Profile up to 38 steroids by LC/MS/MS (24 Δ4 steroids, 8 Δ5 steroids and 6 sulfate-conjugated steroids) in biological matrices such as plasma, serum, tissue, cells, and cell culture media. Sample preparation and extraction of steroids is by either liquid-liquid extraction (LLE) or supported liquid extraction (SLE) followed by an on-line 2D polishing extraction. Target steroid analytes are chromatographically separated on a reverse-phase (RP) column. All analytes and Internal Standards are measured by ESI ionization--pos and/or neg polarity (analyte dependent) on an Agilent 1290 UHPLC coupled with an Agilent 6490 UHPLC-QQQ mass spectrometer with a "Jet-Spray" source and Ion-funnel using MRM methods. The 6490 QQQ with the Jet-Spray and Ion-funnel is up to 10 times more sensitive than a standard electrospray source and results in the achievement of detection limits in the low pM range. Results arereported as total pG/mL (biofluids) or pM (tissues--normalized to wet tissue weight). CV's are generally 10%.
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