Immunonephelometric determination of retinol-binding protein in serum and urine.

1983 ◽  
Vol 29 (5) ◽  
pp. 853-856 ◽  
Author(s):  
M T Parviainen ◽  
P Ylitalo
Keyword(s):  
Binding Protein ◽  
Retinol Binding Protein ◽  
Good Precision ◽  
Serum Samples ◽  
Detection Range ◽  
Analytical Recovery ◽  
Beta 2 Microglobulin ◽  
Urine Specimens ◽  
Serum And Urine

Abstract An immunonephelometric method developed for measurement of retinol-binding protein (RBP) in serum and urine can detect it in concentrations of about 30 micrograms/L, which is in the lower limit of its normal concentration in urine (range 0-0.56 mg/L; mean +/- SD 0.19 +/- 0.15; n = 44). Urinary RBP was increased (range 0.93-29.5 mg/L) in all of 25 urine specimens from 13 subjects being treated with aminoglycoside (tobramycin). Urinary excretion of RBP was correlated (r = 0.83) with the excretion of beta 2-microglobulin. The within-assay and day-to-day precision (CV) was determined over the detection range of 0.03-8 mg/L. Within these limits the corresponding CVs varied from 4 to 27% and from 8 to 30%, respectively. The method had fairly good precision within the optimal measuring range of approximately 0.4 to 4.5 mg/L for both urine and 20-fold diluted serum samples. For various RBP concentrations our analytical recovery was 89-114% of added RBP. Results by this method correlated well (r = 0.96, n = 24) with those by a radial immunodiffusion method.

Micromethod for determination of creatinine in biological fluids by high-performance liquid chromatography.

1978 ◽  
Vol 24 (5) ◽  
pp. 747-750 ◽  
Author(s):  
S J Soldin ◽  
J G Hill
Keyword(s):  
High Performance Liquid Chromatography ◽  
Continuous Flow ◽  
Present Method ◽  
High Performance ◽  
Biological Fluids ◽  
Coefficients Of Variation ◽  
Analytical Recovery ◽  
Urine Specimens ◽  
Serum And Urine

Abstract We describe a procedure for the rapid and specific measurement of creatinine, in which it is separated from other compounds in serum or urine by paired-ion chromatography and is quantified by measuring its absorbance at 200 nm. The procedure can be done on as little as 10 microliter of serum. Between-day precision studies for concentrations of 13 and 62 mg/liter yielded coefficients of variation of 6.9 and 2.2%, respectively. Analytical recovery of various amounts of creatinine added to plasma exceeded 95% in all cases. The proposed procedure was compared with the continuous-flow procedure by analyzing a series of serum and urine specimens by both methods. There was excellent agreement for urine specimens, but with serum the results by the present method were significantly (P less than 0.001) lower.

scholarly journals Determination of Retinol-Binding Protein in Serum by Kinetic Immunonephelometry with Polyethylene Glycol Pretreatment

1984 ◽  
Vol 21 (6) ◽  
pp. 484-487 ◽  
Author(s):  
M J Hallworth ◽  
Jacqueline Calvin ◽  
C P Price
Keyword(s):  
Polyethylene Glycol ◽  
Detection Limit ◽  
Binding Protein ◽  
Regression Line ◽  
Retinol Binding Protein ◽  
Serum Samples ◽  
Coefficients Of Variation ◽  
Method Yield ◽  
Scattering Material

This work describes the use of polyethylene glycol as a pretreatment reagent to remove endogenous light scattering material from serum samples prior to automated immunonephelometric analysis on a centrifugal analyser. An assay system for retinol-binding protein is described, which allows rapid (10 minutes) quantitation of retinol-binding protein in serum samples with a detection limit of 5 mg/L and between-assay coefficients of variation ranging from 2·9% to 4·0%. The assay range is 5–80 mg/L and accuracy comparisons with a Mancini single radial immunodiffusion method yield a regression line y=0·89x+0·52 ( r=0·98, n=22). The problem of analyte precipitation associated with use of pretreatment regimes is discussed.

