Liquid-Phase Microextraction and Gas Chromatographic-Mass Spectrometric Analysis of Antidepressants in Vitreous Humor: Study of Matrix Effect of Human and Bovine Vitreous and Saline Solution

Abstract

In forensic bioanalytical methods, there is a general agreement that calibrators should be prepared by fortifying analytes in matrix-based blank samples (matrix-based). However, in the case of vitreous humor (VH), the collection of blank samples for the validation and for routine analysis would require the availability of many cadavers. Besides the difficulty of obtaining enough blank VH, this procedure could also represent an ethical issue. Here, a study of matrix effect was performed taking into consideration human and bovine vitreous and saline solution (SS) (NaCl 0.9%). Tricyclic antidepressants [amitriptyline (AMI), nortriptyline (NTR), imipramine (IMI) and desipramine (DES)] were used as model analytes and were extracted from samples by means of liquid-phase microextraction and detected by gas chromatography-mass spectrometry. Samples of human and bovine VH and SS were prepared in six different concentrations of antidepressants (5, 40, 80, 120, 160 and 200 ng/mL) and were analyzed. Relative matrix effect was evaluated by applying a two-tailed homoscedastic Student's t-test, comparing the results obtained with the set of data obtained with human VH and bovine VH and SS. No significant matrix effect was found for AMI and NTR in the three evaluated matrices. However, a great variability was observed for IMI and DES for all matrices. Once compatibilities among the matrices were demonstrated, the method was fully validated for AMI and NTR in SS. The method was applied to six VH samples deriving from real cases whose femoral whole blood (FWB) was analyzed by a previously published method. An average ratio (VH/FWB) of ∼ 0.1 was found for both compounds.

Recent Advances in Analytical Techniques for Antidepressants Determination in Complex Biological Matrices: A Review

“…Protein precipitation, 26,48,58,59 LLE, 15,25,29,41,46,53,60 SPE, 12,17,[20][21][22][23]28,32,34,36,42,51,56,59,62,64 and some miniaturized techniques-such as SPME, 33 MEPS, 30,50,54 DLLME, 14 AA-DLLME, 27 BAμE, 24 and LPME 13 are the most utilized procedures for extracting antidepressants from the biological samples.…”

Alternative matrices in forensic toxicology: a critical review

“…LLE has also been reported and explored for VH samples [169][170][171][172][173][174][175]. Other techniques have been less frequently explored such as microwave assisted extraction [176], disposable pipette extraction [177], supercritical fluid extraction [178], dispersive liquid-liquid microextraction [179] and liquid-phase microextraction (LPME) [180].…”

“…Chromatography coupled to mass spectrometry-based methods have been frequently used due to its high sensitivity, selectivity and accuracy [151]. Gas chromatographic techniques used in separation and analysis of VH samples include GC-MS [154,156,163,164,[167][168][169][170][180][181][182], gas chromatography with nitrogen-phosphorus detector (GC-NPD) [170,177] and GC-FID [159,166]. Gas chromatography coupled to tandem ion trap mass spectrometry (GC-MS/MS) has also been applied in the analysis of cocaine in sheep's VH [183].…”

“…Over the years, VH has been successfully used in forensic casework, for illicit drugs and NPS testing. Several classic drugs of abuse have been detected in authentic VH specimens such as the following examples: ethanol [159,181,190], cocaine and its metabolites [155,164,166,172,176,181,[190][191][192], opioids and/or their metabolites [155,167,168,173,176,177,182,185,190], amphetamine and/ or methamphetamine [164,173,190], MDA derivatives [169], ketamine [172], phencyclidine [154,170], LSD and its metabolites [175], γ-hydroxybutyric acid [193], benzodiazepines [171,173,184,190], antidepressants [173,180,194,195] and barbiturates [196]. On the other hand, VH does not seem to be a good biological matrix for investigating cannabinoids [151].…”

Green Approaches to Sample Preparation Based on Extraction Techniques

“…HF-LPME has been utilized as a green microextraction methodology for multiple analytes, such as chlorophenols in water samples [169], organic contaminants in the aquatic environment [170], pesticides and their metabolites in water and soils [171], antidepressants in vitreous humor [172], NSAIDs in sewage sludge [173], amino alcohols in water samples [174], anabolic steroids in urine [175], selective serotonin reuptake inhibitors in sewage sludge [176], basic drugs in water [177], haloacetic acids in water [178], cocaine and its derivatives in hair [179], bisphenol A in river water samples [180], organochlorine pesticides in water [181], pesticides in environmental water [182,183], chloroanilines in water samples [184], aromatic amines in environmental water samples [185], paraben preservatives in cosmetic samples [186], short-chain fatty acids in plasma [187], fungicides in water [188], food analysis [189,190], nortriptyline in biological matrices [191], aromatic hydrocarbon isomers [192] and pesticide residues in food samples [193].…”