浸透限制固定相高压液相色谱法和荧光偏振免疫法测定人血中卡马西平浓度的比较

目的:比较浸透限制固定相高效液相色谱法(RAM-HPLC)与荧光偏振免疫法(FPIA)测定人血中的卡马西平(CBZ)浓度.方法:建立一种直接进样RAM-HPLC法测定病人静脉和指端血浆的CBZ浓度.结果:两种测定方法都不需样品前处理,有好的重现性和近100％的回收率.FPIA测定静脉血样CBZ的结果与RAM-HPLC分别测定静脉和指端血样CBZ的结果有良好的相关性(R=0.989,0.995),但相差显著(P<0.05);而RAM-HPLC法测定静脉和指端血样两组数据间相差不显著(P>0.05).结论:RAM-HPLC和FPIA法均可测定CBZ浓度.在治疗药物监测中,FPIA更适合常规的监测;RAM-HPLC法更适用于相关研究和特殊病例监测,这里我们成功地应用于测定人外周微量血样中CBZ的浓度.     

高效毛细管电泳与荧光偏振免疫法测定癫患者血清卡马西平的方法学比较

（1. ［摘要］目的建立高效毛细管电泳（HPCE）法测定癫患者血清卡马西平浓度,比较HPCE法和荧光偏振免疫方法（FPIA）分析卡马西平含量的差异性。方法HPCE法采用石英毛细管柱（27 cm×75 μm）, 运行电压18 kV,温度30 ℃,紫外检测波长280 nm,以30 mmol&#8226;L 1磷酸氢二钠（pH＝8.0）含75 mmol&#8226;L 1 十二烷基硫酸钠（SDS）为缓冲液,血样经乙酸乙酯提取后氮气吹干,再用运行缓冲液溶解,压力进样10 s。FPIA分析采用标准TDx测定方法。结果HPCE方法测定卡马西平在2.188～100.000 μg&#8226;mL 1浓度范围内线性关系良好（r＝0.998 9）,日内和日间RSD均≤5%。结论HPCE方法准确、简便、快速,与FPIA检测结果差异无显著性,因其监测成本低更适用于常规血药监测。     

比较HPLC和FPIA法测定人血清中卡马西平的血药浓度

目的比较治疗药物监测中高效液相色谱法(HPLC)和荧光偏振免疫法(FPIA)测定血清中卡马西平(CBZ)的浓度。方法收集上海华山医院和北京天坛医院服用CBZ的癫痫患者608例的稳态谷浓度血样,分别用HPLC法和FPIA法进行测定,用回归法和图解法考察2种测定方法的相关程度以及环氧化卡马西平(CBZE)的交叉反应率。结果CBZE能干扰CBZ的测定,CBZ测定值FPIA较HPLC法高4.58% (95%置信区间:-5.7%~17.6%)。CBZE的交叉反应率随CBZE浓度增加而减小。结论 在CBZ治疗药物监测中,对不同测定方法测定差异,应予以关注并作相应调整。     

高效液相色谱法和荧光偏振免疫法测定万古霉素血清浓度的比较

目的:比较高效液相色谱(HPLC)法与荧光偏振免疫(FPIA)法分别测定血清万古霉素浓度的结果,探讨两者的相关性。方法:收集临床检测万古霉素药物浓度的血清43份,分别用HPLC法和FPIA法进行测定,运用配对t检验,Bland-Altman分析和线性回归分析比较2种方法的测定结果。结果:HPLC法和FPIA法测定的万古霉素血清浓度具有良好的相关性, 回归方程为:Y_FPIA=1.103X_HPLC+0.831 5(R²=0.957 2); Bland-Altman评价分析2种方法一致性较好;配对t检验显示2种方法测定结果之间有显著统计学差异(P<0.000 1)。结论:相较于FPIA法,HPLC法测定不受代谢和降解产物干扰,能准确检测血清万古霉素浓度,适合应用于治疗药物监测。     

