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Journal of Analytical Toxicology, Vol.

12, March/April 1988

Confirmation of Marijuana, Cocaine, Morphine,
Codeine, Amphetamine, Methamphetamine,
Phencyclidine by GC/MS in Urine Following
ImmunoassayScreening
S . J . Mul(~* a n d G . A . C a s e l l a

New York State DSAS Testing and Research Laboratory, 80 Hanson Place, Brooklyn, New York 11217 and
Department of Psychiatry, SUNY Health Science Center at Brooklyn, New York 12203

Abstract
Rapid, reliable, sensitive, qualitative, and quantitative
methods using small urine volumes (0.2-0.5 mL) were
developed primarily for confirmation of marijuana, cocaine,
benzoylecgonine, ecgonine methyl ester, morphine, codeine,
amphetamine, methamphetamine, and phencyclidine. Using
capillary gas chromatography/mass spectrometry (GC/MS)
and selected ion monitoring (SIM), mass spectra were
obtained for each analyte. Samples were prepared by
hydrolysis where applicable, organic solvent extraction, and
derivatization where necessary. Confirmation was achieved
by comparing abundance of major ions and retention time of
the total ion current (TIC) of an analyte with those of the
appropriate analytical standard. Quantitation was achieved
and calibration curves derived by obtaining the molecular ion
ratios of that analyte/internal standard (IS) over a
concentration range of 10-300 ng/mL (0.16-4.0 ng total
injected into GC/MS). The overall extraction efficiency for
these analytes ranged from 53~ to 96~ Statistically
significant cut-off values Oo< 0.01) were obtained for each
analyte. The slope, y-intercept, and coefficient of
determination (r 2) were calculated for each analyte. All of the
GC/MS methods were extensively tested against urine
samples determined positive or negative by immunoassay
(IA) and are now used in our laboratory.

Introduction

In the past few years urine drug abuse testing has entered the
workplace thus placing the burden of both scientific validity and
legal defensibility upon the analytical toxicologist. This responsibility has led to the conclusion that gas chromatography coupled
with mass spectrometry ( G C / M S ) offers the best means for unequivocal identification of drugs of abuse or their metabolites
in biological materials (1). Although G C / M S and GC methods
are available for marijuana (2-5), morphine and codeine (6-8),
amphetamine and methamphetamine (2,9), cocaine, benzoylecgonine and ecgonine methylester (2,10-14), and phencyclidine
(15-17), rapid, reliable, and sensitive assays requiring small

* Correspondence should be addressed to S.J. Mul~ at the OSAS address

102

volumes of urine were needed in our laboratory in order to meet
the daily requirements of high volume drug abuse testing. Consequently we d e v e l o p e d methods using capillary gas
chromatography with electron impact mass fragmentography
( E I / M F ) operating in the selected ion monitoring mode (SIM).
In this report we present these methods in detail and demonstrate
quantitative use through the addition of an internal standard
(IS) to the assays.

Materials and Methods
Equipment
A Model 5890A Hewlett-Packard G C with 5970B mass selective detector was used for the analyses. The data system in use
was the HP 310 computer with the H P 9133H Winchester disc
drive. The manufacturer provided tile operating application and
three dimensional display software for use with the HI:' 59970C
M S / M S D Chemstation. The MSD was operated in the electron
impact (El) mode at 70 eV with an ion source temperature of
280~ and an m/z range that varied with the drug or metabolite
analyzed (see below). The instrument was autott, ned daily with
perfluorotributylamine (PFTBA). The G C / M S intcrface temperature was 280~ A fused-silica capillary column (HP-190916-312), 12.5 m • 0.2 m m i.d., consisting of cross-linked
dimethyl silicone was used. The helium flow rate was 0.65
m L / m i n at a linear velocity of 34.7 cm/s. The operating temperature of the column varied with the analytes (see below) and
the injection was splitless.
Urine drug screening
The EMIT d . a . u J ' (Syva Corp.) was utilized for most drugs,
and samples indicating drugs at or above cut-off levels were
analyzed by the G C / M S procedures described below. The
radioimmunoassay used was either the Abuscreen"; (Roche
Diagnostics) or the U r i n e - T H C (lmmunalysis Corp.).
Drug analysis, extraction, derivatization, and GC/MS

Marijuana." ll-nor-gxg-tetrahydrocannabinol-9-carboxylic
acM
(THC-COOH). To a 15-mL silanized centrifuge tube, add 0.5
mL urine and 50 ng (in 0.05 mL ethanol) o f 9-carboxy-llnor-2~%tetrahydrocannabinol-5-"H3 (trideuterated THC-COOH)
(Research Triangle Institute). Alkalinize with 0.5 mL of 5~

