Acute ischemic stroke (AIS) ranks among the most prevalent cerebrovascular conditions. The clinical interest lies in utilizing oxidative stress (OS) markers and antioxidant system indicators for diagnosing and predicting stroke development. Our objective was to identify a precise marker of antioxidant stress ability to assess neurofunctional alterations in AIS patients. A total of 66 AIS patients, 33 of whom also had type 2 diabetes mellitus (T2DM), were selected for our study, and 30 patients without AIS and diabetes were used as controls. The total plasma homocysteine (Hcy) concentrations were determined via an enzymatic cycling assay. The reduced Hcy to oxidised Hcy concentration ratio (Hcy/HHcy) was determined using hollow fibre centrifugal ultrafiltration (HFCF-UF) technique we previously established. The Hcy/HHcy was significantly lower in AIS patients with T2DM compared with both controls (P = 0.0262) and AIS patients without T2DM (P = 0.0468). A positive correlation was found between total Hcy concentration and the National Institutes of Health Stroke Scale (NIHSS) score (P = 0.002, r = 0.561), while no correlation was observed in AIS patients without T2DM (P = 0.850, r=-0.034). In AIS patients, the Hcy/HHcy showed a positive correlation with the NIHSS score in those with T2DM (P = 0.037, r = 0.365) and a negative correlation in those without T2DM (P = 0.003, r=-0.497). In AIS patients with T2DM, the receiver operating characteristic’s area under the curve (ROC-AUC) value for total Hcy concentration was 0.796, compared to 0.600 in those without T2DM. Furthermore, the ROC-AUC values of the Hcy/HHcy were 0.719 and 0.735 in AIS patients with and without T2DM, respectively. Hcy/HHcy is anticipated to serve as a precise indicator for assessing antioxidant stress ability and evaluating neurofunctional alterations in AIS patients.

Background Acute ischemic stroke (AIS) is one of the most common cerebrovascular diseases which accompanied by a disruption of aminothiols homeostasis. To explore the relationship of aminothiols with neurologic impairment severity, we investigated four aminothiols, homocysteine (Hcy), cysteine (Cys), cysteinylglycine (CG) and glutathione (GSH) in plasma and its influence on ischemic stroke severity in AIS patients. Methods A total of 150 clinical samples from AIS patients were selected for our study. The concentrations of free reduced Hcy (Hcy), own oxidized Hcy (HHcy), free reduced Cys (Cys), own oxidized Cys (cysteine, Cyss), free reduced CG (CG) and free reduced GSH (GSH) were measured by our previously developed hollow fiber centrifugal ultrafiltration (HFCF-UF) method coupled with high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The concentration ratio of Hcy to HHcy (Hcy/HHcy), Cys to Cyss (Cys/Cyss) were also calculated. The neurologic impairment severity of AIS was evaluated using National Institutes of Health Stroke Scale (NIHSS). The Spearman correlation coefficient and logistic regression analysis was used to estimate and perform the correlation between Hcy, HHcy, Cys, Cyss, CG, GSH, Hcy/HHcy, Cys/Cyss and total Hcy with NIHSS score. Results The reduced Hcy and Hcy/HHcy was both negatively correlated with NIHSS score in AIS patients with P = 0.008, r=-0.215 and P = 0.002, r=-0.249, respectively. There was no significant correlation of Cys, CG, GSH, HHcy, Cyss, Cys/Cyss and total Hcy with NIHSS score in AIS patients with P value > 0.05. Conclusions The reduced Hcy and Hcy/HHcy, not total Hcy concentration should be used to evaluate neurologic impairment severity of AIS patient.

