Method For Determination Of Creatinine Research Articles

Dried blood spot analysis for the quantification of vancomycin and creatinine using liquid chromatography – tandem mass spectrometry: Method development and validation

BackgroundVancomycin is a widely used antibiotic for the treatment of gram-positive bacterial infections, especially for methicillin-resistant Staphylococcus aureus (MRSA) infections. Due to a small therapeutic range and large inter-patient variability, therapeutic drug monitoring (TDM) of vancomycin is required to minimize toxicity and maximize treatment efficacy. Venous blood sampling is mostly applied for TDM of vancomycin, although this widely used sampling method is more invasive compared to less painful alternatives, such as the dried blood spot (DBS) method, which can be performed at home. MethodWe developed an UPLC-MS/MS method for the quantification of vancomycin and creatinine in DBS. A fast sample preparation and short analysis run time of 5.2 min were applied, which makes this method highly suitable for clinical settings. Validation was performed according to international (FDA and EMA) guidelines. ResultsThe validated concentration range was found linear for creatinine from 41.8 µmol/L to 722 µmol/L and for vancomycin from 3.8 mg/L to 76.6 mg/L (r2 > 0.990) and the inaccuracies, imprecisions, hematocrit effects, and recoveries were < 15 % for both compounds. No significant carryover effect was observed. ConclusionHence, we successfully validated a quantification method for the simultaneous determination of creatinine and vancomycin in DBS.

A novel method for electrochemical determination of creatinine in human urine based on its reaction with 2-nitrobenzaldehyde using a glassy carbon electrode

A novel method for electrochemical determination of creatinine in human urine based on its reaction with 2-nitrobenzaldehyde using a glassy carbon electrode

A new method for selective determination of creatinine using smartphone-based digital image

A new method for selective determination of creatinine using smartphone-based digital image

Dilute-and-Shoot HPLC-UV Method for Determination of Urinary Creatinine as a Normalization Tool in Mycotoxin Biomonitoring in Pigs.

A simple, rapid, and accurate HPLC-UV method was developed for the determination of creatinine in pig urine. Usually, it is determined in urine in biomonitoring of xenobiotics to correct for variations in dilutions of urine samples. The colorimetric method (based on Jaffe reaction), which was mainly used for this purpose in mycotoxin biomonitoring, is not a reliable approach for pig urine. Therefore, a novel and accurate HPLC method for creatinine determination was developed. The sample preparation was based on the dilute and shoot approach. An HPLC separation was performed with a porous graphitic carbon column with an aqueous mobile phase to achieve satisfactory retention time for creatinine. The method has been successfully validated, applied for the determination of creatinine in pig urine, and compared with other methods commonly used for that purpose—a colorimetric method based on Jaffe reaction and commercial ELISA test. The developed HPLC method shows the highest precision and accuracy for pig urine samples. Finally, the method was applied as a normalization tool in LC-MS/MS mycotoxin biomarkers analysis. The standardization to a constant creatinine level (0.5 mg/mL) enables similar matrix effects for eleven mycotoxin biomarkers for pig urine samples with different creatinine levels.

An alternative, single-point method for creatinine determination in urine samples with optoelectronic detector. Critical comparison to Jaffé method

A rapid, single-point method for colorimetric creatinine determination has been developed as an alternative for a well-established Jaffé assay, which lacks analytical specificity. 3,5-dinitrobenzoic acid was employed in an alkaline environment as a chromophore instead of picric acid. Apart from the spectrophotometer, a simple optoelectronic detector was applied as an alternative detection method. The conditions of the reaction were optimized to provide satisfactory analytical parameters. The linear ranges of the proposed assays were 40–4000 and 10–300 µmol L−1 of creatinine for conventional spectrophotometry and a PEDD detector, respectively. A broad spectrum of potential interferents was examined. The presence of compounds such as glucose, urea or uric acid was the source of a significantly smaller bias in the examined method than in the case of the Jaffé method variants. The presented method was validated with the use of 10 human urine samples with the creatinine level determined by the recommended method. Two-tail paired Student's t-tests with 9 degrees of freedom at the 95% confidence level showed the agreement between the proposed and the reference methods.

