Simultaneous determination of norethindrone and ethinyl estradiol in human plasma by high performance liquid chromatography with tandem mass spectrometry—experiences on developing a highly selective method using derivatization reagent for enhancing sensitivity
| Autor: | Weng Naidong, Shaolian Zhou, Yinghe Li, Austin C. Li, Wenkui Li |
|---|---|
| Rok vydání: | 2005 |
| Předmět: |
Clinical Biochemistry
Ethinyl Estradiol Tandem mass spectrometry Sensitivity and Specificity Biochemistry High-performance liquid chromatography Mass Spectrometry Analytical Chemistry Matrix (chemical analysis) chemistry.chemical_compound Drug Stability Liquid chromatography–mass spectrometry Ethinylestradiol medicine Humans Derivatization Chromatography High Pressure Liquid Dansyl Compounds Chromatography Elution Dansyl chloride Reproducibility of Results Cell Biology General Medicine chemistry Norethindrone medicine.drug |
| Zdroj: | Journal of Chromatography B. 825:223-232 |
| ISSN: | 1570-0232 |
| DOI: | 10.1016/j.jchromb.2005.01.012 |
| Popis: | In the present work, for the first time, a liquid chromatographic method with tandem mass spectrometric detection (LC-MS/MS) for the simultaneous analysis of norethindrone, and ethinyl estradiol, was developed and validated over the concentration range of 50-10000pg/ml and 2.5-500pg/ml, respectively, using 0.5 ml of plasma sample. Norethindrone, ethinyl estradiol, and their internal standards norethindrone-(13)C2, and ethinyl estradiol-d4, were extracted from human plasma matrix with n-butyl chloride. After evaporation of the organic solvent, the extract was derivatized with dansyl chloride and the mixture was injected onto the LC-MS/MS system. The gradient chromatographic elution was achieved on a Genesis RP-18 (50 mm x 4.6 mm, 3 microm) column with mobile phase consisted of acetonitrile, water and formic acid. The flow rate was 1.0 ml/min and the total run time was 5.0 min. Important parameters such as sensitivity, linearity, matrix effect, reproducibility, stability, carry-over and recovery were investigated during the validation. The inter-day precision and accuracy of the quality control samples at low, medium and high concentration levels were6.8% relative standard deviation (RSD) and 4.4% relative error (RE) for norethindrone, and 4.2% RSD and 5.9% RE for ethinyl estradiol, respectively. Chromatographic conditions were optimized to separate analytes of interest from the potential interference peaks, arising from the derivatization. This method could be used for pharmacokinetic and drug-drug interaction studies in human subjects. |
| Databáze: | OpenAIRE |
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