MINIATURIZED HPLC-UV METHOD FOR ANALYSIS OF COENZYME Q10 IN HUMAN PLASMA

• Published in: Journal of liquid chromatography & related technologies Vol. 34; no. 20; pp. 2485 - 2494
• Main Authors: Contin, Mario, Lucangioli, Silvia, Martinefski, Manuela, Flor, Sabrina, Tripodi, Valeria
• Format: Journal Article
• Language: English
• Published: Colchester Taylor & Francis Group 01.12.2011; Taylor & Francis; Taylor & Francis Ltd
• Subjects: Biological and medical sciences; coenzyme Q10; CoQ10, coenzyme Q10; EC, electrochemical detection; Evaporation; HPLC, high performance liquid chromatography; Investigative techniques, diagnostic techniques (general aspects); Liquid chromatography; LOD, limit of detection; LOQ, limit of quantitation; Medical sciences; micro fast HPLC; microcolumn; Miscellaneous. Technology; mitochondrial disease; Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques; plasma; UV, ultraviolet; Biological fluid; coenzyme Q10; Chemical analysis; HPLC chromatography; Enzymopathy; Mitochondrial disorder; Blood; mitochondrial disease; Blood plasma; Ubiquinone; Fast HPLC chromatography; microcolumn; Clinical biology; Quantitative analysis; Human; Validation; UV; Healthy subject; Quinone; Metabolic diseases; Routine analysis; Patient; Genetic disease; Coenzyme; Microbore column; Ultraviolet detector; Treatment; Miniaturization; micro fast HPLC; plasma
• ISSN: 1082-6076; 1520-572X
• DOI: 10.1080/10826076.2011.591028

Coenzyme Q10 (CoQ10) is a cofactor in the respiratory chain and a potent endogenous antioxidant. Abnormally low levels of CoQ10 in the circulatory system are often involved with pathological states such as heart failure, mitochondrial and muscular diseases, and cancer. The development of simple, rapid, and highly sensitive methods capable of quantifying low CoQ10 levels in plasma is, therefore, increasingly required. In this work, we developed a miniaturized HPLC-UV system for the determination CoQ10 in human plasma using an XTerra microcolumn (50 mm × 2.1 mm i.d., 3.5 µm particle size), methanol:water (98:2, v/v) as mobile phase, 275 nm, flow rate of 0.3 mL/min, 30 °C column temperature, and 2 µL injection volume. The extraction procedure consists of a plasma precipitation with 1-propanol and evaporation under nitrogen to allow a 1.5-fold enrichment. The chromatographic analysis was accomplished in 7 min. As a result, it was possible to quantitate CoQ10 in small sample volumes down to 0.07 µM and detect as low as 0.02 µM in real plasma, with good accuracy and precision, even in pathological conditions. The proposed method was found to be suitable for routine CoQ10 determination in clinical laboratories.