Alkaline Picrate Research Articles

Evaluation of Some Renal Biomakers in Patients with Benign Prostatic Hyperplasia/Prostate Cancer Attending Nnamdi Azikiwe University Teaching Hospital Nnewi

Benign prostatic hyperplasia (BPH) and prostate cancer (PCa) are among the urological conditions that affect aging men. Patients with this pathology could develop renal impairment. Renal biomarkers like beta-2-microglobulin (BMG), urea, urine creatinine, microalbumin (MALB), and albumin to creatinine ratio (ACR) were assessed with total (tPSA) and free prostate specific antigen (fPSA), systolic blood pressure (SBP), diastolic blood pressure (DBP), height, weight and BMI. This cross sectional study recruited 120 men using convenient sampling technique which comprised 40 BPH, 40 PCa and 40 control attending Urology Clinic of NAUTH Nnewi, Anambra State. Five (5) milliliters of blood/ urine samples were collected from the patients and the necessary data were obtained from clinical records of the patients. The weight (kg) and height (m) were measured using standard beam balance scale and a stadiometer respectively and the BMI calculated. Blood pressure was measured using sphygmomanometer and stethoscope. PSA (ng/ml) was estimated by Enzyme immunoassay technique, BMG (mg/l) and MALB (mg/l) by immunoturbidimetric method, urea and creatinine were determined spectrophotometrically using modified Urease-Berthelot method and Jaffe slot alkaline picrate method respectively. Data analysis was conducted using SPSS version 21.0. The results were presented as median. Kruskal Wallis was used to determine significant differences between the mean values. P<0.05 was considered to be statistically significant. The results showed a significantly higher median values of tPSA, fPSA BMG and MALB in patients with BPH /PCa when compared with the control. MALB had a strong correlation with ACR in all the groups. Despite the significant increase in BMG and MALB in this study, routine indices of renal function remained normal. The increase in the sensitive markers in patients with BPH/PCa is an indication of early renal impairment.

Compliance to specifications in an external quality assurance program: did new biological variation estimates of the European Federation of Laboratory Medicine (EFLM) affect the quality of laboratory results?

The results of external quality assurance schemes are evaluated against specifications generally based on biological variation (BV) data. This study was carried out to determine whether new BV values affected the level of compliance to specifications. Our secondary objective was to identify the conditions that would be compromised as a result of poor analytical performance in disease associated markers. This study was based on the results of the SEQCML External Quality Assurance scheme for the 2015-2022 period. Deviation of the individual result from the target value was estimated. Additionally, we calculated the percentage of results that met the pre-established specification. In 97 of the 133 analytes, the level of compliance was maintained in 80-90 % of the results obtained in the two study periods. In 23 analytes, the level of compliance ranged from 51 to 79 % in the two study periods. In ALT, AST and sodium, the level of compliance was≤50 % of the results obtained in the first study period, with sodium being the only analyte that maintained this poor level of compliance in the second study period. The level of compliance to specifications remained independent from the specification used (SEQCML or EFLM) for the majority of the analytes. The results for sodium ion were below the target value, which may lead to misdiagnosis of hyponatremia. Non-compensated alkaline picrate methods overestimate creatinine, which may produce false information suggestive of kidney failure.

A Microfluidic Paper-based Analytical Device for Simultaneous Measurement of Albumin, Creatinine and Uric Acid in Urine based on Standard Addition Method

This work describes a simple method for the simultaneous measurement of urinary albumin, creatinine and uric acid using a standard addition approach on a microfluidic paper-based analytical device (µPAD). Hydrophobic barrier of the µPAD was created by stamping indelible ink onto a filter paper. The µPAD was designed with a flower-liked configuration. After aliquoting urine to a central sample zone, the sample flowed to ten surrounding channels, namely “the inner channels”, where the blank (water) or standard solutions had been added. The inner channels were connected to circular reagent zones where reagents had been immobilized. Tetrabromophenolphthalein ethyl ester, alkaline picrate, and a mixed solution of ferric chloride and ferric cyanide were used as the chromogenic reagents for the colorimetric detections of albumin, creatinine, and uric acid, respectively. Each reagent zone was linked with a circular detection zone through a second channel, namely “the outer channels”. The blue-, orange- and greenish-blue colored products were observed in the detection zones for the measurement of albumin, creatinine, and uric acid, correspondingly. A digital image of the µPAD was captured with a mobile phone. The color intensities were evaluated by ImageJTM and were employed for the quantitative analyses. The standard addition calibrations were found to be linear (r2 > 0.99) for the spiked analyte concentrations up to 100 mg L-1 albumin, 1000 mg L-1 creatinine, and 50 mg dL-1 uric acid. The paper platform provided high precision (RSD < 5 %) and good analytical recovery (91.8-109.7 %). Under paired t-test, the results obtained by the developed µPAD and the validating methods were not significantly different at 95 % confidence (albumin: tstat = -0.130, tcri = 3.182, creatinine: tstat = -0.133, tcri = 3.182, and uric acid: tstat = 1.119, tcri = 3.182).

