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Determination of Reduced and Oxidized Coenzyme Q10 in Canine Plasma and Heart Tissue by HPLC-ECD: Comparison with LC-MS/MS Quantification

1
Section for Experimental Animal Models, Department of Veterinary and Animal Sciences, University of Copenhagen, Ridebanevej 9, 1870 Frederiksberg C, Denmark
2
Section for Environmental Chemistry and Physics, Department of Plant and Environmental Sciences, Thorvaldsensvej 40, 1870 Frederiksberg C, Denmark
*
Author to whom correspondence should be addressed.
Antioxidants 2019, 8(8), 253; https://doi.org/10.3390/antiox8080253
Received: 19 June 2019 / Revised: 11 July 2019 / Accepted: 25 July 2019 / Published: 30 July 2019
(This article belongs to the Section Methods for Antioxidants Evaluation/Measurement)
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Abstract

Coenzyme Q10 (Q10) plays an important role in mammals for energy production in the mitochondria, and as a potent antioxidant. Oxidation ratio (% oxidized in relation to total Q10) has been proposed as an important biomarker. A sensitive and reproducible HPLC-ECD method was developed for determination of reduced and oxidized Q10 in canine plasma and heart tissue. Chromatographic separation was achieved in 10 min using a Waters Nova-pak C18 column and a mobile phase with lithium perchlorate in ethanol/methanol/2-propanol. The validation showed satisfying results. Excellent linear correlation was found (r2 > 0.9997), intra- and inter-day precisions were below 6.5% (n = 5) and recoveries were between 89 and 109% (n = 5). Sensitivity stated as Lower Limit of Quantification (LLOQ) was 10 nM. Acceptable stability of both extracted and un-extracted samples was observed. The plasma concentration range of total Q10 was found to be between 0.64 and 1.24 µg/mL. Comparison with a developed LC-MS/MS method showed a correlation of r = 0.85 for reduced Q10 and r = 0.60 for oxidized Q10 (N = 17). However, average results were around 30% lower for ubiquinol using the LC-MS/MS method as compared with the HPLC-ECD analysis. The two methods are therefore not considered to be interchangeable. View Full-Text
Keywords: coenzyme Q10; oxidative stress; congestive heart failure; HPLC; LC-MS/MS coenzyme Q10; oxidative stress; congestive heart failure; HPLC; LC-MS/MS
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Schou-Pedersen, A.M.V.; Schemeth, D.; Lykkesfeldt, J. Determination of Reduced and Oxidized Coenzyme Q10 in Canine Plasma and Heart Tissue by HPLC-ECD: Comparison with LC-MS/MS Quantification. Antioxidants 2019, 8, 253.

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