Page 90 - Advances in Forensic Applications of Mass Spectrometry - Jehuda Yinon

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monitored. The LODs ranged from 2 to 30 mg/l in full scan mode and from
0.1 to 3 mg/l in SIM.
Recent publications also contained research concerning the determina-
tion of synthetic or semisynthetic opiates. Gaulier et al. reported a suicidal
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poisoning of a 25-year-old male heroin addict with a high dose of buprenor-
phine. Buprenorphine (BU) and its active metabolite norbuprenorphine
(NBU) were determined in body fluids and organs with LC/ESI/MS after
deproteinization and SPE. In gastric content, only BU was found in concen-
tration of 899 mg/l. The following concentrations were found in selected
matrices: in blood BU 3.3 mg/l, NBU 0.4 mg/l; in bile BU 2035 mg/l, NBU
536 mg/l; and in brain BU 6.4 mg/l, NBU 3.9 mg/l. Besides BU and NBU,
high concentrations of 7-aminoflunitrazepam were found in blood (1.2
mg/l), urine (4.9 mg/l), and gastric content (28.6 mg/l). Polettini and
Huestis developed a LC/ESI/MS/MS method for determination of
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buprenorphine and its metabolites — norbuprenorphine and buprenorphine
glucuronide — in human plasma. SPE extraction with C18 cartridges and
gradient elution was applied. For buprenorphine, and norbuprenorphine, as
well as for deuterated analogs used as internal standards, the protonated
quasi-molecular ions were monitored for the buprenorphine glucuronide
protonated quasi-molecular ion and buprenorphine aglycone. The LOQ was
0.1 mg/l for all compounds. On the basis of the transition m/z 590Æ414,
norbuprenorphine glucuronide was also tentatively detected. The reference
standard of this compound was not available. The authors stated that useful
fragmentation of the buprenorphine molecule was not possible; after the
increase of fragmentation energy this compound dissipated to very small
particles. On the other hand, Moody et al., 53,54, who also used ESI/LC/MS/MS
and fragmentation, and achieved detection at the level of 0.1 mg/l in MRM
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mode. Additionally, Hoja et al. and Bogusz et al., who applied ESI and
APCI/MS, respectively, observed fragmentation of buprenorphine — which
was particularly distinct in the case of the APCI/MS used by Bogusz. This
confirms the need for comparison of both ionization sources (ESI and APCI)
when the method for a particular compound is established.
Naidong et al. published LC/ESI/MS/MS procedures for determination
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of hydrocodone and hydromorphone in plasma. The drugs and deuterated
analogs were extracted with solvent and separated from glucuronides using a
50 mm ¥ 2 mm silica column and a mobile phase consisting of
ACN–water–formic acid (80:20:1). The LOQ was 0.1 mg/l. The same group
determined fentanyl in plasma, using automated 96-well solid-phase extrac-
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tion, straight-phase chromatography, and ESI/MS/MS. The LOQ was 0.05
mg/l in plasma, based on 0.25 ml sample volume. It should be stressed that
the Naidong group is consequently using straight-phase columns and a mobile
phase containing high concentration of organic modifier for opiate agonists.

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