Development of a low-cost, at-home device for early detection of chronic kidney disease

Abstract Chronic kidney disease (CKD) is classified according to patient's kidney state and functionality. Stage 1 represents the initial renal damage, maintaining normal function, while Stage 5 indicates end-stage of CKD, marked by renal insufficiency and the need for dialysis or transplant. Patients in the early stages of CKD may be asymptomatic, but may experience symptoms as fatigue, weakness, and a general feeling of unwellness as the disease progresses, making diagnosis accessibility an essential aim of the health system. A low-cost and home-based device accompanied by a mobile app were developed as an early indicator of CKD, using colorimetry to determine the creatinine levels and chromogens in urine deposited in the device, designed using a solution of picric acid, sodium lauryl sulphate, borate solution, and sodium hydroxide. A range of chromogens was assigned to creatinine concentrations, calibrated by analysing diluted creatinine samples in triplicate, taking 1.2 ml of the sample and adding defined amounts of alkaline picrate according to the calibration curve. The absorbances were recorded at 45, 60, and 180 seconds for each of the creatinine samples combined with alkaline picrate in the creatinine analysis. Finally, the curves corresponding to each time were compared, providing a final absorbance to each creatinine level by subtracting the blank curves’ values. The chromogen is obtained according to the patient's health status, and compared in a database in the application, displaying the possible health status of the kidneys, without providing a diagnosis, but rather an early alert of the kidneys state and approximate glomerular filtration rate. The device has the potential to offer an early diagnosis accessible to all population with a history or risk of CKD. Early detection of CKD may improve patient health through frequent medical appointments, medication, dietary restrictions, and lifestyle changes. Key messages • Early stages of CKD can go unnoticed by patients and physicians, making early diagnosis a valuable tool. • At-home health devices can be designed for the early detection of CKD.

Morphological Characterisation and Cyanide Content Determination in Phaseolus lunatus Linn. (Lima Beans) Accessions

Lima bean (Phaseolus lunatus Linn.) is one of the important legumes in developing countries. The study was carried out to evaluate the morphological characters and determine the amount of cyanide in lima beans. A total of 15 Lima bean accessions were obtained from the genebank of Genetic Resource Centre, International Institute of Tropical Agriculture (IITA) Ibadan, Oyo state, Nigeria and laid in Complete Randomised Design with three replicates. Morphological characters were evaluated based on the descriptors proposed by the International Plant Genetic Resources Institute, while hydrogen cyanide (HCN) content was determined using alkaline picrate colorimetric method. Six of the accessions (2005-014, 2006-003A, 2006-003B, 2006- 005A, PS-16 and TPl-2429) having best yield, moderate yield and lowest yield were selected for cyanide determination. Result showed that the accessions varied morphologically. The highest yielding accessions were TPl-2429, 2005- 014, 2006-003A and 2005-011, while 2006-003B and TPl-178 have moderate yields. Accession 2005-014 had the highest growth characters, followed by 2006-015 and 2006-003A which showed good quality. Lower amounts of cyanide were detected in 2005-014 and PS-16 accessions, while 2006-003A, 2006-003B and 2006-005A accessions did not show cyanide contents. Therefore, TPl-2429, 2006-003A and 2005-011 accessions which showed higher growth and yield with no cyanide content could be further improved in breeding of lima beans.

#3619 CKD EPI 2021 EQUATION FOR EGFR HAS GOOD EFFICACY TO PREDICT CLINICAL OUTCOMES IN A LARGE PROSPECTIVE COHORT OF ADULT KOREANS