Therapeutic drug monitoring (TDM) of imatinib (IM) in cancer therapy offers the potential to improve treatment efficacy while minimizing toxicity. There was a significant correlation between unbound concentration and clinical response and toxicity, compared with total plasma concentrations, and the quantification of unbound IM and its metabolite, N-desmethyl imatinib (NDI) are of interest for TDM. However, traditional unbound drug separation methods have shortcomings, especially are susceptible to non-specific binding (NSB) of drugs to the polymer-constructed components of filter membranes, which are difficult to avoid at present. Hence it is necessary to developed a reliable separation method for the analysis of the unbound fraction of IM and NDI in TDM. We developed and validated an hollow fiber solid phase microextraction (HF-SPME) method coupled with high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) that to measure unbound IM and NDI concentration in human plasma. It used the NSB phenomenon and solve the NSB problem. The preparation procedure only involves a common vortex and ultrasonication without dilution of samples and modification of membrane. A total of 50 chronic myeloid leukemia (CML) patients were enrolled in our study. The relationship between the unbound and total concentrations for IM and NDI, as well as the concentration ratios of NDI to IM in 50 clinical plasma samples were investigated. The extraction recovery is high to 95.5~106% with validation parameters for the methodological results were all excellent. There were both a poor linear relationship between the unbound and total concentrations for IM (r2=0.504) and NDI (r2=0.201) in 50 clinical plasma samples. The unbound concentration ratios of NDI to IM varied widely in CML patients. The determination of unbound IM and NDI concentration is meaningful and necessary. The developed HF-SPME method is simple, accurate and precise that could be used to measure unbound IM and NDI concentration in clinical TDM.

Background: oxidative stress is linked to various human diseases which developed into the idea of "disrupted redox signaling". Osteoporosis (OP) is a chronic skeletal disorder characterized by low bone mineral density and deterioration of bone microarchitecture among which estrogen deficiency is the main cause. Lack of estrogen leads to the imbalance between oxidation and anti-oxidation in patients, and oxidative stress is an important link in the pathogenesis of OP. The ratio of the reduced to the oxidized thiols can characterize the redox status. However, few methods have been reported for the simultaneous determination of reduced forms and their oxidized forms of thiols in plasma. Methods: we developed a hollow fiber centrifugal ultrafiltration (HFCF-UF) method for sample preparation and validated a high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) method to determine two reduced forms of thiols-homocysteine (Hcy), cysteine (Cys) levels and their respective oxidized compounds, homocystine (HHcy) and cystine (Cyss) in rat plasma simultaneously for the first time. Thirty-six female rats were randomly divided into three groups: normal control (NC), oxidative stress (ovariectomy, OVX) and ovariectomy with hydrogen-rich saline administration (OVX + HRS). Results: the validation parameters for the methodological results were within the acceptance criteria. There were both significant differences of Hcy/HHcy (Hcy reduced/oxidized) and Cys/Cyss (Cys reduced/oxidized) in rat plasma between three groups with both p < 0.05 and meanwhile, the p values of malondialdehyde, superoxide dismutase and glutathione peroxidase were all less than 0.01. The value of both Hcy/HHcy and Cys/Cyss were significantly decreased with the change of Micro-CT scan result of femoral neck in OVX group (both the trabecular thickness and trabecular number significantly decreased with a significant increase of trabecular separation) which demonstrate OP occurs. The change of Hcy/HHcy is more obvious and prominent than Cys/Cyss. Conclusions: the Hcy/HHcy and Cys/Cyss could be suitable biomarkers for oxidative stress and especially Hcy/HHcy is more sensitive. The developed method is simple and accurate. It can be easily applied in clinical research to further evaluate the oxidative stress indicator for disease risk factors.

This paper improves the vibration isolation effect of the conventional wave impeding block by double-layer wave impeding block with the same thickness. Based on the theory of wave motion in unsaturated porous medium and elastic medium, the vibration isolation performance of double-layer wave impeding block in unsaturated foundation under SV wave incidence is studied. Based on the Helmholtz principle, the solutions of surface vertical displacements of the unsaturated foundation under SV wave incidence are obtained. The influence of the wave impedance ratio between the layers of double-layer wave impeding block and the wave impedance ratio at the interface between the wave impeding block and the unsaturated soil foundation are firstly studied, and it was found that the optimal vibration isolation effect could be obtained by designing and controlling the wave impedance ratio between the layers of the double-layer wave impeding block and the wave impedance ratio at the interface between the wave impeding block and the unsaturated foundation. Then the physical and mechanical parameters such as incidence angle, incidence frequency, saturation, thickness and burial depth of the double-layer wave impeding block were analyzed to determine the influence on the vibration isolation performance of the double-layer wave impeding block. The results show that the best vibration isolation effect of double-layer wave impeding block is achieved at 5° of S-wave incidence angle. The double-layer wave impeding block has better vibration isolation effect at low frequency, medium frequency and high frequency. The vibration isolation effect of double-layer wave impeding block increases and then decreases with an increase of saturation, increases with an increase of burial depth and thickness, decreases and then increases with an increase of incident frequency.

