SIMULTANEOUS DETERMINATION OF 32 NEW SYNTHETIC CANNABINOIDS IN HUMAN URINE AND HAIR BY LC-MS/MS

Yıl 2017, Cilt: Volume 2 Sayı: İssue 1 (1) - 2.İnternational Congress Of Forensic Toxicology, 86 - 86, 16.02.2017

Chung-feng Wang

Öz

The extraction procedure and detection
methods of new Synthetic Cannabinoids (ex: BB-22, SDB-005, THJ-018,
JZL-195……etc.) for human urine and hair samples are in great need due to these
new drugs are abused severely in recent years all over the world. Highly
sensitive analytical techniques are therefore required for trace-level
identification and quantification of these kinds of drugs. We report a fully
validated method here developed by our team which could simultaneously
determine 32 new Synthetic Cannabinoids in human urine and hair. We believe
this method could be used in our daily analysis for authentic cases in Taiwan,
and could also be adopted in other labs universally.

The aim of our research was to develop
a new method of simultaneous identification of Synthetic Cannabinoids in urine
and hair. And this method must be practical, highly sensitive (down to picogram
level), easily applicable, and carefully validated.

Synthetic Cannabinoids from spiked
urine or hair specimens were analyzed by LC-MS/MS with different pretreatments
as follows:

Urine Specimens

0.2 ml of each urine sample was
adjusted to basic condition with 0.1 ml of 1N sodium hydroxide. After addition
of deuterated internal standards, the urine sample was extracted by 1 ml ethyl
acetate in an ultrasonic bath for 30 mins, and followed by centrifugation at
13,000 rpm for 5 mins. Subsequently, 1 ml of extraction solution was transferred
to a glass tube and evaporated to dryness under a gentle stream of nitrogen at 35 °C. The residue was reconstituted in 0.2
ml of mixture of water/ACN, 70/30 (v/v). Gradient elution was performed by an
Agilent Zorbax Extend-C18 (5 um, 4.6 x150 mm) analytical column for LC-MS/MS instrumental
system consisting of a QTrap 4500 triple-quadrupole linear ion trap mass
spectrometer fitted with a TurboIonSpray interface by Applied Biosystems/Sciex
(Germany) and a 1260 Infinity Quaternary LC system by Agilent (USA) .

Hair Specimens

A 10 mg portion of washed hair sample
was added with 0.2 ng/mg deuterated internal standard and incubated at 40°C for 1 hour with 0.4 ml 1N sodium
hydroxide. Subsequently, a liquid-liquid extraction was performed, the
incubated hair sample was extracted with 1ml of ethyl acetate. The organic
solvent layer was transferred and gently evaporated to dryness in nitrogen at 35°C. The residue was dissolved in 0.2
ml mixture of water/ACN, 70/30 (v/v) and analyzed by the same condition just
mentioned above in urine specimens.

Validation of this new method for
simultaneously qualitative and quantitative analysis of Synthetic Cannabinoids
from spiked urine and hair samples by LC-MS/MS was fully performed. And over 12
authentic urine specimens were analyzed by this new method.

Urine Specimens

32 Synthetic Cannabinoids (SCs) from
spiked urine specimens were analyzed by LC-MS/MS. The limits of detection (LOD)
range from 0.05 ng/mL to 1 ng/mL (dependent on different SCs) and the limits of
quantification (LOQ) range from 0.20 ng/mL to 5 ng/mL. Linearity is in the
range between 0.2 ng/mL and 200 ng/mL for each compound (R2 value above 0.995).
Mean relative errors are between ± 10.0%. Precision variances are all below
15.0%.   

Hair Specimens

32 SCs from spiked hair specimens were
also analyzed by the same method. The limits of detection (LOD) range from 5
pg/mg to 20 pg/mg and the limits of quantification (LOQ) range from 10 pg/mg to
20 pg/mg. Linearity is in the range from 0.01 ng/mg to 2.0 ng/mg for each compound.
Mean relative errors are between ± 10.0%. Precision variances are all below
15.0%.

Highly specific qualitative and quantitative analysis of 32 novel
Synthetic Cannabinoids in urine and hair by LC-MS/MS has been developed and
successfully applied to real samples in Taiwan.

Anahtar Kelimeler

SIMULTANEOUS DETERMINATION OF 32 NEW SYNTHETIC CANNABINOIDS IN HUMAN URINE AND HAIR BY LC-MS/MS

Kaynakça

• Chung-Feng WANG Special Agent, Division of Forensic Science, MJIB, Taiwan