Abstract Background and Aims A new eGFR equation using serum creatinine (CKD-EPI 2021) without race were developed by Chronic Kidney Disease Epidemiology Collaboration in 2021. We projected the changes of eGFR, CKD prevalence, and incidence of end stage renal disease (ESRD) and mortality in a large prospective cohort of adult Koreans who had voluntary health check-ups, using current (CKD-EPI 2009) and new equation (CKD-EPI 2021). Method We included 112,772 adult participants, aged 18 years or older, who had voluntary routine health check-ups including serum creatinine at three medical centers in Korea from 2003 to 2009. We compared the difference of eGFR and predictability of mortality and ESRD between eGFR values calculated by CKD-EPI 2009 and CKD-EPI 2021. Serum creatinine was expressed as the IDMS-traceable creatinine according to the manufacturer's conversion formula from the original value measured by the alkaline picrate Jaffe kinetic method. The incidence of mortality data was extracted from Statistics Korea and the ESRD data from the ESRD registry of the Korean Society of Nephrology. Results At baseline study, there were 60,723 males (53.8%). Age was 48.9 ± 12.0 years. The value of IDMS-traceable serum creatinine was 0.86 ± 0.21 mg/dl. Levels of eGFR at baseline study were 93.5 ± 15.6 ml/min/1.73 m2 by CKD-EPI2009, and 97.1 ± 15.0 ml/min/1.73 m2 by CKD-EPI2021. Values subtracted eGFRs by CKD-EPI2009 from eGFR by CKD-EPEI2021 were 3.6 ± 1.2 (range: -32.9∼6.1) ml/min/1.73 m2. The difference of eGFR was higher in males compared to females (p<0.001), was the highest in the oldest group among patients aged 18–44 years, 45–64 years, and 65 years or more (p<0.001). Patients with diabetes mellitus (DM) or hypertension showed higher difference of eGFR compared patients without DM or hypertension, respectively (each p<0.001). Patients with eGFR ≥120.0 ml/min/1.73 m2 by CKD-EPI 2009 showed the highest difference between two eGFRs compared to patients with the other values. CKD stage was improved in 11,828 (10.5%) participants using eGFR calculated by CKD-EPI 2021 instead of CKD-EPI 2009, however, only 0.63% of participants with eGFR <60 ml/min/1.73 m2 by CKD-EPI equation was reclassified into eGFR ≥60 ml/min/1.73 m2 by CKD-EPI 2021.During 11.6 ± 2.0 years, 3354 (3.00%) subjects were dead and 151 (0.13%) subjects had end stage renal disease (ESRD) before death. Any eGFR or stage of eGFR was an independent risk factors to ESRD or mortality estimated by Cox's hazard proportional model adjusted by related factors. AUC to estimate renal survival by eGFRs was not different between eGFRs by CKD-EPI2021 and CKD-EPI2009 [0.739 (0.687-0.790) vs 0.740(0.688-0.792), p = 0.170]. p<0.001]. AUC to estimate survival by eGFRs calculated through CKD-EPI equation was slightly higher than that by CKD-EPI2021 equation [0.673(0.664-0.682) vs 0.667(0.658-0.676), p<0.001], however, the difference of AUC was negligible [standard error of AUC difference by two eGFRs; 0.96 (95% CI; 0.006-0.007)]. Conclusion The eGFR calculated by CKD-EPI2021 was higher compared to eGFR calculated by CKD-EPI2009. The power to estimate renal survival was not different between eGFRs by CKD-EPI2021 and CKD-EPI2009.

A smartphone-integrated portable rotating platform for estimation of concentration level of plasma-creatinine using whole human blood

The measurement of creatinine concentration is performed to monitor the renal health. The devices available in modern clinical laboratories for measuring creatinine concentration are accurate and provide results rapidly but may be prohibitively expensive for resource-poor settings. Therefore, developing an inexpensive yet accurate device for measuring creatinine concentration is needed. Consequently, we developed a simple, affordable, and portable spinning disc for measuring plasma-creatinine concentration with 10 μL of whole human blood.5 μL of the alkaline picrate solution is loaded into the device and rotated at 1000 rpm to transport this solution to the periphery of the microchannel. Further, 10 μL whole blood is loaded in the same channel and spun at 1300 rpm for 10 min. The creatinine in plasma reacts with alkaline picrate (Jaffe reaction), and the color of the mixture changes to yellow-orange color. The resulting color is captured with a smartphone, and creatinine concentration is estimated using an in-house developed app (CREA-SESE).The value of creatinine measured with the present device and the gold standard device are highly correlated (R2 = 0.998). The bias and standard deviation of the difference between the two measurements are 0.134 mg/dL and 0.143 mg/dL. This study demonstrates the feasibility of a simple, inexpensive, and portable rotating device for measuring creatinine concentration using 10 μL of whole human blood, which can easily be deployed to the underserved population in resource-constrained settings to monitor renal diseases.