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。     

两种检测方法测定甲氨蝶呤血药浓度的比较分析

目的:比较荧光偏振免疫分析法(FPIA)和酶增强免疫法(EMIT)监测甲氨蝶呤(MTX)血浆药物浓度的相关性。方法:收集接受大剂量甲氨蝶呤化疗后的患儿血液样品,分别用FPIA和EMIT法进行测定,考察2种测定方法的相关程度。结果:在MTX血浆浓度≥0.30μmol·L-1时,以FPIA法测定结果(X)和EMIT法测定结果(Y)所作的线性回归方程如下:Y=1.0451 X+0.1153,两组结果间存在较好的相关性(Spearmen相关系数为0.968),EMIT方法的测定结果高于FPIA法所测结果,差异有统计学意义(P<0.05)。结论:MTX血浆药物浓度≥0.30μmol·L-1时,EMIT法测定结果高于FPIA法,且和后者相关性良好,经相互换算后可用于指导四氢叶酸钙(CF)的解救。     

单克隆荧光偏振免疫法和高效液相色谱法测定环孢素A全血浓度的相关性

目的比较单克隆荧光偏振免疫法（FPIA—m法）和高效液相色谱（HPLC）法测定环孢素A（CsA）全血浓度的相关性。方法分别采用HPLC法和FPIA—m法测定42份肾移植患者的CsA血样，测定值用t检验和线性回归方法进行分析。结果FPIA—m法与HPLC法测定值之间相关性好；回归方程为CF=78．12＋1．21CH，r=0．9893（P〈0．001）。结论FPIA—m法和HPLC法同属于特异性分析法，FPIA—m法测定值均高手HPLC法。     

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。     

EMIT与FPIA法测定肾移植术后患者体内环孢素A血药浓度结果比较

目的 比较酶扩大免疫法（EMIT）和荧光偏振免疫法（FPIA）监测环孢素A（CsA）血药浓度的相关性.方法 收集肾移植患者服药后达稳态的谷浓度血样,分别用EMIT法和FPIA法进行测定,考察两种方法测定结果的相关程度.结果 两种方法测定结果高度相关,差异有显著性.结论 EMIT法和 FPIA法测定CsA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.     

HPLC法与FPIA法测定癫痫患者苯巴比妥的血药浓度比较

目的比较治疗药物监测中高效液相色谱法（HPLC）与荧光偏振免疫法（FPIA）测定血浆中苯巴比妥的浓度。方法收集66例服用苯巴比妥的癫痫患者稳态浓度的血浆样品,分别以高效液相色谱法和荧光偏振免疫法进行测定,用回归法考察两种方法测定结果的相关性,比较测定结果。结果2种方法在统计学上无显著性差异,具有良好的相关性（P〉0.05）。以高效液相色谱法测定结果（X）与荧光偏振免疫法测定结果（Y）所作线性回归方程如下：Y（PB）=-0.1270＋0.9637x,相关性分析：r=0.9493。结论高效液相色谱法与荧光偏振免疫法测定苯巴比妥血清浓度具有相关性,两种方法均可用于抗癫痫药物血清浓度的常规监测。     

高效液相色谱法与荧光偏振免疫法在抗癫痫药血清浓度测定中的相关性比较分析

目的探究HPLC与FPIA两种方法在抗癫痫药血清浓度测定中的相关性。方法采集本院收治的经苯妥英钠（PT）、苯巴比妥（PB）、卡马西平（CBZ）治疗的103例癫痫患者血清,分别采用高效液相色谱法、荧光偏振免疫法测定其血药浓度并进行线性回归分析。结果所得的线性回归方程为YPV=-1．421＋1．141X（r=O．963）;YPB=-0．128＋0．956）（（r=0．949）;YcBz=0．183＋0．954X（r=0．944）,且两种方法测得的值呈线性相关;经配对t检验后,两者差异有统计学意义（P〉0．05）。结论高效液相色谱法与荧光偏振免疫法两种方法测定抗癫痫药血清浓度具有相关性。采用HPLC法和FPIA法进行CBZ、PT、PB的血清浓度测定各有利弊,须结合检测药物种类数目、受检人数等合理选择监测方式     

Evaluation of the Ames Seralyzer for the determination of carbamazepine, phenobarbital, and phenytoin concentrations in saliva