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Derivatize sample by adding 200 #L of tetramethylammonium hydroxide (TMAH)/ dimethylsulfoxide (DMSO) (1:1. and 243 for PCP and m/z 180. Evaporate to dryness under N~ in a heated water bath (50-60~ Derivatize sample by adding 50 #L of pentafluoropropionic anhydride (PFPA. extraction. if quantitation is required. and 25 ng (25 #L) of phenylcyclohexylamine(Abbott Labs) as internal standard. The injector temperature is 250~ column temperature 165~ (l. Co.2 mL of blank urine. 118. add 1 mL of distilled water to the tube. To a 15-mL silanized tube. Amphetamine and methamphetamine. To a 15-mL silanized tube add 0.0 mL with distilled water. dissolve residue in 25 #L of ethyl acetate. for codeine m/z 282.L) of nalorphine in 0. v/v) and shake for 10 rain at low speed on an Eberbach shaker.2 mL of urine. Pierce Chem. Centrifuge for 5 min at 2500 rpm. 388.2 mL of blank urine adjusted to l mL with water. Add 3-4 drops of conc. v/v) and shaking for 10 min at low speed on an Eberbach shaker. Cool and bring to final volume of l mL with distilled water. 50 #L of 50~ NaOH.) as internal standard and bring to final volume of 1 mL with distilled water. The injector temperature is 250~ the column temperature 110~ (0. Dissolve residue in 25 #L ethyl acetate and inject 1 #L into GC/MS. and 140.L of ethyl acetate and inject 1 p.2 mL of blank urine. Utilizing the method described for PCP. 473. Pierce Chem. then evaporate to dryness as described.414. (0.385.5 mL of blank urine. Extract sample with 3 mL of chloroform/isopropanol (90:10.) and 25 #L of pentafluoropropanol (PFP. and 25 ng (25 #L) of phenylcyclohexylamine (IS) in 0. Hydrolyze for 15 min in a boiling water bath.). Shake sample for 10 min on Eberbach shaker at low speed.2 mL of blank urine is made alkaline and brought to a final volume of 1. and 445. 242. Co. being very careful not to carry over any of the aqueous phase. Pierce Chem.2 mL of urine and 100 ng (0.3 g of solid NaHCO~/K2COj (2:1) to the sample (pH 9) and mix on a vortex. Dissolve residue in 25 p. and 577. Vol. shake and centrifuge as described. A standard control urine (run concomittantly) containing 25 ng (25 #L) of PCP and 50 ng (50 #L) of ketamine in 0. Transfer lower organic phase to a clean tube and add 25/~L of glacial acetic acid and 50 #L of trifluoroacetic anhydride (TFAA.).0 min) to 225~ at 30~ SIM is used to detect major ion peaks of m/z 272. Extract sample by adding 3 mL of chloroform/isopropanol (90:10. PCR/SCM Chem. A standard control urine (analyzed simultaneously) contains 50 ng (50 ~L) of morphine. Standard control urine (analyzed at the same time) containing 25 ng (25/~L) of amphetamine. and 237 for ketamine (IS). 50 #L of pentafluoropropionic anhydride (PFPA. and 488 were obtained for the MSTFA derivative of THC-COOH (TMS) and m/z 374. 15-mL tube and evaporate to dryness under N2 in a heated water bath (50-60~ Derivatize the sample by adding 25 #L of pentafluoropropanol (PFP). and 603. for methamphetamine at m/z 110.L into the GC/MS. Phencyclidine (PCP). A standard control urine (analyzed simultaneously) should contain 100 ng (100 #L) of cocaine and 100 ng (100 #L) of ecgonine methyl ester in 0. Co. Morphine and codeine. Heat for 15 min in a heating block at 60~ and evaporate to dryness under N2. 15-mL centrifuge tube. Add 3 mL of 0. ll9. Co. 50 ng (50/~L) of ketamine may be added to the urine and extracted 103 . Add 1 mL of water. shake. Evaporate extract to dryness in water bath (50-60~ under nitrogen. An alternative derivatization method is to add to each tube (after hydrolysis.1 mL) of the internal standard scopolamine (Sigma Chem. 476. cap the tube tightly and heat for 15 rain at 60~ allow tube to cool at room temperature.2 mL of urine. both C and EME may be identified in an extract from 0.5 min) to 140~ at 25~ SIM is used to obtain major ion peaks for the TFA derivatives of amphetamine at m/z 115. and centrifuge as described. Pierce Chem. Co. Standard control urine (analyzed concurrently) consisted of 50 ng (50 #L) of benzoylecgonine (Alltech-Applied Science) and 100 ng (100 #L) of scopolamine (IS) in 0. Remove upper aqueous phase. v/v) as described previously and transfer the lower organic phase to a clean. v/v) plus 100 #L of iodopropane. 