Smartphone-based automated estimation of plasma creatinine from finger-pricked blood on a paper strip via single-user step sample-to-result integration

Measuring creatininelevels in blood plasma is one of the most commonly used means of routine monitoring of renal function. Point-of-care devicesfor creatinine level estimation potentially allow rapid measurement, facilitating continuous monitoring of vulnerable patients. However, a majority of such devices currently available is constrained by several factors such as lack of simplicity in fabrication and operation (including plasma separation), a limited regime of efficacy, compromised reagent stability, and elevated costs. Here, we report the invention of a simple-to-fabricate and easy-to-use paper strip for rapid evaluation of plasma creatinine levels (ranging from 0.1 to 15 mg dL-1) in an integrated single-step process. For the testing, one drop of whole blood is first dripped onto the calcium salt-functionalized sample pad of the strip. Subsequently, blood plasma separation occurs due to spontaneous aggregation of red blood cells (RBCs) in the presence of calcium ions. The plasma spontaneously traverses into the reaction zone of the test-strip via capillary action where it reacts with previously dispensed alkaline picrate reagent, resulting in a colorimetric signal. Smartphone-interfaced imaging and image analytics of the reaction spot renders the dissemination of the test result in a turn-around time of about 2–3 min, without necessitating additional device interfacing. High efficacy of performance is established by validating with the results obtained from laboratory-benchmarked biochemical auto-analyzer, exhibiting an accuracy of 94%, and the coefficient of variation ranging from 0.64 to 6.4%. Suitability of use in extreme resource-limited settings is established by favorable stability of the reagents outside controlled laboratory ambience and engagement of unskilled personnel to execute the testing procedure with uncompromised accuracy.

Fast colorimetric detection of albumin-to-creatinine ratio using paper-based analytical devices with alkaline picrate and Bromothymol Blue reagents

Is Cystatin C a powerful Predictor of Cardiovascular Diseases in patients with type 2 Diabetes Mellitus? (Study on Egyptian patients)Hesham Rafat, Sameh H. Soror, Zeinab Hassan, Ashref Ismaail

Assessment of lipid profile in-patients with Nephrotic syndrome in Kano Metropolis

Nephrotic syndrome (NS) is a complex kidney disease associated with numerous complications which can subsequently lead to cardiovascular disease among others. This study was aimed at assessing the lipid profile, serum and urinary proteins of patients with Nephrotic syndrome (NS) in Kano metropolis. A total of 50 NS patients and 25 apparently healthy volunteers(controls) were recruited for the study, made up of 32 males and 18 females with the age range of 4-70 years. Blood and urine sample were collected from the participants. Serum urea and creatinine were determined using urease berthelot’s reaction and Alkaline picrate methods. Serum total protein and albumin were assayed using Biuret and bromocresol green binding method through the manual colorimetrictechnique. Serum lipid profile were measured by an enzymatic spectrophotomeric method and the precipitation enzymatic method was specifically used for evaluating the levels of high-density lipoprotein cholesterol (HDL-C). Urine protein was determined using sulphur salicylic acid test. SPSS software package version 21 was used for the analysis of data. High frequency of NS of 40(80%) was observed in patients of <18 years while patient of >46 years had a lower frequency of 4(8%). Males recorded higher frequency of 32(64%) and the frequency of NS among females was 18(36%), thus the male to female ratio for NS was 1.78:1. The mean values of serum creatinine, urea, urinary protein (UP), total cholesterol (T.C), triglyceride (TG), low density lipoprotein cholesterol (LDL-C), T.C/HDL-C, TG/HDL-C, LDL/HDL-C ratio were significantly higher (p<0.05) in patients with NS than the healthy volunteers. Total protein (TP), albumin (Alb), globulin, HDL-C ratio were significantly lower (p<0.05) in NS when compared to healthy volunteers. There was positive correlation between UP and TC, TG and LDL, however, negative correlation was observed between UP and HDL-C with no statistical significance. Dyslipidemia, decreased serum protein and increase serum creatinine, Urea as well as UP were associated with NS. In conclusion, lipid profile and UP analysis may be a useful tool for diagnosis of NS and early diagnosis can reduce the disease morbidity.

