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 Table of Contents  
Year : 2022  |  Volume : 9  |  Issue : 1  |  Page : 66-70

To compare creatinine estimation by jaffe and enzymatic method

1 Department of Biochemistry, St. John's Medical College and Hospital, Bengaluru, Karnataka, India
2 Department of Nephrology, St. John's Medical College and Hospital, Bengaluru, Karnataka, India
3 Department of Biochemistry, Shri Atal Bihari Vajpayee Medical College and Research Institute, Bengaluru, Karnataka, India

Date of Submission13-Apr-2020
Date of Decision22-Oct-2020
Date of Acceptance29-Jul-2021
Date of Web Publication18-Oct-2022

Correspondence Address:
Girish Konasagara Shanthaveeranna
Department of Biochemistry, Shri Atal Bihari Vajpayee Medical College and Research Institute, Shivajinagar, Bangalore - 560001, Karnataka
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Source of Support: None, Conflict of Interest: None

DOI: 10.4103/cjhr.cjhr_34_20

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Introduction: Creatinine in urine and serum are used in the assessment of renal function. It is commonly estimated by Jaffe's and enzymatic method. In many institutions, serum creatinine is estimated by (POCT) Point of care testing device (Enzymatic method) and follow-up of the patients with creatinine results by other methods, analyzed in the Biochemistry laboratory. If the results of POCT do not correlate with the Jaffe method, it leads to differences in serum creatinine values and wrong treatment decisions during the follow-up of the patients. Hence, this study was done to know the difference between the two methods in a tertiary care hospital. Aim and Objective: To estimate creatinine by Jaffe's and enzymatic method and to compare the serum creatinine values between the two methods. Materials and Methods: It is an observational cross-sectional study, for a period of 17 months from November 2018 to March 2020. Seventy-five samples were analyzed for serum creatinine by Jaffe's method in the Biochemistry laboratory and by enzymatic method in the emergency department in POCT device. Results: Mean differences between Jaffe's and enzymatic method were −0.063 mg/dL, 0.070 mg/dL, 0.198 mg/dL, and 0.0685 in Group I, II, III, and all the groups together. The overall intraclass correlation coefficient including all the three groups (0.995) indicates a very good correlation between the two methods. Conclusion: Our study showed a good agreement and good correlation between the two methods, which is similar to other studies analyzed on same instrument.

Keywords: Creatinine, enzymatic method, Jaffe's method, POCT device

How to cite this article:
Augustin SM, Deshpande RP, Shanthaveeranna GK. To compare creatinine estimation by jaffe and enzymatic method. CHRISMED J Health Res 2022;9:66-70

How to cite this URL:
Augustin SM, Deshpande RP, Shanthaveeranna GK. To compare creatinine estimation by jaffe and enzymatic method. CHRISMED J Health Res [serial online] 2022 [cited 2023 Jan 30];9:66-70. Available from: https://www.cjhr.org/text.asp?2022/9/1/66/358821

  Introduction Top

Diseases of the kidneys are among the most important causes of death and disability in many countries globally.[1],[2] Serum creatinine is one of the most commonly measured parameters to assess renal functions in clinical chemistry laboratories. Currently, there are several methods available for creatinine estimation. Jaffe's method based on the alkaline picrate method, with few modifications to remove interference, is most commonly used for creatinine estimation first described by Jaffe in 1886.[3] The other methods are enzymatic methods and isotope dilution-liquid chromatography-mass spectrometry method (gold standard method).[4] Although several methods have been described, the Jaffe's method is most widely used.[5] Expert professional bodies have recommended that all creatinine methods should be traceable to a reference method based on isotope dilution mass spectrometry (IDMS) to harmonize the results among all the laboratories.[6]

Creatinine is also estimated by point of care testing devices, as there is a need for faster test results for treatment. There are several drawbacks with advantages associated with this point of care testing (POCT) devices. Where whole blood creatinine is estimated on the Radiometer ABL800 FLEX blood gas analyzer, using Amperometric biosensor (electrode) based on the enzymatic conversion of creatinine to sarcosine with the generation of hydrogen peroxide.[7] The enzymatic assay is less prone to nonspecific bias than the Jaffe assay, but it is considerably more expensive. Hence, most laboratories consider creatinine measurement by Jaffe method to minimize the cost of the test.[8]

