西罗莫司全血浓度测定方法及临床应用

目的：评价2种分析方法同时检测中国肾移植患者的全血中西罗莫司浓度的差异。方法用高效液相色谱质谱联用法（LC-MS/MS）和化学发光微粒子免疫分析法（ CMIA）分别测定中国肾移植患者全血西罗莫司浓度,评价2种分析方法的相关性。结果 LC-MS/MS法专属性强、精密度、稳定性好,能满足临床检测要求。2种方法测得的数据相关系数为0.963（ P＜0.05）,2组数据间存在相关性（n＝78）。当西罗莫司浓度在2～10 ng· mL-1时,2种方法测定的平均偏差为2.8 ng· mL-1。结论 LC-MS/MS和CMIA测定西罗莫司全血浓度的分析方法存在相关性;同一份样本用CMIA测定的浓度均值比LC-MS/MS法测定的值高。     

CMIA法和FPIA法监测全血环孢霉素A浓度的比较

目的:比较荧光偏振免疫法(FPIA)和化学发光微粒子免疫法(CMIA)监测全血环孢霉素A(CsA)浓度结果的相关性。方法:收集服用CsA患者的稳态浓度全血样本,分别用FPIA法和CMIA法进行测定,考察2种方法的相关程度;进一步将浓度结果分为低浓度组和高浓度组进行相关性研究。结果:2种方法测定全血CsA浓度结果r=0.992具有显著性差异(P<0.05),CMIA法测定结果偏低。低浓度组结果r=0.961具有显著性差异(P<0.05);高浓度组结果r=0.992亦具有显著性差异(P<0.05)。结论:CMIA法作为新的全血CsA浓度监测方法,测定结果比FPIA法要显著降低,但监测数据更为准确,需要向医师和患者说明。     

HPLC—MS法和mFPIA法测定全血环孢霉素A及其代谢物的比较

目的:对测定全血环孢霉素 A 的 HPLC-MS 法和单克隆荧光偏振免疫分析法(mFPIA 法)进行评价。方法:建立HPLC-MS 测定全血环孢霉素 A 及其代谢物的方法,采用多元线性回归对 HPLC-MS 法和 mFPIA 法测定的结果进行比较。结果:当患者肝、肾功能异常时,mFPIA 法测定结果同环孢霉素 A 及主要代谢物浓度呈线性相关,易导致检测结果偏高。结论:当患者肝、肾功能异常时,mFPIA 法不适宜作为环孢霉素 A 的临床监测方法。     

肾移植患者口服国产环孢素A软胶囊后治疗药物监测指标的选择

目的:探讨肾移植患者口服国产环孢素A(CsA)软胶囊后治疗药物监测的适宜指标。方法:采集10例肾移植术后稳定期患者口服国产CsA软胶囊后12h内不同时间点的血样,以单克隆荧光偏振免疫(mFPIA)法测定血药浓度。计算各采样点CsA浓度与AUC0～12和AUC0～4的相关系数,考察相关系数间的差异性。结果:除0.5、0.75h外,其余各时间点与0h的相关系数两两间检验未显示有统计学意义的差异(P>0.05)。结论:对于肾移植术后稳定期口服国产CsA软胶囊患者,谷浓度(c0)仍然可以作为适宜的治疗药物监测指标之一。     

LC-MS/MS法和EMIT法测定人血清中丙戊酸浓度的对比研究

目的:比较液相色谱-串联质谱(LC-MS/MS)法与酶放大免疫(EMIT)法测定人血清中丙戊酸(VPA)浓度的差异,为临床治疗药物监测提供参考。方法:分别采用LC-MS/MS法和EMIT法对144份住院或门诊患者血清样品中的VPA浓度进行检测,采用配对t检验、Pearson相关性分析和Bland-Altman偏差图等方法对其检测结果的差异进行评价。结果:LC-MS/MS法和EMIT法的检测结果呈正相关(r=0.924,P<0.05);两者的回归方程为c_(EMIT)=0.920 7c_(LC-MS/MS)-1.114 4(r=0.924);LC-MS/MS法和EMIT法测得的VPA平均血药浓度分别为(49.9±21.2)、(54.9±21.3)μg/m L,组间比较差异有统计学意义(P<0.05);EMIT法检测结果比LC-MS/MS法偏高8.3μg/mL,95%置信区间为(-13.6,18.7)。结论:LC-MS/MS法和EMIT法测定人血清中VPA的浓度相关性良好,但检测结果有一定的差异,建议同一患者长期监测应选用同一种方法。     