Biochemical parameters of mild, moderate & severe COVID-19 patients

Background and objectives: Coronavirus disease 2019 (COVID-19) is affecting millions of people world-wide. It is caused by the severe acute respiratory syndrome corona virus 2(SARS-CoV-2). The laboratory findings are very important to assess the progress of the disease. The present study is aimed to discuss the biochemical parameters among mild, moderate and severe COVID-19 patients. Materials and methods: A cross sectional study were conducted in the Department of Physiology, Dhaka Medical College, Dhaka from January 2020 to December 2020. After obtaining ethical clearance, a total of 100 real time-polymerase chain reaction (RT-PCR) COVID-19 positive patients were selected from Dhaka Medical College Hospital. With all aseptic precautions, 10 ml of venous blood was collected from ante-cubital vein. D-dimer, prothrombin time, C-reactive protein (CRP), lactate dehydrogenase (LDH), serum ferritin, random blood glucose (RBG), serum creatinine, serum glutamic-pyruvic-transaminase (SGPT) and serum albumin measured in the Department of Laboratory Medicine, Dhaka Medical College Hospital, Dhaka. CRP was measured by Immunoturbidimetric method, serum ferritin was estimated by Chemiluminescent microparticle immunoassay. STA-neoplastine CI plus used with STA-R analyzer was used for determination of prothrombin time. D-dimer was estimated by Immunofluorescence Assay method. Serum LDH is measured by Dimention clinical chemistry system, serum albumin is measured by bromocresol purple dye binding method, serum creatinine is measured by Jaffe alkaline picrate method and serum SGPT is measured by colourmetric (IFCC 1980) method and RBG is measured by enzymatic colorimetric method (GOD-PAP). Data were recorded in a pre-designed structured data collection form. For statistical analysis, ANOVA followed by Bonferroni test, Chi square test, Spearman’s rho correlation coefficient test was performed as applicable using SPSS for windows version 25.0. Results: By analyzing biochemical parameters of mild, moderate and severe RT-PCR positive 100 COVID-19 patients revealed evaluation of biochemical parameters shows severity of the disease was significantly associated with CRP, SGPT, S. Creatinine, LDH, Ferritin, D-dimer & Prothrombin time. No significant association was found with RBG & S. Albumin. Bonferroni correction following ANOVA was performed to compare between each group. Spearman’s correlation reveals statistically significant strong positive correlation with CRP, Ferritin & D-dimer, moderate positive correlation with S. Creatinine, LDH and mild positive correlation with SGPT & Prothrombin time. Conclusion: This study showed D-dimer, prothrombin time, CRP, LDH, ferritin, serum creatinine and SGPT are significantly associated with the severity of the illness that is higher in severe group in comparison to mild and moderate groups. So, comprehensive analysis of the biochemical parameters will be very helpful for early identification & better management of severe disease.

Bamboo (Bambusoideae) Leaf Application on the Detoxification of Cassava Wastewater for Potential Biogas Production

This work presents report of the detoxification of cassava wastewater using bamboo (Bambusoideae) leaf. The aim of the detoxification process is toreduce the poisonous cyanide content to a permissible level of using cassava effluent for biogas production. Fresh leaves of bamboo and cassava roots used for this experiment were obtained from Enugu, Nigeria. The effluent was obtained from the cassava roots through grinding and dewatering processes, while fine particles of the bamboo leaf were obtained through sun-drying, grinding and sieving processes. Fourier transform infrared (FTIR) spectroscopy aided the determination of the functional groups characteristic to the substances which are present in bamboo leaf. Physicochemical properties of the cassava effluent (pH, BOD, COD and cyanide content) were evaluated. BOD tests were carried out by measuring the amount of dissolved oxygen present in the samples before and after incubation at 20°C for 5 days. Alkaline picrate method was used to determine the concentration of the cyanide. The cyanide content reduction was optimized using response surface methodology. It involved the use of central composite design of Design Expert software. The considered factors were particle size, mass/volume ratio, temperature and time while the response was cyanide content reduction. Analysis of variance (ANOVA) and mathematical modelling of the percentage removal of cyanide content were carried out. From the analysis of the results, the major functional groups responsible for the cyanide content removal werefound to be: C–H deformation, C=O stretching, C–H stretching, C=C=C stretching and O–H stretching. The presence of bamboo leaf reduced the cyanide content to 0.12 mg/L, a permissible level that is harmless to enzymatic reaction of biogas production. Quadratic model adequately described the percentage removal of the cyanide content. Optimum cyanide content removal of 83.6% was obtained at particle size of 0.458 mm, mass/volume ratio of 0.042 g/mL, temperature of 331.4 K, and time of 11.1 h. The model was confirmed by various criteria; correlation coefficient R2 (0.9715), p value (<0.0001) and adequate precision (24.365).

