title: Automated statistical experimental design approach for rapid separation of coenzyme Q10 and identification of its biotechnological process related impurities using UHPLC and UHPLC-APCI-MS.
abstract: A novel ultra high performance liquid chromatography method development strategy was ameliorated by applying quality by design approach. The developed systematic approach was divided into five steps (i) Analytical Target Profile, (ii) Critical Quality Attributes, (iii) Risk Assessments of Critical parameters using design of experiments (screening and optimization phases), (iv) Generation of design space, and (v) Process Capability Analysis (Cp) for robustness study using Monte Carlo simulation. The complete quality-by-design-based method development was made automated and expedited by employing sub-2 μm particles column with an ultra high performance liquid chromatography system. Successful chromatographic separation of the Coenzyme Q10 from its biotechnological process related impurities was achieved on a Waters Acquity phenyl hexyl (100 mm × 2.1 mm, 1.7 μm) column with gradient elution of 10 mM ammonium acetate buffer (pH 4.0) and a mixture of acetonitrile/2-propanol (1:1) as the mobile phase. Through this study, fast and organized method development workflow was developed and robustness of the method was also demonstrated. The method was validated for specificity, linearity, accuracy, precision, and robustness in compliance to the International Conference on Harmonization, Q2 (R1) guidelines. The impurities were identified by atmospheric pressure chemical ionization-mass spectrometry technique. Further, the in silico toxicity of impurities was analyzed using TOPKAT and DEREK software.
authors.affiliation: Department of Pharmaceutical Analysis, National Institute of Pharmaceutical Education & Research, IDPL, R&D Campus, Balanagar, Hyderabad, India. narendra.talluri@gmail.com.
authors.forename: Murali V N Kumar
authors.initials: MV
authors.lastname: Talluri
authors.affiliation: Department of Pharmaceutical Analysis, National Institute of Pharmaceutical Education & Research, IDPL, R&D Campus, Balanagar, Hyderabad, India.
authors.forename: Pradipbhai D
authors.initials: PD
authors.lastname: Kalariya
authors.affiliation: Department of Pharmaceutical Analysis, National Institute of Pharmaceutical Education & Research, IDPL, R&D Campus, Balanagar, Hyderabad, India.
authors.forename: Shireesha
authors.initials: S
authors.lastname: Dharavath
authors.affiliation: Department of Pharmacoinformatics, National Institute of Pharmaceutical Education and Research (NIPER), Sector-67, S.A.S (Mohali) Nagar, Punjab, India.
authors.forename: Naeem
authors.initials: N
authors.lastname: Shaikh
authors.affiliation: Department of Pharmacoinformatics, National Institute of Pharmaceutical Education and Research (NIPER), Sector-67, S.A.S (Mohali) Nagar, Punjab, India.
authors.forename: Prabha
authors.initials: P
authors.lastname: Garg
authors.affiliation: Mass and Analytical Division, Indian Institute of Chemical Technology, Tarnaka, Hyderabad.
authors.forename: Nageswara Rao
authors.initials: NR
authors.lastname: Ramisetti
authors.affiliation: Department of Pharmaceutical Analysis, National Institute of Pharmaceutical Education & Research, IDPL, R&D Campus, Balanagar, Hyderabad, India.|Mass and Analytical Division, Indian Institute of Chemical Technology, Tarnaka, Hyderabad.
authors.forename: Srinivas
authors.initials: S
authors.lastname: Ragampeta
mesh_terms: D001331:Automation / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004340:Drug Contamination; D057230:Limit of Detection; D013058:Mass Spectrometry / Q000379:methods*; D011786:Quality Control; D014451:Ubiquinone / Q000031:analogs & derivatives* / Q000032:analysis
chemical_list: D014451:Ubiquinone; C024989:coenzyme Q10
keywords: Coenzyme Q10; In silico toxicity; Mass spectrometry; Process impurities; Quality by design