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ClinicalForensic
OpiateAnalysis
note
A
pplications
cat.#59576
Figure 2
Compounds
1. meperidine
2. alphaprodine
3. methadone
4. levorphanol (TMS)
5. codeine (TMS)
6. hydrocodone
7. morphine (TMS)
8. hydromorphone (TMS)
9. oxycodone (TMS)
10. oxymorphone (TMS)
11. nalorphine (TMS)
Opiates analysis onan
Rtx
®
-5column.
c-gram #3033
30m, 0.25mm ID, 0.25µmRtx
®
-5 (cat.# 10223). 2.0µl split injection of opiates.
Oven temp.:
200°C to 325°C@ 7°C/min.;
Inj./det. temp.:
250°C/300°C;
Carrier gas:
helium;
Linear velocity:
30cm/sec. set @ 200°C;
Split ratio:
50:1
Ionization:
EI
Mode:
SIM
Opiates or opioids are terms that
classify agroupof compoundswith
morphine-likeactions.Their pharmaco-
logical properties include analgesiaor
pain relief, drowsiness and respiratory
depression.
Figure1
shows the
structure formorphine. Substitutions at
the 3, 6, and17positions produce
compoundswithvaryingdegrees of
potencyandpharmacological activity.
TheNational Institute forDrugAbuse
IonsMonitored
71, 246
172, 187
72
150, 270, 271, 328
178, 196, 234, 371
242, 299
234, 429
356
371, 386
444, 445
414, 455
Figure 1
MorphineStructure
(NIDA) has targetedopiates as a class
tobemonitored inurine for detectionof
drug abuse. Testingguidelines have
been establishedwith a limit of
detectionof 0.3µg/ml formorphine.
Screeningof opiates is commonlydone
byusingenzyme immunoassays.
Enzyme immunoassays have the ability
to cross reactwith a number of
structurally similar opiates including
codeine,hydromorphone,hydrocodone,
levorphanol, andoxycodone. Inorder to
differentiate between all of the possible
substances beingdetectedby enzyme
immunoassay, confirmational analysis
byGC/MS shouldbeperformed.
Chromatographicperformanceof the
opiates is significantly affectedby small
changes in their chemical structure. The
presenceof hydroxyl groups at the3
and6positions produce compounds that
aremorepolar and reactive. Com-
poundswith reactivehydroxyl groups in
their chemical structure can suffer from
adsorption andpeak tailing, leading to
diminished response inchromato-
graphic systems that contain active
sites. Samplepreparationof sensitive
compounds, likeopiates, should take
place in silanizedglassware and
samples shouldbe stored indeactivated
samplevials.Derivatizationof reactive
hydroxyl groups can improvechromato-
graphicperformanceanddetection
limits andprevent sample loss on
glassware and sample vials. Both
trimethylsilyl and fluoroacyl derivatives
of the opiates yield endproducts that
are less polar and/ormore volatile than
theunderivatizedcompound.
For this analysis, trimethylsilyl
derivativeswerepreparedusingBSTFA
with1%TMCS.Derivatizing the
reactivehydroxyl groupwith a less
polar trimethylsilyl group eliminates the
tailingpeaks commonly seenwith
compounds likemorphine.
Figure2
shows the analysis of a selectionof
opiates on anRtx
®
-5 column. Com-
pounds that havebeenderivatizedprior
to analysis are designated asTMS in
the peak list. TheTMSderivatized
1 2
3
4
5
6
7,8
9 10
11
O
HO
HO
N–CH
3
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
