Miniaturized HPLC-UV method for analysis of Coenzyme Q10 in human plasma

CONTIN M.,; LUCANGIOLI S.,; MARTINEFSKI M.,; FLOR S.,; TRIPODI V.

JOURNAL OF LIQUID CHROMATOGRAPHY AND RELATED TECHNOLOGIES

TAYLOR & FRANCIS INC

Lugar: Boca Raton; Año: 2011 p. 2485 - 2494

1082-6076

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. It is increasingly required the development of simple, rapid and highly sensitive methods capable to quantify low CoQ10 levels in plasma. In this work, we developed a miniaturized HPLC-UV system for the determination CoQ10 in human plasma using an XTerra microcolumn (50 mm x 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 ºC 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.