Research Information System
61th Annual Meeting TIAFT 2024, Sankt Gallen, Switzerland, 2 - 06 September 2024, pp.1-374, (Summary Text)
Background & Aims: Drug and illicit substance abuse during pregnancy is a significant public health problem. This
is because in utero substance exposure can affect the development of the newborn and therefore the child’s
long-term behavioral, cognitive and developmental abilities. There is little data on drug and illicit substance use
during pregnancy. The most common methods used to detect drugs and illicit substance use during pregnancy
are maternal interviews or surveys. In our country, according to our knowledge, no data has been found regarding
the determination of drugs and illegal substance in meconium. In this study, we aimed to develop and validate an
appropriate extraction method for anxiolytics (bromazepam, lorazepam, oxazepam, alprazolam, flunitrazepam,
diazepam, nordiazepam, medazepam, flurazepam, chlordiazepoxide, moclobemide), anticonvulsants (clonazepam,
7-aminoclonazepam, carbamazepine, oxcarbazepine, clobazam, nitrazepam, gabapentin, pregabalin), and
illegal drugs in meconium by liquid chromatography /tandem mass spectrometry.
Methods: Meconium sample (250±10 mg) was weighed into a 16x100 mm glass tube, followed by the addition of 2
mL of methanol and homogenized by ultrasonication for 30 min until uniform, then centrifuged at 3500 rpm for 10
min. The supernatant was transfer to another tube.
A solid-phase cartridge (OASIS MCX 60 mg, 3 cc) was used for sample extraction. Firstly, an SPE cartridge was conditioned
with 2 mL of methanol and 2 ml of deionized water. The sample was passed through the cartridge and washed
with 2 mL of 2% formic acid in pure water and 2 mL methanol:water: formic acid (47.5:47.5:5). The cartridge
was dried under vacuum for 15 min. Eluent was subsequently extracted with 2 mL of a mixture of dichlorometan:2-
propanol: ammonium hydroxide (20:78:2, v/v). After the extract was dried under nitrogen, it was reconstituted in
250μL methanol. A sample of 20 μL was injected into the LC- MS/MS. The LC-MS/MS analysis was performed on a
Shimadzu Nexera chromatography system equipped with an electrospray interface (ESI) in MRM mode. The chromatographic
separation was performed with a pentafuorophenylpropyl (PFPP) column (Allure 50×2.150 mm i.d., 5
μm, Restek, Bellefonte, PA, USA) using a gradient binary with 10 mM ammonium formate (A) in ultrapure water and
methanol (B). The run time is 20 min.
Results & Discussion: The method was validated in terms of limits of detection (LODs) (0.14–7.28 ng/g) and quantifcation
(LOQs) (0.44–22.06 ng/g), extraction recovery (ER%) (65.0–110.0%), matrix effect (ME%) (12.2-99.9 %),
linearity (r2> 0.99), intra- and interday precision (CV˂ 20%) and carryover. In this study, we compared two different
extraction cartidge (HLB and MCX) in meconium. In many routine laboratories, a single extraction procodure is
preferred to reduce laboratory costs and HLB cartridge is widely used. However, it was seen in our study that gabapentin
and pregabalin were detected with HLB cartidge only in very high concentrations.
Conclusion: A simple, cost-effective, and accurate LC/MS–MS method has been developed and validated for anxiolytics,
anticonvulsants and classic illegal drugs. It has been suggested that the MCX cartridge should be preferred
in meconium for these drugs.