A sensitive and selective LC-differential mobility-mass spectrometric analysis of allopregnanolone and pregnanolone in human plasma. In process uri icon

Overview

abstract

  • A sensitive and selective method was developed to quantitate allopregnanolone and its 5β isomer pregnanolone in human plasma using liquid chromatography-differential mobility separation combined with MS/MS detection. The method employed a simple liquid-liquid extraction of 100 μL plasma with hexane/ethyl acetate. After extraction, the sample was derivatized using a quaternary aminooxy reagent. Separation of allopregnanolone, pregnanolone, and their 3β epimers (epiallopregnanolone and epipregnanolone) was achieved using a Phenomenex Kinetex C18 2.1 × 100-mm 2.6-μm column. A linear calibration curve was obtained over the concentration range from 10 to 25,000 pg/mL, and the inter- and intra-day accuracy of the quality control samples were between 90 and 110 % with the inter- and intra-day precision less than 10 %. The lower limit of quantitation is 50 fg (157 amol) on column for both allopregnanolone and pregnanolone which is 100-fold less than the underivatized compounds. The recovery is above 95 %, and the extracted samples are stable for at least 6 days when stored at 4 °C. Plasma samples from normal, pregnant, and postpartum women were analyzed using this method.

publication date

  • November 2013

Research

keywords

  • In press

Identity

Language

  • eng

PubMed Central ID

  • PMC3984615

Digital Object Identifier (DOI)

  • 10.1007/s00216-013-7391-2

PubMed ID

  • 24121434

Additional Document Info

start page

  • 9497

end page

  • 9508

volume

  • 405

number

  • 29