液相色谱-串联质谱技术在临床检验中的应用研究进展

液相色谱-串联质谱（LC-MS/MS）技术具有高灵敏度、高特异性、高分辨率和高效率的优点。近年来随着仪器灵敏度的提高，LC-MS/MS在常规临床检验中显示出极大的潜力，并在疾病早期预防和诊断中发挥着不可替代的作用。本文对LC-MS/MS在新生儿疾病筛查、维生素D检测、内分泌激素检测、肽类和蛋白质定量分析等临床检验方面的研究进展进行综述，并讨论了未来面临的挑战。     

液相色谱-串联质谱法测定豆类中左旋多巴

文章建立一种豆类中左旋多巴的液相色谱-串联质谱测定方法。样品经0.3 mol/L乙酸水溶液超声提取,Agilent Proshell 120 SB-C18(2.1 mm×150 mm,2.7μm)色谱柱分离,以0.1%甲酸水-甲醇为流动相进行梯度洗脱,多反应监测(MRM),内标法定量。结果表明,左旋多巴在0.1~5.0 mg/L范围内线性关系良好,相关系数为0.9994,检出限为1.5 mg/kg,定量限为5.2 mg/kg,方法的平均加标回收率为96.9%~105.5%,日内精密度和日间精密度均小于4%,具有操作简单、灵敏度高、准确度高、重现性好等特点,能够很好满足豆类中左旋多巴含量的检测需要。     

诺氟沙星片在中国健康受试者空腹和餐后状态下生物等效性研究#br#

目的：采用随机、开放、两周期、自身交叉、单次给药试验设计比较浙江医药股份有限公司新昌制药厂生产的诺氟沙星片与原研产品BACCIDAL在中国健康人体中的生物利用度，并评价两种制剂的生物等效性。方法：分别在空腹和餐后条件下，健康受试者随机交叉单剂量口服诺氟沙星片受试制剂或参比制剂100 mg，采用液相色谱-质谱串联（LC-MS/MS）法测定受试者服药前后不同时间点血浆内药物浓度，采用WinNonlin 7.0软件计算主要药代动力学参数，并评价两种制剂的生物等效性。结果：空腹试验共有28例受试者入组并完成试验，诺氟沙星受试制剂与参比制剂的Cmax分别为（607.62±125.24）ng/mL和（552.01±134.11）ng/mL；AUC0-t分别为（2 551.66±509.08）ng·mL-1·h和（2 429.98±460.47）ng·mL-1·h；AUC0-∞分别为（2 675.40±523.04）ng·mL-1·h和（2 557.68±485.43）ng·mL-1·h；t1/2分别为（6.07±0.69）h和（6.18±0.92）h；两种制剂的Cmax、AUC0-t和AUC0-∞几何均值比的90%置信区间分别为101.45%～121.94%、98.96%～111.27%、98.82%～110.76%。餐后试验共有28例受试者入组并完成试验，诺氟沙星受试制剂与参比制剂的Cmax分别为：（256.54±58.87）ng/mL和（300.80±94.67）ng/mL；AUC0-t分别为（1 314.74±349.92）ng·mL-1·h和（1 278.60±314.77）ng·mL-1·h；AUC0-∞分别为（1 413.73±361.98）ng·mL-1·h和（1 374.98±321.62）ng·mL-1·h；t1/2分别为（6.66±1.23）h和（6.66±1.34）h；两种制剂的Cmax、AUC0-t和AUC0-∞几何均值比的90%置信区间分别为81.42%～93.56%、99.61%～105.58%、99.80%～105.21%。结论：浙江医药股份有限公司新昌制药厂生产的诺氟沙星片与原研产品BACCIDAL在中国健康受试者空腹和餐后服用的状态下等效且安全性良好，临床上可以替换使用。     

Uncertainties in the determination of pyrrolizidine alkaloid levels in naturally contaminated honeys and comparison of results obtained by different analytical approaches

The contamination of honey with hepatotoxic pyrrolizidine alkaloids (PAs) is a well-known hazard for food safety. While management strategies and controls of the honey industry aim to reduce the PA levels, uncertainties remain with regard to the safety of regionally produced and marketed honey. In addition, a previous study showed large differences of results obtained after various periods of storage and apparent differences between the analytical results of different laboratories. Therefore, this study aimed at examining these uncertainties by monitoring the impact of storage on the PA and PA N-oxide (PANO) content of two freshly harvested honeys and on possible demixing effects caused by pollen settling. Additionally, three analytical approaches – target analysis with matrix-matched calibration or standard addition and a sum parameter method – were applied for a comparative analysis of 20 honeys harvested in summer 2016. All samples originated from Schleswig-Holstein in Northern Germany where the PA plant Jacobaea vulgaris is currently observed on a massive scale. The results of the time series analyses showed that PANO levels markedly decreased within a few weeks and practically reached the LOD 16 weeks after harvest. Tertiary PAs, by contrast, remained stable and did not increase as a consequence of PANO decrease. The experiments on a putative demixing, which may result in a heterogeneous distribution of PAs/PANOs, revealed that there was no such effect during storage of up to 12 weeks. A comparison of the PA/PANO levels obtained by different analytical approaches showed that in some cases the sum parameter method yielded much higher levels than the target approaches, whereas in other cases, the target analysis with standard addition found higher levels than the other two methods. In summary, the results of this study highlight uncertainties regarding the validity and comparability of analytical results and consequently regarding health risk assessment.     

