and the split ratio of 30∶1; The carrier gas was nitrogen at the flow rate of 1. 0 mL/min with the injection volume of 1 μL for automatic injection. The specificity, limit of detection(LOD), accuracy, reproducibility, linear range and robustness of the method were verified, and the method was used to detect the residual amount of BPL in two batches of stock solution and semi-finished products of freeze-dried rabies vaccines for human use(Vero cells).Results Using acetonitrile as the diluent and an isothermal injection program, the resolution of the mixture of BPL standard and vaccine stock solution was more than 1. 5, and the retention time of BPL peak was the same as that of standard solution. After injecting six portions of the BPL standard solution, the RSDs of the peak area, retention time and recovery rate were all less than 5%. BPL had a good linear relationship within the range of 10 to 200 μg/mL. The regression equation was y = 0. 241 6 x + 0. 024 7 with the correlation coefficient(r) of 0. 999 93. The LOD was 2 μg/mL. Under different column temperature conditions, the RSDs of the test results, peak area and retention time of the BPL standard solution were all less than 10. 0%. BPL was not detected in either the vaccine stock solution or the semi-finished products.Conclusion A gas chromatographic method for the determination of BPL residues in the stock solution and semi-finished products of freeze-dried human rabies vaccine(Vero cells) was developed with good specificity, accuracy, reproducibility and durability, and can be used for the determination of BPL residues in the vaccine stock solution and semi-finished products.