Choosing the Optimal Method for Measuring Glomerular Filtration Rate in Pediatric Intensive Unit

The measurement of glomerular filtration rate (GFR) in intensive care unit patients is strategically important and determines the further treatment tactics. The article provides a literature review on choosing the various methods for assessing the glomerular filtration rate in children with kidney injury. We identified the main limitations of creatinine as a marker for measuring GFR; the features of laboratory methods for creatinine determination were detected.

Pegylated liposomal doxorubicin (Caelyx®) interference with the spectrophotometric Jaffe method for quantitative determination of creatinine in human plasma

• We investigated the in vitro and in vivo pegylated liposomal doxorubicin interference on plasma creatinine determination. • The kinetic Jaffe methods showed PLD negative interference on creatinine measurement. • The plasma creatinine values decreased of 85% ( n = 6) within two days after PLD infusion in patients.

Chemiluminescence of creatinine/H2O2/Co(2+) and its application for selective creatinine detection.

Creatinine is an important biomarker in clinical diagnosis and biomonitoring programs as well as urinary metabolomic/metabonomics research. Current methods are either nonselective, time consuming or require heavy and expensive instruments. In this study, chemiluminescence of creatinine with hydrogen peroxide has been reported for the first time, and its chemiluminescence is remarkably enhanced in the presence of cobalt ions. By utilizing these phenomena, we have developed a sensitive and selective chemiluminescence method for creatinine determination by coupling with flow injection analysis. The calibration curve is linear in the range of 1×10(-7)-3×10(-5)mol/L with a limit of detection (S/N=3) of 7.2×10(-8)mol/L, which is adequate for detecting creatinine in the clinically accepted range. The relative standard deviation for seven measurements of 3×10(-5)mol/L creatinine is 1.2%. The chemiluminescence method was then utilized to detect creatinine in human urine samples after simple dilution with water. It takes less than 1min each measurement and the recoveries for spiked urine samples were 100-103%. The interference study demonstrates that some common species in urine, such as amino acids, ascorbic acid and creatine, have negligible effects on creatinine detection. The present method does not use expensive instruments, enzymes and separation technique. This method has the advantages of sensitivity, selectivity, simplicity, rapidity, and low cost. It holds great promise for basic or comprehensive metabolic panel, drug screening, anti-dopping, and urinary metabolomic/metabonomics research.

Validation of a multi-analyte HPLC-DAD method for determination of uric acid, creatinine, homovanillic acid, niacinamide, hippuric acid, indole-3-acetic acid and 2-methylhippuric acid in human urine

During the last decades exposure sciences and epidemiological studies attracts more attention to unravel the mechanisms for the development of chronic diseases. According to this an existing HPLC-DAD method for determination of creatinine in urine samples was expended for seven analytes and validated. Creatinine, uric acid, homovanillic acid, niacinamide, hippuric acid, indole-3-acetic acid, and 2-methylhippuric acid were separated by gradient elution (formate buffer/methanol) using an Eclipse Plus C18 Rapid Resolution column (4.6mm×100mm). No interfering signals were detected in mobile phase. After injection of blank urine samples signals for the endogenous compounds but no interferences were detected. All analytes were linear in the selected calibration range and a non weighted calibration model was chosen. Bias, intra-day and inter-day precision for all analytes were below 20% for quality control (QC) low and below 10% for QC medium and high. The limits of quantification in mobile phase were in line with reported reference values but had to be adjusted in urine for homovanillic acid (45mg/L), niacinamide 58.5(mg/L), and indole-3-acetic acid (63mg/L). Comparison of creatinine data obtained by the existing method with those of the developed method showing differences from -120mg/L to +110mg/L with a mean of differences of 29.0mg/L for 50 authentic urine samples. Analyzing 50 authentic urine samples, uric acid, creatinine, hippuric acid, and 2-methylhippuric acid were detected in (nearly) all samples. However, homovanillic acid was detected in 40%, niacinamide in 4% and indole-3-acetic acid was never detected within the selected samples.

