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, Vladimir G. Kukes.

Journal Name: Current Pharmaceutical Analysis

Volume 15 , Issue 7 , 2019

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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.

[1]
Saini, R. Coenzyme Q10: The essential nutrient. J. Pharm. Bioallied Sci., 2011, 3(3), 466-467.
[2]
Bhagavan, H.N.; Chopra, R.K. Coenzyme Q10: absorption, tissue uptake, metabolism and pharmacokinetics. Free Radic. Res., 2006, 40(5), 445-453.
[3]
Sharma, A.; Fonarow, G.C.; Butler, J.; Ezekowitz, J.A.; Felker, G.M. Coenzyme Q10 and heart failure: A state-of-the-art review. Circ Heart Fail, 2016, 9(4)e002639
[4]
Garrido-Maraver, J.; Cordero, M.D.; Oropesa-Avila, M.; Vega, F.A.; de la Mata, M.; Delgado Pavon, A.; de Miguel, M.; Perez, C.C.; Villanueva, P.M.; Cotan, D.; Sanchez-Alcazar, J.A. Coenzyme q10 therapy. Mol. Syndromol., 2014, 5(3-4), 187-197.
[5]
Nohl, H.; Gille, L.; Staniek, K. The biochemical, pathophysiological, and medical aspects of ubiquinone function. Ann. N. Y. Acad. Sci., 1998, 854, 394-409.
[6]
Siciliano, G.; Volpi, L.; Piazza, S.; Ricci, G.; Mancuso, M.; Murri, L. Functional diagnostics in mitochondrial diseases. Biosci. Rep., 2007, 27(1-3), 53-67.
[7]
Garrido-Maraver, J.; Cordero, M.D.; Oropesa-Avila, M.; Vega, A.F.; de la Mata, M.; Pavon, A.D.; Alcocer-Gomez, E.; Calero, C.P.; Paz, M.V.; Alanis, M.; de Lavera, I.; Cotan, D.; Alcazar, J.A. Clinical applications of coenzyme Q10. Front. Biosci. (Landmark Ed), , 2014, 19, 619-633.
[8]
Groneberg, D.A.; Kindermann, B.; Althammer, M.; Klapper, M.; Vormann, J.; Littarru, G.P.; Doring, F. Coenzyme Q10 affects expression of genes involved in cell signalling, metabolism and transport in human CaCo-2 cells. Int. J. Biochem. Cell Biol., 2005, 37(6), 1208-1218.
[9]
Wada, H.; Hagiwara, S.; Saitoh, E.; Ieki, R.; Okamura, T.; Ota, T.; Iguchi, M.; Yuasa, K.; Kodaka, T.; Koishi, T.; Yamamoto, Y.; Goto, H. Increased oxidative stress in patients with Chronic Obstructive Pulmonary Disease (COPD) as measured by redox status of plasma coenzyme Q10. Pathophysiology, 2006, 13(1), 29-33.
[10]
Quinzii, C.M.; Lopez, L.C.; Von-Moltke, J.; Naini, A.; Krishna, S.; Schuelke, M.; Salviati, L.; Navas, P.; DoMauro, S.; Hirano, M. Respiratory chain dysfunction and oxidative stress correlate with severity of primary CoQ10 deficiency. FASEB J., 2008, 22(6), 1874-1885.
[11]
Adarsh, K.; Kaur, H.; Mohan, V. Coenzyme Q10 (CoQ10) in isolated diastolic heart failure in hypertrophic cardiomyopathy (HCM). Biofactors, 2008, 32(1-4), 145-149.
[12]
Celik, T.; Iyisoy, A. Coenzyme Q10 and coronary artery bypass surgery: What we have learned from clinical trials. J. Cardiothorac. Vasc. Anesth., 2009, 23(6), 935-936.
[13]
Rosenfeldt, F.L.; Haas, S.J.; Krum, H.; Hadj, A.; Ng, K.; Leong, J.Y.; Watts, G.F. Coenzyme Q10 in the treatment of hypertension: a meta-analysis of the clinical trials. J. Hum. Hypertens., 2007, 21(4), 297-306.
[14]
Golbidi, S.; Ebadi, S.A.; Laher, I. Antioxidants in the treatment of diabetes. Curr. Diabetes Rev., 2011, 7(2), 106-125.
[15]
Roffe, L.; Schmidt, K.; Ernst, E. Efficacy of coenzyme Q10 for improved tolerability of cancer treatments: A systematic review. J. Clin. Oncol., 2004, 22(21), 4418-4424.
[16]
Contin, M.; Flor, S.; Martinefski, M.; Lucangioli, S.; Tripodi, V. New analytical strategies applied to the determination of Coenzyme Q10 in biological matrix. Methods Mol. Biol., 2015, 1208, 409-420.
[17]
Andrade, S.C.; Guine, R.P.F.; Roseiro, L.C.P. In: 9th Baltic Conference on Food Science and Technology “Food for consumer wellbeing”, Jelgava, Latvia, May 8-9, 2014, Rakcejeva, T.; Hampshire, J.; Hansen, A.S.; Karklina, D.; Lantto, R.; Shleikin, A.; Straumite, E.; Talou T.; Venskutonis, R.; Vokk, R.; Yalcin, E., Eds.; Riga, Latvia, 2014, 51-52.
[18]
Leray, C.; Andriamampandry, M.D.; Freund, M.; Gachet, C.; Cazenave, J.P. Simultaneous determination of homologues of vitamin E and coenzyme Q and products of alpha-tocopherol oxidation. J. Lipid Res., 1998, 39(10), 2099-2105.
[19]
Tang, P.H.; Miles, M.V.; DeGrauw, A.; Hershey, A.; Pesce, A. HPLC analysis of reduced and oxidized coenzyme Q(10) in human plasma. Clin. Chem., 2001, 47(2), 256-265.
[20]
