DETECTABUSE™ "NO VACUUM" GRAVITY SERIES GV-65 METHOD FOR THE ANALYSIS OF
PHENCYCLIDINE (PCP) IN URINE, SERUM OR ORAL FLUID BY GC-MS
Although this method has performed well in our laboratory, it must be validated by your laboratory before it is used to report patient values. We would appreciate your comments on its performance and welcome your suggestions for improvements or enhancements.
Revised: January 2003 (* TEA Elution Solvent Stability)
SAMPLE PREPARATION - (Please see Notes and Supplemental Information before proceeding)
1. Pipet 2.0 mL of sample into a
16 x 100 mm disposable borosilicate glass tube with an inert screw cap top. Notes: When adding an internal standard dissolved in an organic
solvent to a sample, the solvent volume must not exceed 3% of the buffered
sample volume. Higher solvent concentrations may produce extraction losses.
HARDWARE SETUP - (Please refer to the Detectabuse Hardware Setup
Instructions) >
COLUMN CONDITIONING
1. Wash column with 1.0 mL of Methanol. Allow to flow by gravity. 1. Pour samples onto preconditioned column. Allow to flow by gravity.
Samples will flow through the column at a rate of 1-2 mL/min. SAMPLE ELUTION
1. Sample elution is done outside of the vacuum box. Note: If liquids do not elute freely by gravity flow, there
is probably air trapped within the column bed or frits. Tapping the column
mounting plate onto the vacuum box should initiate flow.
RECONSTITUTION
1. To each dried extract add 100 µL Ethyl Acetate, vortex mix, then
flush with nitrogen or argon. SUPPLEMENT - When using an automated robotic system all liquids
may be allowed to flow unassisted through the column or may be pulled
through the column with vacuum or pushed through with positive pressure.
2. Add 25 ng of PCP-d5 per mL of sample as internal standard..
3. Add 2.0 mL, 1% HCl in Deionized Water.Vortex mix.
4. If adjusted sample is turbid or precipitated centrifuge for 3 minutes
at 3000 RPM.
2. Add 1.0 mL of a Sodium Bisulfite solution to each column. Prepare
by dissolving 25 grams of Sodium Bisulfite in 100 mL of a (1:1)
mixture of H2O:0.25M Phosphate Buffer, pH 6.0. Prepare monthly.
3. Proceed to Sample Extraction within 60 min. of column conditioning.
2. Wash column with 3.0 mL of Deionized Water. Allow to flow by gravity.
3. Wash column with 2.0 mL of Methanol. Allow to flow by gravity.
4. Wash column with 1.0 mL Ethyl Acetate. Allow to flow by gravity. Proceed
to Sample Elution.
2. Place the column mounting plate on the elution rack loaded with an
appropriate number of 12 x 75 mm or 15 x 85 mm borosilicate glass test
tubes. Make sure that the hole pattern on the plate matches the hole pattern
on the rack.
3. Add 1.5 mL of n-Butylchloride:Ethyl Acetate (80:20) w/4% Triethylamine
(TEA)* to each column and allow solvent to flow through the columns by
gravity into the test tubes.
4. Dry under N2 or argon at less than 50°C.
* Elution solvent with 4% TEA, (4 mL TEA is added to 96 mL of n-Butyl
Chloride:Ethyl Acetate 80:20) is stable for approximately one week stored
in a glass bottle with a Teflon or polypropylene lined cap. Close bottle
tightly when not in use. A white residue begins to appear in the dried
down eluate when the TEA begins to deteriorate. Artifacts from this process
may interfere with "fast" GC/MS methods.
3. Mix the tube contents, and cap the tube or transfer contents into 100
µL reaction vials and seal.
4. Inject 2.0 µL.
Assisted flow parameters may be set as follows:
Column Conditioning - Pass through column in approximately 20 seconds
(± 20%).
Sample, Sample Washes, and Elution Solvent - Pass through column in approximately
60 seconds (± 20%).