比较高效液相色谱法和液质联用法测定化疗患者血浆中甲氨蝶呤浓度的相关性

目的 比较高效液相色谱(HPLC)法和液质联用(LC-MS/MS)法测定人血浆中甲氨蝶呤(MTX)浓度的相关性.方法 收集接受大剂量MTX化疗后的患者的血液样品,分别用HPLC、LC-MS/MS法进行MTX血药浓度测定,通过Wilcoxon检验或配对t检验来比较2种测定方法在不同浓度范围的差异,通过压轴回归分析法建立回...     

EMIT法和HPLC法监测环孢菌素A血药浓度比较研究

目的:比较酶免疫增强法(EMIT)和高效液相色谱法(HPLC)监测环孢菌素A血药浓度的相关性.方法:收集肾脏移植患者服药后的稳态谷浓度血样,分别用EMIT法和HPLC法进行测定,考察2种测定方法的相关程度.结果:以HPLC法测定结果(X)与EMIT法测定结果(Y)所做线性回归方程如下:Y=0.8168X+34.699(r=0.9550),EMIT法测定全血中环孢菌素A浓度较HPLC法为高.结论:EMIT法和HPLC法测定环孢菌素A血药浓度结果差异具有统计学意义,在CsA治疗药物监测中应予以关注并作相应调整.     

肝脏、心脏移植病人的全血环孢素监测方法

为把肾毒性降至最低并优化免疫抑制,应对全血环孢素(cyclosporinA,CsA)浓度进行定期监测。HPLC法具有高选择性,但费时;采用单克隆抗体的荧光极化免疫分析(mFPIA)、酶倍增免疫测定技术(EMIT)和放射免疫分析(RIA)等方法几乎已取代了HPLC法。本文评价了两种免疫分析方法mFPIA和EMIT测定全血CsA母药浓度过高估算的幅度,及这种过高估算在心脏和肝脏移植受者肝功能减弱情况下,对治疗范围内CsA血药浓度的影响。3例心脏移植、8例肝脏移植病人在移植后4周期间接受监测。每日上午服CsA前采集EDTA抗凝全血样品,冷冻于冰箱,1周内分析。…     

EMIT法与HPLC法测定人血浆中环孢素A浓度的比较研究

目的:比较酶放大免疫(EMIT)法和高效液相色谱(HPLC)法测定人血浆中环孢素A(CsA)浓度的区别与相关性。方法:收集肾移植患者服药后的稳态谷浓度血样20份,分别用EMIT法和HPLC法进行测定,考察2种测定方法的区别和相关程度。结果:HPLC法和EMIT法检测的CsA血药浓度差异有统计学意义(P<0.05),EMIT法测定血浆中CsA浓度较HPLC法高26.4ng·mL-1,通过Passing-Bablok回归分析2种方法具有良好的相关性(r=0.9940)。结论:EMIT法和HPLC法测定CsA血浆药物浓度结果差异具有统计学意义,在临床进行CsA药物浓度监测中应予以关注并作相应调整。     

HPLC—MS／MS法与MEIA法在监测器官移植患者他克莫司全血浓度中的比较

目的：比较HPLC-MS／MS法与MEIA法在监测器官移植患者他克莫司全血浓度中的应用。方法：建立和确讧可行的HPLC-MS／MS法,并与ME—M法分别测定他克莫司相同样本的浓度,对测试结果进行统计分析,评价两种方法。结果：HPLC-MS肌S法测定他克莫司平均浓度为（4．86±0．46）ng／mL,MEIA法测定浓度平均为（5．52±0．43）ng／mL,两种方法测定结果相关系数r平均为0．8771,两种方法相关性较强。LC-MS／MS法测定浓度值／MEn法测定浓度值平均值为（90．34-5．3）％。结论：HPLC—MS／MS法测定浓度为他物在体内的准确浓度,更适用于日常治疗药物监测（TDM）工作。     

Relationship between cyclosporine concentrations obtained using the Roche Cobas Integra and Abbott TDx monoclonal immunoassays in pre-dose and two hour post-dose blood samples from kidney transplant recipients

