Measurement of catecholamines in rat and mini-pig plasma and urine by liquid chromatography-tandem mass spectrometry coupled with solid phase extraction

J Chromatogr B Analyt Technol Biomed Life Sci. 2015 Aug 1:997:154-61. doi: 10.1016/j.jchromb.2015.05.014. Epub 2015 Jun 5.

Abstract

A tandem mass spectrometry method combined with an ion-pair chromatographic separation after weak cation exchange solid phase sample extraction for epinephrine (E), norepinephrine (NE) and dopamine (DA) has been developed. Two surrogate matrixes for plasma and urine as well as stable isotope labeled internal standards were utilized for quantitation. The observed dynamic range of E, NE and DA was 0.025-100ng/ml for plasma, and 0.25-1000ng/ml for urine with a r(2) regression coefficient >0.99. Extraction recoveries were greater than 60% and the lower limit of quantitation was 25pg/ml for all three analytes in plasma. This method provided excellent sensitivity and selectivity for use with small sample volumes (≤25uL), enabling high-throughput pharmacodynamic animal model development and screening of adverse effects.

Keywords: Catecholamines; Dopamine; Epinephrine; High performance liquid; Norepinephrine; Pharmacodynamics; Pharmacokinetics; Solid phase; Tandem mass spectrometry; chromatography; extraction.

MeSH terms

  • Animals
  • Blood Glucose / analysis
  • Catecholamines / blood*
  • Chromatography, High Pressure Liquid / methods*
  • Linear Models
  • Rats
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Solid Phase Extraction / methods*
  • Swine
  • Swine, Miniature
  • Tandem Mass Spectrometry / methods*

Substances

  • Blood Glucose
  • Catecholamines