A validated liquid chromatographic/tandem mass spectrometric method for the determination of phencyclidine in microliter samples of rat serum

Abstract

A liquid chromatographic/tandem mass spectrometric method is described for the determination of phencyclidine (PCP) in small volumes of rat serum (e.g. 50 microl). Samples were extracted using a mixed-mode strong cation-exchange column and then separated isocratically using a narrow-bore (2.1 mm i.d.) 3 microm Hypersil phenyl column and a mobile phase consisting of an ammonium formate buffer (pH 2.7) with 60% (v/v) methanol. Detection was accomplished using positive ion electrospray ionization in the multiple reaction monitoring mode. Mass spectra were obtained and peaks were observed at an m/z (% abundance) of 244 (100), 159 (25), and 86 (89). Tandem mass spectra were also obtained from the m/z 244 precursor ion with peaks observed at m/z 159 (100), 86 (96), and 91 (11). Optimum serum PCP sensitivity and precision were obtained at a transition of m/z 244 --> 159. Matrix-associated ion suppression did not significantly affect the accuracy (100-112%) or precision (CV < or =8%) of the assay. The lower limit of quantitation was 1 ng ml(-1) in 50 microl of serum. The method was used to study the serum pharmacokinetics of PCP in rats after an intravenous bolus dose of PCP.

The Fate and Function of Therapeutic Antiaddiction Monoclonal Antibodies across the Reproductive Cycle of Rats

“…PCP was extracted from serum by liquidphase extraction, using a simple trichloroacetic acid treatment as described previously for METH (Hendrickson et al, 2006). PCP concentrations (ng/ml) in serum samples were determined by liquid chromatography with tandem mass spectrometry detection (LC-MS/ MS), as described previously (Hendrickson et al, 2005).…”

“…PCP was extracted from thawed tissue homogenates by solid-phase extraction as described previously (Hendrickson et al, 2005). Calibration standards (1, 5, 10, 50, 100, 500, and 1000 ng/ml PCP in NRS), quality-control standards for serum samples (2, 80, and 800 ng/ml PCP in NRS), and qualitycontrol standards for brain samples (100 and 400 ng/ml PCP in brain homogenate from untreated male rats) were treated identically to experimental samples.…”

“…Calibration standards (1, 5, 10, 50, 100, 500, and 1000 ng/ml PCP in NRS), quality-control standards for serum samples (2, 80, and 800 ng/ml PCP in NRS), and qualitycontrol standards for brain samples (100 and 400 ng/ml PCP in brain homogenate from untreated male rats) were treated identically to experimental samples. LC-MS/MS procedures were performed by using a Pursuit C8 100 ϫ 2.0-mm liquid chromatography column (Varian Inc., Palo Alto, CA) and Quattro LC triple-quadrupole mass spectrometer with a Z-Spray ion interface and a Mark II source (Waters, Milford, MA) as described previously (Hendrickson et al, 2005). Calibration and control sample intraday/interday variability were less than 20% at the upper and lower limits of quantitation and ϳ6% at common sample concentrations (Hendrickson et al, 2005).…”

“…LC-MS/MS procedures were performed by using a Pursuit C8 100 ϫ 2.0-mm liquid chromatography column (Varian Inc., Palo Alto, CA) and Quattro LC triple-quadrupole mass spectrometer with a Z-Spray ion interface and a Mark II source (Waters, Milford, MA) as described previously (Hendrickson et al, 2005). Calibration and control sample intraday/interday variability were less than 20% at the upper and lower limits of quantitation and ϳ6% at common sample concentrations (Hendrickson et al, 2005). Brain tissue PCP concentrations were corrected for blood PCP content as described previously (Laurenzana et al, 2003a).…”

Detection of phencyclidine in human oral fluid using solid-phase extraction and liquid chromatography with tandem mass spectrometric detection

“…Phencyclidine (PCP) is included in the proposed United States Federal regulations for saliva drug testing in the workplace, and the suggested cut-off concentration is 10 ng/mL of neat oral fluid. Surprisingly, there are no published procedures for the determination of PCP in oral fluid, using liquid chromatography with tandem mass spectrometry, however, there is one method for its analysis in rat serum [1]. Other methods for the determination of PCP in blood [2], urine [3], hair [4] and meconium [5] have been reported, which incorporate the more standard gas chromatography-mass spectrometry instrumentation, usually found in forensic laboratories.…”