Determination of creatinine and creatine by capillary electrophoresis : a thesis presented in partial fulfilment of the requirements for the degree of Master of Science in Chemistry at Massey University

The assessment of creatinine and creatine in biological fluids is important in the evaluation of renal and muscular functions. For routine creatinine determinations in the clinical laboratory, the most frequently used method is the spectrophotometric one based on the Jaffé reaction. However, this reaction is not specific for creatinine. For this reason, several methods have been proposed, but the elimination of interferences in the determination of creatinine has still not been achieved in some of these methods; others solved this problem either with expensive equipment that does not suit routine analysis or necessitates time-waste procedures. In this thesis capillary electrophoresis was the new tool investigated. It was applied in an attempt to achieve both the separation of creatinine from the non-creatinine 'Jaffé- reacting' chromogens and the determination of creatine in serum. Capillary zone electrophoresis was performed with detection at wavelength 480 nm to separate creatinine from the non-creatinine 'Jaffé-reacting' chromogens in urine. The principle was based upon the different migration times due to the different molecule weights, molecular sizes and charges under the applied high voltage. The picric acid was employed as part of the running buffer to allow reaction of creatinine and picrate to take place after the sample injection. This procedure eliminated the negative influence of the reaction time that is controlled manually in the common Jaffé reaction method. Therefore, compared to the Jaffé reaction method, the new method achieved more accuracy and precision in the determination of creatinine. Determination of creatinine in serum and urine were studied at a new wavelength 417 nm, which gave a higher sensitivity of detection than at 480 nm. This wavelength shift made the determination of creatinine in serum possible by capillary zone electrophoresis without the non-creatinine 'Jaffé-reacting' chromogens interfering. In this method, serum only needed a simple filtration before the analysis. Creatine was discovered to have absorption at 417 nm in alkaline medium. Moreover, specific sample stacking was introduced in this method. The sample was dissolved in a mixture of two-volumes acetonitrile and one-volume 3 % ammonium chloride to give a 10-fold enhancement of detection sensitivity

The Comparison of Creatinine and Cystatin C Value in Preeclampsia Severity and Neonatal Outcome

Objectives: to compare the levels of creatinine and cystatin C with the severity of preeclampsia, and assess neonatal outcomes.Materials and Methods: Creatinine, cystatin C, and neonatal outcomes were assesed in 17 normotensive samples, 17 samples of mild preeclampsia and 17 samples of severe preeclampsia. Analysis of data with statistical tests of ANOVA and t test differences between 2 proportions.Results: The mean levels of creatinine in the normotensive group, mild preeclampsia, severe preeclampsia are 0.56 mg/dL, 0.67 mg/ dL, and 0.75 mg/dL, p=0.138; While on cystatin C are 0.82 mg/L, 1.03 mg/L and 1.32 mg/L, p=0.000. The adverse neonatal out-come wasn't found in the normotensive group. In mild pre-eclampsia obtained 1 preterm birth and 1 intrauterine fetal death (IUFD), whereas in severe preeclampsia obtained 3 babies born preterm, 1 IUFD, and 1 intrauterine growth restriction (IUGR).Conclusion: levels of cystatin C was increased significantly in line with increased severity of preeclampsia, whereas creatinine was not increased significantly. Cystatin C is better than crea-tinine as a marker of renal dysfunction in preeclampsia patients. There was an increase in adverse neonatal outcomes in the group of preeclampsia

Prospective observational study of point-of-care creatinine in trauma.

Background:Patients with trauma are at risk for renal dysfunction from hypovolemia or urological injury. In austere environments, creatinine values are not available to guide resuscitation. A new portable device, the Stat Sensor Point-of-care (POC) Whole Blood Creatinine Analyzer, provides accurate results in <30 s and requires minimal training. This device has not been evaluated in trauma despite the theoretical benefit it provides. The purpose of this study is to determine the clinical impact of the POC device in trauma. Methods:40 patients with trauma were enrolled in a prospective observational study. One drop of blood was used for creatinine determination on the Statsensor POC device. POC creatinine results were compared to the laboratory. Turnaround time (TAT) for POC and laboratory methods was calculated as well as time elapsed to CT scan if applicable. Results:Patients (n=40) were enrolled between December 2014 and March 2015. POC creatinine values were similar to laboratory methods with a mean bias of 0.075±0.27 (p=0.08). Mean analytical TATs for the POC measurements were significantly faster than the laboratory method (11.6±10.0 min vs 78.1±27.9 min, n=40, p<0.0001). Mean elapsed time before arrival at the CT scanner was 52.9±34.2 min. Conclusions:The POC device reported similar creatinine values to the laboratory and provided significantly faster results. POC creatinine testing is a promising development for trauma practice in austere environments and workup of a subset of stable patients with trauma. Further study is warranted to determine clinical impact, both in hospital-based trauma and austere environments