316. Cap and hydrolyze 30 min at 100~ in a heating block or boiling water bath. and mix by vortexing. Dissolve residue in 25 #L of ethyl acetate and inject 1/~L into the GC/MS. Evaporate to dryness under N~ in a heated water bath (40-50~ Reconstitute residue in 0. and for phenylcyclohexylamine (IS) at m/z 189. Add 0.IN KOH plus 3 mL of hexane and shake for 5 min. and 491 for the MSTFA derivative of the deuterated THCCOOH (IS). Extract with 3 mL of chloroform/isopropanol (90:10. 50 ng (50 #L) of codeine. The injector temperature is 250~ the column temperature 165~ (1 min) to 240~ at 30~ Using SIM.217.2 mL blank urine is made alkaline and brought to a final volume of 1. Transfer the lower organic layer with a Pasteur pipette to a clean. and 271. The injector temperature is 250~ column temperature 165~ (30 s) to 250~ at 50~ Selected ion monitoring (SIM) is utilized to detect major ion peaks of m/z 341.3 g of solid NaHCO3/K2CO3 (2:1).).). the major ion peaks for the PFP derivatives of morphine are m/z 357.OH and add 50 #L TFAA by mixing on vortex.2M TMAH in methanol. v/v). Major ion peaks of m/z 371. To a 15-mL silanized tube add 0. Centrifuge 5 rain. March/April 1988 NaOH by mixing on a vortex. 250 ng (25 ttL) of nalorphine as the internal standard. and 2-3 drops of cone. and 431 for the TMAH derivative of the deuterated THC-COOH (IS).) and bring to final volume of 1 mL with distilled water.2 mL of urine. Cap the tube tightly and heat for 15 min at 60~ (heating block or hot water bath). and 250 ng (25 p. then carefully transfer the lower organic phase (being careful not to carry over the aqueous phase) to a clean. Extract the sample by adding 3 mL of chloroform/isopropanol (90:10. v/v) by shaking for 10 min at low speed on an Eberbach shaker and centrifuge 5 min at 2500 rpm.L directly into the GC/MS. DMSO and iodopropane. 118.2 mL of urine. 138. 15-mL tube. and 421 for the PFP derivative of benzoylecgonine and m/z 108.). 300. 15-mL silanized tube and evaporate to dryness under N~. and for nalorphine (IS) m/z 357. Alternatively. Cocaine (C) and ecgonine methyl ester (EME). and inject 1 #L into GC/MS. and inject 1 p. then transfer upper organic phase to a clean. 15-mL tube and evaporate under N2 in a heated water bath (50-60~ Dissolve residue in 25 #L of ethyl acetate and inject 1 #L into the GC/MS.5 mL of CHCL3/C3H.440. and 449 for the PFP derivative of scopolamine. Regis Chemical Co. Bring to final volume of 1 mL with distilled water. 209. add 0. place in freezer to cool..Journal of Analytical Toxicology. 25 ng (25 ~L) of methamphetamine. and evaporation) 50 #L of n-methyl-n-trimethylsilyltrifluoroacetamide (MSTFA. Allow to cool at room temperature. add about 0. 388.0 mL with distilled water. Cocaine (benzoylecgonine). HCI to lower pH (<2) and add 3 mL of hexane/ethyl acetate (7:1. 50 #L of 50~ NaOH. heat for 15 min in a heating block at 90~ evaporate to dryness under N2. 12. Standard control urine (analyzed simultaneously) consisted of 50 ng (50 #L) of THC-COOH (Research Triangle Institute) plus 50 ng of deuterated THC-COOH (50 #L) as the internal standard in 0. Sigma Chem. Centrifuge at 2500 rpm for 5 min to separate the aqueous and organic phases and transfer the upper organic phase to a clean. Co. 200. The injector temperature is 250~ the column temperature 100~ (1 min) to 190~ at 20~ The major ion peaks using SIM are m/z 186. HCI. and 428 for the TMAH-catalyzed derivative of THC-COOH (dipropyl ester) and m/z 344. To a 15-mL silanized tube add 0. Centrifuge 5 min at 2500 rpm and then siphon off upper aqueous phase. and 50 ng (50 #L) of ketamine (Alltech-Applied Sci. and 154.

