Development and validation of a high-sensitivity liquid chromatography/tandem mass spectrometry (LC/MS/MS) method with chemical derivatization for the determination of ethinyl estradiol in human plasma
An ultra‐sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for the analysis of oral contraceptive ethinyl estradiol (EE) was developed and validated over the curve range of 2.5–500 pg/mL using 1 mL of human plasma sample. Ethinyl estradiol and the internal standard, ethinyl...
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| Published in | Biomedical chromatography Vol. 18; no. 7; pp. 414 - 421 |
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| Main Authors | , , |
| Format | Journal Article |
| Language | English |
| Published |
Chichester, UK
John Wiley & Sons, Ltd
01.09.2004
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| Subjects | |
| Online Access | Get full text |
| ISSN | 0269-3879 1099-0801 |
| DOI | 10.1002/bmc.329 |
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| Abstract | An ultra‐sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for the analysis of oral contraceptive ethinyl estradiol (EE) was developed and validated over the curve range of 2.5–500 pg/mL using 1 mL of human plasma sample. Ethinyl estradiol and the internal standard, ethinyl estradiol tetra‐deuterated (EE‐d4), were extracted from the plasma matrix with methyl t‐butyl ether, derivatized with dansyl chloride and then back‐extracted into hexane. The hexane phase was evaporated to dryness, reconstituted and injected onto the LC/MS/MS system. The chromatographic separation was achieved on a Luna C18 column (50 × 2 mm, 5 µm) with an isocratic mobile phase of 20:80 (v/v) water:acetonitrile with 1% formic acid. The offline derivatization procedure introduced the easily ionizable tertiary amine function group to EE. This greatly improved analyte sensitivity in electrospray ionization and enabled us to achieve the desired lower limit of quantitation at 2.5 pg/mL. This high sensitivity method can be used for therapeutic drug monitoring or supporting bio‐equivalence and drug–drug interaction studies in human subjects. Copyright © 2004 John Wiley & Sons, Ltd. |
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| AbstractList | An ultra-sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for the analysis of oral contraceptive ethinyl estradiol (EE) was developed and validated over the curve range of 2.5-500 pg/mL using 1 mL of human plasma sample. Ethinyl estradiol and the internal standard, ethinyl estradiol tetra-deuterated (EE-d4), were extracted from the plasma matrix with methyl t-butyl ether, derivatized with dansyl chloride and then back-extracted into hexane. The hexane phase was evaporated to dryness, reconstituted and injected onto the LC/MS/MS system. The chromatographic separation was achieved on a Luna C(18) column (50 x 2 mm, 5 micro m) with an isocratic mobile phase of 20:80 (v/v) water:acetonitrile with 1% formic acid. The offline derivatization procedure introduced the easily ionizable tertiary amine function group to EE. This greatly improved analyte sensitivity in electrospray ionization and enabled us to achieve the desired lower limit of quantitation at 2.5 pg/mL. This high sensitivity method can be used for therapeutic drug monitoring or supporting bio-equivalence and drug-drug interaction studies in human subjects. An ultra‐sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for the analysis of oral contraceptive ethinyl estradiol (EE) was developed and validated over the curve range of 2.5–500 pg/mL using 1 mL of human plasma sample. Ethinyl estradiol and the internal standard, ethinyl estradiol tetra‐deuterated (EE‐d4), were extracted from the plasma matrix with methyl t ‐butyl ether, derivatized with dansyl chloride and then back‐extracted into hexane. The hexane phase was evaporated to dryness, reconstituted and injected onto the LC/MS/MS system. The chromatographic separation was achieved on a Luna C 18 column (50 × 2 mm, 5 µm) with an isocratic mobile phase of 20:80 (v/v) water:acetonitrile with 1% formic acid. The offline derivatization procedure introduced the easily ionizable tertiary amine function group to EE. This greatly improved analyte sensitivity in electrospray ionization and enabled us to achieve the desired lower limit of quantitation at 2.5 pg/mL. This high sensitivity method can be used for therapeutic drug monitoring or supporting bio‐equivalence and drug–drug interaction studies in human subjects. Copyright © 2004 John Wiley & Sons, Ltd. An ultra-sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for the analysis of oral contraceptive ethinyl estradiol (EE) was developed and validated over the curve range of 2.5-500 pg/mL using 1 mL of human plasma sample. Ethinyl estradiol and the internal standard, ethinyl estradiol tetra-deuterated (EE-d4), were extracted from the plasma matrix with methyl t-butyl ether, derivatized with dansyl chloride and then back-extracted into hexane. The hexane phase was