FPIA法与HPLC法测定苯妥英钠血药浓度的对比

目的比较荧光偏振免疫法（FPIA）与高效液相色谱法（HPLc）测定血浆中苯妥英钠的浓度。方法收集服用苯妥英钠的患者稳态浓度的血浆样品,分别以荧光偏振免疫法和高效液相色谱法进行测定,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）。以荧光偏振免疫法测定结果（X）与高效液相色谱法测定结果（Y）所作线性回归方程如下：Y=-0．1093＋0．9812x,相关性分析：r=0．9517。结论荧光偏振免疫法与高效液相色谱法测定苯妥英钠血药浓度具有相关性．2种方法均可用于血药浓度的常规监测。     

限进介质-HPLC法与荧光偏振免疫法测定人血浆中卡马西平浓度的比较

目的：建立以限进介质-高效液相色谱（RAM—HPLC）法测定患者血浆中卡马西平（CBZ）浓度的方法,并与荧光偏振免疫（FPIA）法测定结果进行比较。方法：血浆样品离心后,静脉血浆用FPIA法和RAM—HPLC法测定,指端血浆仅用RAM—HPLC法测定。其中,色谱柱为Hisep shielded hydrophobic phase分析柱,流动相为醋酸铵缓冲液-乙腈（88：12）,流速为1．5mL·min^-1,检测波长为280nm,进样量为20μL,柱温为室温。结果：FPIA法测定静脉血浆CBZ的结果与RAM—HPLC法分别测定静脉和指端血浆CBZ的结果有良好的相关性（r分别为0．989、0．995）,但有显著性差异（P〈0．05）;RAM—HPLC法测定静脉和指端血浆CBZ的结果亦有良好相关性（r=0．998）,但无显著性差异（P〉0．05）。结论：RAM—HPLC法和FPIA法均可测定CBZ浓度,FPIA法更适合常规监测,RAM—HPLC法更适合相关研究和特殊病例监测。2种测定方法均不需样品前处理,有良好的重现性。     

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.     

HPLC法和FPIA法测定苯妥英钠血药浓度的比较

目的　研究用HPLC和FPIA两种方法测定苯妥英钠血药浓度的相关性。方法　建立苯妥英钠血药浓度的HPLC测定方法。分别采用HPLC法和FPIA法测定32例癫痫患者的苯妥英钠血药浓度,比较两种方法的测定结果。结果　HPLC与FPIA法测定的苯妥英钠血药浓度的回归方程为:Y =0 . 90 75X 1 792 3(r=0 . 930 0 ) ,两种方法在统计学上无显著性差异(P >0 . 0 5 )。结论　HPLC法测定苯妥英钠血药浓度灵敏度高,成本低,测定结果与FPIA法相关性良好,适用于临床苯妥英钠血药浓度的常规测定。     

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。     

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。     

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.     

HPLC法和FPIA法测定苯妥英钠血药浓度的比较

目的研究用HPLC和FPIA两种方法测定苯妥英钠血药浓度的相关性.方法建立苯妥英钠血药浓度的HPLC测定方法.分别采用HPLC法和FPIA法测定32例癫痫患者的苯妥英钠血药浓度,比较两种方法的测定结果.结果 HPLC与FPIA法测定的苯妥英钠血药浓度的回归方程为:Y= 0.9075X 1.7923(r=0.9300),两种方法在统计学上无显著性差异(P＞0.05).结论 HPLC法测定苯妥英钠血药浓度灵敏度高,成本低,测定结果与FPIA法相关性良好,适用于临床苯妥英钠血药浓度的常规测定.     

HPLC法测定人血清中去甲万古霉素浓度

目的建立人血清中去甲万古霉素浓度的高效液相色谱测定方法及临床应用。方法以万古霉素为内标,采用10%高氯酸溶液直接沉淀蛋白后进样。色谱柱为SHIMADZUVP-ODS(150 mm×4.6 mm,5μm),流动相为磷酸二氢钾缓冲液-乙腈(92∶8),流速1 mL.min-1,柱温40℃,检测波长236 nm。结果去甲万古霉素血清浓度Y与内标峰面积比X进行线性回归,回归方程为Y=32.544 56X-3.147 39,r=0.999 3;线性范围2.5-80.0μg.mL-1;日内RSD小于3.0%;日间RSD小于5.7%;去甲万古霉素低、中、高浓度的平均方法回收率分别为97.36%、99.51%、101.17%;平均提取回收率分别为86.82%、89.84%、92.68%。结论方法操作简便快速,回收率稳定,适于临床血药浓度监测及人体药动学研究,尤其适于ICU或NICU患者接受去甲万古霉素治疗时的血药浓度监测。     

浸透限制液相色谱、毛细管电泳和荧光偏振免疫法测定人血中苯巴比妥浓度的比较

目的:比较浸透限制高效液相色谱法(RAM-HPLC)、荧光偏振免疫法(FPIA)和高效毛细管电泳法(HPCE)测定人血中苯巴比妥(PB)浓度的异同。方法:建立一种直接进样 RAM-HPLC 法测定血中 PB 的浓度,并与 FPIA 和 HPC E法相比较。结果:三种方法测定 PB 血样浓度的结果有良好的相关性,相关系数分别为0.979(RAM-HPLC 对 FPIA),0.985(HPCE 对 FPIA)和0.989(HPCE 对 RAM-HPLC)。FPIA 法测定 PB 血样的值略高于 RAM-HPLC 和 HPCE 法。结论:RAM-HPLC 优点是准确度高和省去了样品前处理;HPCE 是分辨率高和污染少;FPIA 是简单和快速。三种方法都可用于PB 的治疗药物监测。     

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.     

