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A completely automated SPE/LC/MS method for the analysis of cocaine and metabolites in whole blood

Summary As laboratories are called upon to develop novel, fast, and sensitive methods, here we present a completely automated method for the analysis of cocaine and its metabolites (benzoylecgonine, ecgonine methyl ester, ecgonine and cocaethylene) from whole blood. This method utilizes an online solid-phase extraction [SPE] with high performance liquid chromatographic separation and tandem mass spectrometric detection. Pretreatment of samples involve only protein precipitation and ultracentrifugation. An efficient online solid-phase extraction [SPE] procedure was developed using Hysphere MM anion sorbent. A gradient chromatography method with a Gemini C6-Phenyl [50 mm x 3.00 mm i.d., 5 [µm] column was used for the complete separation of all components. Analysis was by positive ion mode electrospray ionization tandem mass spectrometry, using multiple reaction monitoring [MRM] to enhance the selectivity and sensitivity of the method. For the analysis, two MRM transitions are monitored for each analyte and one transition is monitored for each internal standard. With a 30-µL sample injection, linearity was analyte dependent but generally fell between 8 and 500ng/mL. The limits of detection [LODs] for the method ranged from 3 to 16 ng/mL and the limits of quantitation [LOQs] ranged from 8 to 47ng/mL. The bias and precision were determined using a simple analysis of variance [ANOVA: single factor}. The results demonstrate bias as <7%, and %precision as <9% for all components at each QC level.
Category 4. Symbiosis User Applications in collaboration with Spark
These are applications that have been made by Symbiosis customers and are based on the SPE and LC method principles as used by Spark. They are validated and have been published. These are 'ready to use applications'
Matrix Whole blood, blood
Market Forensic
Cartridge HySphere MM-Anion
Detection MS
Compounds Cocaine, Ecgonine, Ecgonine methyl ester, Benzoylecgonine, Cocaethylene, BZE
Compound Group Cocaine, DOA, Drugs of Abuse
Sample_volume 30 ÁL
Contact Eshwar Jagerdeo, Madeline A. Montgomery, Marc A. Lebeau, Martin Sibum (Spark)
Account Federal Bureau of Investigation Laboratory, 2501 Investigation Parkway, Quantico, VA 22135, United States
Published Journal of Chromatography B, 874 (2008), 15-20
Date 8-8-2008
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Remarks We have developed and validated an analytical method to quantitatively measure Cocaine and four metabolites in whole blood
This fully automated intrumental method eliminates many of the time consuming, manual sample processing stepsused by other spe methods, therefore minimizing the change of errors.
This method demonstrates excellent accuracy and precision, a wide linear range and a good lowe rlimit of detection.
Status Finished