In several tertiary hospitals, creatinine is estimated by enzymatic method available in POCT device in emergency setting and treatment decision are considered based on that. In condition where there is doubt regarding results by POCT device, samples are sent for creatinine estimation by Jaffe method to Biochemistry laboratory, for further follow-up and treatment. This leads to repeat test, add on cost to the patient, and delay in treatment. To avoid this, our study was designed to estimate serum creatinine by Jaffe's method in the Biochemistry laboratory and enzymatic method by POCT device and to compare creatinine results between the two methods to check the differences.

  Materials and Methods Top

The study was conducted in tertiary care hospital, Bangalore, after obtaining Institutional ethical review board clearance. Sixty-two samples were required for the study as per the statistical analysis. It was an observational cross-sectional study for a period of 17 months from November 2018 to March 2020. The patients who have undergone ABG analysis in the emergency department and also serum creatinine estimation at the same time in the Biochemistry laboratory are included in the study. The data of all the above patients were collected from the records of various Departments of Nephrology, Emergency Medicine, and Biochemistry Departments. Hemolyzed, wrong labeling, icteric, lipemic samples, and the creatinine results from the instruments due to analytical error were excluded from the study. Finally, 75 samples were included in the study.

Serum creatinine was estimated by Jaffe method on Abbot Architect ci 8200 auto analyzer, in Biochemistry department, in which creatinine reacts with picric acid in alkaline medium and produces orange creatinine picrate color which is measured at 500 nm. Whole blood creatinine was estimated in Arterial blood Gas analyzer (ABL 800 FLEX) by enzymatic method, where creatinine in whole blood is acted upon by creatinine amidohydrolase, finally reducing to hydrogen peroxide and further to oxygen, hydrogen, and electrons. Amperometry principle is used to measure the amount of creatinine in the sample. The creatinine results between the two methods were compared to check the agreement between them using Bland-Altman plot. Statistical analysis was done using the Statistical Package for Social Sciences (SPSS) software version 23.

  Results Top

This study was conducted in tertiary care hospital. The creatinine results by both the methods were divided into three groups based on clinical decision limits as per the nephrologist opinion, to see the agreement at different levels. The groups are Group I comprises 25 samples (<2 mg/dL), Group II comprises 25 samples (2–6 mg/dL), and Group III comprises 25 samples (>6 mg/dL).

The mean, standard deviation, mean difference, and interclass correlation between whole blood (Radiometer ABL 800 FLEX) and serum creatinine (Abbott Architect ci 8200) is shown in [Table 1]. Intraclass correlation coefficients (ICCs) for the agreement between the two methods including all the groups are 0.995. Method comparison between the enzymatic creatinine method and Jaffe's method by linear regression for all the groups together (n = 75) showed a coefficient correlation R of 0.9893 [Figure 1]d. [Figure 1] and [Figure 2] explain the Bland – Altman plot for agreement and scatterplot for correlation between the two methods in each group and all the groups together.
Table 1: Mean, standard deviation and mean difference for Jaffe and enzymatic method in the four group

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Figure 1: Correlation graph showing the association between the two methods (a) Group I – 0–2 mg/dL with correlation coefficient of 0.908. (b) Group II - 2-6 mg/dL with correlation coefficient of 0.943 (c) Group III – 6 mg/dl with correlation coefficient of 0.962. (d) All the groups with correlation coefficient of 0.989

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Figure 2: Bland Altman plot between Jaffe's and Enzymatic Method (a) - showing the results for 0–2 mg/dL of creatinine, with mean difference of − 0.063 (95% confidence interval − 0.318, 0.192). (b) Group II - 2-6 mg/dl with mean difference of 0.070 (95% confidence interval - 0.54 to 0.681). (c) Group III – 6 mg/dl with mean difference of 0.198 (95% confidence interval - 1.287 to 1.683). (d) All the groups with mean difference of 0.0685 (95% confidence interval -0.881 to 1.018)