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     

HPLC法与荧光偏振免疫法检测血样中环孢素A浓度的比较研究

目的:比较高效液相色谱(HPLC)法与荧光偏振免疫法检测血样中环孢素A(CsA)浓度的结果,评价HPLC法测定CsA血药浓度的临床实用性。方法:在HPLC系统和荧光偏振免疫分析仪上检测20对血样,对比检测结果。结果:HPLC系统和荧光偏振免疫分析仪检测CsA血药浓度有显著性差异,HPLC系统测定的结果偏低,但相关性良好(r=0.9952)。结论:HPLC系统和荧光偏振免疫分析仪检测CsA结果间的换算公式有助于临床了解方法改变后检测结果的差异,但HPLC法的检测值要对比相应的参考值才有较强的临床指导作用。     

Evaluation of fluorescence polarization immunoassay for determination of cyclosporin in plasma

The fluorescence polarization immunoassay (FPIA) method for determination of cyclosporin in plasma was evaluated and compared with the high-performance liquid chromatography (HPLC) and the radioimmunoassay (RIA) methods. The coefficients of variation for the within-run and between-run precision were less than 5 and less than 8%, respectively, for samples ranging in concentration from 50 to 600 ng/ml. Recoveries were determined by adding cyclosporin at concentrations from 25 to 1,000 ng/ml to patient plasma; they were, on average, 98.5%. The calibration curve was stable throughout a 10-week study period. There was no clinically significant interference due to hemolysis, icterus, lipemia, or other commonly used drugs. There was considerable variation of the ratio of the FPIA result to the HPLC result, whereas there was a good correlation between the FPIA and the RIA results (r = 0.975, n = 25, y = 1.2x - 36.4), when evaluated using specimens from renal transplant patients receiving cyclosporin orally. It was concluded that the FPIA is an appropriate, rapid method for patient cyclosporin analysis in plasma and serves as a practical alternative to the RIA.     

Measuring endogenous digoxin-like substance and exogenous digoxin in the serum of low-birth-weight infants

The interference with three serum digoxin assay methods of endogenous digoxin-like substance (EDLS) in the serum of low-birth-weight (LBW) infants was assessed. The serum from 5-mL blood samples obtained from each of 19 LBW infants was divided into four 0.5-mL portions. Each portion was spiked with 10 microL of a distilled water-ethanol solution with or without digoxin to produce final digoxin concentrations of 0 (control), 0.49, 0.98, or 1.96 ng/mL. Each portion in each patient was then analyzed by radioimmunoassay (RIA), fluorescence polarization immunoassay (FPIA), and radial partition immunoassay (RPIA) using the control portions to measure EDLS. Serum digoxin concentrations measured by each assay method were calculated by subtracting the EDLS concentrations in the control portions from the measured digoxin concentrations in the spiked samples. The mean +/- S.D. concentrations of EDLS measured by RIA and FPIA were 0.26 +/- 0.13 ng/mL and 0.33 +/- 0.16 ng/mL, respectively. Of the 19 control samples assayed by RPIA, 18 had EDLS concentrations less than 0.1 ng/mL; one sample reflected an apparent concentration of 0.11 ng/mL. Mean recovered digoxin concentrations by RIA at each spiked digoxin concentration were significantly different from those obtained by FPIA and RPIA. A low but significant correlation was noted between EDLS concentrations in serum samples assayed by RIA and FPIA. The RPIA method appears to be preferred over the RIA and FPIA methods used in this study for serum digoxin analysis in LBW infants because of acceptable accuracy and minimal interference by EDLS.     

Analysis of carbamazepine serum by differential pulse voltammetry (DPV) and comparison with fluorescence polarization immunoassay (FPIA): an animal study

Few studies have investigated the value of using an electrochemical method with dropping mercury electrodes to determine carbamazepine level. However, the toxic mercury can result in environmental pollution. In our previous study, we successfully developed a differential pulse voltammetric (DPV) method using a glassy carbon electrode instead of a dropping mercury electrode for the determination of carbamazepine level in standard solutions. In this study, we used the DPV method to determine the serum level of carbamazepine in rabbits. Blood samples were obtained from rabbits which had been fed carbamazepine. The serum concentration of carbamazepine from DPV using glassy carbon electrode and fluorescence polarization immunoassay (FPIA) technique was compared using a correlation test. In addition, DPV and FPIA techniques in carbamazepine detection were evaluated for precision, linearity, and detection limits using a standard solution. The correlation between the carbamazepine concentrations from DPV compared with those by FPIA was good (RSQ?=?0.998). In addition, the coefficient of variation (CV) for the DPV technique and the FPIA technique were low. The precision and detection limit for both methods were satisfactory. The DPV method using a glassy carbon electrode can be a potential method for the determination of carbamazepine in serum.     