March/April 1968 as described..4 ~03)(06)<10) N Bi ~ (zo) (3~0) MSTFA-THC-COOH 9 SLOPE:O.: 001. The calibration curve for phencyclidine (PCP) over the concentration range of 10-1(90 ng/mL (0.4) C)------r--TOTALng Figure 2.6) 250 = (ZO) 9 n0/r~ TOTALng Figure 3. Each data point is the mean value ol at leasl 3 determinations.2-2..O 0 10 20 30 40 50 60 (0.2-2 ng total).98094. / ~ o ~ TFA-AMPHETAMINE E6PE./~ SLOPE:O. The sensitivity level (cut-off) was equal to 25 ng/mL for amphetamine and methamphetamine at select ion ratios of m / z 140/271 and m / z 154/271.. Injector temperature is 250~ and column temperature 100~ (1 min) to 190~ at 20~ hold 8.4) (0. The ratio of the molecular ion of the analyte to the molecular ion of the internal standard (except for amphetamines) was then determined and plotted against the concentration (ng/mL or ng total) of the analyte.2) (03) (04) (oS) ((16) 80 (06] 7 Q .c.1) (0.2)(0.4 ng total). Linearity with these drugs was obtained over the range of 25-250 ng/mL (0. ng/mL of analyte added to urine (total ng analyzed). Results and Discussion Although these assays were primarily developed to confirm positive results of screening by immunoassay.99325 and for methamphetamine 0. Select ion ratio of m/z 154/271 for methamphetamine/IS (phenylcyclohexylamine) vs.o 0. respectively. 104 T'FA-METM/ I f ~'k~HETAMINE SLOPE~O.4 7 INTERCEPT=-0.0] TMAH-THC-COOH SLOOIE:O0045 1 . The coefficient of determination for morphine was 0.. then to 225~ at 50~ The abundant ion peaks monitored (SIM) for cocaine are m / z 82.1-1 ng total) appear in Figure 1B with linearity achieved using the molecular ion ratio m / z 488/491. 12. the primary metabolite of ~ PFI~PHINE ~S'E-OPE=O. ng/mL of amphetamine added to urine (total ng analyzed). Excellent linearity was obtained with the molecular ion ratios m / z 428/431.1730 (4o~ 75 1OO 125 150 2. 96.o..8) (12) (Z0) (2.99491. ng/mL of methampbetamine added to urine (total ng analyzed).99067 and for the MSTFA derivative was 0. Molecular ion ratio of m/z 445/603 for codeine/IS (nalorphine) vs. .O015 A ~.o. (A) Calibration curve for TMAH-catalyzed derivative of 11-norAg-THC-9-carboxylicacid OHC-COOH).Journal of Analytical Toxicology.O012 . Each data point is the mean value of at least 3 determinations.6) (08) (10) (t2) 200 (1. ng/mL of codeine added to urine (total ng analyzed)..o ~ ~ ~" - o. 182. In Figure 2 appear the calibration curves and the slope and y-intercepts for the PFP derivatives of morphine and codeine.O037 v. Linearity was achieved with both of these drugs over the concentration range of 25-300 ng/mL (0. The latter value for the TMAH derivative was 0. ng/mL of morphine added to urine (total ng analyzed).0~ . and 303 and for ecgonine methyl ester are m / z 82. with phenylcyclohe• amine (PCHA) as internal standard.8 ng total) appears in Figure 4A. Calibration curve for PFP derivative of morphine and codeine with molecular ion ratio of m/z 577/603 for morphine/IS (nalorphine) vs.3 ng total).. The sensitivity of this assay (cut-off) was equal to or greater than 15 ng/mL (0.O001 J..99724 and for codeine was 0. Such calibration curves were obtained by adding known quantities of each analyte to control urines along with the appropriate internal standard. Each data point on each curve is the mean value of at least 3 determinations.08-0. and 199. Calibration curve for the TFA derivative of amphetamine with select ion ratio of m/z 140/271 for amphetamine/IS (phenylcyclohexylamine) vs. Vol. at least 50 ng/mL (0. in Figure 4B appears the calibration curve for the PFP derivative of benzoylecgonine.0234 INTIERCEPT=O.5 min. The results obtained with the TMAH-catalyzed derivative of THC-COOH (15-200 ng/mL. Molecular ion ratio of m/z 428/431 for analyte/IS (deuterated THC-COOH) vs. including their slopes and y-intercepts. 0. The slopes and the y-intercepts as well as the coefficient of determination (rz) were calculated. Molecular ion ratio of m/z 488/491 for analyte/IS (deuterated THC-COOH) vs. The results obtained with the MSTFA derivative of THC-COOH (10-100 ng/mL.^ "/ - "~ 9 ~j - ol o_o T I I 02550 I I I I 250 300 4~--ng/ml 100 150 (0.3-4 ng total) appear in Figure IA.NTERCEPT=O. they may also be quantitated by the addition of an internal standard (IS) to each analyte assay.. Linearity was achieved over this range with a maximum cut-off sensitivity of I 0 ng/mL using molecular ion ratios of m / z 243/237 for phencyclidine and the internal standard ketamine. ng/mL of analyte added to urine (total ng analyzed).. (B) Calibration curve for MSTFA derivative of THC-COOH.0207 E ~ 6.o2o i INTERCEPT0 0624 1QO~ n g / ~ (10( ~TOTALnQ 50 Figure 1. 0. The sensitivity (cut-off) of ~he GC/MS assays for both analytes was ~ ~ 1.O5~0- ~ ~ ~ ~ 4.4 ng total) with molecular ion ratios of m / z 577/603 for morphine and m / z 445/603 for codeine with nalorphine as the internal standard for both drugs.1 ng total) of the MSTFA derivative TMS-THC-COOH. The coefficient of determination for amphetamine was 0. In Figure 3 appear the quantitative calibration curves for the TFA derivatives of amphetamine and methamphetamine.e ~ O D E I N E /.o. ~- . The sensitivity (cutoff) obtained was equal to or greater than 10 ng/mL (0..$ (O2) (04) (0.99439.s- z~O. A i~o.

