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

[ Vol. 15 , Issue. 7 ]

Author(s):

Vladlena I. Zozina*, Evgeniy S. Melnikov, Olga A. Goroshko, Liudmila M. Krasnykh and Vladimir G. Kukes   Pages 795 - 807 ( 13 )

Abstract:


Background: CoQ10 is a very important compound which is found in every tissue of our organism. It participates in the processes of cellular respiration and ATP production. Also, it acts as a strong antioxidant. In an 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: This study developed and compared two methods of 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 an HPLC/MS/MS system utilizing Electrospray 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 was 0.50 µg/ml. HPLC-MS/MS method sample preparation was based on protein precipitation. The LLOQ was 0.10 µg/ml.

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

Keywords:

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

Affiliation:

Department 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 of the Russian Federation, Moscow, A.P. Arzamastsev 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 of the Russian Federation, Moscow, Federal State Budgetary Institution “Scientific Centre for Expert Evaluation of Medical Products” of the Ministry of Health of the Russian Federation, Moscow, Federal State Budgetary Institution “Scientific Centre for Expert Evaluation of Medical Products” of the Ministry of Health of the Russian Federation, Moscow, Department 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 of the Russian Federation, Moscow

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