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A rapid fluorescence polarization immunoassay (FPIA) has been developed for the determi-nation of aflatoxins in samples of naturally-contaminated herbal teas. The tracers were synthesized by chemical method and determined by thin layer chromatography (TLC) and mass spectroscopy (MS). Fluorescence polarization was evaluated by the detection of polarized light. The results showed that the limit of detection (LOD) of FPIA for aflatoxins was 20 ng·mL-1, the IC50 was 371.80 ng·mL-1, and the linear range of the developed FPIA was 92.76-252.32 ng·mL-1. Compared with conventional HPLC methods, the FPIA developed in this study has the advantages of short analysis time and low cost. This method may be suitable for high- throughput screening of aflatoxins in herbal teas.
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Objective To investigate the comparation of sodium valproate concentration in peripheral blood monitoring by fluorescence polarization immunoassay method(FPIA)and high performance liquid chromatography method(HPLC)in epilepsy children.Methods 87 cases of epilepsy children received Sodium valproate treatment in our hospital from February 2014 to June 2016 were selected,fasting venous blood of elbow vein were collected the next morning after last medication, blood concentrations of Sodium valproate in serum samples were detected by FPIA method and HPLC method respectively,the correlation and consistency of results of the two methods were observed and compared.Results The intra day and inter day RSD of Sodium valproate concentration in peripheral blood in epilepsy children detected by FPIA were <5%,the recovery rate was 90%-110%,the precision and accuracy were high;The intra day and inter day RSD of Sodium valproate concentration in peripheral blood in epilepsy children detected by HPLC were <5%,the recovery rate was 90%-110%,the precision and accuracy were high; the linear regression equation between determination value of HPLC method (X) and determination value of FPIA method (Y) was:Y=0.8355X+1.8231,correlation coefficient r=0.914,the detection results were positively related;the Sodium valproate blood concentration detected by FPIA was significantly lower than that detected by HPLC method, the difference was statistically significant (P<0.05);Bland-Altman deviation chart results show that determination of blood drug concentration by HPLC method was higher than that of FPIA method by 7.2μg/mL.Conclusion The precision and accuracy of sodium valproate concentration in peripheral blood monitoring by FPIA method and HPLC method were all high, and the correlation was good, but the detection results of the two methods were significantly different,the detection result of HPLC method was higher than that of FPIA method,need to choose and judge according to the clinical situation.
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Objective To evaluate the correlation and difference of reversed phase high performance liquid chromatography (RP-HPLC) and fluorescence polarization immunoassay (FPIA) on determining serum concentration of carbamazepine.Methods Fifty serum samples were collected,both RP-HPLC and FPIA methods were employed to determine the concentration of carbamazepine.The results were analyzed by paired t test,Bland-Altman and Deming regression methods,respectively.Results The results of measuring 50 samples by the two methods showed that FPIA datas were significantly higher than RP-HPLC datas,and there was statistically significant difference(P<0.05) and poorer consistency between two methods;There was good correlation between carbamazepine concentrations determined by the two methods.Deming regression equation was CFPIA=1.195 3 CRP-HPLC-0.144 0,and Pearson correlation coefficient was 0.968 5.Conclusion Clinicians should pay more attention to the difference of carbamazepine concentration determination by different methods when carbamazepine individualized dosage regimen was adjusted according to therapeutic drug monitoring.