We developed a hollow fiber centrifugal ultrafiltration (HFCF-UF) method to study the change of plasma levels of free glycine (Gly) in patients with acute ischemic stroke (AIS). Twenty-four patients with AIS confirmed by diffusion-weighted imaging (DWI) were enrolled. During the study period, the patients did not receive any supplemental amino acids therapy that could affect the obtained results. Our results showed that although AIS patients adopted different methods of treatment (thrombolytic and non-thrombolytic), the clinical NIHSS score of AIS showed a downward trend whereas Gly concentration showed increased trend. Moreover, plasma free Gly concentration was positively correlated with ASPECTS score. The correlation between Gly levels and infarct volume showed a statistical significance. That is to say, higher Gly level predicted smaller infarct size. Thus, the change of free Gly level in plasma could be considered as a potential biomarker of AIS.

A fluorescence probe based on carbon dots (CDs) coated with silica molecularly imprinted polymer (MIPs) was synthesized for selective and sensitive determination of cetirizine (CTZ). Green source carbon dots were firstly derived from orange peels through a microwave method, and had the merits of eco-friendly and low toxicity. Then a thin silica film was formed on the surface of CDs by reverse microemul-sion technique, and molecularly imprinted polymer coated on silica-carbon dots. In this scene, CTZ, 3-aminopropyltriethoxysilane (APTES) and tetraethoxysilane (TEOS) were employed as a template, a func-tional monomer and cross linker, respectively. The obtained CDs-MIPs can selectively bind CTZ through the specific interaction between recognition sites and template, and obey photoinduced electron transfer fluorescence quenching mechanism. Fluorescence dropped linearly in the range of 0.5-500 ng mL-1, under the optimal conditions, with a detection limit of 0.41 ng mL-1. Furthermore, the proposed method was successfully intended for the determination of trace CTZ in human saliva and urine samples without the interference of other molecules and ions. And recoveries ranged from 95.8% to 99.8% with relative standard deviation less than 3.0%. (c) 2021 Elsevier B.V. All rights reserved.

目的 建立一种超高效液相色谱法(UPLC),同时分析人血清中拉莫三嗪(LTG)、奥卡西平(OXC)及其代谢物10,11-二氢-10-羟基卡马西平(MHD)浓度,并用于本院治疗药物监测.方法 血清样本经乙醚萃取,采用Waters UPLC BEH C18柱(50.0 mm×2.1 mm,1.7μm),流动相为乙腈-磷酸缓冲盐(pH 4.5)=26:74;流速:0.2 mL·min-1;柱温:30℃;检测波长:220 nm,进样量:2μL,内标为非那西丁,考察该方法的专属性、标准曲线、精密度、准确度、稳定性,收集临床癫痫患者血清样本进行方法验证.结果 血清中LTG、OXC和MHD分离良好,不受血清中内源性物质干扰,色谱分析时间仅需5 min,血清中LTG和OXC在0.25～25.00μg·mL-1和MHD在0.50～50.00μg·mL-1范围内线性关系均良好,方法回收率为88.37％～105.00％,LTG、MHD和OXC的绝对回收率分别可达78.45％,62.50％和75.76％以上,内标的提取回收率为61.41％.批内、批间RSD均不大于7.0％,室温放置16 h、冻融循环3次和冰冻30 d的稳定性良好.所建方法在本院临床采用LTG治疗癫痫的患者10例和服用OXC治疗癫痫患者7例中进行验证.结论 所建方法简便准确,提高了分析效率,适宜于LTG、OXC和MHD的临床常规血药浓度监测,并成功用于本院治疗药物监测.

Aim: To establish a simple and accurate method to explore the correlation between free and total concentrations of lamotrigine (LTG) and the active oxcarbazepine metabolite monohydroxy derivative (MHD) (10,11-dihydro-10-hydroxycarbamazepine) in clinical patients. Materials & methods: Serum samples were prepared by hollow-fiber centrifugal ultrafiltration and then injected into UPLC for analysis. Results: Absolute recovery was as high as approximately 90.1-98.6% with excellent precision (relative standard deviation <6.7%). Analysis time was reduced to 5 min. There were significant individual differences in the protein binding rates of both LTG and MHD that were probably due to the use of different clinical patients. Conclusion: Free concentrations of LTG and MHD cannot be estimated by total concentration in specific clinical patients. Free drug monitoring of LTG and MHD in clinical therapeutic drug monitoring is important and essential.