Monoclonal antibody-based immunoenzymometric assays of retinol-binding protein

1993 ◽  
Vol 39 (3) ◽  
pp. 472-476 ◽  
Author(s):  
A B Pereira ◽  
S K Nishida ◽  
J G Vieira ◽  
M T Lombardi ◽  
M S Silva ◽  
...  
Keyword(s):  
Binding Protein ◽  
Retinol Binding Protein ◽  
Good Precision ◽  
Routine Work ◽  
One Step ◽  
High Concentrations ◽  
The One ◽  
Convoluted Tubules ◽  
Good Agreement

Abstract Retinol-binding protein (RBP) is a low-molecular-mass protein (21 kDa), easily filtered in renal glomeruli and very efficiently reabsorbed by the proximal convoluted tubules (PCTs). In PCT dysfunction, high concentrations of RBP are found in urine. Several methods have been used to determine RBP in serum or urine. We describe the production, selection, labeling, and utilization of anti-RBP monoclonal antibodies in two- or one-step immunoenzymometric assays for the determination of RBP. The one-step assay has good precision, with within-run and between-run CVs < 6.6% and 5.9%, respectively. Comparison with radial immunodiffusion (x) showed good agreement: y = 0.068 mg/L + 0.899x (n = 24). Comparison between the one-step (y) and two-step (x) versions of the assay also showed a very good correlation: y = 212 micrograms/L + 0.910x. The one-step assay has been adopted for routine work; it detects transthyretin-bound as well as free RBP and may have clinical usefulness in evaluating the functional status of PCTs.

Automated enzymic determination of ethanol in blood, serum, and urine with a miniature centrifugal analyzer.

1976 ◽  
Vol 22 (6) ◽  
pp. 802-805 ◽  
Author(s):  
T P Hadjiioannou ◽  
S I Hadjiioannou ◽  
J Avery ◽  
H V Malmstadt
Keyword(s):  
Blood Serum ◽  
Reaction Rates ◽  
Rate Of Change ◽  
Coefficients Of Variation ◽  
Analytical Recovery ◽  
Rate Method ◽  
Relative Errors ◽  
Urine Specimens ◽  
Serum And Urine

Abstract We describe an automated spectrophotometric reaction-rate method for determination of ethanol in serum and urine with a miniature centrifugal analyzer. The theanol is selectively oxidized in the presence of alcochol dehydrogenase and NAD+ to form NADH, which is measured by the rate of change of its absorbance. Reaction rates are determined automatically, and unknown concentrations are calculated from a computer-generated working curve based on aqueous ethanol standards. Blood, serum, or urine specimens need not be deproteinized. The method permits duplicate analysis of at least 30 samples per hour. Coefficients of variation and relative errors are about 2-3% for ethanol concentrations of 0.3-3.0 mug per 2 mul of sample. Analytical recovery of ethanol added to serum is 92-103% (average, 98.5%). Comparisons with distillation-oxidation, gas-chromatographic, and conventional enzymic procedures give satisfactory agreement.

An Immunoassay Method for the Determination of Rat Retinol-Binding Protein in Serum and Urine

2015 ◽  
pp. 164-168 ◽  
Author(s):  
C. Biagini ◽  
R. Alinovi ◽  
E. Bergamaschi ◽  
A. Mutti ◽  
R. Berni ◽  
...  
Keyword(s):  
Binding Protein ◽  
Retinol Binding Protein ◽  
Serum And Urine

Latex immunoassay of retinol-binding protein.

1982 ◽  
Vol 28 (5) ◽  
pp. 1167-1171 ◽  
Author(s):  
A M Bernard ◽  
D Moreau ◽  
R R Lauwerys
Keyword(s):  
Latex Particle ◽  
Binding Protein ◽  
Biological Fluids ◽  
Correlation Coefficients ◽  
Retinol Binding Protein ◽  
Coated Particles ◽  
Standard Curve ◽  
The Mean ◽  
Beta 2 Microglobulin