Gold nanoparticles-based colorimetric and visual creatinine assay

We demonstrate a selective and sensitive method for determination of creatinine using citrate-stabilized gold nanoparticles (AuNPs) as a colorimetric probe. It is based on a direct cross-linking reaction that occurs between creatinine and AuNPs that causes aggregation of AuNPs and results in a color change from wine red to blue. The absorption peak is shifted from 520 to 670 nm. Under the optimized conditions, the shift in the absorption peak is related the logarithm of the creatinine concentration in the 0.1 to 20 mM range, and the instrumental detection limit (LOD) is 80 μM. This LOD is about one order of magnitude better than that that of the Jaffe method (720 μM). The assay displays good selectivity over interfering substances including various inorganic ions, organic small compounds, proteins, and biothiols. It was successfully employed to the determination of creatinine in spiked human urine.

Fast vaporization solid phase microextraction and ion mobility spectrometry: A new approach for determination of creatinine in biological fluids

In this work a rapid and simple method for creatinine determination in urine and plasma samples based on aqueous derivatization of creatinine and complete vaporization of sample (as low as 10µL), followed by ion mobility spectrometry analysis has been proposed. The effect of four important parameters (extraction temperature, total volume of solution, desorption temperature and extraction time) on ion mobility signal has been studied. Under the optimized conditions, the quantitative response of ion mobility spectrometry for creatinine was linear in the range of 0–500mgL−1 with a detection limit of 0.6mgL−1 in urine and 0–250mgL−1 with a detection limit of 2.6mgL−1 in plasma sample. The limit of quantitation of creatinine was 2.1mgL−1 and 8.7mgL−1 in urine and plasma samples, respectively. The relative standard deviation of the method was found to be 13%. The method was successfully applied to the analysis of creatinine in biological samples, showing recoveries from 92% to 104% in urine and 101–110% in plasma samples.

Evaluating an Alternative Method for Rapid Urinary Creatinine Determination

Creatinine (CR) is an endogenously produced chemical that is routinely assayed in urine specimens to assess kidney function and sample dilution. The industry-standard method for CR determination, known as the kinetic Jaffé (KJ) method, relies on an exponential rate of a colorimetric change, and can therefore require automated processing equipment for moderate- to high-throughput analysis (hundreds to thousands of samples per day). This study evaluates an alternative colorimetric method, the “plateau Jaffé” (PJ) method, which utilizes the chemistry of the KJ method, a commercially available kit, and a multipoint calibration curve. This method is amenable to moderate-throughput sample analysis and does not require automated processing equipment. Thirty-two spot urine samples from healthy adult volunteers were analyzed for creatinine concentration (CRc) using the KJ and PJ methods. Samples were also analyzed using a liquid chromatography time-of-flight mass spectrometry (LC-TOF/MS) method, which acted as an analytical control. Replicate measurements of spot samples (natural log-transformed values) were used to estimate method precision, and linear regression models were used to evaluate method accuracy (LC-TOF/MS measurements were considered the analytical benchmark). Measurement precision was comparable across all three methods, with coefficent of variation estimates ranging from 3 to 6%. Regression models generally showed good agreement across methods with R2 estimates ranging from .996 to .998, slope estimates ranging from .944 to .986, and y-intercept estimates ranging from 0.111 to 0.303. Minor bias (between 2 and 16%) was observed across methods at the tails of the measurement distributions. The provided regression equations can be used to adjust for this bias and to improve CR measurement comparisons across studies employing different methods. Considering these results, the PJ method is a suitable alternative to the industry standard KJ method for urinary CRc determination. It can be implemented for moderate-throughput sample analysis using modest and commonly available lab instrumentation and manual sample preparation techniques.

A novel reversed-phase HPLC method for the determination of urinary creatinine by pre-column derivatization with ethyl chloroformate: comparative studies with the standard Jaffé and isotope-dilution mass spectrometric assays

Creatinine is an important biomarker for renal function diagnosis and normalizing variations in urinary drug/metabolites concentration. Quantification of creatinine in biological fluids such as urine and plasma is important for clinical diagnosis as well as in biomonitoring programs and urinary metabolomics/metabonomics research. Current methods for creatinine determination either are nonselective or involve the use of expensive mass spectrometers. In this paper, a novel reversed-phase high-performance liquid chromatographic (HPLC) method for the determination of creatinine of high hydrophilicity by pre-column derivatization with ethyl chloroformate is presented. N-Ethyloxycarbonylation of creatinine significantly enhanced the hydrophobicity of creatinine, facilitating its chromatographic retention as well as quantification by HPLC. Factors governing the derivatization reaction were studied and optimized. The developed method was validated and applied for the determination of creatinine in rat urine samples. Comparative studies with isotope-dilution mass spectrometric method revealed that the two methods do not yield systematic differences in creatinine concentrations, indicating the HPLC method is suitable for the determination of creatinine in urine samples.

