Abstract:
A rapid, reproducible and accurate high-performance liquid chromatographic (HPLC) method for the quantitative determination of sphingomyelin in rat brain was developed and validated using normal-phase silica gel column, acetonitrile-methanol-water (65:18:17 (v-v)) at a flow rate of 1 ml-min, isocratic elution, UV detection at 207 nm and 1,2-dimyristoyl-sn-glycero-3-phosphocholine as an internal standard. Total run time was 10.0 min. The calibration curve was linear over the range of 0.025-0.4 mg-ml sphingomyelin (R2 andgt; 0.99). The intra-day coefficient of variation ranged from 1.4percent to 2.2percent. The average inter-day coefficient of variation over a period of 4 days was 3.1percent. The practical limit of detection was 0.005 mg-ml with a quantification limit of 0.01 mg-ml. © 2007 Elsevier B.V. All rights reserved.