Abstract Latex immunoassay, a sensitive method based on latex particle agglutination, is used here for the determination of retinol-binding protein in human biological fluids. The assay, similar to that described previously for beta 2-microglobulin (Clin. Chem. 27:832-837, 1981), consists of incubating the sample at 37 degrees C for 30 min with antibody-coated particles, then quantifying the resulting agglutination by particle-counting or turbidimetry. Latex particles coated with antibody are stabilized just before use by dispersing them in a solution of bovine serum albumin at pH 10. The standard curve ranges from 0.5 to 32 micrograms/L; recovery averages 102% in urine and 93.5% in serum; between- and within-assay CVs range from 5.1 to 11.7%. The correlation coefficients of latex immunoassay with rocket immunoelectrophoresis for analysis of retinol-binding protein in 26 urines and with radial immunodiffusion in 30 sera are 0.99 and 0.91, respectively. In healthy subjects, the mean urinary excretion of retinol-binding protein is 52.5 micrograms/g of creatinine (SD = 59.2 micrograms/g of creatinine; n = 150) and the concentration in serum averages 46 mg/L (SD = 10.4 mg/L, n = 22).

Determination of β2-Microglobulin in Serum by a Microparticle-Enhanced Nephelometric Immunoassay

1992 ◽  
Vol 38 (12) ◽  
pp. 2464-2468 ◽  
Author(s):  
J A Viedma ◽  
S Pacheco ◽  
M D Albaladejo
Keyword(s):  
Linear Regression Analysis ◽  
Reference Interval ◽  
Serum Samples ◽  
Standard Curve ◽  
Analytical Recovery ◽  
Beta 2 ◽  
Beta 2 Microglobulin ◽  
Scattering Signal ◽  
Sample Dilution

Abstract This fully automated nephelometric immunoassay to quantify beta 2-microglobulin in human serum measures the light-scattering signal produced by agglutination of commercially available latex microparticles (diameter 0.1 micron) coated with specific F(ab')2 against beta 2-microglobulin. The calibration curve, generated by serial dilutions of a beta 2-microglobulin standard of known concentration, is used to calculate beta 2-microglobulin concentrations in serum samples by the logit-log function and linear-regression analysis. The assay range (sample dilution 400-fold) extends from 0.3 to 40.0 mg/L. No antigen excess appears at beta 2-microglobulin concentrations up to 320 mg/L. Within-run CVs ranged from 1.0% to 3.4%, and between-days from 1.2% to 2.8%. Total imprecision (CV) was < 5%. Analytical recovery averaged 99.5% +/- 2.8%. Rheumatoid factor, complement, bilirubin (up to 340 mumol/L), and hemoglobin (up to 2.0 g/L) do not interfere. Strongly turbid lipemic samples must be cleared before analysis. Standard curve linearity was very good in samples covering the clinical useful range of concentrations. Results of the method correlated well with those of radioimmunoassay and microparticle enzyme-linked immunoassay (r = 0.979 and 0.975, respectively). The reference interval (nonparametric estimation) in apparently healthy adults (n = 303) was 0.87 (0.80-0.94) to 2.42 (2.28-2.45) mg/L; the median value was 1.54 mg/L.

PSV-4 Determination of Deoxynivalenol (DON) Content in Biological Samples as an Indicator of DON Intake in Grower-finisher Pigs

2021 ◽  
Vol 99 (Supplement_1) ◽  
pp. 206-207
Author(s):  
Michael O Wellington ◽  
Michael A Bosompem ◽  
Veronika Nagl ◽  
Daniel A Columbus
Keyword(s):  
Mass Spectrometry ◽  
Biological Samples ◽  
High Performance ◽  
Tandem Mass ◽  
Serum Samples ◽  
Blood Samples ◽  
Swine Diets ◽  
Direct Quantification ◽  
Serum And Urine

Abstract Due to difficulties in obtaining consistent and/or reliable measures of deoxynivalenol (DON) in complete swine diets, we investigated whether measuring DON in biological samples could be used as an indicator of DON ingestion in pigs. In this study, graded levels of DON (1, 3, or 5 ppm) were fed to grower-finisher pigs for a period of 77-d. On d 35 and 77 of the study, urine samples were quantitatively collected over a 24-h period and blood samples were collected between 3 – 4 h after the morning meal on each of those days for serum DON analysis. For direct quantification of DON in urine, high-performance liquid chromatography with tandem mass spectrometry was performed. For serum samples, indirect quantification of DON was performed via enzymatic hydrolysis. We observed that DON content in urine increased linearly as intake of DON increased (Fig.1A; P < 0.05). Analysis of DON in serum follow a similar trend, where serum DON content was increased as DON intake increased (Fig.1B; P < 0.05). An average of 30% of DON ingested was recovered as DON in urine over a 24-h period. In summary, there was a linear relationship between DON intake and DON content in both urine and blood serum, therefore, analyzing DON concentration in serum and urine could be used as a tool to estimate for DON exposure in pigs under controlled conditions.