evaporated to dryness, reconstituted and injected onto the LC/MS/MS system. The chromatographic separation was achieved on a Luna C(18) column (50 x 2 mm, 5 micro m) with an isocratic mobile phase of 20:80 (v/v) water:acetonitrile with 1% formic acid. The offline derivatization procedure introduced the easily ionizable tertiary amine function group to EE. This greatly improved analyte sensitivity in electrospray ionization and enabled us to achieve the desired lower limit of quantitation at 2.5 pg/mL. This high sensitivity method can be used for therapeutic drug monitoring or supporting bio-equivalence and drug-drug interaction studies in human subjects.An ultra-sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for the analysis of oral contraceptive ethinyl estradiol (EE) was developed and validated over the curve range of 2.5-500 pg/mL using 1 mL of human plasma sample. Ethinyl estradiol and the internal standard, ethinyl estradiol tetra-deuterated (EE-d4), were extracted from the plasma matrix with methyl t-butyl ether, derivatized with dansyl chloride and then back-extracted into hexane. The hexane phase was evaporated to dryness, reconstituted and injected onto the LC/MS/MS system. The chromatographic separation was achieved on a Luna C(18) column (50 x 2 mm, 5 micro m) with an isocratic mobile phase of 20:80 (v/v) water:acetonitrile with 1% formic acid. The offline derivatization procedure introduced the easily ionizable tertiary amine function group to EE. This greatly improved analyte sensitivity in electrospray ionization and enabled us to achieve the desired lower limit of quantitation at 2.5 pg/mL. This high sensitivity method can be used for therapeutic drug monitoring or supporting bio-equivalence and drug-drug interaction studies in human subjects. An ultra‐sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for the analysis of oral contraceptive ethinyl estradiol (EE) was developed and validated over the curve range of 2.5–500 pg/mL using 1 mL of human plasma sample. Ethinyl estradiol and the internal standard, ethinyl estradiol tetra‐deuterated (EE‐d4), were extracted from the plasma matrix with methyl t‐butyl ether, derivatized with dansyl chloride and then back‐extracted into hexane. The hexane phase was evaporated to dryness, reconstituted and injected onto the LC/MS/MS system. The chromatographic separation was achieved on a Luna C18 column (50 × 2 mm, 5 µm) with an isocratic mobile phase of 20:80 (v/v) water:acetonitrile with 1% formic acid. The offline derivatization procedure introduced the easily ionizable tertiary amine function group to EE. This greatly improved analyte sensitivity in electrospray ionization and enabled us to achieve the desired lower limit of quantitation at 2.5 pg/mL. This high sensitivity method can be used for therapeutic drug monitoring or supporting bio‐equivalence and drug–drug interaction studies in human subjects. Copyright © 2004 John Wiley & Sons, Ltd. |
| Author | Naidong, Weng Shou, Wilson Z. Jiang, Xiangyu |
| Author_xml | – sequence: 1 givenname: Wilson Z. surname: Shou fullname: Shou, Wilson Z. email: wilson.shou@covance.com organization: Covance Laboratories Inc., 3301 Kinsman Boulevard, Madison, WI 53704, USA – sequence: 2 givenname: Xiangyu surname: Jiang fullname: Jiang, Xiangyu organization: Covance Laboratories Inc., 3301 Kinsman Boulevard, Madison, WI 53704, USA – sequence: 3 givenname: Weng surname: Naidong fullname: Naidong, Weng organization: Covance Laboratories Inc., 3301 Kinsman Boulevard, Madison, WI 53704, USA |
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APQ1249) publication-title: Pharmaceutical Research – volume: 43 start-page: 16 year: 1993 ident: e_1_2_1_14_1 publication-title: Arzneimittelforschung – ident: e_1_2_1_7_1 doi: 10.1016/0002-9378(90)90553-J – volume: 20 start-page: 10 year: 1997 ident: e_1_2_1_11_1 article-title: Analytical methods for measurement of contraceptive steroids in human plasma publication-title: Journal of Clinical Ligand Assay – ident: e_1_2_1_3_1 doi: 10.2165/00003088-199018060-00004 |
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| Snippet | An ultra‐sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for the analysis of oral contraceptive ethinyl estradiol (EE) was developed... An ultra-sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for the analysis of oral contraceptive ethinyl estradiol (EE) was developed... |
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| SubjectTerms | Chromatography, Liquid - methods Dansyl Compounds - chemistry Ethinyl Estradiol - blood ethinyl estradiolLC/MS/MSchemical derivatization Humans Indicators and Reagents - chemistry Reference Standards Sensitivity and Specificity Spectrometry, Mass, Electrospray Ionization - methods |
| Title | Development and validation of a high-sensitivity liquid chromatography/tandem mass spectrometry (LC/MS/MS) method with chemical derivatization for the determination of ethinyl estradiol in human plasma |
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