Session: SUN 130-162-Neuroendocrinology
Poster Board SUN-157
The challenges for LC-MS/MS analysis of allopregnanolone are (i) its poor ionization efficiency, and (ii) the presence of numerous isobaric interferences in biological samples including, but not limited to, its isomer pregnanolone. To overcome these challenges, ion mobility separation was combined with conventional LC-MS/MS detection using a highly sensitive AB SCIEX Triple Quad™ 6500 mass spectrometer equipped with the SelexION™ ion mobility device. The method employed liquid-liquid extraction of 100µL serum or plasma. After extraction, the sample was derivatized using a commercially available quaternary aminooxy reagent.
Separation of allopregnanolone and its isomer pregnanolone was achieved using a Phenomenex Kinetex C18 2.1x100 mm column. The calibration range was from 5 pg/mL to 100 ng/mL in serum or plasma, with inter- and intra-day precision less than 10% and inter- and intra-day accuracy between 90-110%. The recovery is 98%, and the limit of detection is 5 pg/mL for allopregnanolone and pregnanolone. Plasma samples from ‘normal’, pregnant, and postpartum women were analysed using this method.
Disclosure: MJYJ: Employee, AB SCIEX. WJ: Employee, AB SCIEX. Nothing to Disclose: MA
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