A novel benzylbenzimidazolium based dicationic ionic liquids modified magnetic nanoparticles (MNP@BDILs) was successfully synthesized by a simple one-step reaction at mild condition and employed as adsorbent for determination of polycyclic aromatic hydrocarbons (PAHs) by magnetic solid-phase extraction (MSPE) coupled with high performance liquid chromatography (HPLC). Main parameters affecting the MSPE efficiency, including extraction time, extraction solvent, desorption solvent, desorption time and the amount of sorbent were studied. Under the optimized conditions, the proposed method provided good linearity with determination coefficients over 0.9991, and excellent reproducibility. Finally, for the determination of six PAHs in soil, milk and black wolfberry, satisfactory spiked recoveries were between 91.2% and 104.1% with the relative standard deviations below 6.5%. The results demonstrated that MNP@BDILs synthesized by the simple one-step reaction was a promising strategy for the application in sample preparation.

目的 建立一种测定人血浆中卡马西平游离浓度的中空纤维离心超滤(HFCF-UF)-超高效液相色谱(UPLC)法.方法 血浆样本经中空纤维离心超滤技术预处理.色谱柱:Waters UPLC BEH C18柱(50.0 mm×2.1 mm,1.7μm),流动相:乙腈-水=35:65,流速:0.2 mL·min-1,柱温:30℃,进样量:2μL.考察该方法的专属性、标准曲线与定量下限、精密度与回收率、稳定性.结果 血浆中游离卡马西平在0.2～50.0μg·mL-1内,线性关系良好,标准曲线为y=2.04×104x-4.61×102(r=0.9989).游离浓度分析方法的回收率为96.86％～104.89％,绝对回收率达到98.40％～102.40％,批内、批间RSD均小于3.00％.结论 本方法简便、准确,适用于血浆中卡马西平游离浓度的分析.

建立一种结合便携式高分辨离子迁移谱(IMS)快速简便萃取唾液中的苯海拉明(DPH)的分散液-液微萃取法(DLLME).取2 mL唾液,采用10％(w:V)硫酸锌沉淀蛋白,以2 mL二氯甲烷为萃取剂对唾液中的苯海拉明进行萃取,萃取时间5 min,离心(3500 r/min,20 min),取二氯甲烷层,45℃真空干燥,甲醇复溶后进行IMS分析.结果表明,该方法可在短时间内完成对DPH的萃取分离及分析,DPH在39.4～1080 ng/mL范围内线性关系良好,标准曲线方程为I=3.62 × 103C-0.754(R2 =0.978),检出限和定量限分别为11.8和39.4 ng/mL,平均回收率为89.0％～112％,RSD ≤ 1.5％.该方法可实现对人体唾液中苯海拉明的快速测定.

: In order to ensure the safety of new energy vehicles, wireless communication and network facilities should be added to the old electronic control system of new energy vehicles based on cloud platform, so that vehicles can be controlled at any time, and vehicle faults can be analyzed and calibrated remotely, thus improving driving safety. Based on the cloud platform, this paper briefly introduces the SAEJ1939 communication protocol standard, and implements the SAEJ1939 communication protocol in the form of protocol stack. The protocol stack includes hardware abstraction layer, data link layer, transport layer, network management layer, application layer and diagnosis. Among them, the hardware abstraction layer directly calls the CAN module driver of the hardware system, which facilitates the transplantation of SAEJ939 communication protocol stack in different hardware systems. In this way, we can fully understand the actual situation of the vehicle.

