Development and validation of a liquid chromatography–mass spectrometric assay for simultaneous determination of tacrolimus and 13-O-desmethyl tacrolimus in rat kidney tissue

Tatian Kirresh, Sony Tuteja, D. Russo, P. D. Brophy, Daryl J Murry

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

A sensitive and robust LC–MS/MS method has been developed and validated to determine the concentrations of tacrolimus and its major metabolite 13-O-desmethyl tacrolimus (13-ODMT) in kidney tissue from rats who received tacrolimus intra-peritoneally at doses of 0.5 mg/kg and 2 mg/kg. The samples were prepared by a liquid-liquid extraction procedure using ethyl ether as the extraction solvent and ascomycin as the internal standard. Chromatographic separation was achieved using Phenomenex Kinetex column (2.6 μm C18 100 Å, 100 × 2.1 mm, Phenomenex, Torrance CA) and a gradient mobile phase of water and methanol-acetonitrile (50:50, v/v) both containing 0.1% formic acid. The limit of quantification was 0.25 ng/ml and the calibration curves covered a concentration range from 0.25 to 50 ng/ml. Intra-and inter-assay precision and accuracy for both tacrolimus and 13-ODMT were all within FDA guidelines for bioanalysis. Extraction efficiency for tacrolimus ranged from 67.00 to 74.90% and from 66.70 to 78.40% for 13-ODMT. Several challenges interfering with the performance of the method such as phospholipid build-up have also been addressed. Kidney tissue samples from six rats receiving either 0.5 or 2 mg/kg dose were analyzed and resulted in a median concentration of 11.54 and 0.72 ng/ml for tacrolimus and 13-ODMT, respectively, for the lower dose level, and a median concentration of 8.89 ng/ml and 1.50 ng/ml for tacrolimus and 13-ODMT, respectively, at the higher dose level.

Original languageEnglish (US)
Pages (from-to)32-37
Number of pages6
JournalJournal of Pharmaceutical and Biomedical Analysis
Volume136
DOIs
StatePublished - Mar 20 2017
Externally publishedYes

Fingerprint

Tacrolimus
Rats
Assays
Tissue
Kidney
Liquids
formic acid
Liquid-Liquid Extraction
Solvent extraction
Metabolites
Ether
Calibration
Methanol
Phospholipids
Guidelines

Keywords

  • 13-O-desmethyl tacrolimus
  • IT-TOF
  • Mass spectroscopy
  • Nephrotoxicity
  • Organ rejection
  • Tacrolimus

ASJC Scopus subject areas

  • Analytical Chemistry
  • Pharmaceutical Science
  • Drug Discovery
  • Spectroscopy
  • Clinical Biochemistry

Cite this

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title = "Development and validation of a liquid chromatography–mass spectrometric assay for simultaneous determination of tacrolimus and 13-O-desmethyl tacrolimus in rat kidney tissue",
abstract = "A sensitive and robust LC–MS/MS method has been developed and validated to determine the concentrations of tacrolimus and its major metabolite 13-O-desmethyl tacrolimus (13-ODMT) in kidney tissue from rats who received tacrolimus intra-peritoneally at doses of 0.5 mg/kg and 2 mg/kg. The samples were prepared by a liquid-liquid extraction procedure using ethyl ether as the extraction solvent and ascomycin as the internal standard. Chromatographic separation was achieved using Phenomenex Kinetex column (2.6 μm C18 100 {\AA}, 100 × 2.1 mm, Phenomenex, Torrance CA) and a gradient mobile phase of water and methanol-acetonitrile (50:50, v/v) both containing 0.1{\%} formic acid. The limit of quantification was 0.25 ng/ml and the calibration curves covered a concentration range from 0.25 to 50 ng/ml. Intra-and inter-assay precision and accuracy for both tacrolimus and 13-ODMT were all within FDA guidelines for bioanalysis. Extraction efficiency for tacrolimus ranged from 67.00 to 74.90{\%} and from 66.70 to 78.40{\%} for 13-ODMT. Several challenges interfering with the performance of the method such as phospholipid build-up have also been addressed. Kidney tissue samples from six rats receiving either 0.5 or 2 mg/kg dose were analyzed and resulted in a median concentration of 11.54 and 0.72 ng/ml for tacrolimus and 13-ODMT, respectively, for the lower dose level, and a median concentration of 8.89 ng/ml and 1.50 ng/ml for tacrolimus and 13-ODMT, respectively, at the higher dose level.",
keywords = "13-O-desmethyl tacrolimus, IT-TOF, Mass spectroscopy, Nephrotoxicity, Organ rejection, Tacrolimus",
author = "Tatian Kirresh and Sony Tuteja and D. Russo and Brophy, {P. D.} and Murry, {Daryl J}",
year = "2017",
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doi = "10.1016/j.jpba.2016.12.034",
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TY - JOUR

