Influence of phenobarbital on morphine metabolism and disposition

LC-MS/MS determination of morphine (M) and morphine-3-glucuronide (M3G) in Wistar-Kyoto rat serum, bile, and urine

Yazen Alnouti, Melinda K. Shelby, Chuan Chen, Curtis D. Klaassen

Research output: Contribution to journalReview article

8 Citations (Scopus)

Abstract

A simple LC-MS/MS method has been developed and validated for the simultaneous determination of morphine (M) and morphine-3-glucuronide (M3G) in rat serum, bile, and urine. Deuterated D3-M and D3-M3G were used as internal standards (IS) for M and M3G, respectively. Serum samples were processed by acetonitrile precipitation. Bile samples were prepared by solid-phase extraction (SPE) using Oasis MCX cartridges. Urine samples were directly analyzed after dilution with mobile phase. Chromatography was performed using a Luna C18 column (5 μm, 150 × 2.1 mm I.D.). The mobile phase consisted of acetonitrile (ACN) and 7.5 mM ammonium formate (pH 9.3) delivered from separate pumps with a simple gradient. The method was validated to quantify M in the range of 1-1000 ng/ml in bile and serum, and 0.025-25 μg/ml in urine. M3G was quantified in the range of 1-1000 ng/ml in serum, 0.1-100 μg/ml in bile, and 0.05-25 μg/ml in urine. The method was applied to study the pharmacokinetics and disposition of M and M3G in Wistar-Kyoto (WKY) rats, and the effect of phenobarbital (PB) on M and M3G disposition. M is metabolized to M3G at a lower rate in male than female rats leading to higher M levels and lower M3G levels in serum, urine, and bile of male than female rats. PB administration induces M glucuronidation to M3G in male, but not female WKY rats, and abolishes the gender differences in M and M3G pharmacokinetics.

Original languageEnglish (US)
Pages (from-to)79-89
Number of pages11
JournalCurrent Drug Metabolism
Volume8
Issue number1
DOIs
StatePublished - Jan 1 2007

Fingerprint

Inbred WKY Rats
Phenobarbital
Metabolism
Bile
Morphine
Rats
Urine
Serum
formic acid
Pharmacokinetics
morphine-3-glucuronide
Solid Phase Extraction
Chromatography
Dilution
Pumps

Keywords

  • CAR
  • HPLC
  • Induction
  • Mass spectrometry
  • Morphine
  • Morphine glucuronide
  • Phenobarbital
  • UGT
  • Wistar-Kyoto rats

ASJC Scopus subject areas

  • Pharmacology
  • Clinical Biochemistry

Cite this

Influence of phenobarbital on morphine metabolism and disposition : LC-MS/MS determination of morphine (M) and morphine-3-glucuronide (M3G) in Wistar-Kyoto rat serum, bile, and urine. / Alnouti, Yazen; Shelby, Melinda K.; Chen, Chuan; Klaassen, Curtis D.

In: Current Drug Metabolism, Vol. 8, No. 1, 01.01.2007, p. 79-89.

Research output: Contribution to journalReview article

@article{0a6518144a964307be825f58a4c6bfe1,
title = "Influence of phenobarbital on morphine metabolism and disposition: LC-MS/MS determination of morphine (M) and morphine-3-glucuronide (M3G) in Wistar-Kyoto rat serum, bile, and urine",
abstract = "A simple LC-MS/MS method has been developed and validated for the simultaneous determination of morphine (M) and morphine-3-glucuronide (M3G) in rat serum, bile, and urine. Deuterated D3-M and D3-M3G were used as internal standards (IS) for M and M3G, respectively. Serum samples were processed by acetonitrile precipitation. Bile samples were prepared by solid-phase extraction (SPE) using Oasis MCX cartridges. Urine samples were directly analyzed after dilution with mobile phase. Chromatography was performed using a Luna C18 column (5 μm, 150 × 2.1 mm I.D.). The mobile phase consisted of acetonitrile (ACN) and 7.5 mM ammonium formate (pH 9.3) delivered from separate pumps with a simple gradient. The method was validated to quantify M in the range of 1-1000 ng/ml in bile and serum, and 0.025-25 μg/ml in urine. M3G was quantified in the range of 1-1000 ng/ml in serum, 0.1-100 μg/ml in bile, and 0.05-25 μg/ml in urine. The method was applied to study the pharmacokinetics and disposition of M and M3G in Wistar-Kyoto (WKY) rats, and the effect of phenobarbital (PB) on M and M3G disposition. M is metabolized to M3G at a lower rate in male than female rats leading to higher M levels and lower M3G levels in serum, urine, and bile of male than female rats. PB administration induces M glucuronidation to M3G in male, but not female WKY rats, and abolishes the gender differences in M and M3G pharmacokinetics.",
keywords = "CAR, HPLC, Induction, Mass spectrometry, Morphine, Morphine glucuronide, Phenobarbital, UGT, Wistar-Kyoto rats",
author = "Yazen Alnouti and Shelby, {Melinda K.} and Chuan Chen and Klaassen, {Curtis D.}",
year = "2007",
month = "1",
day = "1",
doi = "10.2174/138920007779315026",
language = "English (US)",
volume = "8",
pages = "79--89",
journal = "Current Drug Metabolism",
issn = "1389-2002",
publisher = "Bentham Science Publishers B.V.",
number = "1",

