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Analytical Method Development for Coq10 Determination in Human Plasma Using HPLC-UV and HPLC/MS/MS

Author(s):

Vladlena I. Zozina*, Evgeniy S. Melnikov, Olga A. Goroshko, Liudmila M. Krasnykh and Vladimir G. Kukes   Pages 1 - 14 ( 14 )

Abstract:


Background: CoQ10 is a very important compound which is found in every tissue of our organism. It participates in processes of cellular respiration and ATP production. Also it acts like a strong antioxidant. In organism it is represented in two forms: oxidized (ubiquinone) and reduced (ubiquinol). Its low blood level may be a signal for a list of diseases.

Materials and Methods: There were developed and compared two methods of the CoQ10 determination in order to find the fastest and the most convenient one. The first one involved HPLC-UV with the wavelength of ubiquinone determination equivalent to 290 nm and 275 nm for ubiquinol, respectively. The second one was carried out on a HPLC/MS/MS system utilizing electro spray ionization (ESI) and triple quadrupole mass analyzer for quantification in MRM positive mode.

Results: Two methods of ubiquinol and ubiquinone determination were developed and validated. HPLC-UV included sample preparation based on liquid-liquid extraction. The LLOQ constituted 5 µg/ml. HPLC-MS/MS method sample preparation was based on protein precipitation. The LLOQ constituted 0.10 µg/ml.

Conclusion: During the investigation it was drawn a conclusion that HPLC-UV method it too insensitive for simultaneous determination of ubiquinol and ubiquinone. Furthermore, ubiquinol is very unstable and during exogenous factors’ exposure it rapidly turns into ubiquinone. HPLC-MS/MS method turned out to be sensitive, selective, rapid and it provides accurate determination of both forms of CoQ10 in spiked human plasma.

Keywords:

CoQ10, ubiquinone, ubiquinol, HPLC-UV, HPLC-MS/MS.

Affiliation:

Depatment of Clinical Pharmacology and Propaedeutics of Internal diseases, Federal State Autonomous Educational Institution of Higher Education I.M. Sechenov First Moscow State Medical University of the Ministry of Health,, Department of Pharmaceutical and Toxicological Chemistry, I.M. Sechenov First Moscow State Medical University (Sechenov University); Department of Health I. V. Davydovsky Municipal Clinical Hospital, Federal State Budgetary Institution “Scientific Centre for Expert Evaluation of Medical Products” of the Ministry of Health,, Federal State Budgetary Institution “Scientific Centre for Expert Evaluation of Medical Products” of the Ministry of Health,, Federal State Budgetary Institution “Scientific Centre for Expert Evaluation of Medical Products” of the Ministry of Health,, Depatment of Clinical Pharmacology and Propaedeutics of Internal diseases, Federal State Autonomous Educational Institution of Higher Education I.M. Sechenov First Moscow State Medical University of the Ministry of Health



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