title: A high performance liquid chromatographic method for the measurement of total carnitine in human plasma and urine.
abstract: Total carnitine in plasma and urine can be measured by high performance liquid chromatography (HPLC) using the novel fluorescent derivatisation reagents 6'-methoxynaphthacyl trifluoromethanesulfonate and 2'-phenanthrenacyl trifluoromethanesulfonate. Sample preparation for total carnitine analysis involves: extraction of plasma and urine in methanol, the optional addition of serine betaine as an internal standard, saponification of acyl carnitines with calcium hydroxide, followed by derivatisation with 6'-methoxynaphthacyl trifluoromethanesulfonate or 2'-phenanthrenacyl trifluoromethanesulfonate. The derivatives were separated using an alumina column and measured by fluorescence detection. The coefficient of variation was below 5% using internal standard calibration, and recoveries of acyl carnitines after saponification were over 90%. The total carnitine method was shown to be linear at biological levels for plasma (over the range 30-130 micromol/l) and urine (over the range 80-180 micromol/l). Advantages of this method include good precision, accuracy and linearity, the use of fluorescence to gain sensitivity, the small sample volume required and a relatively low sample preparation time.
authors.affiliation: Biochemistry Unit, Canterbury Health Laboratories, P.O. Box 151, Christchurch, New Zealand. chris.mcentyre@cdhb.govt.nz
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: McEntyre
authors.forename: Michael
authors.initials: M
authors.lastname: Lever
authors.forename: Malina K
authors.initials: MK
authors.lastname: Storer
mesh_terms: D002331:Carnitine / Q000097:blood* / Q000302:isolation & purification / Q000652:urine*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D019937:Diagnostic Techniques and Procedures / Q000592:standards; D006801:Humans; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D002331:Carnitine