Development, validation and clinical evaluation of a dried urine spot method for determination of hippuric acid and creatinine

ObjectivesThe purpose of this study was to develop and validate a HPLC–DAD method for determination of creatinine and hippuric acid in dried urine spots (DUS), evaluating its clinical applicability. Design and methodsSample preparation was based on a simple one step extraction with water. Analysis was performed in a reversed phase column Hypersil Gold® C18 (150×4.6mm, 3μm) with isocratic elution. Mobile phase was a mixture of triethylammonium phosphate buffer 5mM, pH3.3 and acetonitrile (90:10, v/v). ResultsTotal analytical run time was 6min. Precision assays presented CV% lower than 11.8%. Accuracy was 96.0–103%. The lower limit of quantitation was 50mgL−1. Mean extraction yields were 99% for CR and 79.3% for HA. Analytes were stable in DUS up to 11days at 40°C. The method was tested in 49 paired DUS and urine samples, with acceptable agreement. HA–CR concentration ratios in DUS were in the range of 0.06 to 2.74, being in average 100.7% from those values found in urine. ConclusionsDUS samples are a useful alternative for biological monitoring of toluene occupational exposure, especially in Developing Countries where sample logistics could be an important concern.

A simple, rapid and reliable high performance liquid chromatography method for the simultaneous determination of creatinine and uric acid in plasma and urine

A simple, rapid and reliable method for the simultaneous determination of creatinine (Cr) and uric acid (UA) in plasma and urine has been developed using high performance liquid chromatography with ultraviolet detection. Separation was achieved by an Agilent ZORBAX SB-C18 column and an optimized mobile phase consisting of 20 mmol L−1 sodium acetate, 30 mmol L−1 acetic acid and 1% methanol. The total time of analysis was 4 min. Good linear responses were found with correlation coefficient (r) >0.999 for both Cr and UA within the concentration range of relevant levels. The recoveries of Cr and UA were 102 ± 1% and 99.3 ± 2.1% for plasma; and 100 ± 2% and 102 ± 4% for urine. CVs of repeatability and intermediate precision of all analytes were less than 5%. The limit of detection of the developed method was: Cr, 0.09 μmol L−1 and UA, 0.04 μmol L−1. The proposed HPLC method could be used for not only reference purpose but also routinely in clinical laboratory testing.

Determination of Creatinine in Human Urine with Flow Injection Tandem Mass Spectrometry

Background/Aims: Excretion of urinary compounds in spot urine is often estimated relative to creatinine. For the growing number of liquid chromatography-tandem mass spectrometry (LC-MS/MS) assays of urine-excreted molecules, a fast and accurate method for determination of creatinine is needed. Methods: A high-throughput flow injection tandem mass spectrometry method for exact quantitation of creatinine in urine has been developed and validated. Sample preparation used only two-step dilution for protein precipitation and matrix dilution. Flow injection analysis without chromatographic separation allowed for total run times of 1 min per sample. Creatinine concentrations were quantitated using stable isotope dilution tandem mass spectrometry. Selectivity and coelution-free quantitation were assured by qualifier ion monitoring. Results: Method validation revealed excellent injection repeatability of 1.0% coefficient of variation (CV), intraday precision of 1.2% CV and interday precision of 2.4% CV. Accuracy determined from standard addition experiments was 106.1 ± 3.8%. The linear calibration range was adapted to physiological creatinine concentrations. Comparison of quantitation results with a routinely used method (Jaffé colorimetric assay) proved high agreement (R² = 0.9102). Conclusions: The new method is a valuable addition to the toolbox of LC-MS/MS laboratories where excretion of urinary compounds is studied. The ‘dilute and shoot’ approach to isotope dilution tandem mass spectrometry makes the new method highly accurate as well as cost- and time-efficient.

