Simple and sensitive dilute-and-shoot analysis of carotenoids in human plasma

Carotenoids in human plasma are used as biomarkers of vegetable and fruit intake. Large sample volumes and intensive sample processing make measurement of these species cumbersome. We developed a dilute-and-shoot method for the quantitation of alpha-carotene, beta-carotene, beta-cryptoxanthin, lycopene and lutein/zeaxanthin using 10 mu L of plasma. Plasma was injected into methanol containing internal standard and deproteinized by centrifugation. The carotenoids in the supernatant were separated using a YMC C-30 column and quantified by tandem mass spectrometry. The linearity for carotenoids ranged from sub-fmol to approximately 300 fmol on-column. Spike recovery experiments were used to correct for matrix effects. The method was validated using the human plasma standard NIST SRM 968e. Over 400 sample analyses were performed using the same guard and analytical columns. This method represents an improvement over current techniques because of the small sample size needed, ease of sample preparation, and improvement in the determination of carotenoid content.