Determination of creatinine in serum and urine by a rapid liquid-chromatographic method

1990 ◽  
Vol 36 (6) ◽  
pp. 830-836 ◽  
Author(s):  
R Paroni ◽  
C Arcelloni ◽  
I Fermo ◽  
P A Bonini
Keyword(s):  
Signal To Noise Ratio ◽  
Liquid Chromatographic Method ◽  
Analytical Recovery ◽  
Internal Standardization ◽  
Assay Precision ◽  
Fuller’S Earth ◽  
Liquid Chromatographic ◽  
Specific Evaluation ◽  
Serum And Urine

Abstract We describe an HPLC ion-pair procedure for rapid and specific evaluation of creatinine in serum and urine. We used a 15 cm X 4.6 mm ODS column with a 50/50 (by vol) mixture of sodium decanesulfonic acid (10 mmol/L, pH 3.2) and methanol and measured absorbance at 236 nm. Serum (100 microL) or 30-fold-diluted urine (100 microL) was added to 400 microL of acetone. After centrifugation, the supernates (300 microL) were dried, reconstituted with the mobile phase, and injected into the HPLC. Assay precision was tested for concentrations of 10, 29, and 130 mg/L and yielded, respectively, 3.1%, 2.1%, and 1.1% for within-day CV and 2.8%, 2.1%, and 2.2% for total CV. Analytical recovery was 102 (+/- 6.7%). Linearity was demonstrated in the 0-200 mg/L range for serum and 0-3.5 g/L range for urine (r greater than or equal to 0.999). The detection limit for creatinine (signal-to-noise ratio = 3) was 0.5 mg/L. We used cimetidine for internal standardization. Correlation was good between this procedure and the Jaffé kinetic, the enzymatic (creatinine amidohydrolase), and the Fuller's earth alkaline picrate methods.

scholarly journals Direct Injection Liquid Chromatographic Technique for Simultaneous Determination of Two Antihistaminic Drugs and Their Main Metabolites in Serum

2007 ◽  
Vol 90 (2) ◽  
pp. 384-390 ◽  
Author(s):  
Samy Emara ◽  
Alaa El-Gindy ◽  
Mostafa K Mesbah ◽  
Ghada M Hadad
Keyword(s):  
Human Serum ◽  
Simultaneous Determination ◽  
Chromatographic Technique ◽  
Simple Liquid ◽  
Good Precision ◽  
Active Metabolites ◽  
Serum Samples ◽  
Antihistaminic Drugs ◽  
Liquid Chromatographic

Abstract A very simple liquid chromatographic technique was developed and validated for the simultaneous determination of 2 antihistaminic drugs, loratadine (LT) and terfenadine (TR), and their major active metabolites, desloratadine (DL) and fexofenadine (FX), respectively, in human serum. LT, DL, TR, and FX from directly injected serum samples were enriched on a protein-coated RP8 silica precolumn (10 4.6 mm id) while serum constituents, such as proteins and salts, were eluted to waste. Using an online column-switching system, the drugs and their metabolites were quantitatively transferred and separated on a second analytical column (Shim-pack 5 μm particle size cyanopropyl, 250 × 4.6 mm id) followed by ultraviolet detection at 243 nm for LT and DL and 220 nm for TR and FX. Very good precision, accuracy, and linearity were obtained over the range of 101000 ng/mL for LT and DL, 10500 ng/mL for TR, and 103000 ng/mL for FX in human serum. High extraction recoveries from serum ranging from 96.03 to 98.19, 95.44 to 97.26, 95.61 to 98.17, and 95.60 to 97.89 for LT, DL, TR, and FX, respectively, were obtained.

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