and 488 with percentage abundancein reference to the major ion m/z 371 (100). These two drugs at a concentration of 20 ng in 1 #L were each scanned over the range m / z 40-250..D Ion 385.O8 amu.2) m~n) . In Figure 5C appear the mass fragmentation spectra of the TMAHcatalyzed derivative of T H C . A TIC retention time of 2. The slopes and y-intercepts of these analytes also appear in Figure 4.o 7.1 ~a . In order to avoid this problem. of which 20 ng (1 #L) was injected into the G C / M S and scanned from m / z 60 to 450.4 ng total) with molecular ion ratios of m / z 421/449 for benzoylecgonine and the internal standard scopolamine. as well as the base peak ion rn/z 282..488 (m + ) after extraction of 50 ng from 0. and 428 with percentage abundance ions in reference to major ion m/z 341 (100).527 min was obtained for amphetamine and 3.395 ~~ 4.. For methamphetamine (Figure 7D). A retention time of 7...2-1. were monitored after extraction of 50 ng codeine from 0. Linearity of response was obtained over the concentration range 25-125 n g / m L (0.0 ng total). v/v) with a subsequent derivatization with TFAA as described and 1 #L injected into the G C / M S (0. March/April 1988 cocaine. For the TIC.345 mln) or V3:TNC. 12.gg amu. from V3:TTHC3U.99749. (D) The selected ion current profile for TMAH-THC-COOH of m/z 341..C O O H (500 ng). (A) Mass spectra of TMS-THC-COOH. The codeine ion current profile appears in Figure 6D.3 Time (mln. .5 7+4 200 L0. The selected ions monitored for the TMAH-catalyzed derivatives of THC-COOH were m / z 341.5 ng of methamphetamine was extracted into chloroform/ isopropanol (90:10. The pentafluoropropionic anhydride derivative of morphine (40 ng in 1 #L) was injected into the GC/MS and scanned over the range m / z 70-625 for the mass spectra (Figure 6A)./ .345 min was obtained for the total ion current (TIC). Molecular ion ratio of mlz 2431237 for PCP/IS (ketamine) vs. respectively.2 (0.414 (base peak). The assay was sensitive (cut-off) at 50 n g / m L (0.5 ng of each analyte) and scanned for selected ions.2 mL of urine with 2 ng in 1 #L injected into the GC/MS.J 400 +. and 154 (base peak).395 was obtained. (A) Calibrationcurve for phencyclidine (PCP).! .687 min for methamphetamine.C O O H (500 ng).5 and 17. .5E4 t 10000] % 7.I .593 min for the TIC of codeine was observed. 300 400 M&~s/Charge Ion 32t.D // s.. In Figure 5A appear the mass fragmentation spectra of the MSTFA derivative of T H C .. the dipropyl derivative of THC-COOH was prepared with TMAH and iodopropane.324 min was observed for the TIC of morphine.OmS INTERCEPT=O. it unfortunately readily contaminated the ion source of the instrument.385.Journal of Analytical Toxicology. a retention time of 10.01 II [i~!l 413 299 / S~ " .OO74 INTERCEPT:-O.. of which 10 ng (1 ~L) was injected into the G C / M S and scanned from m / z 70 to 500. In Figure 7B appear the ion current profiles obtained at rn/z 115. ... Molecularion ratio of mlz 4211449 for benzoylecgoninellS(scopolamine) vs..5 ~= o.D Ion 42Bl~[~. ng/mL of analyte added to urine (total ng analyzed).7.s C ~2.O TO'[~I_i~ Figure 4.5 ng of amphetamine plus 12.98588. 100 ~ ++i 6000] 5000] z~m-m 2 .. I 300 H~g~/Charge ]on 34). (B) PFP derivative of benzoylecgonine. Although the MSTFA derivative of THC-COOH was easy to prepare.. and 140 (base peak) for amphetamine..~ imu. The mass fragmentation spectra for the trifluoroacetic anhydride derivative of amphetamine and methamphetamine appear in Figure 7A and 7C. Lg. 118. . 10. They appear as ion currents in Figure 5D.~[4] 0.. from V]:TTHC3U..~ .4) 10. 118.2 .2 mL of urine. 105 ..~ (mln I . 27.(~1 (0.. The selected ion current profiles are illustrated in Figure 5B showing the abundant ions for TMS-THC-COOH of m / z 371.61 (0. The mass fragmentation spectra for the PFP derivative of codeine (40 ng in 1 #L) over the range scanned m / z 55-450 appear in Figure 6C. and 577 (m + ) (Figure 6B). The methamphetamine molecular ion 245 was barely detectable and therefore of little quantitative value.G of V3:TTHC... and 18.mu from V3:TTHC3UD 2000~-' 1000 7. Figure 5. .o+++/ t++ ~+t ..eE4~ 62 O.(C) Mass spectra of the TMAH-catalyzedTHC-COOH.5 mL urine and injection of 1 ng in 1 #L into the GC/MS. ~'t ~\ 7. of which 2 ng in 1 #L was scanned in the SIM.~. Vol.1. A retention time of 5. From 0..81 (1. (B) The selected ion current profile for TMS-THC-COOHof m/z 371. respectively.. Morphine (50 ng) extracted from 0. A retention time of 5. major peaks and abundances were obtained at m l z 110.. 12. In Figure 8A appear the mass fragmentation spectra for the PFP derivative of benzoylecgonine. revealed three major ion peaks at m / z 357.D 207 / 7..2 mL of urine. } Sczm 63 (IO. of which 20 ng (1 #L) was Scam 03 (7..11 . and for benzoylecgonine was 0.473.4 Time (0. even though additional steps were required. Each data point on each curve is the mean value of at least 3 determinations.~ tl. The molecular ion m / z 445 and 388. 26.473. The molecular ion (m + ) of 231 is barely detectable and therefore of little use in quantitation or identification.mE+i P+ +.~ 2 " O ~ q l ! PHENCYCMO~E(PCP) SLO~=O..068 9 . The coefficient of determination calculated for PCP was 0.| 100 B +. respectively. .1 H . ng/mL of PCP added to urine (total ng analyzed). ?i 290 .D 2-gE4 t 1.O811 t \ tO. 0 ~ ! 147 193 \ \ / ~ SLOPIE-'O. Either derivative of THC-COOH may be used effectively to identify and confirm marijuana use.4. 385. Crem V3:T U/HC... and 428 (m + ).