The presence of reduced aminothiols, including homocysteine (Hcy), cysteine (Cys), cysteinyl-glycine (CG), and glutathione (GSH), is significantly increased in the pathological state. However, there have been no reports on the relationship between reduced aminothiols (Hcy, Cys, CG, and GSH) and different genders, ages, and drug combinations in human blood. The accurate quantification of these reduced thiols in biological fluids is important for monitoring some special pathological conditions of humans. However, the published methods typically not only require cumbersome and technically challenging processing procedures to ensure reliable measurements, but are also laborious and time-consuming, which may disturb the initial physiological balance and lead to inaccurate results. We developed a hollow fiber centrifugal ultrafiltration (HFCF-UF) method for sample preparation coupled with a high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method and used it to determine four reduced aminothiols (Hcy, Cys, CG, and GSH) in human blood for the first time. A total of 96 clinical patients were enrolled in our study. The influence of different genders, ages, and drug combinations on the levels of four reduced thiols in human blood was also discussed by SPSS 24.0. The sample preparation was simplified to a single 5 min centrifugation step in a sealed system that did not disturb the physiological environment. The validation parameters for the methodological results were excellent. The procedure was successfully applied to monitoring the concentrations of four reduced aminothiols (Hcy, Cys, CG, and GSH) in 96 clinical blood samples. There were no significant differences in Hcy, Cys, CG, or GSH for the different genders, ages, or combinations with methotrexate or vancomycin (P > 0.05). However, there was a significant increase in Hcy concentration in patients treated with valproic acid who were diagnosed with epilepsy (p=0.0007). It is advisable to measure reduced Hcy level in patients taking valproic acid. The developed HFCF-UF method was simple and accurate. It can be easily applied in clinical research to evaluate oxidative stress in further study.

目的:建立简单、快速的中空纤维离心超滤-高效液相色谱法(HFCF-UF-HPLC)测定人血浆中5-氟尿嘧啶血药浓度的方法.方法:将加入少量释放剂的血浆样本经中空纤维离心超滤预处理后,直接取20μL超滤液进行HPLC分析,采用Ve-nusil MP C18 色谱柱(250 mm×4.6 mm,5 μm),流动相:甲醇-0.01 mol.L-1 磷酸二氢钾缓冲液(2∶98,V∶ V),流速:1.0 mL·min-1,检测波长:265 nm.结果:5-氟尿嘧啶在0.05～50.1 μg·mL-1的范围内线性关系良好(R2=0.999 9),绝对回收率为99.5％～104.2％,RSD＜1.8％,检测限为0.03 μg·mL-1,定量限0.05 μg·mL-1,室温放置24 h,循环冻融3次和冰冻3个月的稳定性良好.结论:所建立的方法避免了由于疾病所引起的血浆白蛋白水平的变化对其血药浓度测量结果的影响,为临床中5-氟尿嘧啶的血药浓度监测提供了简单、快速、灵敏的样品前处理方法.

High-dose methotrexate (HD-MTX) can be highly effective as well as extremely toxic. Many drug molecules can bind to plasma proteins to different extents in vivo, whereas only the free drug can reach the site of action to exert a pharmacological effect and cause toxicity. However, free MTX concentrations in plasma have not been reported. Traditional analyses of free drugs are both cumbersome and inaccurate. We collected 92 plasma samples from 52 children diagnosed with ALL or NHL or other lymphomas that were treated with HD-MTX. The hollow fiber centrifugal ultrafiltration (HFCF-UF) was used to prepare plasma samples for analysis of the free MTX concentration. Protein precipitation was employed to measure the total MTX concentration. The HFCF-UF is a simple method involving a step of ordinary centrifugation; the validation parameters for the methodological results were satisfactory and fell within the acceptance criteria. A linearity coefficient r 2 of 0.910 was obtained for the correlation between the free and total MTX plasma concentrations in 92 plasma samples. However, the free and total MTX concentrations was only weakly correlated in 16 clinical plasma specimens with total MTX concentrations >2 μmol L−1 (r 2 = 0.760). Both the free and total MTX concentrations at 42 h were negatively correlated with the creatinine clearance (CCr) level (P = 0.023, r = −0.236 for total MTX and P = 0.020, r = −0.241for free MTX, respectively). The free MTX concentration could not be accurately estimated from the total MTX concentration for patients with high MTX levels which are conditions under which toxic reactions are more likely to occur. High plasma MTX levels could become a predictor of the occurrence of MTX nephrotoxicity to draw people's attention. The proposed HFCF-UF method is a simple and accurate way to evaluate efficacy and toxicity in clinical therapeutic drug monitoring.

Laboratory biosafety has become a core focus in biological analysis, owing to the frequent occurrence of laboratory-acquired infections caused by the leakage of pathogenic microorganisms. For this purpose, the authors developed a safe pretreatment device combining a sealing technique with a direct injection technique. In this study, several bacteria and viruses were used to validate the filtration effect of the invention. Data show that the new device can completely filter bacteria and that the filtration rates for hepatitis B virus and hepatitis C virus reached 94% and 96%, respectively. The results show that the new preparation device can effectively block these pathogens and can improve biological safety and provide powerful protection for technicians.