}

TY - JOUR

T1 - Influence of phenobarbital on morphine metabolism and disposition

T2 - LC-MS/MS determination of morphine (M) and morphine-3-glucuronide (M3G) in Wistar-Kyoto rat serum, bile, and urine

AU - Alnouti, Yazen

AU - Shelby, Melinda K.

AU - Chen, Chuan

AU - Klaassen, Curtis D.

PY - 2007/1/1

Y1 - 2007/1/1

N2 - A simple LC-MS/MS method has been developed and validated for the simultaneous determination of morphine (M) and morphine-3-glucuronide (M3G) in rat serum, bile, and urine. Deuterated D3-M and D3-M3G were used as internal standards (IS) for M and M3G, respectively. Serum samples were processed by acetonitrile precipitation. Bile samples were prepared by solid-phase extraction (SPE) using Oasis MCX cartridges. Urine samples were directly analyzed after dilution with mobile phase. Chromatography was performed using a Luna C18 column (5 μm, 150 × 2.1 mm I.D.). The mobile phase consisted of acetonitrile (ACN) and 7.5 mM ammonium formate (pH 9.3) delivered from separate pumps with a simple gradient. The method was validated to quantify M in the range of 1-1000 ng/ml in bile and serum, and 0.025-25 μg/ml in urine. M3G was quantified in the range of 1-1000 ng/ml in serum, 0.1-100 μg/ml in bile, and 0.05-25 μg/ml in urine. The method was applied to study the pharmacokinetics and disposition of M and M3G in Wistar-Kyoto (WKY) rats, and the effect of phenobarbital (PB) on M and M3G disposition. M is metabolized to M3G at a lower rate in male than female rats leading to higher M levels and lower M3G levels in serum, urine, and bile of male than female rats. PB administration induces M glucuronidation to M3G in male, but not female WKY rats, and abolishes the gender differences in M and M3G pharmacokinetics.

AB - A simple LC-MS/MS method has been developed and validated for the simultaneous determination of morphine (M) and morphine-3-glucuronide (M3G) in rat serum, bile, and urine. Deuterated D3-M and D3-M3G were used as internal standards (IS) for M and M3G, respectively. Serum samples were processed by acetonitrile precipitation. Bile samples were prepared by solid-phase extraction (SPE) using Oasis MCX cartridges. Urine samples were directly analyzed after dilution with mobile phase. Chromatography was performed using a Luna C18 column (5 μm, 150 × 2.1 mm I.D.). The mobile phase consisted of acetonitrile (ACN) and 7.5 mM ammonium formate (pH 9.3) delivered from separate pumps with a simple gradient. The method was validated to quantify M in the range of 1-1000 ng/ml in bile and serum, and 0.025-25 μg/ml in urine. M3G was quantified in the range of 1-1000 ng/ml in serum, 0.1-100 μg/ml in bile, and 0.05-25 μg/ml in urine. The method was applied to study the pharmacokinetics and disposition of M and M3G in Wistar-Kyoto (WKY) rats, and the effect of phenobarbital (PB) on M and M3G disposition. M is metabolized to M3G at a lower rate in male than female rats leading to higher M levels and lower M3G levels in serum, urine, and bile of male than female rats. PB administration induces M glucuronidation to M3G in male, but not female WKY rats, and abolishes the gender differences in M and M3G pharmacokinetics.

KW - CAR

KW - HPLC

KW - Induction

KW - Mass spectrometry

KW - Morphine

KW - Morphine glucuronide

KW - Phenobarbital

KW - UGT

KW - Wistar-Kyoto rats

UR - http://www.scopus.com/inward/record.url?scp=33846004452&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33846004452&partnerID=8YFLogxK

U2 - 10.2174/138920007779315026

DO - 10.2174/138920007779315026

M3 - Review article

VL - 8

SP - 79

EP - 89

JO - Current Drug Metabolism

JF - Current Drug Metabolism

SN - 1389-2002

IS - 1

ER -