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Detection of Morphine and Morphine-Glucuronide in Saliva or Urine with a Rapid and Simple Method Without Derivatization on the Varian 500-MS

in only five minutes. Due to the complexity of the final samples injected, a tandem MS technique is employed to increase the method selectivity; the MS/MS conditions are shown in the previous table. Figures 2 and 3 are the calibration curves for morphine and morphine-glucuronide, respectively. Excellent linearity can be demonstrated from 20 to 1000 pg (six points, triplicate runs) for morphine and 40 to 1000 pg (five points, triplicate runs) for morphine-glucuronide. The on column Limits of detection (LOD) and quantification (LOQ) are 10 and 20 pg (respectively) for morphine and 40 and 60 pg (respectively) for morphine-glucuronide. These limits are well within the limits required by the National Institute on Drug Abuse (NIDA) limits for opiates in saliva and urine: 40 ng/mL and 300 ng/mL, respectively.

Figure 4 shows the separation of morphine that was added to a sample of saliva taken with the OraTube sampler. Panel 1 shows the separation of morphine (Arrow marked B) from the salts and matrix interfering peaks (Arrow A). These peaks are now removed when compared to a saliva sample further spiked with morphine (to confirm its identity) but run with the automated 6-port valve of the 500-MS plumbed as a diverter valve.

The diverter valve is an important tool which allows direct injection of urine samples into the ESI source, preventing contamination of the ESI source with urinary salts and extending the maintenance cycle time. To demonstrate this, we have placed a UV detector in-line between the HPLC column and the MS source so that the sample was analyzed by the UV detector prior to the MS. Figure 5 shows the detection of morphine-glucuronide in urine as detected by the 500-MS with the diverter valve, the expanded region on top shows the same sample as detected by UV/Vis at 214 nm.

The bottoming out of the chromatogram (at approximately 0.6 min, UV detection) is the sample v
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