液相色谱-串联质谱法测定腐乳中的生物胺

建立了一种使用液相色谱-串联质谱（LC-MS/MS）技术检测腐乳中9种生物胺的方法。样品经5%三氯乙酸水溶液提取后，进行液相色谱-串联质谱法测定，采用外标法定量。对检测方法进行方法学验证，9种生物胺在质量浓度为10～200 ng/mL范围内线性相关系数R2＞0.999，方法检出限为0.03 mg/kg，定量限为0.1 mg/kg，回收率在85%～101%之间，相对标准偏差（RSD）在0.9%～4.8%之间。该方法结果精密、准确、重复性好、操作简单，适用于腐乳中9种生物胺的测定。     

液相色谱-串联质谱法测定畜禽肌肉和表皮组织中松香酸、脱氢松香酸和胡椒酸

目的 建立了畜禽肌肉和表皮组织中松香酸、脱氢松香酸和胡椒酸的液相色谱-四极杆/串联质谱(LC-MS/MS)的检测方法。方法 均质样品中加入乙腈和水分别提取目标物,经涡旋分散、震荡提取、离心和固相萃取柱净化。提取液以乙酸铵-甲醇为流动相梯度洗脱C18色谱柱分离,5 mmol的乙酸铵和甲醇为流动相进行梯度洗脱,三重四级杆质谱ESI+/ESI-切换模式检测,以特征离子峰和保留时间定性,定量离子峰面积定量。结果 松香酸、脱氢松香酸的测定低限为5.0 μg/kg、胡椒酸20.0 μg/kg,线性范围分别为5.0~200.0 μg/kg和20.0~500.0 μg/kg,相关系数(r)在0.999以上。分别添加定量限1、2、4倍的浓度验证加标回收率,三个浓度水平的回收率为68.5% ~ 107.6%,相对标准偏差(RSD)为6.8%~14.2%(n=6)。结论 该方法快速、准确、灵敏,适用于监管部门对市售畜禽食品中工业松香残留的检测。     

神经性贝类毒素免疫亲和柱的制备与应用

通过纯化PbTX-2单克隆抗体,以琼脂糖凝胶(sepharose 4B)为载体制备神经性贝类毒素(NSP)免疫亲和柱,并建立NSP的免疫亲和柱净化-液相色谱-串联质谱分析方法。样品以含80%甲醇水溶液提取,PBS缓冲液稀释,经过NSP免疫亲和柱净化和富集后进行LCMS/MS测试分析。同时对上样液、淋洗液、洗脱液等亲和柱参数进行优化。结果表明:NSP免疫亲和柱对贝类样品有很好的净化作用,降低了贝类基质中脂肪、蛋白质、色素、分泌黏液等杂质干扰,提高了样品回收率。在40~800μg/kg范围内样品加标平均回收率为82.95%~112.95%,相对标准偏差为1.27%~2.94%。该NSP免疫亲和柱净化效果强、灵敏度高且受基质干扰小,适用于贝类中神经性贝类毒素的分析测定。     

快速灵敏的LC-MS/MS法测定人血浆中阿莫西林并应用于两种制剂的一致性评价

目的: 建立快速灵敏的LC-MS/MS法测定人血浆中阿莫西林浓度，并用于两种阿莫西林胶囊的一致性评价。方法: 采用岛津公司LCMS-8060型LC-MS/MS仪，以MRM模式测定阿莫西林（m/z 366.00/114.00）的浓度，d4-阿莫西林作内标（m/z 370.10/114.05），离子源为ESI源。色谱柱选用Waters ACQUITY BEH C18（2.1×50 mm，1.7 μm），梯度洗脱。血浆样本加入内标，经甲醇沉淀蛋白后取上清液进样检测。结果: 所建方法经验证，其线性、准确度、精密度、最低定量限、提取回收率、特异性、基质效应、稳定性等各项指标均符合CFDA的指导原则及最新核查标准要求，并较文献报道中的方法有处理简单、灵敏度高、色谱峰形好的优点。结论: 所建方法快速、灵敏，适用于人血浆中阿莫西林浓度的检测。用于一致性评价的样本实测，两种制剂生物等效。     

Comparison of two Analytical Methods (ELISA and LC-MS/MS) for Determination of Aflatoxin B1 in Corn and Aflatoxin M1 in Milk

The aim of this paper is to assess the closeness of agreement between results of ELISA and LC-MS/MS methods for determination of aflatoxin B₁ in corn and aflatoxin M₁ in milk. Samples of corn (n=100) and milk (n=250) were simultaneously analyzed using ELISA and LC-MS/MS methods, after the severe drought that affected Serbia in summer 2012 resulting in occurrence of aflatoxin B1 in corn and aflatoxin M₁ in milk. Regression analysis showed higher level of agreement between aflatoxin B₁ samples (R²=0.994), compared to aflatoxin M₁ samples (R²=0.920). However, both techniques were satisfactory in meeting the requirements for official control purposes.