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We also analyzed few samples which had flags for creatinine results in the ABG analyzer, which are used to treat the patients. The mean difference and ICC is −0.220 mg/dL and 0.894, respectively between the two methods as shown in [Table 2]. [Figure 3]a and [Figure 3]b is the Bland – Altman plot showing higher mean difference and regression analysis with correlation coefficient R of 0.7995 indicating lower correlation in comparison with the samples having no flags.
Table 2: Mean, standard deviation, mean difference and intraclass correlation coefficient for Jaffe and enzymatic method for the flagged samples

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Figure 3: (a) Correlation graph between the two methods for the results with flags on arterial blood gas analyzer showing correlation coefficient of 0.7995. (b) Bland - Altman plot between the two methods for the results with flags on arterial blood gas analyzer showing mean difference of − 0.220 (95% confidence interval -4.530 to 4.091)

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  Discussion Top

Acute or chronic renal disease is a major health problem all over the world. Estimation of serum/whole blood creatinine levels is required to assess renal function.[9] The availability of a rapid creatinine assay by POCT device with inaccurate values may allow differential treatment decisions to be made for patients at risk of developing renal failure.[10] Clinical studies are needed to demonstrate whether whole blood creatinine assays at POCT devices are accurate enough to provide reliable estimates of renal function in their settings.

In our study, the mean difference between Jaffe's and enzymatic method in all the group together is 0.0685 mg/dL. ICC of 0.995 between the two methods showed very good correlation [Table 1]. Marakala et al. evaluated 167 normal samples (Group 1) and did not find a significant difference (−0.042) and the ICC also showed a very good correlation (0.995) between the two methods.[11] All the measurements were performed using an Olympus AU 400 analyzer.

Malukar et al. showed that enzymatic method has greater linearity (up to 60 mg/dL) than Jaffe's (up to 20 mg/dL) method.[12] This study was done on six groups, each group comprises 66 samples and both the methods were done using ROBONIK Semi auto analyzer. The first three groups compared creatinine between the two methods and the other three groups were compared for interferences such as glucose, bilirubin and both together. There was a very good correlation for serum creatinine (ICC 0.931, 0.998, 0.986 in Group I, II, III respectively) between the two methods in all the three groups.

Anne-Sophie et al. proved that in comparison of three POCT devices (ABL 800, i-STAT, Stat sensor), ABL 800 creatinine presented excellent agreement with the IDMS-traceable Roche enzymatic assay with a small bias.[7] The i-STAT creatinine overestimates whereas the Stat Sensor underestimates plasma creatinine in comparison to the IDMS-traceable Roche enzymatic assay.[13],[14] ABL 800 FLEX is the instrument used in our setting as POCT device, where the results had good agreement with the Jaffe method. Similar studies were not found which compared creatinine between Jaffe method in fully automated analyzer and enzymatic method by POCT device.

In our study in the Group III, the mean difference is high (0.198 mg/dL) as compared to the other two groups (−0.063 mg/dL, 0.070 mg/dL). This may be due to higher range of serum creatinine values in Group III.[15] We also had few serum creatinine comparisons, where the samples were measured at different points of time with the samples drawn at the same time between the two methods. These values were included, as the study by Ford and Berg concludes that delay in the separation of clotted blood samples have shown increase in creatinine is significant after only a 16-h delay in sample separation.[16]

We also had few samples which flag in creatinine results in the ABG analyzer, which was not included in the above data and analyzed separately. This was analyzed, because in emergency conditions, these values were used for treatment. Hence the results of this would help the clinician for better decision when required. The mean difference in this group was 0.894 and ICC 0.795, where the mean difference is very high, and low correlation compared to the samples without flags. This guides the treating physician, to not consider the results which have flags.

  Conclusion Top

We found that the whole blood creatinine levels on the Radiometer ABL 800 FLEX without flags, have good agreement and good correlation with the serum creatinine level on the Architect Abbot Ci8200 auto analyzer. The clinicians can use the results without flags from the POCT device by enzymatic method and no need to repeat the test by Jaffe's method. This leads to decrease in cost, early clinical decision, and treatment of the patient in the emergency department.


Department of Biochemistry, Department of Nephrology, Department of Emergency Medicine, Department of Statistics St Johns Medical college and Hospital.

Financial support and sponsorship


Conflicts of interest

There are no conflicts of interest.

