Determination of a method for extraction of coenzyme Q10 in human plasma: optimization of the use of surfactants and other variables.
OBJECTIVE
To establish a routine for the extraction of the total levels of CoQ10 in human plasma through the Ultra High Performance Liquid Chromatography (UHPLC).


METHODS
Two extraction protocols were tested: a) methanol: hexane and b) 1-propanol. The following parameters were analyzed: extraction temperature (19ºC and 4ºC), extraction tubes (glass and polypropylene), and surfactants (SDS, Triton X-100, Tween-20) at different concentrations, i.e., 1%, 3%, 5% and 10%.


RESULTS
The results showed that the method of extraction of CoQ10 in a sample of human plasma at 4ºC, using solvents methanol: hexane (85:15, v/v) in the presence of surfactant Tween-20 at 3% and polypropylene tubes showed better efficiency and reproducibility when compared to the method with 1-propanol.


CONCLUSIONS
By the analyses performed, it was possible to observe that the addition of the surfactant Tween-20 promoted an increase in the recovery of CoQ10 by the methanol:hexane extraction method. This method showed good reproducibility, with a low coefficient of variation and high sensitivity, since CoQ10 was detected in samples of plasma of a control individual using a UV-type detector. The use of UHPLC equipment allowed a total analysis with total run time of 3.5 minutes, enabling the rapid achievement of results, considered mandatory for laboratory routines.
Hospital Israelita Albert Einstein, Instituto do Cérebro, São Paulo, SP, Brazil.
Claudia Cristina
CC
Ferreiro-Barros
Eduardo Kinio
EK
Sugawara
Livia Rentas
LR
Sanches
D002851:Chromatography, High Pressure Liquid; D057075:Enzyme Assays / Q000379:methods*; D006801:Humans; D011136:Polysorbates / Q000494:pharmacology; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet; D013501:Surface-Active Agents / Q000494:pharmacology*; D013696:Temperature; D014451:Ubiquinone / Q000031:analogs & derivatives* / Q000097:blood / Q000302:isolation & purification
D011136:Polysorbates; D013501:Surface-Active Agents; D014451:Ubiquinone; C024989:coenzyme Q10