Rapid quantitative determination of fat-soluble vitamins and coenzyme Q-10 in human serum by reversed phase ultra-high pressure liquid chromatography with UV detection.
We are presenting the first ultra-high pressure LC (UHPLC) method for rapid quantitative measurement of vitamin A, E (alpha- and gamma-tocopherol), beta-carotene and CoQ(10) from human serum. The chromatography was performed on Shield RP(18) UHPLC column with UV detection. The method was validated based on linearity, accuracy, matrix effects study, precision and stability. The calibration was linear over the following range: 0.09-10.0 for retinol and gamma-tocopherol, 0.05-5 for beta-carotene, 0.9-100 for alpha-tocopherol and 0.14-15 mg/L for CoQ(10). The limit of detection and quantitation for retinol, gamma-tocopherol, beta-carotene, alpha-tocopherol and CoQ(10) were as follows 0.07/0.024, 0.018/0.06, 0.004/0.12, 0.078/0.261, 0.008/0.028 mg/L. The recoveries were above 85%. The inter- and intra-assay precision was below 10%. Reference intervals were established for children and adults. Because of its low cost, extremely short analysis time (2 min) and excellent chromatographic reproducibility this UHPLC method can easily be adopted for high-throughput clinical and pharmacokinetics studies.
Paliakov
Ekaterina M
EM
Metametrix Clinical Laboratory, 3425 Corporate Way, Duluth, GA 30096, USA. epaliakov@metametrix.com
Crow
Brian S
BS
Bishop
Michael J
MJ
Norton
Dean
D
George
Joe
J
Bralley
J Alexander
JA
D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005223:Fats / Q000737:chemistry*; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012995:Solubility; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*; D014451:Ubiquinone / Q000031:analogs & derivatives* / Q000097:blood; D014815:Vitamins / Q000097:blood*
D005223:Fats; D014815:Vitamins; D014451:Ubiquinone; C024989:coenzyme Q10