Liquid chromatography with tandem mass spectrometric method for determination of 52 antibiotics in human whole blood and urine and application to forensic cases

IF 2.8 4区医学 Q2 TOXICOLOGY

Forensic Toxicology Pub Date : 2024-04-20 DOI:10.1007/s11419-024-00688-y

Qianwen Shi, Gege Wang, Shuhui Wang, Chao Zhang, Zhiwen Wei, Zhongyuan Guo, Dan Zhang, Keming Yun, Shanlin Fu

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Abstract

Purpose

A rapid and reliable method was developed and validated for the simultaneous analysis of 52 antibiotics (cephalosporins, penicillins, carbapenems, lincosamides, quinolones, nitroimidazoles, macrolides, sulfonamides, tetracyclines, glycopeptide) in urine and whole blood by high-performance liquid chromatography–tandem mass spectrometry (HPLC-MS/MS).

Method

Analytes were extracted by dilution or protein precipitation and analyzed on an Agilent 1260 HPLC system coupled to an Agilent 6470 Triple Quadrupole Mass Spectrometer.

Results

The method attended method validation criteria. The limits of detection were equal or lower than 2.0 ng/mL, whereas the limits of quantification ranged from 0.1 to 10.0 ng/mL, from 0.1 to 5.0 ng/mL, in urine and whole blood, respectively. For all analytes, the bias and intra- and inter-day precision values were less than 14.7%. The ranges of recovery values of all antibiotics were 76.5–124.5% in whole blood and 76.3–121.8% in urine, values of the effects were lower than 25% in two matrices. No evidence of carryover was observed. The study of sample stability showed that almost all analytes were stable at 24 °C for 24 h, all analytes were stable at −20 °C for 14 days and at −80 °C for 30 days. Freeze–thaw cycles stability showed that antibiotics were stable except for imipenem. Autosampler stability study showed that all analytes were stable for 24 h, except for imipenem and amoxicillin. Applicability was proven by analyzing authentic whole blood (n = 86) and urine (n = 79) samples from patients under antibiotics treatment. Therefore, this method was applied to the analysis 3 forensic allergy cases, which were positive for at least one analyte.

Conclusions

A simple, sensitive and high-throughput method for the simultaneous determination of different classes of antibiotics in urine and whole blood samples was developed and applied. This sensitive method was successfully applied to forensic cases.

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液相色谱-串联质谱法测定人体全血和尿液中 52 种抗生素的含量及在法医案件中的应用

目的建立了一种快速、可靠的高效液相色谱-串联质谱（HPLC-MS/MS）同时分析尿液和全血中52种抗生素（头孢菌素类、青霉素类、碳青霉烯类、林可霉素类、喹诺酮类、硝基咪唑类、大环内酯类、磺胺类、四环素类、糖肽类）的方法。结果该方法符合方法验证标准。尿液和全血中的检出限等于或低于2.0纳克/毫升，定量限分别为0.1至10.0纳克/毫升和0.1至5.0纳克/毫升。所有分析物的偏差以及日内和日间精密度值均小于 14.7%。所有抗生素在全血和尿液中的回收率范围分别为 76.5%-124.5%和 76.3%-121.8%，在两种基质中的影响值低于 25%。没有观察到携带的迹象。样品稳定性研究表明，几乎所有的分析物在 24 °C 下 24 小时内都是稳定的，所有分析物在 -20 °C 下 14 天内和 -80 °C 下 30 天内都是稳定的。冻融循环稳定性表明，除亚胺培南外，其他抗生素都很稳定。自动进样器稳定性研究表明，除亚胺培南和阿莫西林外，所有分析物在 24 小时内均稳定。通过分析抗生素治疗患者的真实全血（86 份）和尿液（79 份）样本，证明了该方法的适用性。结论开发并应用了一种简单、灵敏、高通量的方法，用于同时测定尿液和全血样本中不同种类的抗生素。这种灵敏的方法成功地应用于法医案件。

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来源期刊

Forensic Toxicology TOXICOLOGY-

CiteScore

5.80

自引率

9.10%

发文量

审稿时长

3 months

期刊介绍： The journal Forensic Toxicology provides an international forum for publication of studies on toxic substances, drugs of abuse, doping agents, chemical warfare agents, and their metabolisms and analyses, which are related to laws and ethics. It includes original articles, reviews, mini-reviews, short communications, and case reports. Although a major focus of the journal is on the development or improvement of analytical methods for the above-mentioned chemicals in human matrices, appropriate studies with animal experiments are also published. Forensic Toxicology is the official publication of the Japanese Association of Forensic Toxicology (JAFT) and is the continuation of the Japanese Journal of Forensic Toxicology (ISSN 0915-9606).