Development and validation of a spectrophotometric method for quantification of residual cyclodextrin (DIMEB; Heptakis) in pertussis antigens. (February 2023)

• Record Type: Journal Article
• Authors: Shinde, Bharat; Patil, Dadasaheb; Nandre, Vinod; Gautam, Manish; Doshi, Pooja; Gairola, Sunil
• Is Part Of: Biologicals. Volume 81(2023)
• Publication Date: 2023-02
• Subjects: DIMEB -- Acellular pertussis -- Spectrophotometric -- Heptakis -- Anthrone reagent; Biological products -- Standards -- Periodicals; Biological Products -- Periodicals; Biological Products -- standards -- Periodicals; Produits biologiques -- Normes -- Périodiques; Biological products -- Standards; Periodicals; 615.37
• DOI: 10.1016/j.biologicals.2023.101663
• Languages: English
• ISSNs: 1045-1056

Abstract

Abstract: Methylated derivatives of cyclodextrins such as DIMEB (2, 6-di-O-methyl)-β-cyclodextrin or Heptakis is commonly used as culture medium modifier in manufacturing of pertussis antigens for promoting the growth of bacteria. We report here development and validation of a spectrophotometric method for estimation of DIMEB in different product matrices of pertussis vaccine antigens i.e. Filamentous haemagglutinin (FHA), Pertactin (PRN) and Pertussis toxin (PT). The detection is based on characteristic reaction of hydrolyzed sugars derivatives from DIMEB i.e., furfural derivatives with anthrone reagent to form colored complexes which could be quantified at 625 nm. Method showed excellent linearity with correlation coefficient (R 2 ) > 0.995 over the concentration of 5.0–80.0 μg. LOD and LOQ of 1.47 μg and 4.46 μg respectively was reported. The overall precision (repeatability and intermediate precision) showed % RSD for DIMEB content <10.0% for all the matrices. % Recoveries for DIMEB after three different spike levels (low, middle and high) were within 90%–113%. The method was successfully applied for determination of residual DIMEB in different product matrices of FHA, PRN and PT protein antigens. This can be used to monitor residual DIMEB levels during manufacturing of acellular pertussis antigens.