G005895

LC/MS/MS Analysis of Steroid Hormones in Plasma on Ascentis^® Express C18 after Sample Prep using HybridSPE^®-Phospholipid

application for HPLC

Properties

• technique(s): HPLC: suitable
• test parameters: SPE tube/cartridge: HybridSPE^®-Phospholipid, 96-well plate, 50 mg/well (575656-U); sample addition: To each well add 100 μL of plasma followed by 300 μL of precipitation solvent (1% formic acid or 0.5% citric acid in acetonitrile). (Agitate via vortex for 4 minute, place on vacuum manifold and apply 10" Hg vacuum for 4 minutes. Collect filtrate and analyze directly.); column: Ascentis^® Express C18, 10 cm x 2.1 mm I.D., 2.7 μm particles (53823-U); gradient: 60% B for 3 min, 60% B to 95% B in 5 min, held at 95% B for 2 min; mobile phase: [A] 5 mM ammonium formate pH 4.0 with formic acid; [B] methanol; flow rate: 0.3 mL/min; column temp.: 50 °C; detector: ESI+, MRM; injection: 2 μL
• suitability: application for HPLC
• application(s): clinical

Description

Analysis Note

This application shows the separation of steroid hormones in plasma using LC/MS/MS. HybridSPE-Phospholipid removed endogenous phospholipids and precipitated proteins, thereby allowing rapid, sensitive analysis by LC/MS/MS using an Ascentis Express C18 column. The highest grade LC-MS solvents were used to supply low background interference and low particulate contaminants for robust, trouble-free operation. Cerilliant CRMs provided reliable identification and quantification.

Legal Information

Ascentis is a registered trademark of Merck KGaA, Darmstadt, Germany

HybridSPE is a registered trademark of Merck KGaA, Darmstadt, Germany

Analyte

Aldosterone solution, 100 μg/mL in acetonitrile, ampule of 1 mL, certified reference material, Cerilliant^®
Corticosterone solution, 1.0 mg/mL in methanol, ampule of 1 mL, certified reference material, Cerilliant^®
11-Deoxycorticosterone solution, 100 μg/mL in methanol, ampule of 1 mL, certified reference material, Cerilliant^®
Testosterone solution, 1.0 mg/mL in acetonitrile, ampule of 1 mL, certified reference material, Cerilliant^®
17α-Methyltestosterone solution, 1.0 mg/mL in 1,2-dimethoxyethane, ampule of 1 mL, certified reference material, Cerilliant^®
Progesterone solution, 1.0 mg/mL in acetonitrile, ampule of 1 mL, certified reference material, Cerilliant^®