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ABSTRACT The emergence of chikungunya virus in the Americas means the affected population is at risk of developing severe, chronic, rheumatologic disease, even months after acute infection. Accurate diagnostic methods for past infections are essential for differential diagnosis and consequence management. This study evaluated three commercially-available chikungunya Immunoglobulin G immunoassays by comparing them to an in-house Enzyme-Linked ImmunoSorbent Assay conducted by the Centers for Disease Control and Prevention (Atlanta, Georgia, United States). Results showed sensitivity and specificity values ranging from 92.8% - 100% and 81.8% - 90.9%, respectively, with a significant number of false-positives ranging from 12.5% - 22%. These findings demonstrate the importance of evaluating commercial kits, especially regarding emerging infectious diseases whose medium and long-term impact on the population is unclear.(AU)
RESUMEN Como consecuencia de la aparición del virus del chikungunya en las Américas, la población afectada corre el riesgo de padecer reumatismos crónicos graves, aun meses después de la infección aguda. Es fundamental contar con métodos precisos para diagnosticar los antecedentes de la infección a fin de elaborar un diagnóstico diferencial y abordar las manifestaciones de la fase crónica. Se han estudiado tres inmunoensayos comercializados de detección de inmunoglobulinas G para el diagnóstico del chikungunya, comparándolos con el enzimoinmunoanálisis de adsorción (ELISA) propio. Los resultados señalan valores de sensibilidad del 92,8% al 100% y de especificidad del 81,8% al 90,9%, así como un número significativo de falsos positivos, de entre el 12,5% y el 22%.(AU)
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Humans , Reagent Kits, Diagnostic , Immunoglobulin G , Chikungunya virus/isolation & purification , Fluorescence Polarization Immunoassay , Immunoenzyme Techniques , Chikungunya Fever/diagnosis , Americas , Caribbean RegionABSTRACT
Drug addiction is a chronic disease with a potential for fatality if not treated. The drugs with potential for abuse are mostly psychoactive drugs. Serious widespread medical and health consequences associated with drug abuse involve neurotoxicity, cardiovascular complications, impairment of the immune system function, and many other physiological effects. Illicit drug use remains the second most common mode of HIV infection. Various analytical techniques and number of biological matrices has been used for the detection of drug of abuse in cases such as drug addiction, driving under influence of drugs, neonatal drug exposure in case of drug abuse by pregnant women etc. Urine and blood sample remain the most widely used conventional biosample for the detection of drug of abuse. Various other alternative biological matrices such as saliva, hair, nails, tears and meconium have also been used for the same purpose. Number of analytical techniques such as liquid chromatography with mass spectrometry (LC-MS) and LC with tandem MS (LC-MS2), enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), electrospray ionization Time-of- Flight mass spectrometry (ESI-TOF), combination of ultra-performance liquid chromatography (UPLC) and TOF, fluorescence polarization immunoassay (FPIA) and enzyme multiplied immunoassay technique (EMIT) have been used for the detection of drugs of abuse in above mentioned biosamples. This review summarizes the conventional as well as alternative biological matrices and various analytical techniques used for the determination of drugs of abuse.
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Objective To explore the matrix effect on cyclosporine A (CsA) test by fluorescence polarization immunoassay (FPIA) and enzyme-multiplied immunoassay technique (EMIT), explain the discrepancy of external quality control results between these two methods and find the corrective action.Methods One hundred whole blood samples with various concentrations were adopted and CsA levels were detected by FPIA and EMIT.The results were compared with each other.Moreover, the influence of residual metal ions upon immunoreactions was assessed by adding Cu2+ and Zn2+.The effect of non-whole blood matrix on extraction efficiency for quality control materials and CsA calibrator was evaluated by adding identical volume of Hb-rich reagents followed with re-extraction.Results There is good correlation between results measured with FPIA(X) and EMIT(Y) methods ( Y=0.926 8X -8.115,R2 =0.996 9).Neither FPIA nor EMIT was affected by residual metal ions ( P ＞ 0.05 ). Non-whole blood matrix decreased the extraction efficiency of two methods, but it could be corrected by supplementation of the Hb-rich reagents (≥30 g/L).Conclusions Non-whole blood matrix may be the main reason for the inconsistent results measured by FPIA and EMIT methods.It could be corrected by using Hb-rich reagents.In addition,we should consider the influence of low lib on CsA test,espocially for organ transplant patients with lower Hb ( ＜30 g/L).
