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Simultaneous Determination of Hydrocodone, and Its Two Metabolites in Human Plasma by HPLC–MS–MS
A rapid, sensitive and specific assay method has been developed to simultaneously determine human plasma concentrations of hydrocodone and its metabolites, norhydrocodone, hydromorphone, using high-performance liquid chromatography with an electrospray ionization (ESI) tandem mass spectrometry (HPLC...
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Published in: | Chromatographia 2011-10, Vol.74 (7-8), p.567-574 |
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Main Authors: | , , , , , , , , |
Format: | Article |
Language: | English |
Subjects: | |
Citations: | Items that this one cites Items that cite this one |
Online Access: | Get full text |
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Summary: | A rapid, sensitive and specific assay method has been developed to simultaneously determine human plasma concentrations of hydrocodone and its metabolites, norhydrocodone, hydromorphone, using high-performance liquid chromatography with an electrospray ionization (ESI) tandem mass spectrometry (HPLC–MS–MS). Hydrocodone, its metabolites, and internal standard, hydrocodone-
d
3
, norhydrocodone-
d
3
, hydromorphone-
d
3
, were separated from human plasma using solid-phase extraction (Empore MPC-SD Solid Phase Extraction Disk). The eluate was dried, reconstituted and injected into the LC–MS–MS system. Chromatographic separation was performed on a Kromasil 100-5SIL-Dimensions C
18
column (100 × 2.1 mm, 5.0 μm, Thermo Hypersil-Keystone, USA) using a gradient mobile phase with 20 mmol L
−1
ammonium formate in water with 0.2% formic acid and 0.1% formic acid in acetonitrile. Detection and quantitation were performed by MS/MS using electrospray ionization and multiple reactions monitoring in the positive ion mode. The calibration curves were linear over the concentration ranges 0.05–50 ng mL
−1
for hydrocodone (
r
2
= 0.9991) and norhydrocodone (
r
2
= 0.9990), and 0.01–10 ng mL
−1
for hydromorphone (
r
2
= 0.9990). The limit of quantification was 0.05 ng mL
−1
for hydrocodone and norhydrocodone, and 0.01 ng mL
−1
for hydromorphone. The extraction recovery was above 64.36, 68.51 and 71.78% for hydrocodone, norhydrocodone and hydromorphone. The accuracy was higher than 99.06, 97.70 and 100.07% for hydrocodone, norhydrocodone and hydromorphone. The intra- and inter-day precisions were |
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ISSN: | 0009-5893 1612-1112 |
DOI: | 10.1007/s10337-011-2118-z |