Designing and evaluating analytical parameters to adapt siemens urinary creatinine enzymatic method to open system analysers

Urinary creatinine is measured to assess kidney function and also as part of sample validity testing in drugs of abuse. Creatinine methods based on alkaline picrate Jaffe’s reaction require extra cleaning on chemistry analysers to minimise any interference from picric acid and sodium hydroxide on other reagents on board. Enzymatic methods reagents are not as invasive and more specific. Siemens enzymatic method is more sensitive and specific when compared to Thermo Fisher alkaline picrate method but there were no available analytical parameters to setup the Siemens enzymatic method on Beckman-Coulter AU5800 analyser as an open system analyser. Emulating setup parameters from Siemens chemistry analysers did not work. The analytical parameters were developed through systematic testing using different settings to adopt and optimise the method. A full correlation was performed using the developed parameters with alkaline picrate method. The Deming regression weighted analysis for 438 samples showed a good correlation at a 95% confidence interval. Slope is 1.029 to 1.041, Y-intercept when X=0.0 is -0.9156 to -0.7481 and correlation coefficient (r) is 0.998. Alkaline picrate method Mean and SD is 12.059 and 7.248 respectively and for the Enzymatic method is 11.653 and 7.504 respectively. Many interferences from drugs and other substances are eliminated when using the enzymatic method and the stability of other reagents on-board improved because the reagents used in the enzymatic method are less invasive.

Paper-Based Sensors for Point-of-Care Kidney Function Monitoring

The ratio of protein to creatinine (PCR) in urine is a measure of kidney health of a human body. We report the development of a point-of-care (POC) kidney function test (KFT) kit, which is composed of a paper-based PCR sensor, an opto-electrochemical measurement unit, a display, and a facility to transfer data to a smartphone. For the total protein sensor, the pieces of filter papers were coated with a mixture of alcoholic bromophenol blue and citric buffer leading to a chrome-yellow coloration. While for the creatinine sensor the papers were coated with alkaline picrate solution leading to a lemon-yellow coloration of the surface. Dispensing protein (creatinine) solution of known concentration on the respective sensors led to the conversion of chrome-yellow to bluish-green coloration (lemon-yellow to orange coloration) for the total protein (creatinine) sensor. The intensity of such colorations varied with the concentration of the analytes in the solutions. The variations in the intensities of colors with analyte concentration were quantified by integrating a light emitting diode (LED) at one side of each sensor and a light intensity sensor (LIS) on the other side. The intensity of the transmitted rays, passing through the sensors, was found to vary the electrical output of the LIS monotonically with the loading of the total protein or creatinine in the analyte. This helped in obtaining the calibration plots for the PCR sensors. Further, the sensors were employed for the on-spot detection of the unknown PCR in the real human urine samples.

Effects of CST3 Gene G73A Polymorphism on Cystatin C in a Prospective Multiethnic Cohort Study

Background/Aims: G73A polymorphism in the CST3 gene of cystatin C has been associated with Alzheimer’s disease, age-related macular degeneration, and cardiovascular disease. However, studies investigating the influence of this genetic variability on serum cystatin C and cystatin-based renal function estimate are limited. Therefore, the aim of this study is to investigate the possible association of single-nucleotide polymorphism (rs1064039) of the CST3 gene on the serum cystatin C level and cystatin C-based estimated glomerular filtration rate (eGFR). Methods: Study subjects include patients with various levels of renal function recruited from the nephrology clinic and wards of a tertiary hospital. The blood samples collected were analyzed for serum cystatin C and creatinine levels by particle-enhanced turbidimetric immunoassay and kinetic alkaline picrate method, respectively. DNA was extracted using a commercially available kit. ­Polymerase chain reaction results were confirmed by direct DNA Sanger sequencing. Results: The genotype percentage (G/G = 73%, G/A = 24.1%, and A/A = 2.9%) adhere to the Hardy-Weinberg equilibrium. The dominant allele found in our population was CST3 73G allele (85%). The regression lines’ slope of serum cystatin C against creatinine and cystatin C-based eGFR against creatinine-based eGFR, between G and A allele groups, showed a statistically significant difference (z-score = 3.457, p < 0.001 and z-score = 2.158, p = 0.015, respectively). Patients with A allele had a lower serum cystatin C level when the values were extrapolated at a fixed serum creatinine value, suggesting the influence of genetic factor. Conclusion: Presence of CST3 gene G73A polymorphism affects serum cystatin C levels.

Study of various glycolytic inhibitors for preservation of blood glucose and clinical biochemical parameters