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A distrofia muscular dos cães Golden Retriever (GRMD), uma miopatia degenerativa causada pela ausência da distrofina é geneticamente homóloga à distrofia muscular de Duchenne que acomete humanos, portanto, estes cães são considerados modelos experimentais para estudos em terapia celular. Seu sucesso depende da imunossupressão adequada. A ciclosporina A (CsA) é indicada para tal e a monitorização de suas concentrações sangüíneas e efeitos adversos são essenciais para viabilizar a terapia. Foram estudados cães GRMD, e normais da mesma raça, submetidos a terapia com CsA, associada, nos GRMD, ao transplante de células tronco. Foram avaliados as concentrações sangüíneas do fármaco através de amostras coletadas a cada dois ou três dias e analisadas pelo método do imunoensaio por fluorescência (FPIA). Como resultado observamos que as concentrações de CsA oscilaram muito, em seis dos oito animais. Concluímos que as doses variam individualmente sendo de maior importância avaliar a concentração do fármaco no sangue e sua viabilização no uso da terapia celular
The muscular dystrophy of Golden Retriever (GRMD) is a degenerative miopaty caused by the absence of dystrophy and it is genetically homologue of the Duchenne muscular dystrophy in humans, so, these dogs are considerably experimental models for studies on cellular therapy. Their successful depends of the adequate immunosuppression. Cyclosporin A (CsA) is indicated for that and the monitoring of the blood concentration and adverse effects are essential to viabilise the therapy. It was studied GRMD dogs, and normal dogs from the same breed, submitted for therapy with CsA, associated, on GRMD, of cell transplantation. It was evaluated blood concentration of the drug, between two or tree days using the method of FPIA. In our results we found that the CsA blood concentrations oscillated too much on six than eight of our animals. We concluded that the doses varieties Braz. J. vet. Res. anim. Sci., São Paulo, v. 45, n. 2, p. 131-137, 2008 individually and the correct dosage as to important as the evaluation of the blood concentration of the drug and became viable for cell therapy
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Animals , Dogs , Muscular Dystrophy, Animal/blood , Fluorescent Antibody Technique/methods , Immunosuppression Therapy/adverse effectsABSTRACT
OBJECTIVE:To explore quality control of plasma concentration monitoring for sodium valproate and to improve the quality of pharmaceutical monitoring.METHODS:The statistical and continuous analysis of sodium valproate control data from our hospital of 2008 were conducted using fluorescence polarization immunoassay(FPIA).RESULTS:Mean recovery and RSD were 98.37% and 1.94% for low control samples,99.33% and 2.88% for medium control samples,98.17% and 4.10% for high control samples.The RSD of sample was lower than 5% and in line with the requirement for biological sample determination in Chinese Pharmacopoeia.CONCLUSION:FPIA is a comparatively accurate and stable method for plasma concentration monitoring of sodium valproate.
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85 mL/min),CL=6.0?(WT/60)~(0.52).④The increased volume of peripheral distribution (V_2) was observed when norvancomycin was co-administered with diuretics;④Reduced drug clearance,prolonged t_(1/2),and increased values of AUC_(24) were found in elderly patients.Conclusions Renal function impairment and age have significant impact on PK parameters of norvancomycin.Dosing regimens of norvancomycin were finally established for different patients on the basis of important PPK parameters generated from different groups of patients.
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Objective To study the effect of folic acid on decreasing level of plasma total homocysteine(tHcy)in patients with sudden sensorineural hearing loss(SSHL) and the optimal dosage of folic acid. Methods Ten randomized controlled trials involving treatment data on 210 patients with SSHL were retrospectively studied.They were divided into seven groups according to the daily dosage of folic acid: group A to group G,0.2 mg,0.4 mg,0.8 mg,2.0 mg,5.0 mg,10.0 mg and 15.0 mg,respectively.Besides oral administration of folic acid,Vitamine B6 and B12were supplemented,and other routine treatment were performed.Fluorescence polarization immunoassay was employed to detect the plasma tHcy before and 3 months after the treatment.And the data of plasma tHcy of 210 patients without SSHL were collected and served as controls.The levels of plasma tHcy were statistically analysed between the SSHL group and control group and among group A to group G. Results The level of plasma tHcy in the SSHL group was significantly higher than that in the control group,(18.07?1.58)?mol/L vs(13.63?1.33) ?mol/L(P0.05). Conclusion The levels of plasma tHcy are significantly increased in SSHL.Folic acid may play an important role in decreasing the levels of tHcy in patients with SSHL,and a dosage of 10 mg/d for oral adminstration is well suggested.
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OBJECTIVE: To analyze the therapeutic drug monitoring results of valproic acid( anti- epileptic drug) . METH-ODS: The concentrations of valproic acid in 226 cases in our hospital were determined by fluorescence polarization immunoassay. RESULTS: The results showed that 47. 79% were within the normal referenced concentration range, with average concentration at( 66. 19? 12. 13) ? g? mL- 1, 46. 90% were below the lower limit and 5. 31% were above the upper limit. CONCLUS-ION: The monitoring results are far from satisfactory. It is advisable to adopt individualized administration so as to achieve the goal of treatment.
