LC-MS/MS quantification of salvinorin A from biological fluids.
Michael J Caspers, Todd D Williams, Kimberly M Lovell, Anthony Lozama, Eduardo R Butelman, Mary Jeanne Kreek, Matthew Johnson, Roland R. Griffiths, Katherine A Maclean, Thomas E Prisinzano
Analytical methods : advancing methods and applications December 21, 2013 DOI: 10.1039/C3AY40810H (opens in new tab)
Study at a glance
AI-extracted from the abstract| Characteristics | Method development and validation Peer reviewed |
|---|---|
| Population | Rhesus monkeys and healthy human research participants |
| Topics | Salvia divinorum |
| Keywords | Bioanalytical method development Bioanalysis Drug analysis Quantification method Detection technique Advanced instrumentation Sample preparation Sensitivity Precision Accuracy Measurement Robust approach Optimized method Hallucinogen Psychoactive compound Potent compound Pharmacokinetics Drug levels Drug disposition Toxicology Brain fluid analysis Blood analysis Drug impact Active doses Biomonitoring Body fluid analysis Compound levels In vivo analysis |
| Citations | 6 |
| Key points | A validated LC-MS/MS method quantifies salvinorin A in CSF and plasma at low ng/mL levels with high precision and accuracy. |
Abstract
A facile method for quantifying the concentration of the powerful and widely available hallucinogen salvinorin A (a selective kappa opioid agonist) from non-human primate cerebrospinal fluid (CSF) and human plasma has been developed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in positive electrospray ionization (ESI) mode. With CSF solid phase extraction can be avoided completely by simply diluting each sample to 10 % (v/v) acetonitrile, 1 % (v/v) formic acid and injecting under high aqueous conditions for analyte focusing. Extensive plasma sample preparation was investigated including protein precipitation, SPE column selection, and plasma particulate removal. Human plasma samples were centrifuged at 21,000 × gravity for 4 minutes to obtain clear particulate-free plasma, from which 300 μl was spiked with internal standard and loaded onto a C18 SPE column with a 100 mg mL-1 loading capacity. Guard columns (C18, hand packed 1 mm × 20 mm) were exchanged after backpressure increased above 4600psi, about 250 injections. A shallow acetonitrile/water gradient was used, 29 to 33% CH3CN over 8 minutes to elute salvinorin A. Reduction of chemical noise was achieved using tandem mass spectrometry with multiple reaction monitoring while sensitivity increases were observed using a 50 μL injection volume onto a small bore analytical column (C18, 1 mm ID × 50 mm) thus increasing peak concentration. Limits of quantification were found to be 0.0125 ng mL-1 (CSF) and 0.05 ng mL-1 (plasma) with interday precision and accuracy below 1.7 % and 9.42 % (CSF) and 3.47 % and 12.37 % (plasma) respectively. This method was used to determine the concentration of salvinorin A from an in vivo Rhesus monkey study and a trial of healthy human research participants, using behaviorally active doses.