Introduction: Sample for plasma glucose estimation is taken in fluoride Vacutainer and that is to prevent glycolysis and so that accurate glucose estimation can be done. Objectives: The objective of the study is to study various glycolytic inhibitors for preservation of blood glucose and other common clinical biochemical parameters. Materials and Methods: This prospective study was conducted at new civil hospital Surat. Study includes total 100 participants. We have prepared 5 mmol/L DL Glyceraldehydes containing Vacutainer. Blood sample from all participants were taken in 3 Vacutainer like plain Vacutainer, fluoride Vacutainer and DL Glyceraldehydes containing Vacutainer. Various biochemical investigations were performed from all above collected blood samples to see the difference of significant among them by calculating p value. P value less than 0.05 was considered as a difference of significant. Results: There is no significant changes in results of glucose between fluoride and glycrealdehyde containing Vacutainer. Biochemical parameter like SGPT, Creatinine, Total Bilirubin, Albumin, cholesterol, Total protein, Uric acid, electrolytes also does not show any significant difference between DL-Glyceraldehyde and plain Vacutainer. The Result of Serum Creatinine was found to be high from Glyceraldehyde containing Vacutainer as compared to plain Vacutainer and difference among them is highly significant.(p Conclusion: From our study we would like to conclude that. DL -Glyceraldehyde containing vial for many biochemistry related parameter analysis is better option as it is save additional use of vaccutte, except for serum creatinine (by alkaline picrate method). So DL Glycrealdehyde cannot be solely used as alternative for plain and fluoride containing Vacutainer. Keywords: DL-Glyceraldehyde, Glycolysis, Fluoride, Creatinine, Vacutainer.

Investigation of Some Elements and Cyanide Content of the Cassava Specie (Accra) Cultivated in Ekowe, Bayelsa State, Nigeria (P10-044-19)

ObjectivesCassava carbohydrate has become an important source of energy, as it is a staple food for many of the 374 ethnic groups in Nigeria. Little is known of the variety grown and consumed in Ekowe community, Bayelsa State; as such specie “Accra” which is thought to belong to the variety of sweet cassava (manihot palmata). We will determine the processing effects and evaluate the nutrient and anti nutritional/toxic content of the various segments (epicarp, endocarp, mesocarp and the fermented pulp) of the cassava specie. MethodsThe researcher used atomic absorption spectrophotometric and alkaline picrate methods for the elemental and cyanide analysis respectively. ResultsThe minerals; Magnesium (Mg), Calcium (Ca), and Iron(Fe), levels were low in all segments, and on the other the toxic chemicals components such as Lead (Pb), Cadmium (Cd) and Cyanide (CN) content levels of the cassava segments were within the consumption level, recommended by the world health organization. There is no significant difference in the means at P < 0.05. Expectedly, the processing of the cassava pulp (endocarp) by fermentation appeared to lower the toxic levels of the samples. The mineral and chemical concentrations of the various segments in ppm are as follows: EPICARP: Mg 0.23 ± 0.00a; Ca 7.56 ± 0.06a; Fe 0.12 ± 0.01a; Pb 0.48 ± 0.01a and Cd 0.00 ± 0.00 ENDOCARP: Mg 0.24 ± 0.00a; Ca 6.98 ± 0.01a; Fe 0.09 ± 0.01a; Pb 0.00 ± 0.00 and Cd 0.00 ± 0.00 MESOCARP: Mg 0.06 ± 0.00a; Ca 0.00 ± 0.00; Fe 0.00 ± 0.00; Pb 0.23 ± 0.00a and Cd 0.00 ± 0.00 FERMENTED PULP: Mg 0.06 ± 0.00a; Ca 4.29 ± 0.01a; Fe 0.03 ± 0.00a; Pb 0.00 ± 0.00 and Cd 0.00 ± 0.00The cyanide concentration in ppm of the cassava segments (Accra specie) are as follows: EPICARP:1.23 ± 0.01cENDOCARP:1.88 ± 0.01cMESOCARP: 1.50 ± 0.01cFERMENTED PULP: 1.13 ± 0.01cKEY: Cd (cadmium); Fe (iron); Ca (calcium); Pb (lead); Mg (magnesium). ConclusionsThe segments have a nutrient content level below the recommended daily requirement prescribed by the world health organization (WHO). Because of the low content or even the near absence of the toxic chemicals such as Lead, Cadmium, and cyanide, and low content of mineral elements Accra cassava specie is recommended as safe for consumption. however, it should not be used as a nutrient supplement, but its products should be consumed along with other foods rich in magnesium, iron, and calcium. Funding Sourcesfunding was done individually.

Simultaneous and direct determination of urea and creatinine in human urine using a cost-effective flow injection system equipped with in-house contactless conductivity detector and LED colorimeter