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OBJECTIVE:To observe the efficacy and safety of the effective blood concentration of cyclosporin A in patients after bone marrow transplantation.METHODS:By fluorescence polarization immunoassay(FPIA),the whole blood trough concentration of CsA in 14 patients after undergoing bone marrow transplantation was monitored for 216 times in total from day 1 to day 466,and the monitoring results were analyzed.RESULTS:The effective blood concentrations of CsA in patients with chronic myelocytic leukemia(CML),acute non lymphocytic leukemia(AML),acute lymphoblastic leukemia (ALL),Mediterranean disease(THAL),and pancytopenia were(50～450) ng?mL~(-1),(100～450) ng?mL~(-1),(100～350) ng?mL~(-1)(200～500) ng?mL~(-1),and(250～500) ng?mL~(-1),respectively.Rejection reaction appeared in 7 cases after bone marrow transplantation,with whole blood trough concentration of CsA ranged from 67.4 ng?mL~(-1) to 189.34 ng?mL_(-1) (125.44?39.56 ng?mL~(-1)on average).Adverse drug reaction appeared in 3 cases,with trough concentration of CsA ranged from 412.5 ng?mL~(-1) to 548.62 ng?mL~(-1)(481.39?68.08 ng?mL~(-1) on average).CONCLUSION:Timely monitoring of plasma concentration of CsA and modifying of the dosage regimen can avoid the occurrence of rejection reaction and adverse drug reaction,which is of great significance for patients' medication safety and efficacy.
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OBJECTIVE:To study the optimal monitoring indexes of plasma concentration of cyclosporine A(CsA) in patients receiving hematopoietic stem cell transplantation.METHODS:The lowest CsA concentration(C0) and the highest CsA concentration(C2) in 23 hematopoietic stem cell transplant recipients were detected by FPIA.All the data were analyzed statistically.RESULTS:Within 6 months after hematopoietic stem cell transplanttion,C0,C2,C0+C2 and C2/C0 in the recipients were(228.84?142.48) ?g?L-1,(741.50?294.42) ?g?L-1,(970.34?391.18) ?g?L-1 and(3.88?1.94) ?g?L-1,respectively.CONCLUSION:As reasonable monitoring indexes for the plasma concentration of CsA,C0+C2 and C2/C0 can comprehensively reflect the exposure of drug in body and monitor the toxicity of CsA in liver and kidney.
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60a,24cases)according to their age,the blood concentrations of the2groups in different time were compared.RESULTS:Under the circumstance of close oral dosage of CsA,the blood concentration of CsA in the older-aged group were found remarkably higher than that in the young&middle-aged group(P
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BACKGROUND: It was purposed to estimate correlation between fluorescence polarization immunoassay (FPIA) and high performance liquid chromatography (HPLC), and precision of individual methods. It was also objected to describe distribution of plasma total homocysteine in Korean adults. METHODS: The subjects were 100 adults admitted to Inha University Hospital during the month of October, 1998. The total plasma homocysteine concentration was measured by FPIA (IMx analyzer, Abbott Laboratories, IL, USA) and by HPLC (ACCLAIM Biogenic Amines Testing System, Bio-Rad Laboratories, CA, USA) using Bio-Rad Homocysteine. RESULTS: Plasma homocysteine levels (mean+/-SD) from Korean healthy adults by FPIA and HPLC were 9.75+/-3.80micromol/L, 7.72+/-3.36micromol/L, respectively. Plasma homocysteine levels according to sex by FPIA were 11.79micromol/L for male, 7.71micromol/L for female, and those by HPLC were 9.47micro mol/L for male, 5.98micromol/L for female, respectively. Intra-assay coefficient variations (CVs) of low, medium, and high concentration by FPIA are 1.83%, 0.47%, and 1.66%, and those by HPLC are 5.53%, 5.37%, and 4.56%, respectively. Inter-assay CVs of low, medium, and high concentration by FPIA are 2.28%, 1.44%, and 1.29%, and by HPLC are 7.23%, 5.54%, and 4.95%, respectively. CONCLUSION: Plasma homocysteine levels from male were significantly higher than female in Korean. Plasma homocysteine levels were increased according to increment of age. FPIA was more convenient, automatic, rapid, and reproducible than HPLC and also excellently correlated with HPLC. It is concluded that FPIA will potentially benefit for quantifying homocysteine in clinical laboratories.
