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HPLC Analysis of Dermorphin in Horse Plasma on Ascentis® Express F5 after SPE with HybridSPE®

HPLC Analysis of Dermorphin in Horse Plasma on Ascentis® Express F5 after SPE with HybridSPE®

Conditions

sample preparation SPE (Solid Phase Extraction)
sample/matrix horse plasma spiked at 10 µg/mL
SPE tube/cartridge [TM"HybridSPE"]-Phospholipid Plate, 50mg/96-Well (575656-U)
sample addition 3:1, (1% formic acid in acetonitrile); plasma
column Ascentis Express F5, 10 cm x 2.1 mm I.D., 2.7 µm particles (53569-U)
column temp. 35 C
mobile phase [A] acetonitrile [B] 20 mM ammonium formate; (90:10, A:B); pH 4.5 with formic acid, after mixing A & B
flow rate 0.4 ml/min
pressure 1305 psi (90 bar)
injection 1 µl
detector MS, ESI(+), TIC, m/z 100-1000

Description

Analysis Note Dermorphin is a natural heptapeptide opioid. Because of poor recovery from horse plasma following protein precipitation, alternate methods of extraction are sought. While the HybridSPE protocol successfully removes proteins and phospholipids, dermorphin solubility in plasma remains a limitation. Dermorphin is effectively retained on the pentafluorophenyl phase under aqueous normal phase conditions.
Categories Analytical Chromatography, Forensics and Toxicology, SPE Applications, Drugs of Abuse, analgesics, Analgesics, drugs of abuse, Peptides, peptides, Drugs of abuse
Featured Industry Forensics and Toxicology
Legal Information Ascentis is a registered trademark of Sigma-Aldrich Co. LLC, HybridSPE is a registered trademark of Sigma-Aldrich Co. LLC
suitability application for SPE, application for HPLC

Materials

     
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