Komajda Kamila, Przygodzka Dominika
Chair and Department of Forensic Medicine, Medical University of Lublin, Poland.
Arch Med Sadowej Kryminol. 2025;74(4):296-307. doi: 10.4467/16891716AMSIK.24.023.21222.
The aim of this study was to develop and validate a quantitative analysis procedure for four benzodiazepines (alprazolam, diazepam, flunitrazepam, clonazepam) and one Z-drug (zolpidem), applied to cotton material and Whatman® 903 cards in the form of blood spots.
The extraction was conducted using a mixture of phosphate-carbonate buffer pH=6, acetonitrile, and methanol. The blood spots were analyzed using high-performance liquid chromatography coupled with triple quadrupole mass spectrometry.
All calibration curves maintained linearity (R²>0.995) in the concentration ranges of 1-50 ng/ml on cotton material and Whatman ® 903 cards. The Limit of Detection (LOD) and Limit of Quantitation (LOQ) for the analyzed substances were in the ranges of 0.04- 0.40 ng/ml (LOD) and 0.12-1.08 ng/ml (LOQ) isolated from Whatman® 903 cards, and 0.06-1.15 ng/ml (LOD) and 0.17-2.04 ng/ml (LOQ) isolated from cotton. Accuracy for concentrations of 5 ng/ml, 10 ng/ml, and 50 ng/ml was comparable in both groups, ranging from 76-101% for Whatman® 903 cards and 73-117% for cotton. No significant matrix effect was observed on the quantitative results of the analyzed benzodiazepines.
In the presented study, a qualitative and quantitative analysis procedure for four xenobiotics from the benzodiazepine group and zolpidem, applied as dried blood spots on cotton material and Whatman® 903 cards, was developed and validated. The extraction method presented focused on isolating xenobiotics from blood spots of known volume. Sample extraction using a simple liquid-liquid extraction (LLE) technique creates opportunities for applying the described method to real samples secured on cotton material or stored on Whatman® 903 cards.
本研究的目的是开发并验证一种针对四种苯二氮䓬类药物（阿普唑仑、地西泮、氟硝西泮、氯硝西泮）和一种Z类药物（唑吡坦）的定量分析方法，该方法适用于以血斑形式存在于棉质材料和Whatman® 903滤纸上的样本。
使用pH = 6的磷酸盐 - 碳酸盐缓冲液、乙腈和甲醇的混合物进行提取。采用高效液相色谱 - 三重四极杆质谱联用技术对血斑进行分析。
在棉质材料和Whatman® 903滤纸上，所有校准曲线在1 - 50 ng/ml的浓度范围内均保持线性（R²>0.995）。从Whatman® 903滤纸上分离出的分析物的检测限（LOD）和定量限（LOQ）分别在0.04 - 0.40 ng/ml（LOD）和0.12 - 1.08 ng/ml（LOQ）范围内，从棉质材料上分离出的分别在0.06 - 1.15 ng/ml（LOD）和0.17 - 2.04 ng/ml（LOQ）范围内。两组中5 ng/ml、10 ng/ml和50 ng/ml浓度的准确度相当，Whatman® 903滤纸组为76 - 101%，棉质材料组为73 - 117%。在所分析的苯二氮䓬类药物的定量结果上未观察到显著的基质效应。
在本研究中，开发并验证了一种针对苯二氮䓬类药物组中的四种外源性物质和唑吡坦的定性和定量分析方法，这些物质以干血斑形式存在于棉质材料和Whatman® 903滤纸上。所提出的提取方法着重于从已知体积的血斑中分离外源性物质。使用简单液 - 液萃取（LLE）技术进行样品提取为将所述方法应用于保存在棉质材料上或储存在Whatman® 903滤纸上的实际样品创造了机会。