and 87~ for PCP. .6 and 6. from DATk:A-URMPH. 65. Ion 4 t 4 ~ amu. and 243 (m + ).01 level of significance. 73% for the TMAH derivative of THC-COOH. . (B) The selected ion current profile for TFA-amphetamine of m/z 140..51 "-= 2. deuterated standards of the parent drug or metabolite make the Of DATA:H2 D G. In Figure 8C appear the mass fragmentation spectra for phencyclidine (PCP) over the range m / z 70-300 for 10 ng in 1 /~L. (5) with the aforementioned methods were 84~ for TMSTHC-COOH.. [ I ' I 88 SCan 128 ( 3 .j #tom [JI4Tr 4 i / . and 118 with percentage abundance ions in reference to the base peak of m/z 154 (100). / 5~ (/ 4gg 500 60 #rom DAT~:H4 9 8 D 225 Ion Ion 1/65 \ 268 ' 341 O 68081 4800 ]" 28Of. 61~ for TFAamphetamine. D ~LI~. may also affect the signal to background ratio.8 8~s~. Paul et al. respectively. V o l . The methods employed to extract and identify PCP may also be utilized to extract cocaine and its metabolite ecgonine methyl ester (EME). 55. (B) The ions in the selected ion current profile for PFP-morphine were m/z 414. Using the methods described in this report. from DRTIq~R-URHPH.qTA:R HETN. S '"'-.8. .5 Figure 7. o# ~ '118 9l gg~Jg . Final concentration of PCP injected into the G C / M S was 0. (D) Major ions in the SICP for TFA-methamphetamine were m/z 154.2 mL of urine (Figure 8B). M a r c h / A p r i l acetate (7:1.Oe Ion 1 1 8 .3808 ] 3 80981 1. 20~ . (A) Mass spectra for TFA-amphetamine.353 min for the TIC. The selected ion current peaks monitored (Figure 8D) for PCP were m / z 186. 182 (base peak). ) 490 / / " 27. and benzoylecgonine where the value was 50 ng/mL at the p<0.0[5 \ . v/v) for all other analytes. (C) Mass spectra of the PFPcodeine. 12. PCP was analyzed in the SIM mode by extracting 0.1. 65~ for PFP-benzoylecgonine.2XL"g4.Journal injected into the G C / M S and scanned from m / z 70 to 450. .01) was the lowest concentration analyzed (Figures 1-4) except for morphine.f DATR:H2. Two primary extracting solvents were used: hexane/ethyl Scan L3 ( 5 .4FI 8 . The sensitivity of these assays may be increased in some cases by increasing the volume of urine extracted 2. 20. and 199 (m + ). 50 ng of benzoylecgonine was extracted and derivatized with PFPA. 300 (base peak). . '~ 1 Ion ll(~. selected major ion peaks were m / z 82 (base peak).8 ng in 1-#L. ] 1.. v/v) for THC-COOH and chloroform/isopropanol (90:10. with percentage abundance ions in reference to the base peak of m/z 414 (100).0 Time ( m l n . B . A retention time of 4.0. Selected ion monitoring for cocaine provided values o f m / z 82. . 96% for PFP-morphine.Charge Ion 282 t~O ainu.004 "-1 445 2~8 ]g(] klass . m:~ ~.4 2.5. 53% for TFA-methamphetamine. A retention time of 4. gE5 114 " / \ 0 302 .3 and 31. [JRTA:H4 9 / 8 .[. 388 5. A cursory examination revealed that the stability of the PFP and TFA derivatives as well as the THC-COOH derivatives normally did not appear to exceed 72 h at . . increasing the volume injected into the G C/ MS (2-3 p. and 421 ( m + ) . ( ~ amu. respectively. oo . . r~me I m l n l 42 LIg rain) Of 2.to 3-fold. codeine. gg5 ~RTRI r4 R~!PN L) l 2. . .OE58OES] Toxicology. A . respectively.~ D.g Tlme [mln. . and 388 with percentage abundance ions in reference to the base peak of m/z 282 (100).D I 1988 . 577.9 and 9. 110. and 303 (m + ) at a retention time of 10.6 and 9.0 ].341 min for EME total ion current was also obtained. . 8 8 a~:~. One microliter (2 ng) was injected into the GC/MS and scanned to obtain selected ion current profiles of m / z 272.. 6 8 ? !" 1282 t/19 14g .28 Figure 6. oc5 i(~08~i .150 min (TIC) was observed for PCP. however. (A) Mass spectra of the PFP-morphine. and 115 with percentage abundance ions in reference to the base peak of m/z 140 (100).. ) 4.L). 200 (base peak).6g . Obviously. or decreasing the volume of ethyl acetate used to dissolve the residue following extraction and derivatization of the analyte. .oo . 106 C /I !. 242.~2 188 120 14~1 168 tO0 tOO Has~/Charge 14~.$8 / 22D '. 91 ~ for PFP-codeine. 2. A retention time of 8.445. from DRTA:H4 9.o 4 80~ 0 I D "~\ / . For ecgonine methyl ester. . . (D) The ions in the SICP for PFP-codeine were m/z 282. 118.o. the statistically significant cut-off value for each analyte Lo<0.2 mL of urine containing 20 ng of PCP. 3 2 4 mln) of Analytical Scan 39 ( 2 5 2 7 4. 30~ Has~/Charge 577 ..o. .~ Scan 3~ 15597 ~Inl . Quantitation may be achieved for these analytes by adding ketamine (50 ng) as an internal standard and determining the ratios of the molecular ion of cocaine or EME metabolite to ketamine.D..e ~ Ion 4 4 5 . (C) Mass spectra for TFA-methamphetamine.oEsI ecs] .5 4. 96. the primary purpose being quantitation with confirmation of the positive sample. 352 . . ~rom D~TA:ALJI9 4 ~) 1. /. 33. . These changes. . frou.2 0 ~ The internal standards used in these assays were chosen for functional ease of application. From 0. The extraction efficiencies or recoveries obtained using the method described by B. .~O ainu. .543 min (TIC) was observed. J I 5.D /\ 1808 ~\ 3. 316. and 357. / - 5. respectively.