Current evidence suggests that cyclosporine (CsA) concentration in blood samples taken 2 hours after Neoral microemulsion (Novartis Pharmaceuticals; East Hanover, NJ) administration (C2) predicts clinical events in transplant patients better than the pre-dose (trough) concentration (C0). Similarly, previous findings have shown that the metabolites/CsA ratio is substantially lower in C2 than in C0 samples; however the between-monoclonal immunoassay differences for C2 samples have received little attention in the literature. In 56 C samples and 60 C samples from renal transplant patients, CsA levels were determined using the monoclonal fluorescence polarization immunoassay (mFPIA) from Abbott (Abbott Park, IL) and the homogeneous enzyme immunoassay technique (HEIT) from Roche Diagnostics (Basel, Switzerland). In both cases a high correlation coefficient between the results was obtained (r > or = 0.971), with a linear regression for C0 samples: mFPIA = 1.47 HEIT + 22.0 and for C2 samples: mFPIA = 1.11 HEIT + 71.96. The difference between the linear regression slopes was statistically significant (P < 0.001), and the mFPIA/HEIT ratio was significantly higher for C than for C samples (P < 0.001).     

Simultaneous and rapid analysis of cyclosporin A and creatinine in finger prick blood samples using liquid chromatography tandem mass spectrometry and its application in C2 monitoring

A simple and rapid liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the simultaneous analysis of cyclosporin A (CsA) and creatinine using capillary blood has been developed. Venous and capillary blood samples were taken predose and at C2 from 65 heart and lung transplant recipients (65 x 4 samples). For comparisons, serum creatinine and blood CsA concentrations were measured by the Jaffe and EMIT methods, respectively, using an Olympus AU600 analyzer. For the LC-MS/MS assay, samples were prepared in a 96 x 700-microL well block by adding 10 microL of blood (or serum) to 40 microL of 0.1 mol/L zinc sulphate solution containing deuterated creatinine internal standard. Proteins were precipitated by adding 100 microL acetonitrile containing ascomycin internal standard. After vigorous mixing and centrifugation, 5 microL of the supernatant was injected into the LC-MS/MS system. A Waters 2795 high-performance liquid chromatography (HPLC) system was used to elute a C18 cartridge (3 mm x 4 mm) at 0.6 mL/min with a step gradient of 50-100% methanol containing 2 mmol/L ammonium acetate and 0.1% (v/v) formic acid. The column was maintained at 55 degrees C, and the retention times were creatinine, 0.4 minutes; ascomycin, 0.98 minutes; and CsA, 1.2 minutes. Cycle time was 2.5 minutes, injection to injection. The analytes were monitored using a Quattro microtandem mass spectrometer operated in multiple reaction monitoring mode using the following transitions: creatinine, m/z 114>44; d3-creatinine (IS), m/z 117>47; ascomycin (IS), m/z 809>756; and CsA, m/z 1,220>1,203. Assay characteristics were CsA intraassay CV, 3.6-3.0% (33-1,500 microg/L); CsA interassay CV, 6.7-2.5% (10-5,000 microg/L); LC-MS/MS capillary [CsA] = 0.99 x LC-MS/MS venous [CsA] - 4.2, R = 0.98; and LC-MS/MS venous [CsA] = 0.93 x EMIT venous [CsA] + 2.9, R = 0.98. Creatinine intraassay CV, 6.6-2.5% (20-720 micromol/L); interassay CV, 5.7-3.3% (80-590 micromol/L); LC-MS/MS capillary [creatinine] = 0.99 Jaffe plasma [creatinine] -42.6, R = 0.87. Total time for the preparation and analysis of 30 samples was approximately 2 hours. This assay will provide a flexible, robust, and cost-effective solution for the monitoring of CsA and creatinine in transplant recipients with potential applications in pediatric medicine and pharmacokinetic studies, in which frequent sampling is required.     