Prognostic value of preoperatively obtained clinical and laboratory data in predicting survival following orthotopic liver transplantation

Twenty‐seven clinical and laboratory data and the subsequent clinical course of 93 consecutive adult patients who underwent orthotopic liver transplantation for various chronic advanced liver diseases were analyzed retrospectively to assess the risk factors of early major bacterial infection and death after the procedure. Forty‐one patients (44%) had early major bacterial infection during hospitalization for orthotopic liver transplantation. The mortality rate was 70.7% in patients with early major bacterial infection and was 7.7% in patients without early major bacterial infection (p < 0.001). Total serum bilirubin, total white blood cell count and polymorphonuclear cell count, IgG (all p < 0.05) and plasma creatinine level (p < 0.001) were higher in patients that developed early major bacterial infection than in those who did not. By step‐wise discriminant analysis, the strongest risk factor for early major bacterial infection was the serum creatinine level, which achieved an accuracy of 69% for a creatinine level greater than 1.58 mg per dl. Seven variables (ascites, hepatic encephalopathy, elevated white blood and polymorphonuclear cell count, decreased helper to suppressor T cell ratio and elevated plasma creatinine and bilirubin levels) were associated with a significant increased risk for death. A step‐wise discriminant analysis of these seven factors resulted in the demonstration of serum creatinine as the greatest risk factor for mortality. A preoperative serum creatinine either less than or greater than 1.72 mg per dl accurately predicts survival or death, respectively, in 79% of cases. These data suggest that the baseline preoperative serum creatinine level provides the best indication of the short‐term prognosis after liver transplantation than does any other preoperatively obtained index of the patient's status. Copyright © 1986 American Association for the Study of Liver Disease

Creatinine, diet, micronutrients, and arsenic methylation in West Bengal, India.

BackgroundIngested inorganic arsenic (InAs) is methylated to monomethylated (MMA) and dimethylated metabolites (DMA). Methylation may have an important role in arsenic toxicity, because the monomethylated trivalent metabolite [MMA(III)] is highly toxic.ObjectivesWe assessed the relationship of creatinine and nutrition--using dietary intake and blood concentrations of micronutrients--with arsenic metabolism, as reflected in the proportions of InAS, MMA, and DMA in urine, in the first study that incorporated both dietary and micronutrient data.MethodsWe studied methylation patterns and nutritional factors in 405 persons who were selected from a cross-sectional survey of 7,638 people in an arsenic-exposed population in West Bengal, India. We assessed associations of urine creatinine and nutritional factors (19 dietary intake variables and 16 blood micronutrients) with arsenic metabolites in urine.ResultsUrinary creatinine had the strongest relationship with overall arsenic methylation to DMA. Those with the highest urinary creatinine concentrations had 7.2% more arsenic as DMA compared with those with low creatinine (p &lt; 0.001). Animal fat intake had the strongest relationship with MMA% (highest tertile animal fat intake had 2.3% more arsenic as MMA, p &lt; 0.001). Low serum selenium and low folate were also associated with increased MMA%.ConclusionsUrine creatinine concentration was the strongest biological marker of arsenic methylation efficiency, and therefore should not be used to adjust for urine concentration in arsenic studies. The new finding that animal fat intake has a positive relationship with MMA% warrants further assessment in other studies. Increased MMA% was also associated, to a lesser extent, with low serum selenium and folate

Risk factors for deterioration of renal function after coronary artery bypass grafting

Abstract Objective: Various definitions of impairment of renal function after coronary artery bypass grafting (CABG) are used in the literature. Depending on the definition, several risk factors are identified. We analysed our data to determine the risk factors for postoperative deterioration of the creatinine clearance of 10% or more. Methods: All patients undergoing isolated coronary surgery in a single centre between January 1998 and December 2007 are included. Clinical data, including demographics and renal risk factors, were prospectively collected in our database. The most recent preoperative serum creatinine level and the maximum serum creatinine level within the first week postoperatively were used to calculate the creatinine clearance. A deterioration of 10% or more was considered to be an endpoint for this study. Results: In 10 098 out of a total of 10 626 patients, the preoperative as well as the postoperative creatinine clearance could be calculated. In 1053 patients, the deterioration of the creatinine clearance was 10% or more. We could identify the following risk factors: advanced age, diabetes, chronic obstructive pulmonary disease, peripheral vascular disease, emergency operation, previous cardiac surgery, low preoperative haemoglobin level, high preoperative C-reactive protein level, perioperative myocardial infarction, re-exploration and the number of blood transfusions. Conclusions: Risk factors for the deterioration of renal function after revascularisation have been confirmed in this study. In addition, we found peripheral vascular disease, previous cardiac surgery, low preoperative haemoglobin, increased preoperative C-reactive protein level, perioperative myocardial infarction and the number of blood transfusions to be risk factors that have not been described earlier