  References Top

Draczevski L, Teixeira L. Evaluation of per filbiochemical and parameters haematological in patients subjected to dialysis. Rev Saud Pesq 2011;4:15-22.  Back to cited text no. 1
Verma M, Khadapkar R, Sahu PS, Das BR. Comparing age-wise reference intervals for serum creatinine concentration in a “Reality check” of the recommended cut-off. Indian J Clin Biochem 2006;21:90-4.  Back to cited text no. 2
Jaffe M. About the precipitate that picric acid produces in normal ears and about a new reaction of creatinine. Z Physiol Chem 1886;10:391-400.  Back to cited text no. 3
Liu WS, Chung YT, Yang CY, Lin CC, Tsai KH, Yang WC, et al. Serum creatinine determined by Jaffe, enzymatic method, and isotope dilution-liquid chromatography-mass spectrometry in patients under hemodialysis. J Clin Lab Anal 2012;26:206-14.  Back to cited text no. 4
Syal K, Banerjee D, Srinivasan A. Creatinine estimation and interference. Indian J Clin Biochem. 2013;28:210-1. doi: 10.1007/s12291-013-0299-y. Epub 2013 Jan 26. PMID: 24426213; PMCID: PMC3613509.  Back to cited text no. 5
Vaishya R, Arora S, Singh B, Mallika V. Modification of Jaffe's kinetic method decreases bilirubin interference: A preliminary report. Indian J Clin Biochem 2010;25:64-6.  Back to cited text no. 6
Korpi-Steiner NL, Williamson EE, Karon BS. Comparison of three whole blood creatinine methods for estimation of glomerular filtration rate before radiographic contrast administration. Am J Clin Pathol 2009;132:920-6.  Back to cited text no. 7
Schmidt RL, Straseski JA, Raphael KL, Adams AH, Lehman CM. A risk assessment of the Jaffe vs. enzymatic method for creatinine measurement in an outpatient population. PLoS One 2015;10:e0143205.  Back to cited text no. 8
Schweiger MJ, Chambers CE, Davidson CJ, Blankenship J, Bhalla NP, Block PC, et al. Prevention of contrast induced nephropathy: Recommendations for the high risk patient undergoing cardiovascular procedures. Catheter Cardiovasc Interv 2007;69:135-40.  Back to cited text no. 9
Briguori C, Airoldi F, D'Andrea D, Bonizzoni E, Morici N, Focaccio A, et al. Renal insufficiency following contrast media administration trial (REMEDIAL). Circulation 2007;115:1211-7.  Back to cited text no. 10
Vijaya M, Avinash SS, Shivashankara AR, Malathi M, Arun Kumar.Serum Creatinine assay: Enzymatic Vs Kinetic Jaffe's method. J Evol Med Dent Sci 2012:1;328-34.  Back to cited text no. 11
Malukar NR, Kanazariya SB, Bapure SK, Purohit J. Comparison of Modified Jaffe's Kinetic Method and Enzymatic Method of Serum Creatinine Estimation for Precision, Linearity and Effect of Interferent. Int J Res Med 2017;6;27-34.  Back to cited text no. 12
Nichols JH, Bartholomew C, Bonzagi A, Garb JL, Jin L. Evaluation of the IRMA TRU point and i-STAT creatinine assays. Clin Chim Acta 2007;377:201-5.  Back to cited text no. 13
Shephard M, Peake M, Corso O, Shephard A, Mazzachi B, Spaeth B, et al. Assessment of the Nova Stat Sensor whole blood point-of-care creatinine analyser for the measurement of kidney function in screening for chronic kidney disease. Clin Chem Lab Med 2010;48:1113-9.  Back to cited text no. 14
Bargnoux AS, Kuster N, Cavalier E, Piéroni L, Souweine JS, Delanaye P et al. Serum creatinine: advantages and pitfalls. J Lab Precis Med 2018;3:71.  Back to cited text no. 15
Ford L, Berg J. Delay in separating blood samples affects creatinine measurement using the Roche kinetic Jaffe method. Ann Clin Biochem 2008;45:83-7.  Back to cited text no. 16


  [Figure 1], [Figure 2], [Figure 3]

  [Table 1], [Table 2]


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