Cyclosporin monitoring in Australasia: survey of laboratory practices in 2000

Therapeutic drug monitoring of cyclosporin has been established as part of the routine clinical management of patients following organ transplantation for some 20 years. The current practices of laboratories in Australia and New Zealand have been surveyed for the year 2000 to update previous similar surveys in the light of considerable changes in CsA formulation (now exclusively Neoral throughout Australasia), assay methods, and blood sampling strategies. The results, representing 93% of CsA laboratories in Australasia (n = 44), found that there was still a plethora of approaches adopted within each organ type for monitoring the established trough (C0) CsA concentration. There was a considerable uptake of 2-hour post-dose sampling (C2) monitoring practices, as demonstrated by assay requests to the responding laboratories, particularly in renal and liver transplantation (46% of centers). Most of these laboratories were also assaying C0 and/or to a much lesser extent, so-called limited sampling strategy AUC (lssAUC) samples at this time. The mFPIA (on TDx and AxSYM analyzers) were still strongly represented (54%) (relatively consistent with international data from the UKQAS proficiency testing scheme at this time) despite on-going concerns expressed in the literature about these methods. However, the Cedia assays had made considerable impact so soon after their introduction with 22% of the laboratories using Cedia or Cedia Plus (Microgenics or Roche Diagnostics; Sydney, Australia) methods. There were a wide variety of dilution protocols adopted in many centers for measuring samples above calibration ranges (such as C2 samples), and hence there was scope for improvement to fully validated techniques. Few centers (16%) made any attempt at interpretation of their results, many seeing their role as purely "measuring" (i.e., an analytical role), not "monitoring" (i.e., including assay and result interpretation). Despite many detailed attempts at providing international or national guidelines for CsA monitoring, etc., there is still considerable scope for improving the quality of the laboratory services offered in this complex as well as expensive area. Several respondents volunteered their support for further Australasian CsA monitoring consensus guidelines.     

肾移植术后患者体内病理生理因素对环孢素血药浓度的影响

目的：探讨肾移植术后患者环孢素（CsA）血药浓度与体内病理生理因素的相关性。方法：采用回顾性调查方法在空军总医院收集58例肾移植术后使用CsA的患者病历资料,记录患者性别、年龄、体重、CsA剂量、血药浓度及相应生化检验值。应用SAS软件（6.04版）对数据进行多元线性回归分析。结果：CsA单位剂量血药浓度与总胆红素（TBil）、用药时间（Time）和年龄（Age）之间存在明显正相关性（P〈0.05）。结论：患者体内总胆红素、年龄与用药时间对CsA血药浓度有显著影响。     

FPIA法与RP-HPLC法检测肾移植患者环孢素血药浓度的比较研究

目的比较反相高效液相色谱法(RP-HPLC)法与荧光偏振免疫(FPIA)法检测肾移植患者环孢素血药浓度结果,评价二者的相关性。方法分别采用RP-HPLC和FPIA方法测定20份血样,并对结果进行统计学处理。结果两种测定方法所得结果差异无统计学意义(P>0.05)。结论RP-HPLC和FPIA方法应用于肾移植患者环孢素血药浓度监测真实可信。     

Relationship between development of nephrotoxicity and blood concentration of cyclosporine A in bone-marrow transplanted recipients who received the continuous intravenous infusion

This study was undertaken to investigate the relationship between blood concentration of cyclosporine A (CsA), administered intravenously by a 24-h continuous infusion, and drug-induced nephrotoxicity or hepatotoxicity. It was investigated retrospectively in 8 patients who had received an allogeneic bone marrow transplant (BMT). The correlation between daily doses and blood concentration of CsA was not significant. Then, the data of blood concentration of CsA and renal or liver function test result were divided into 5-d periods from the date of transplantation, and the mean value for each period was calculated. The maximum values of blood urea nitrogen (BUN) and serum creatinine (SCr) were consistently observed only after the period when the 5-d mean CsA concentration reached the peak level: the maximum BUN and SCr values were witnessed at Periods 2 to 10 and at Periods 1 to 9, respectively. On the other hand, no consistent correlation was found between the 5-d mean CsA concentrations and liver function test result. We also investigated the relationship between renal function and the cumulative dose or AUC of CsA. The parameters of renal function tests reached peak levels at the cumulative dose of 4000 to 10000 mg and at the cumulative AUC of 280000 to 660000 ng/ml.h. These results suggest that: 1) a deterioration of renal function occurs usually after the peak blood concentration of CsA is attained, and 2) the monitoring of the blood concentration of CsA is useful in predicting renal dysfunction in post-BMT patients.