Development and validation of a spectrophotometric method for quantification of residual cyclodextrin (DIMEB; Heptakis) in pertussis antigens
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 est...
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| Published in | Biologicals Vol. 81; p. 101663 |
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| Main Authors | , , , , , |
| Format | Journal Article |
| Language | English |
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01.02.2023
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| 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 (R2) > 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. |
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| AbstractList | 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 (R2) > 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. 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 ) > 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. |
| ArticleNumber | 101663 |
| Author | Patil, Dadasaheb Gairola, Sunil Doshi, Pooja Nandre, Vinod Gautam, Manish Shinde, Bharat |
| Author_xml | – sequence: 1 givenname: Bharat surname: Shinde fullname: Shinde, Bharat organization: Serum Institute of India Pvt. Ltd, Hadapsar, Pune, 411028, Maharashtra, India – sequence: 2 givenname: Dadasaheb surname: Patil fullname: Patil, Dadasaheb organization: Serum Institute of India Pvt. Ltd, Hadapsar, Pune, 411028, Maharashtra, India – sequence: 3 givenname: Vinod surname: Nandre fullname: Nandre, Vinod organization: Department of Chemistry, Savitribai Phule Pune University, Pune, 411007, Maharashtra, India – sequence: 4 givenname: Manish surname: Gautam fullname: Gautam, Manish organization: Serum Institute of India Pvt. Ltd, Hadapsar, Pune, 411028, Maharashtra, India – sequence: 5 givenname: Pooja surname: Doshi fullname: Doshi, Pooja organization: Department of Chemistry, Savitribai Phule Pune University, Pune, 411007, Maharashtra, India – sequence: 6 givenname: Sunil surname: Gairola fullname: Gairola, Sunil email: sunil.gairola@seruminstitute.com organization: Serum Institute of India Pvt. Ltd, Hadapsar, Pune, 411028, Maharashtra, India |
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| Cites_doi | 10.1007/s10311-018-0779-7 10.1016/j.biologicals.2007.09.001 10.3390/molecules23051161 10.1248/bpb1978.5.741 10.1016/0008-6215(95)00071-Z 10.1016/j.ijpharm.2017.11.018 10.3390/pharmaceutics13030409 10.1007/BF00656816 10.1016/S0003-2697(02)00704-2 10.1016/j.ijpharm.2020.119689 10.1021/ac902664x 10.1016/j.chroma.2006.03.019 10.3390/biom9120801 10.1365/s10337-009-1022-2 10.1126/sciadv.aax9318 10.4049/jimmunol.1402027 10.1016/j.chroma.2007.10.049 10.1007/s13346-020-00725-4 |
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| Keywords | Spectrophotometric Heptakis Anthrone reagent DIMEB Acellular pertussis |
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parenteral drug carrier publication-title: J Incul Phenamen doi: 10.1007/BF00656816 contributor: fullname: Szejtli – volume: 44 start-page: 969 year: 2000 ident: 10.1016/j.biologicals.2023.101663_bib12 publication-title: WHO Food Addit Ser contributor: fullname: Abbott – volume: 17 issue: 5 year: 1983 ident: 10.1016/j.biologicals.2023.101663_bib8 article-title: Heptakis (2,6-Dimethyl). 8-Cyclodextrin: a novel growth stimulant for Bordetella pertussis phase I publication-title: J Clinical Mico contributor: fullname: Imaizumi – volume: 315 year: 2003 ident: 10.1016/j.biologicals.2023.101663_bib19 article-title: A microtitermodication of the anthrone sulfuric acid colorimetric assay for glucose-based carbohydrates publication-title: Anal Biochem doi: 10.1016/S0003-2697(02)00704-2 contributor: fullname: Laurentin – volume: 588 year: 2020 ident: 10.1016/j.biologicals.2023.101663_bib23 article-title: The lord of the nano rings: cyclodextrins and the battle against SARS-CoV-2 publication-title: Int J Pharm doi: 10.1016/j.ijpharm.2020.119689 contributor: fullname: Garrido – volume: 82 start-page: 1786 year: 2010 ident: 10.1016/j.biologicals.2023.101663_bib20 article-title: Automation of the anthrone assay for carbohydrate concentration determination publication-title: Anal Chem doi: 10.1021/ac902664x contributor: fullname: Turula – volume: 1116 start-page: 76 year: 2006 ident: 10.1016/j.biologicals.2023.101663_bib17 article-title: Novel stationary phases for high-performance liquid chromatography analysis of cyclodextrin derivatives publication-title: J Chromatogr A doi: 10.1016/j.chroma.2006.03.019 contributor: fullname: Szeman – volume: 9 start-page: 801 year: 2019 ident: 10.1016/j.biologicals.2023.101663_bib11 article-title: Cyclodextrins: emerging medicines of the new millennium publication-title: Biomolecules doi: 10.3390/biom9120801 contributor: fullname: Braga – volume: 69 start-page: 1089 year: 2009 ident: 10.1016/j.biologicals.2023.101663_bib16 article-title: Analytical method for quantification β-Cyclodextrin in milk, cream and butter by LC publication-title: Chromatographia doi: 10.1365/s10337-009-1022-2 contributor: fullname: Lopezl – volume: 6 year: 2020 ident: 10.1016/j.biologicals.2023.101663_bib4 article-title: Modified cyclodextrins as broad-spectrum antivirals publication-title: Sci Adv doi: 10.1126/sciadv.aax9318 contributor: fullname: Jones – volume: 194 year: 2015 ident: 10.1016/j.biologicals.2023.101663_bib6 article-title: Hydroxy-β-cyclodextrin spikes local inflammation that induces Th2 cell and T follicular helper cell response to the Coadministered antigen publication-title: J Immunol doi: 10.4049/jimmunol.1402027 contributor: fullname: Onishi – volume: 1189 start-page: 385 year: 2008 ident: 10.1016/j.biologicals.2023.101663_bib18 article-title: Characterisation of complex amphiphilic cyclodextrin mixtures by high performance liquid chromatography and mass spectrometry publication-title: J Chromatogr A doi: 10.1016/j.chroma.2007.10.049 contributor: fullname: Kieken – volume: 10 start-page: 678 year: 2020 ident: 10.1016/j.biologicals.2023.101663_bib7 article-title: Optimization of an mRNA vaccine assisted with cyclodextrin-polyethyleneimine conjugates publication-title: Drug Deliv Transl Res doi: 10.1007/s13346-020-00725-4 contributor: fullname: Tan |
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| SubjectTerms | Acellular pertussis Anthrone reagent Antibodies, Bacterial Bordetella pertussis Cyclodextrins DIMEB Heptakis Humans Pertussis Toxin Pertussis Vaccine Spectrophotometric Whooping Cough - prevention & control |
| Title | Development and validation of a spectrophotometric method for quantification of residual cyclodextrin (DIMEB; Heptakis) in pertussis antigens |
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