Obviously. pp. fPOm DRTR:P PCPU. and T. 8. Anal.242. J. Anal.E Domino.u.~ ~mu.c*~ ~ f . .0. 5. Mell.s 4. 5 BE4J 62 I-.D. E. D o :i:iii 0--" e.3 e. One trained technician working through a normal shift may prepare and submit about 25 urine samples to G C / M S analysis.B Ion 242.. J. J. Anal Toxicol.~ ~mu\. A. Meyers.7 5. Toxicol. M.H. A major metaboiite of cocaine. 7.I.D. P. Toxicol. 5-198. J..B.F. the deuterated standards are very expensive and are controlled. if available and if in pure form. f r o m DFII'FhAI. Cone. Anal. J. Anal Toxicol. J. 10:175-77 (1986). and R. Friedel. These methods are sensitive.~ . Simultaneous identification and quantitation of codeine and morphine in urine by capillary gas chromatography and mass spectroscopy. Anal.0E41 4 .~)u.) 9. Toxicol. 1986. Mell.V. R. Vol. and currently in routine use in our laboratory.A. l ][ .J.M. 2. 223:331-39 (1981). from DRTA~AI. Harrison.0C*1 . 9 : 2 2 2 . Irving. Biomed. Gas chromatography-chemical ionization mass spectrometry of cocaine and its metabolites in biological fluids. R. 3. Anal 7oxicol. 4. B. . Quantitation of phencyclidine. Anal Toxicol. J. Chinn. Lin. and 13. 50 5.. E. (A) Mass spectra for PFP-benzoylecg0nine. to Mr. Simultaneous determination of phencyclidine and monohydroxylated metabolites in urine of man by mass fragmentography with methane chemical ionization. Toxicol.M. Fentiman. NIDA Research Monograph No.D.5. . its metabolites and derivatives by gas chromatography with nitrogen-phosphorus detection: Application for in vivo and in vitro biotransformation studies. Mitchell. (2) selected ions monitored with an acceptable variance o f +_20~ in the ion ratios. DePace. Wu Chen.S.4.G. and A. References 1.. 300 4e0 i. Kogan. 13. J.S. Toxicol. L.8 4. 0 0 Ion 4 2 1 f I ~ 00~101 . Foltz. 9.R. J.5 Scan 4. Toxicol.A.. 9" 241-45 (1985). 1987. 1980. M.. 6. D..0s 4. DHHS publication number (ADM) 87-1481. Government Printing Office. M.L. . respectively. Detection of O~-monoacetylmorphine in urine samples by GC/MS as evidence for heroin use. Foltz. J. Owens. Schaffer. N.5 9 (1984). 200 H~ss/Ch~rge IOB Ion 3 0 0 . 14.J. R. 1987.I. Urine Testing for Drugs of Abuse. J. Ruo.1 ~2. Rapid method for the GC/MS confirmation of 11-nor-9-carboxy-Ag-tetrahydrocannabinol in urine. Meggs. 34. Mul$. L. Jennison.D ~oa I 243 z. and (3) computer match quality of usually 98~ or greater. GC/MS Assays For Abused Drugs in Body Fluids.Journal of Analytical Toxicology. Arthur Kramer for statistical analysis o f the data. r DRTR:P PCPU. McCurdy.. 73. and 272 with percentageabundanceions in referenceto the base peak of m/z 300 (100). Anal. 16.S.N Chiang.J. the number of technicians.J. and S.W. Detection and Quantitation of urinary 11-nor-delta-9-tetrahydrocannabinol9-carboxylic acid. Mayersohn. Our criteria for confirmation of a positive urine sample by comparison with an extracted standard analyte are (1) a retention time for the total ion current (TIC) with an acceptable variance of less than _ ! 