This work presents a cost-effective and simple flow injection analysis (FIA) system for simultaneous and direct determination of urea and creatinine in human urine. The FIA system comprises two in-house detectors, a contactless conductivity detector and a light emitting diode (LED) detector. The contactless detector was built as a flow-through detection cell with axial electrodes, commonly known as capacitively coupled contactless conductivity detector (C4D) and the diode detector was fabricated based on the concept of paired emitter detector diodes (PEDD). With appropriate dilution of urine, the sample is directly injected into a stream of glycine-NaOH buffer pH 8.8 (the gas donor stream) and is carried by the carrier through a urease minicolumn for on-line enzymatic hydrolysis. The generated NH3 diffuses from the carrier stream through a porous polytetrafluoroethylene (PTFE) membrane into a stream of deionized water (the acceptor stream) leading to an increase in the signal at the C4D due to the NH3 dissolution in the water. In this system, creatinine is determined based on the Jaffé reaction by merging a stream of alkaline picrate with the gas donor stream. The change in color is detected using the PEDD equipped with two green LEDs. Under the optimum condition, the linear range of urea and creatinine were 30–240 mg L−1 and 10–500 mg L−1, with limits of detection of 9.0 mg L−1 and 0.9 mg L−1, respectively. The proposed system provides satisfactorily good precision (RSD < 3%), with sample throughput of 31 sample h−1 for the two analytes. The FIA system tolerates potential interference commonly found in human urine. The system was successfully applied and validated with selected reference methods.

Disposable Nonenzymatic Uric Acid and Creatinine Sensors Using μPAD Coupled with Screen-Printed Reduced Graphene Oxide-Gold Nanocomposites.

Uric acid (UA) and creatinine are the imperative biological substance for clinical monitoring and diagnosis. Measuring the ratio between uric acid and creatinine in urine helps differentiate acute uric acid nephropathy from the hyperuricemia that secondarily occurs to renal failure. In general, the ratio is greater than 0.9 in acute uric acid nephropathy and less than 0.7 in hyperuricemia. In this work, disposable nonenzymatic screen-printed reduced graphene oxide-gold nanocomposites electrodes were firstly developed for the quantitative analysis of uric acid. Our sensors were also coupled with the paper-based colorimetric sensor of the determination of creatinine. Hence, an alternative high-throughput screening test for the uric acid to creatinine ratio with high sensitivity, specificity, simplicity, and rapidity was developed. Under the optimum conditions, our disposable nonenzymatic screen-printed electrode for the determination of uric acid shows the acceptable analytical performance in a wide range of linearity (2.5-1,000 μM) with a low detection limit of 0.74 μM. Our electrodes also showed no interference from common physiologic compound in urine. The determination of creatinine has been developed using Jaffé reaction between the creatinine and picric acid in alkaline condition. The alkaline picrate color on μPAD changed from yellow to orange in the presence of creatinine and the orange intensity is directly proportional to the creatinine amount in a linearity range of 0.20-6.0 mM as a detection limit of 180 μM. Finally, our device has been utilized to determine uric acid and creatinine simultaneously in control urine samples with acceptable result.

Agreement between Creatinine and Cystatin C-Based Equations in the Estimation of Glomerular Filtration Rate among Malaysian Patients with Renal Impairment

Abstract: Background: Cystatin C-based equations have been proposed as an alternative for creatinine-based equations in the estimation of Glomerular Filtration Rate (eGFR). Limited studies were available with regards to the Asian population to evaluate the agreement of eGFR measured based on both biomarkers. This study aimed to evaluate the agreement between various cystatin C- and creatinine-based eGFR equations. Methods: Patients were recruited from the Nephrology Clinic Universiti Kebangsaan Malaysia Medical Centre. Serum cystatin C and creatinine levels were analysed by particle-enhanced immunoturbidimetry assay and kinetic alkaline picrate method, respectively. Various cystatin C-based (Hoek, Larsson, Grubb, Flodin, cystatin C-based Chronic Kidney Disease Epidemiology (CKD-EPI)) and creatinine-based eGFR equations (Cockcroft-Gault, Modification of Diet in Renal Disease, creatinine-based CKD-EPI) were compared by using the Pearson’s correlation and Bland-Altman plot. Results: A total of 118 patients included in this study have a mean age of 61±15 years and 57.6% were female. The mean serum cystatin C and creatinine levels were 2.00±1.06 mg/L and 2.21±1.63 mg/dL, respectively. The correlation between serum cystatin C and creatinine level was significant (r=0.78, P<0.01). All cystatin C-based eGFR correlated significantly with the creatinine-based eGFR equations. The strongest correlation was between creatininebased and cystatin C-based CKD-EPI equation (r=0.93, P<0.01). The Bland-Altman plot shows that cystatin C-based CKD-EPI equation had the least mean difference when compared with creatinine-based CKD-EPI equation. Conclusion: Significant correlation and good agreement were demonstrated between various cystatin C-based and creatininebased eGFR equations. The cystatin C-based equations are appropriate alternative for GFR measurement among Malaysian patients with renal impairment. Key words: Cystatin C, Creatinine, Estimated Glomerular Filtration Rate, Renal Function, Renal Elimination, Chronic Kidney Disease.