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Adult , Female , Humans , Male , Biogenic Amines , Chromatography, High Pressure Liquid , Chromatography, Liquid , Fluorescence Polarization Immunoassay , Fluorescence Polarization , Fluorescence , Homocysteine , PlasmaABSTRACT
In the present study a new method for selectively determining parent cyclosporine in whole blood, a clone enzyme donor immunoassay (CEDIA; Boehringer Mannheim), was compared with a fluorescence polarization immunoassay (FPIA; TDxAbbott). A total of 429 samples were collected, comprising 371 renal, 34 cardiac, 14 bone marrow and 10 corneal transplant recipients. Regression equations in 429 samples is CEDIA cyclosporine (ng/mL) = 0.999 x FPIA (ng/mL) + 1.684, (r=0.997). The linearity analysis was done from 0 – 1,000 ng/mL, which gave a good analytical range of between 15-600 ng/mL. The new CEDIA method also has within-rin CV = 1.77 – 3.69 percent which is better than the FPIA method (2.20 – 6.10 percent). The advantages of the new CEDIA method are the lower cost of the reagents and the fact that there is no need to purchase a new automated clinical chemistry analyser since it can be applied to routine chemistry instruments immediately after the reagents become available to the laboratory.
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BACKGROUND: The penetraton in vivo of topically applied substances can be assessed by physiological or pharmacalogical signs or analysed by chemical or histological techniques. In vitro absorption can be commonly quantitated by measuring the passage of a radioisotope-labelled substance across skin that has been mounted in a diffusion chamber. OBJECTIVE: Fluorescence polarization immunoassay technique has made the possible rapid growth of therapeutic drug nonitoring. We applied this methodology in measuring percutaneous absorption in a diffusion chamber. METHODS: We utilized sheets of whole epidermis prepared from the circumcised prepuce. Some epidermal sheets were treated with 2 ml of acetone for 2 minutes, and others not. The epidermal sheet was mounted in a diffusion chamber between the donor compartment for the penetrant and the receptor compartment containing saline. Lidocaine HC1(10 microgram/cm2) in vehicle(propylene glycol:ethanol; 7:3, vol/vol) was applied to the donor compartment for the penetrant. With flow rate of about 3 ml/h all of the receptor phase collected during 2 hours interval were quantitated for 10 hours by the fluorescence polarization immunoassay. RESULTS: Total absorption of lidocaine HC1 in the acetone-untreated group was 2.14+/-0.74% of the applied dose. Total absorption in the acetone-treated group showed no substantial difference (2.09+/-1.25%) compared to those of acetone-untreated group. The amount of lipid extracted from a epiderrnal sheet with acetone was 19+/-2.97%. CONCLUSION: Fluorescence polarization immunoassay may be a useful method in measuring percutaneous absorption in vitro.
Subject(s)
Humans , Absorption , Acetone , Diffusion , Epidermis , Fluorescence Polarization Immunoassay , Fluorescence Polarization , Fluorescence , Histological Techniques , Lidocaine , Skin , Skin Absorption , Tissue DonorsABSTRACT
OBJECTIVE:To establish an optimal therapeutic window concentration of CsA trough levels in renal transplant recipients on triple immunosuppressants regimen METHODS:A total 1 874 samples from 268 renal transplant recipients were measured by fluorescence polarization immunoassay(FPIA) According to the duration after operation and clinical diagnoses ,the whole blood CsA trough levels were compared among subgroups RESULTS:The optimal therapeutic window concentration of CsA was 300～400?g/L(within 1 month after operation),250～350?g/L(2nd～3rd month),150～250?g/L(4th～6th month),100～200?g/L(7th～12th month)and 100～150?g/L(more than 12 months) CONCLUSION:The above mentioned therapeutic window concentratin of CsA trough levels was ideal for renal transplant recipients with no marked acute toxic effects and rejection reaction