The performance of the dry-phase apoenzyme reactivation immunoassay system (ARIS) for the measurement of carbamazepine (CBZ), phenobarbital (PB), and phenytoin (PHT) concentrations in saliva was compared with fluorescence polarization immunoassay (FPIA). Blood and saliva samples were collected from 163 adult and pediatric epilepsy patients, then analyzed using both methods. Regressions between ARIS saliva CBZ, PB, and PHT concentrations, and FPIA unbound and total serum concentrations were highly correlated, but the ARIS technique was somewhat less precise than the FPIA. Valproic acid co-medication did not affect the relationships between ARIS and FPIA saliva concentrations and unbound serum concentrations of PHT, but did disrupt the relationship between ARIS and FPIA saliva PHT and total serum PHT. The sensitivity, specificity, predicted value positive (PV+) of a therapeutic concentration, and predicted value negative (PV-) of a concentration outside the therapeutic range for the ARIS saliva technique compared very well with FPIA for CBZ, PB, and PHT. The ARIS technique for CBZ, PB, and PHT saliva determination provides acceptable accuracy, precision, and sensitivity for therapeutic monitoring. In practice, the benefits of the ARIS saliva technique, including ease of collection, safety, patient/parent acceptance, and short analysis time, are striking.     

Comparison of bioassay, high-performance liquid chromatography, and fluorescence polarization immunoassay for quantitative determination of vancomycin in serum

This investigation was designed to compare three assay techniques, the traditional bioassay (agar diffusion), and two more recent techniques, high-performance liquid chromatography (HPLC) and fluorescence polarization immunoassay (FPIA), for the determination of vancomycin concentrations in serum. One hundred clinical samples obtained from patients receiving vancomycin were assayed by each method. The results from each assay were compared using linear regression analysis. The resultant correlation coefficients were as follows: 0.9996 for the HPLC versus FPIA, 0.7773 for the FPIA versus bioassay, and 0.7779 for HPLC versus bioassay. The FPIA technique was the easiest and fastest of the three methods; FPIA and HPLC were the most accurate.     

Establishment of norvancomycin fluorescence polarization immunoassay for therapeutic drug monitoring

To establish a rapid and simple fluorescence polarization immunoassay method for determination of norvancomycin serum concentration, we collected 300 serum samples from the patients receiving norvancomycin in the hospitals localized in Shanghai, China. The drug concentrations were measured by the established HPLC method and FPIA with vancomycin kit. A FPIA algorithm for the determination of norvancomycin concentration was established according to the correlation between the FPIA and HPLC results. The methods and algorithm were validated in another 70 clinical samples. HPLC determination showed a good linear correlation within the range of 0.5-100?mg?l(-1) of norvancomycin concentrations. The method was validated via extraction recovery, intra- and inter-day methodological recovery and stability of norvancomycin in serum. Correlation analysis between the measurements of HPLC and FPIA in 300 serum samples gave the linear regression equation: (concentration by HPLC)=0.760 × (concentration by FPIA)-0.577 (P<0.001, R(2)=0.982). An algorithm was derived from this correlation for measuring the serum norvancomycin concentrations with FPIA. When it was validated in additional 70 serum samples from patients, 'FPIA algorithm' showed good accuracy versus HPLC: 'FPIA algorithm'=0.93 (HPLC)+0.63, R(2)=0.962, and 94.3% of the results from FPIA algorithm fell within the range of -20%/+20% of HPLC. This algorithm developed in this study can be easily used for determination of norvancomycin using TDx analyzer with vancomycin kit indirectly. It may also be useful for norvancomycin therapeutic drug monitoring.     

Determination of N-desmethylmethsuximide serum concentrations using enzyme-multiplied and fluorescence polarization immunoassays