T1 - Development and validation of a liquid chromatography–mass spectrometric assay for simultaneous determination of tacrolimus and 13-O-desmethyl tacrolimus in rat kidney tissue

AU - Kirresh, Tatian

AU - Tuteja, Sony

AU - Russo, D.

AU - Brophy, P. D.

AU - Murry, Daryl J

PY - 2017/3/20

Y1 - 2017/3/20

N2 - A sensitive and robust LC–MS/MS method has been developed and validated to determine the concentrations of tacrolimus and its major metabolite 13-O-desmethyl tacrolimus (13-ODMT) in kidney tissue from rats who received tacrolimus intra-peritoneally at doses of 0.5 mg/kg and 2 mg/kg. The samples were prepared by a liquid-liquid extraction procedure using ethyl ether as the extraction solvent and ascomycin as the internal standard. Chromatographic separation was achieved using Phenomenex Kinetex column (2.6 μm C18 100 Å, 100 × 2.1 mm, Phenomenex, Torrance CA) and a gradient mobile phase of water and methanol-acetonitrile (50:50, v/v) both containing 0.1% formic acid. The limit of quantification was 0.25 ng/ml and the calibration curves covered a concentration range from 0.25 to 50 ng/ml. Intra-and inter-assay precision and accuracy for both tacrolimus and 13-ODMT were all within FDA guidelines for bioanalysis. Extraction efficiency for tacrolimus ranged from 67.00 to 74.90% and from 66.70 to 78.40% for 13-ODMT. Several challenges interfering with the performance of the method such as phospholipid build-up have also been addressed. Kidney tissue samples from six rats receiving either 0.5 or 2 mg/kg dose were analyzed and resulted in a median concentration of 11.54 and 0.72 ng/ml for tacrolimus and 13-ODMT, respectively, for the lower dose level, and a median concentration of 8.89 ng/ml and 1.50 ng/ml for tacrolimus and 13-ODMT, respectively, at the higher dose level.

AB - A sensitive and robust LC–MS/MS method has been developed and validated to determine the concentrations of tacrolimus and its major metabolite 13-O-desmethyl tacrolimus (13-ODMT) in kidney tissue from rats who received tacrolimus intra-peritoneally at doses of 0.5 mg/kg and 2 mg/kg. The samples were prepared by a liquid-liquid extraction procedure using ethyl ether as the extraction solvent and ascomycin as the internal standard. Chromatographic separation was achieved using Phenomenex Kinetex column (2.6 μm C18 100 Å, 100 × 2.1 mm, Phenomenex, Torrance CA) and a gradient mobile phase of water and methanol-acetonitrile (50:50, v/v) both containing 0.1% formic acid. The limit of quantification was 0.25 ng/ml and the calibration curves covered a concentration range from 0.25 to 50 ng/ml. Intra-and inter-assay precision and accuracy for both tacrolimus and 13-ODMT were all within FDA guidelines for bioanalysis. Extraction efficiency for tacrolimus ranged from 67.00 to 74.90% and from 66.70 to 78.40% for 13-ODMT. Several challenges interfering with the performance of the method such as phospholipid build-up have also been addressed. Kidney tissue samples from six rats receiving either 0.5 or 2 mg/kg dose were analyzed and resulted in a median concentration of 11.54 and 0.72 ng/ml for tacrolimus and 13-ODMT, respectively, for the lower dose level, and a median concentration of 8.89 ng/ml and 1.50 ng/ml for tacrolimus and 13-ODMT, respectively, at the higher dose level.

KW - 13-O-desmethyl tacrolimus

KW - IT-TOF

KW - Mass spectroscopy

KW - Nephrotoxicity

KW - Organ rejection

KW - Tacrolimus

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U2 - 10.1016/j.jpba.2016.12.034

DO - 10.1016/j.jpba.2016.12.034

M3 - Article

VL - 136

SP - 32

EP - 37

JO - Journal of Pharmaceutical and Biomedical Analysis

JF - Journal of Pharmaceutical and Biomedical Analysis

SN - 0731-7085

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