Aydoğdu Melike, Ertaş Hasan, Ertaş Fatma Nil, Akgür Serap Annette
Institute on Drug Abuse, Toxicology and Pharmaceutical Science, Ege University, Bornova, Izmir 35100, Türkiye.
Chemistry of Department, Faculty of Science, Ege University, Bornova, Izmir 35100, Türkiye.
J Anal Toxicol. 2025 Jan 20;49(1):26-35. doi: 10.1093/jat/bkae081.
This study focused on the simultaneous detection of amphetamine, 3,4-methylenedioxy methamphetamine, morphine, benzoylecgonine, and 11-nor-9-carboxy-tetrahydrocannabinol in whole blood and dried blood spot (DBS). It is aimed to select a solvent mixture for liquid-liquid extraction technique employing liquid chromatography-tandem mass spectrometry (LC-MS-MS). The obtained DBS results were compared with the whole blood samples results. A simple, rapid, and reliable LC-MS-MS method was developed and validated for all analytes in whole blood and DBS. LC was performed on a Hypersil Gold C18 column with an initial gradient of 0.01% formic acid, 5 mM ammonium format buffer in water, and acetonitrile at 0.3 ml/min with 7.5 min runtime. A methanol:acetonitrile (40:60 v/v) mixture was selected for both matrices. Limit of quantitation (LOQ) values were 10-25 ng/mL; linear ranges were LOQ-500 ng/ml for all analytes; correlation coefficients were greater than 0.99, and all calibrator concentrations were within 20%. Analytical recovery in blood and DBS ranged from 84.9% to 113.2% of the expected concentration for both intra- and inter-day. Analytes were stable for 1, 10, and 30 days after three freeze/thaw cycles. It was determined that the variances of the results obtained with the two matrices in the comparison study were equal for each analyte, and the results were highly correlated (r = 0.9625). A sensitive, accurate, and reliable chromatographic method was developed to determine amphetamine, 3,4-methylenedioxy methamphetamine, morphine, benzoylecgonine, and cannabis, by performing the same preliminary steps with whole blood and dried blood spots. It was observed that the results obtained in these two matrices were compatible and interchangeable when statistically compared.
本研究聚焦于全血和干血斑（DBS）中苯丙胺、3,4-亚甲基二氧基甲基苯丙胺、吗啡、苯甲酰爱康宁和11-去甲-9-羧基-四氢大麻酚的同时检测。目的是为采用液相色谱-串联质谱法（LC-MS-MS）的液-液萃取技术选择一种溶剂混合物。将获得的DBS结果与全血样本结果进行比较。针对全血和DBS中的所有分析物，开发并验证了一种简单、快速且可靠的LC-MS-MS方法。液相色谱在Hypersil Gold C18柱上进行，初始梯度为0.01%甲酸、5 mM甲酸铵缓冲液（溶于水）和乙腈，流速为0.3 ml/min，运行时间为7.5分钟。两种基质均选择甲醇:乙腈（40:60 v/v）混合物。定量限（LOQ）值为10 - 25 ng/mL；所有分析物的线性范围为LOQ - 500 ng/ml；相关系数大于0.99，且所有校准物浓度在20%以内。血样和DBS中日内和日间分析物的分析回收率为预期浓度的84.9%至113.2%。经过三次冻融循环后，分析物在1天、10天和30天内保持稳定。在比较研究中确定，两种基质获得的结果对于每种分析物的方差相等，且结果高度相关（r = 0.9625）。通过对全血和干血斑进行相同的预处理步骤，开发了一种灵敏、准确且可靠的色谱方法来测定苯丙胺、3,4-亚甲基二氧基甲基苯丙胺、吗啡、苯甲酰爱康宁和大麻。经统计比较发现，这两种基质获得的结果具有兼容性且可互换。