Purpose: To perform therapeutic drug monitoring (TDM) of total and free plasma valproic acid (VPA) concentrations in pediatric patients with epilepsy and to analyze related factors. Patients and Methods: Pediatric epileptic patients treated in 2015-2019 in our hospital were assessed. Total and free plasma VPA concentrations were obtained by UPLC and LC-MS/MS, respectively. Regression analysis was performed to examine the associations of free plasma VPA with total plasma VPA and plasma protein binding rate. The impacts of individual situation, CYP2C9 genotype, and drug combination on VPA concentration were examined. Results: Of the 251 patients, 81 had lower total concentrations than effective therapeutic levels; 86 and 31 patients had infections and central nervous system dysplasia, respectively. VPA's daily doses and free drug concentrations were significantly lower in the CYP2C9 *3/*3 genotype group versus the CYP2C9 *1/*3 and CYP2C9 *1/*1 groups (P<0.05). Free and total VPA concentrations were linked by Y = 0.0004 X-2 + 0.042 X + 0.3035 (r=0.6981); VPA plasma protein binding rate and free VPA concentration were related by Y = 0.0003 X-2 - 0.0127 X + 0.9777 (r=0.8136). Both total and free VPA concentrations were significantly decreased in patients simultaneously administered phenobarbital, meropenem and biapenem (P<0.05), with therapeutic failure after meropenem/biapenem co-administration. Conclusion: Free VPA amounts have nonlinear relationships with total VPA amounts and plasma protein binding rate in epileptic children. Additionally, CYP2C9 *3/*3 expression affects VPA metabolism. Since phenobarbital affects VPA metabolism, TDM is recommended. Meanwhile, carbapenem-co-administration with VPA should be prohibited.

In this paper, a simple and rapid sample pretreatment device integrating ultrasonication, centrifugation and ultrafiltration (UCU) was reported for preparation of trace analytes in complex matrices. The UCU device was composed of two parts, A and B. The sample and extraction solvent were put into Part B for ultrasonic extraction. Subsequently, Part A and Part B were integrated and sealed for centrifugation and ultrafiltration. Finally, the ultrafiltrate in Part A was taken out for subsequent detection. After optimization, the device was applied to rapid on–site screening of five illegally added chemical components in heat–clearing and detoxicating Chinese patent medicines by combining with electrospray ionization–ion mobility spectrometry (ESI–IMS). The method showed good performance in terms of linearity with correlation coefficients (R2) above 0.9976 and limits of detection (LODs) in the range of 0.049–0.391 μg mL–1. The recoveries were from 96.5 % to 100.8 %. The whole analysis process was within 11 min. The proposed method was further compared with other methods reported in the literature and the advantages and considerations were also explored. The results demonstrated that it was a simple, fast and accurate technique. The establishment of this method not only greatly improved the experimental efficiency but also avoided potential sample pollution brought by multiple sample transfer, and could provide a powerful means for rapid on–site analysis of trace analytes in complex matrices.

目的 建立一种高效液相色谱串联质谱法(HPLC-MS/MS)分析人血浆中甲氨蝶呤(MTX)的浓度,并用于临床治疗药物监测.方法 血浆样本200μL经乙腈600μL沉淀蛋白处理,用Waters System C18柱(150 mm×4.6 mm,3.5μm),流动相为乙腈(10％异丙醇)-水(0.1％甲酸)梯度洗脱;流速:0.8 mL·min-1;柱温:40℃.ESI正离子模式,MTX定量离子对m/z 455.2→m/z 308.1,内标甲硝唑定量离子对m/z 172.2→m/z 128.2.结果 人血浆中MTX在0.1～10μmol·L-1线性关系良好(r2=0.9942),方法回收率分别为99.2％～103.4％,MTX绝对回收率达到71.99％,基质效应101.9％～107.1％,批内、批间RSD均不大于8％,在室温放置12 h、冻融循环3次和-40℃冰冻30 d的稳定性良好.成功用于我院48例使用甲氨蝶呤化疗患者体内MTX的浓度测定.结论 所建方法简便、准确、灵敏,适用于临床MTX治疗药物监测.