DEVELOPMENT AND COMPARISON OF HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHICMETHODS WITH TANDEM MASS-SPECTROMETRIC AND ULTRAVIOLET ABSORBENCY DETECTION FOR THE DETERMINATION OF CYCLOBENZAPRINE IN HUMAN PLASMA AND URINE
M. Constanzer et al., DEVELOPMENT AND COMPARISON OF HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHICMETHODS WITH TANDEM MASS-SPECTROMETRIC AND ULTRAVIOLET ABSORBENCY DETECTION FOR THE DETERMINATION OF CYCLOBENZAPRINE IN HUMAN PLASMA AND URINE, Journal of chromatography B. Biomedical applications, 666(1), 1995, pp. 117-126
Citations number
22
Categorie Soggetti
Chemistry Analytical
Journal title
Journal of chromatography B. Biomedical applications
Sensitive assays for the determination of cyclobenzaprine (I) in human
plasma and urine were developed utilizing high-performance liquid chr
omatography (HPLC) with tandem mass spectrometric (MS-MS) and ultravio
let (UV) absorbance detections. These two analytical techniques were e
valuated for reliability and sensitivity, and applied to support pharm
acokinetic studies. Both methods employed a liquid-liquid extraction o
f the compound from basified biological sample. The organic extract wa
s evaporated to dryness, the residue was reconstituted in the mobile p
hase and injected onto the HPLC system. The HPLC assay with MS-MS dete
ction was performed on a PE Sciex API III tandem mass spectrometer usi
ng the heated nebulizer interface. Multiple reaction monitoring using
the parent --> daughter ion combinations of mit 276 -->, 215 and 296 -
-> 208 was used to quantitate I and internal standard (II), respective
ly. The HPLC-MS-MS and HPLC-UV assays were validated in human plasma i
n the concentration range 0.1-50 ng/ml and 0.5-50 ng/ml, respectively.
In urine, both methods were validated in the concentration range 10-1
000 ng/ml. The precision of the assays, as expressed as coefficients o
f variation (C.V.) was less than 10% over the entire concentration ran
ge, with adequate assay specificity and accuracy. In addition to bette
r sensitivity, the HPLC-MS-MS assay was more efficient and allowed ana
lysis of more biological fluid samples in a single working day than th
e HPLC-UV method.