Urinary biomarker concentrations of captan, chlormequat, chlorpyrifos and cypermethrin in UK adults and children living near agricultural land

There is limited information on the exposure to pesticides experienced by UK residents living near agricultural land. This study aimed to investigate their pesticide exposure in relation to spray events. Farmers treating crops with captan, chlormequat, chlorpyrifos or cypermethrin provided spray event information. Adults and children residing ≤100 m from sprayed fields provided first-morning void urine samples during and outwith the spray season. Selected samples (1–2 days after a spray event and at other times (background samples)) were analysed and creatinine adjusted. Generalised Linear Mixed Models were used to investigate if urinary biomarkers of these pesticides were elevated after spray events. The final data set for statistical analysis contained 1518 urine samples from 140 participants, consisting of 523 spray event and 995 background samples which were analysed for pesticide urinary biomarkers. For captan and cypermethrin, the proportion of values below the limit of detection was greater than 80%, with no difference between spray event and background samples. For chlormequat and chlorpyrifos, the geometric mean urinary biomarker concentrations following spray events were 15.4 μg/g creatinine and 2.5 μg/g creatinine, respectively, compared with 16.5 μg/g creatinine and 3.0 μg/g creatinine for background samples within the spraying season. Outwith the spraying season, concentrations for chlorpyrifos were the same as those within spraying season backgrounds, but for chlormequat, lower concentrations were observed outwith the spraying season (12.3 μg/g creatinine). Overall, we observed no evidence indicative of additional urinary pesticide biomarker excretion as a result of spray events, suggesting that sources other than local spraying are responsible for the relatively low urinary pesticide biomarkers detected in the study population

Urinary Neutrophil Gelatinase-associated Lipocalin as a Marker for Identification of Acute Kidney Injury and Recovery in Dogs with Gentamicin-induced Nephrotoxicity.

BackgroundAcute kidney injury (AKI) is associated with high mortality rates in dogs, which may be a consequence of late recognition using traditional diagnostic tests. Neutrophil gelatinase-associated lipocalin (NGAL) is a protein-induced during kidney injury that may identify AKI earlier than traditional tests.Objectives/hypothesisTo evaluate urinary NGAL (uNGAL) and uNGAL-to-urinary creatinine ratio (UNCR) as early markers of kidney injury and recovery in an AKI model in dogs. It was hypothesized that these markers would document AKI earlier than serum creatinine concentration.AnimalsFive purpose-bred dogs.MethodsProspective study. Acute kidney injury, defined as a &gt; 50% increase in serum creatinine concentration above baseline, was induced in dogs by gentamicin administration (8-10 mg/kg SC q8h). Blood and urine collected for biochemical analyses and uNGAL and urinary creatinine concentrations, respectively, during AKI induction and recovery.ResultsAcute kidney injury was diagnosed significantly earlier based on a 7-fold increase in UNCR compared to a &gt; 50% increase in serum creatinine concentration (day 8; range, 2-10 mg/dl vs day 16; range, 14-19 mg/dl; P = .009). During recovery, the initial decrease in UNCR preceded the decrease in serum creatinine concentration by a median of 2 days. The uNGAL changes paralleled UNCR changes, but the increase in uNGAL was triphasic; the initial peak occurred earlier than UNCR (median, day 11 versus median, day 19).Conclusions and clinical importanceThe UNCR was early marker of gentamicin-induced AKI and its decrease documented onset of renal recovery. Additional studies are needed to validate this marker in dogs with naturally occurring renal injury

Renal and Hepatic Dysfunction in Malaria Patients in Minna, North Central Nigeria

Information on kidney and liver involvement in malaria in Africa is still very scanty. Kidney and liver functions were assessed in 70 malaria patients using serum levels of creatinine and urea and urinary protein levels as test indicators of kidney function and serum levels of bilirubin, aspartate aminotranferase (AST or SGOT), alanine aminotransferase (ALT or SGPT), and alkaline phosphatase (ALP) as indicators of liver function. Descriptive analysis of results obtained showed that 67.14% of patients had creatinine level above the 126µmole/L which is considered the upper limit of the normal range. Three cases (4.29%) had creatinine levels well above 265µmoles/L. The serum concentrations of creatinine, urea, protein, conjugated and total bilirubin, AST, ALT, and ALP in malaria patients were significantly higher (p<0.05) than those of malaria free individuals. We conclude that renal dysfunction, acute renal failure and liver dysfunction are clinical features of malaria in Minna, North Central Nigeri

Increasing the sensitivity of the Jaffe reaction for creatinine

Study of analytical procedure has revealed that linearity of creatinine calibration curve can be extended by using 0.03 molar picric acid solution made up in 70 percent ethanol instead of water. Three to five times more creatinine concentration can be encompassed within linear portion of calibration curve