Acworth, I.N.; Ullucci, P.A.; Gamache, P.H. Determination of oxidized and reduced CoQ10 and CoQ9 in human plasma/serum using HPLC-ECD. Methods Mol. Biol., 2008, 477, 245-258.
[21]
Rodriguez-Estrada, M.T.; Poerio, A.; Mandrioli, M.; Lercker, G.; Trinchero, A.; Tosi, M.R.; Tugnoli, V. Determination of coenzyme Q10 in functional and neoplastic human renal tissues. Anal. Biochem., 2006, 357(1), 150-152.
[22]
Karpinska, J.; Mikoluc, B.; Motkowski, R.; Piotrowska-Jastrzebska, J. HPLC method for simultaneous determination of retinol, alpha-tocopherol and coenzyme Q10 in human plasma. J. Pharm. Biomed. Anal., 2006, 42(2), 232-236.
[23]
Abdul-Rasheed, O.F.; Farid, Y.Y. Development of a new high performance liquid chromatography method for measurement of coenzyme Q10 in healthy blood plasma. Saudi Med. J., 2009, 30(9), 1138-1143.
[24]
Mosca, F.; Fattorini, D.; Bompadre, S.; Littarru, G.P. Assay of coenzyme Q(10) in plasma by a single dilution step. Anal. Biochem., 2002, 305(1), 49-54.
[25]
Teshima, K.; Kondo, T. Analytical method for ubiquinone-9 and ubiquinone-10 in rat tissues by liquid chromatography/turbo ion spray tandem mass spectrometry with 1-alkylamine as an additive to the mobile phase. Anal. Biochem., 2005, 338(1), 12-19.
[26]
Claessens, A.J.; Yeung, C.K.; Risler, L.J.; Phillips, B.R.; Himmelfarb, J.; Shen, D.D. Rapid and sensitive analysis of reduced and oxidized coenzyme Q10 in human plasma by ultra performance liquid chromatography-tandem mass spectrometry and application to studies in healthy human subjects. Ann. Clin. Biochem., 2016, 53(Pt 2), 265-273.
[27]
Ruiz-Jimenez, J.; Priego-Capote, F.; Mata-Granados, J.M.; Quesada, J.M.; Luque de Castro, M.D. Determination of the ubiquinol-10 and ubiquinone-10 (coenzyme Q10) in human serum by liquid chromatography tandem mass spectrometry to evaluate the oxidative stress. J. Chromatogr. A, 2007, 1175(2), 242-248.
[28]
Matuszewski, B.K.; Constanzer, M.L.; Chavez-Eng, C.M. Strategies for the assessment of matrix effect in quantitative bioanalytical methods based on HPLC-MS/MS. Anal. Chem., 2003, 75(13), 3019-3030.
[29]
Kumar, A.; Kaur, H.; Devi, P.; Mohan, V. Role of coenzyme Q10 (CoQ10) in cardiac disease, hypertension and Meniere-like syndrome. Pharmacol. Ther., 2009, 124(3), 259-268.
[30]
Gvozdjakova, A.; Kucharska, J.; Mizera, S.; Braunova, Z.; Schreinerova, Z.; Schramekova, E.; Pechan, I.; Fabian, J. Coenzyme Q10 depletion and mitochondrial energy disturbances in rejection development in patients after heart transplantation. Biofactors, 1999, 9(2-4), 301-306.
[31]
Kucharska, J.; Gvozdjakova, A.; Mizera, S.; Braunova, Z.; Schreinerova, Z.; Schramekova, E.; Pechan, I.; Fabian, J. Participation of coenzyme Q10 in the rejection development of the transplanted heart: a clinical study. Physiol. Res., 1998, 47(6), 399-404.
[32]
Manzoli, U.; Rossi, E.; Littarru, G.P.; Frustaci, A.; Lippa, S.; Oradei, A.; Aureli, V. Coenzyme Q10 in dilated cardiomyopathy. Int. J. Tissue React., 1990, 12(3), 173-178.
[33]
Soongswang, J.; Sangtawesin, C.; Durongpisitkul, K.; Laohaprasitiporn, D.; Nana, A.; Punlee, K.; Kangkagate, C. The effect of coenzyme Q10 on idiopathic chronic dilated cardiomyopathy in children. Pediatr. Cardiol., 2005, 26(4), 361-366.
[34]
Chen, H.H.; Yeh, T.C.; Cheng, P.W.; Ho, W.Y.; Ho, C.Y.; Lai, C.C.; Sun, G.C.; Tseng, C.J. Antihypertensive potential of coenzyme Q10 via free radical scavenging and enhanced akt-nNOS signaling in the nucleus Tractus solitarii in rats. Mol. Nutr. Food Res., 2019, 63(6)1801042
[35]
Hajihashemi, P.; Askari, G.; Khorvash, F. The effects of concurrent Coenzyme Q10, L-carnitine supplementation in migraine prophylaxis: A randomized, placebo-controlled, double-blind trial. Cephalalgia, 2019, 39(5), 648-654.
[36]
Aksoy, A.; Kurnaz, S.Ç. An investigation of oxidative stress and coenzyme Q10 levels in patients with head and neck squamous cell carcinomas. Eur. Arch. Otorhinolaryngol., 2019, 276(4), 1197-1204.
[37]
Gurkan, A.S.; Dundar, O.B. Coenzyme Q10. J. Fac. Pharm. Ankara, 2005, 34(2), 129-154.


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Article Details

VOLUME: 15
ISSUE: 7
Year: 2019
Page: [795 - 807]
Pages: 13
DOI: 10.2174/1573412915666190328215854
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