Rapid Determination of Creatinine in Human Urine by Microchip Electrophoresis with LED Induced Fluorescence Detection

In clinical medicine, urine creatinine concentration is an important marker in the evaluation of renal function and muscular dysfunctions. Herein, we reported a novel method for rapid determination of creatinine in urine by microchip electrophoresis with light-emitting diode induced fluorescence detection. Creatinine was derivatized by fluorescein isothiocyanate, and then quantitatively detected by the developed microchip LED induced fluorescence detection system. The excitation and emission wavelengths were 490 and 523 nm, respectively. The urine samples were analyzed after centrifuge and filtration. A baseline separation was obtained in <30 s using 10 mM borate buffer (pH 9.0, containing 45 mM sodium dodecylsulfate), with separation voltage of 1.5 kV. Good linearity was obtained (r2 = 0.9978) in the concentration range of 10.0–2.00 × 103 μM, and the limit of detection was 2.87 μM (S/N = 3). The recovery was 96.0–107 %, and the interday precision was <4.5 % (n = 6). To validate assay results, we compared the present method with the Jaffe’s colorimetric assay by measuring real urine samples. The method was reliable, sensitive, high-speed, low-cost and suitable for the routine analysis of creatinine in biofluids.

Vérification initiale d'une nouvelle méthode de dosage de la créatinine plasmatique sur UniCel® DxC 600 selon les recommandations du LAB GTA 04

The accreditation process, according to NF EN ISO 15189, implies a prior evaluation of the new reagent on-site for the implementation of each new assay technique. Thus, our new standardized method for determination of creatinine (non compensated method) in plasma or serum on UniCel DxC 600 (Beckman Coulter) has been tested according to LAB GTA 04 protocol. The reagent meets the quality criteria recommended by Valtec protocol, except fidelity with the low concentration standard (50 micromol/L). Besides there is no problem of results transferability with the two other techniques used in the laboratory (Jaffe compensated and enzymatic methods performed on Cobas Integra 800).

GC–MS determination of creatinine in human biological fluids as pentafluorobenzyl derivative in clinical studies and biomonitoring: Inter-laboratory comparison in urine with Jaffé, HPLC and enzymatic assays

In consideration of its relatively constant urinary excretion rate, creatinine in urine is a useful biochemical parameter to correct the urinary excretion rate of endogenous and exogenous biomolecules. Assays based on the reaction of creatinine and picric acid first reported by Jaffé in 1886 still belong to the most frequently used laboratory approaches for creatinine measurement in urine. Further analytical methods for creatinine include HPLC–UV, GC–MS, and LC–MS and LC–MS/MS approaches. In the present article we report on the development, validation and biomedical application of a new GC–MS method for the reliable quantitative determination of creatinine in human urine, plasma and serum. This method is based on the derivatization of creatinine (d 0-Crea) and the internal standard [ methyl-trideutero]creatinine (d 3-Crea) with pentafluorobenzyl (PFB) bromide in the biological sample directly or after dilution with phosphate buffered saline, extraction of the reaction products with toluene and quantification in 1-μl aliquots of the toluene extract by selected-ion monitoring of m/ z 112 for d 0-Crea-PFB and m/ z 115 for d 3-Crea-PFB in the electron-capture negative-ion chemical ionization mode. The limit of detection of the method is 100 amol of creatinine. In an inter-laboratory study on urine samples from 100 healthy subjects, the GC–MS method was used to test the reliability of currently used Jaffé, enzymatic and HPLC assays in clinical and occupational studies. The results of the inter-laboratory study indicate that all three tested methods allow for satisfactory quantification of creatinine in human urine. The GC–MS method is suitable for use as a reference method for urinary creatinine in humans. In serum, creatine was found to contribute to creatinine up to 20% when measured by the present GC–MS method. The application of the GC–MS method can be extended to other biological samples such as saliva.

A rapid MEKC method for the simultaneous determination of creatinine, 1‐ and 3‐methylhistidine in human urine

The present report describes the use of MEKC in the presence of 35 mM sodium tetraborate, pH 9.0, containing 60 mM SDS for the complete separation and identification of creatinine, 1-methylhistidine (1-MeH) and 3-MeH in human urine. Their simultaneous quantification in urine of healthy controls and subjects submitted to elective surgery to their lower limbs allowed to use 3-MeH as a reliable measure of skeletal protein breakdown. Simplicity, sensitivity and low running costs are the main advantages of this method that is suitable for the routine analysis in clinical laboratories of a large number of samples per day.