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.     

Determination of salivary digoxin with a dry strip immunometric assay.

Analysis of salivary digoxin using a rapid dry chemistry, enzyme-labeled immunometric assay (ELIA) was compared with fluorescence polarization immunoassay (FPIA). Saliva and serum samples were obtained from 40 hospitalized patients who were taking digoxin chronically and from 8 patients just prior to treatment with digoxin. Unstimulated saliva samples were collected from 20 patients; however, saliva volumes from 10 pediatric patients were inadequate to permit analysis by FPIA, and 1 other had unmeasurable concentrations by both methods. Stimulated saliva was collected by having patients chew a small piece of Parafilm for 1-2 min. Salivary digoxin was analyzed using the same procedure recommended for serum digoxin by each manufacturer. There were no significant differences found between ELIA and FPIA determinations of unstimulated or stimulated salivary digoxin, serum digoxin, or saliva/serum concentration ratios. The saliva/serum ratio of the unstimulated group was approximately twice that of the stimulated group (p less than 0.01) by both methods, suggesting that salivary digoxin concentration decreases with increased saliva production rate. Excellent correlations were found between ELIA and FPIA salivary digoxin concentrations and between stimulated saliva and serum concentrations by both assays. Weaker correlations were observed between unstimulated saliva and serum concentrations. There was no evidence of assay interference with either method in eight nondigitalized patients, each taking an average of 6.5 medications. The ELIA appears to provide equivalent results compared with the FPIA for the determination of salivary digoxin concentration. Further investigations are needed before salivary digoxin concentration monitoring can be recommended as an acceptable alternative to serum monitoring.     

Teicoplanin measurement in patients with renal failure: comparison of fluorescence polarization immunoassay, microbiological assay, and high-performance liquid chromatographic assay.

Characterization of antibiotic pharmacokinetics in patients with renal insufficiency may be complicated by interfering substances within the assay. We compared three different assays for teicoplanin in serum and dialysate of 10 hemodialysis and six continuous ambulatory peritoneal dialysis (CAPD) patients. The microbiological assay (micro) had a within-run and between-run coefficient of variation (% CV) of less than 7.5% for concentrations ranging from 0.2 to 96 micrograms/ml. The high-performance liquid chromatographic assay (HPLC) within- and between-run %CV was less than 8% for concentrations ranging from 1 to 80 micrograms/ml. The fluorescence polarization immunoassay (FPIA) within- and between-run %CV was less than 7% for concentrations ranging from 5 to 100 micrograms/ml. In serum of hemodialysis patients FPIA results were slightly higher than HPLC results: FPIA = 1.11 HPLC + 2.37 (r = 0.975, n = 202), and FPIA concentrations in serum were also slightly higher than those measured by micro (FPIA = 1.21 micro - 1.57, r = 0.972, n = 161). The HPLC and micro serum results were also comparable in hemodialysis patients: micro = 0.92 HPLC + 2.89, r = 0.953, n = 160. However, in CAPD patients micro results were lower than HPLC results in serum (micro = 0.82 HPLC + 0.49, r = 0.981, n = 262). In peritoneal dialysate, HPLC values were approximately 60% of the micro values. Thus, FPIA may be the optimal technique for therapeutic monitoring of teicoplanin in the clinical setting due to its simplicity, specificity, and good correlation to HPLC and micro.     

Validation of a high-pressure liquid chromatography and fluorescence polarization immunoassay for the determination of methadone in canine plasma

Methadone is a synthetic opiate derivative that possesses analgesic activity. A modified fluorescence polarization immunoassay (FPIA) method and a high-pressure liquid chromatography (HPLC) method with UV detection were compared for measurement of concentrations of methadone in canine plasma following intravenous and oral methadone administration. The mean+/-SD for accuracy (deviation from actual concentration) and precision (coefficient of variation) when methadone-fortified canine plasma was evaluated with the FPIA method were 3.9+/-3.2% and 4.4+/-2.9%, respectively. The accuracy and precision of the HPLC method were 6.2+/-5.2% and 7.7+/-3.9%, respectively. The limit of quantification for the FPIA and HPLC methods were 25 and 20 ng/mL, respectively. The coefficient of determination (r) between FPIA and HPLC analysis was 0.94 when plasma from dogs dosed with methadone was evaluated. FPIA provides a rapid, sensitive, and specific measurement of methadone in canine plasma following oral and intravenous administration.