N-Desmethylmethsuximide (NDM), the active metabolite of the antiepileptic agent methsuximide, has been analyzed by gas-liquid chromatography and high-performance liquid chromatography (HPLC) in the past. This study compares methods using two commercially available immunoassays for ethosuximide, the enzyme multiplied immunoassay technique (EMIT) and fluorescence polarization immunoassay (FPIA), with an HPLC method for the measurement of NDM concentrations in serum. Within-day precision studies, utilizing low therapeutic (15.0 mg/L) and toxic (45.0 mg/L) NDM concentrations (n = 20), resulted in coefficients of variation (CVs) of 4.6 and 4.2%, respectively, for EMIT and 5.4 and 3.2%, respectively, for FPIA. Day-to-day precision studies (n = 10) resulted in CVs of 7.6 and 5.5%, respectively, for EMIT and 3.5 and 2.4%, respectively, for FPIA. No interference was observed from toxic concentrations of acetaminophen, caffeine, carbamazepine, methsuximide, phenobarbital, phensuximide, phenytoin, primidone, salicylate, and valproic acid in the EMIT and FPIA procedures. There was good linear correlation between EMIT and HPLC NDM determinations of 50 patient samples (r = 0.970; y = 0.96 x + 0.03), and a similar correlation between FPIA and HPLC NDM determinations in 48 patient samples (r = 0.975; y = 0.91 x + 1.24). Using ethosuximide reagents, both EMIT and FPIA systems can be adapted to reliably measure NDM serum concentrations.     

Comparison of high pressure liquid chromatography and fluorescence polarization immunoassay methods in a theophylline pharmacokinetic study

High pressure liquid chromatography (HPLC) and fluorescence polarization immunoassay (FPIA) were compared in a theophylline pharmacokinetic study. Eight healthy subjects received single 600-mg oral doses of two different sustained-release theophylline formulations. Fourteen blood samples were collected over 57 h after each dose, and the serum was analyzed for theophylline using both HPLC and FPIA methods. In comparing the two formulations using HPLC, there was no statistical difference in the area under the curve (AUC), terminal rate constant (k), or time of peak. However, there was a 13% difference in peak theophylline concentration (p less than 0.05). The same statistical conclusions were made for all parameters when using FPIA. When comparing the kinetic parameters determined with each assay, the AUC was 12% greater and the k was 17% smaller with FPIA (p less than 0.05). Orthogonal regression of all serum theophylline concentrations showed that FPIA = 1.04 HPLC + 0.20; r = 0.987, p less than 0.001. Stratification of serum theophylline concentrations into different ranges showed that FPIA overestimated the HPLC results in each range, but the percentage of overestimation was greater at lower concentrations (p less than 0.05). The use of FPIA seems appropriate in comparative studies of theophylline pharmacokinetics; however, the calculated kinetic parameters may differ slightly from those obtained with HPLC.     

高效液相色谱法与荧光偏振免疫法测定3种抗癫痫药血清浓度的相关性比较

目的比较2种方法测定3种抗癫痫药血清浓度的相关性。方法76例癫痫患者服药后采用高效液相色谱法和荧光偏振免疫法分别测定其血清浓度,并比较测定结果。结果以高效液相色谱法测定结果(X)与荧光偏振免疫法测定结果(Y)所作线性回归方程如下Y卡马西平=0.1851+0.9550X(r=0.9459);Y苯妥英钠=—1.418+1.136X(r=0.9562);Y苯巴比妥=—0.1273+0.9656X(r=0.9512),2种方法在统计学上无显著性差异(P>0.05)。结论高效液相色谱法与荧光偏振免疫法测定3种抗癫痫药血清浓度具有相关性,2种方法均可用于3种抗癫痫药血清浓度的常规监测。     

Comparison of fluorescence polarization immunoassay and high performance liquid chromatography for the quantitative determination of phenytoin, phenobarbitone and carbamazepine in serum

The Abbott TDx fluorescence polarization immunoassay (FPIA) system has been evaluated and compared with well-established high performance liquid chromatography (HPLC) for the determination of three anticonvulsant drugs: phenytoin, phenobarbitone and carbamazepine. These assays were evaluated for precision, calibration curve stability, specificity and accuracy. Within-run precision studies using control samples (n = 15) in the subtherapeutic, therapeutic, and toxic concentrations, resulted in coefficients of variation in the range of 1.79-3.99% (FPIA) and 1.16-2.52% (HPLC), respectively. Between-run precision ranged from 2.32-6.34% for FPIA and from 2.04-3.38% for HPLC. Comparison of 122 patient samples assayed with both methods indicated an extremely good analytical correlation (r = 0.96) for all three comparisons. The FPIA method offers significant advantages in calibration curve stability while maintaining accuracy and precision comparable with those of established HPLC procedures.