Development and verification of RP-HPLC for determination of benzalkonium bromide content in human interferon α1b spray
10.13200/j.cnki.cjb.004466
- VernacularTitle:人干扰素α1b喷雾剂中苯扎溴铵含量反相高效液相色谱检测方法的建立及验证
- Author:
LIU Linlin
- Publication Type:Journal Article
- Keywords:
Reversed-phase high performance liquid chromatography(RP-HPLC);
Human interferon α1b(hIFNα1b);
Spray;
Benzalkonium bromide
- From:
Chinese Journal of Biologicals
2025;38(04):467-472
- CountryChina
- Language:Chinese
-
Abstract:
Objective To develop a reversed-phase high performance liquid chromatography(RP-HPLC) method for the determination of benzalkonium bromide content in human interferon α1b(hIFNα1b) spray, and to verify and preliminarily apply the method in order to provide a reference for the dosage control of benzalkonium bromide in hIFNα1b spray.Methods The conditions of liquid chromatography were as follows: ZORBAX 300SB-C18 column(4. 6 mm × 250 mm, 5 μm); mobile phase was acetonitrile and 5 mmol/L ammonium acetate solution at a volume ratio of 65∶35; detection time was 30 min at flow rate of 1. 0 mL/min and column temperature of room temperature; UV detection wavelength was 262 nm; injection volume was 20 μL; isocratic elution. The system adaptability, specificity, linearity, accuracy, precision, limit of quantitation(LOQ),limit of detection(LOD), and robustness of the method were verified, and the content of benzalkonium bromide in three batches of hIFNα1b spray was determined by using this method.Results The method demonstrated good system applicability and specificity for benzalkonium bromide detection. A good linear relationship was observed within the concentration range of 10-100 μg/mL(R2 = 0. 999 7). The overall mean spiked recovery rate of accuracy verification was 97. 2% with an RSD of1. 61%. The RSDs of reproducibility and intermediate precision verification were 3. 36% and 1. 20%, respectively. The LOQ and LOD were 0. 02 and 0. 008 μg/mL, respectively. Minor variations in flow rate, mobile phase pH, and column temperature within specified ranges showed no significant impact on the analytical results. Quantitative analysis of three batches of hIFNα1b spray showed benzalkonium bromide contents of 45. 4, 45. 0, and 45. 2 μg/mL, respectively, all conforming to the labeled specification range of 40-60 μg/mL.Conclusion The RP-HPLC method has good specificity, accuracy, precision and robustness, and can be used for the determination of benzalkonium bromide in hIFNα1b spray.