070. Toxicol.2. 30. Quantitative determination of benzoylecgonine and cocaine in human biofluids by gas-liquid chromatography. . 9: 273-74 (1985). . Yousefnejad. Acknowledgments The authors are extremely grateful to Ms.M. Woodworth. and FLJ. to Mr. J. Quantitation of biologically important primary amines as their iosthiocynate derivatives by gas chromatography using nitrogen detector and validation by selected ion monitoring. Lewellen. A. and to Mrs. Anal. 243. B. 49:1965-69 (1977).5. 12.2 9 (1982). J. J. Eds. Ecgonine methyl ester. Government Printing Office. Novak. 16.D [On I ~ D R T R r P 2 P C P U . Anal. Ambre and 1". Ambre. Toxicol. Evaluation of the ion trap detector for the detection of 11-norL@-THC-9-carboxyfic acid in urine after extraction by bonded-phase adsorption. March/April 1988 best possible G C / M S internal standards.. the availability of G C / M S instruments.D I B 4. Clin. Eds. 32.(B) The selected ion current profile for PFP-benzoylecgoninewas m/z 300. reproducible. L. Chem. Resnick.6. 6:231-34 (1982). Elizabeth McLeod for typing the manuscript. J. Narasinhachari and R.9 142 (e. BE41 100 150 200 Hi~/Ch~rge Ion 200. Fehn and G. 15. Peat. DHHS Publication number (ADM) 80-1014.k. (C) Mass spectra for phencyclidine(PCP). and D. Acta 110:235-43 (1981). 38.B. and S.S. 10:107-15 (1986).J. T. Appl.00 ~ .C. Paul. 30-42. and 186 with percentageabundanceions in reference to the basepeak of m/z 200 (100). Hawks and C. Use of DB-1 capillary columns in the GC/FID analysis of benzoylecgonine.C. thus adding more controls to an already heavily regulated process. 8 : 2 .J. 8 : 2 5 5 .8. 11 : 1-5 (1987). J. . J. Verebey. NIDA Research Monograph No.D. 316. Folk and B. McKinley. Harry. Jed Shaw for the illustrations. 11. N. 107 . Unfortunately. H. Ann DePace for technical assistance. J.J.4 Figure 8. Halsztynska and E. so.2 Time (~ln. The urinary excretion of cocaine and metabolites in humans: A kinetic analysis of published data.L. J.z ! 41 Gu. Anal. Baker.15g mlnl of D~Tg:PCP. and R.S. A metabolite of tetrahydrocannabinol by capillary gas chromatography and electron impact mass fragmentography. Russell. Manuscript received June 2. Chromatogr. Quantitative analysis of phencyclidine and metabolites by capillary column gas chromatography. from DRTR:P:PCPU.2 6 (1985).D. . Chim. respectively. L. 4:37-42 (1980).M. and the use of automatic sampler devices will enormously enhance the daily confirmation capability. J. BE4] C p o . K. Anal. Crouch.t e. pp. 17. M. and R. 12_ J. and 21..D imu. B.J. Callhan. Buchwald.L. Stein. 10. 421.M. Sirnuttaneous quantitation of morphine and codeine in biological samples by electron impact mass fragmentography.4.. Childs. and P. Finkle. R. . Paul. revision received October 19.D IOn 243. D.B. W. 9:134-38 (1985).M. rapid. 6 : 2 6 . U. J.6 (1984). U. Mitchell.S.. (D) The major ions in the SICP for PCP were m/z 200.