LC-MS/MS Method for Serum Creatinine: Comparison with Enzymatic Method and Jaffe Method

Abstract

Accurate quantification of creatinine (Cre) is important to estimate glomerular filtration rate (GFR). Differences among various methods of Cre quantification were previously noted. This study aims to develop a liquid chromatography tandem mass spectrometry (LC-MS/MS) method for serum Cre and compare this method with clinical routine methods. LC-MS/MS analysis was performed on API 4000 triple quadrupole mass spectrometer coupled with an Agilent 1200 liquid chromatography system. After adding isotope-labeled Cre-d3 as internal standard, serum samples were prepared via a one-step protein precipitation with methanol. The LC-MS/MS method was compared with frequently used enzymatic method and Jaffe method. This developed method, with a total run time of 3 min, had a lower limit of quantification of 4.4 μmol/L, a total imprecision of 1.15%–3.84%, and an average bias of 1.06%. No significant matrix effect, carryover, and interference were observed for the LC-MS/MS method. The reference intervals of serum Cre measured by LC-MS/MS assay were 41–79 μmol/L for adult women, and 46–101 μmol/L for adult men. Using LC-MS/MS as a reference, the enzymatic method showed an average bias of -2.1% and the Jaffe method showed a substantial average bias of 11.7%. Compared with the LC-MS/MS method, significant negative bias was observed for the enzymatic and Jaffe methods in hemolytic and lipimic samples. We developed a simple, specific, and accurate LC-MS/MS method to analyze serum Cre. Discordance existed among different methods.

Serum Metabolic Signatures of Chronic Limb-Threatening Ischemia in Patients with Peripheral Artery Disease

“…Moreover, control charts for the three recovery standards used in MSI–CE–MS (10 μM F-Phe and NMS) and MSI–NACE–MS (50 μM 14:0-d27) added to all serum samples, prior to ultrafiltration or MTBE extraction, further demonstrate the acceptable intermediate precision (mean CV < 15%) with few outliers (<2%, n = 57 total runs) exceeding warning limits (±2 s) ( Figure 2 C–E). An inter-laboratory method comparison of the serum creatinine concentrations measured independently from 56 participants was also performed by MSI–CE–MS relative to the Jaffé colorimetric method, which was used for the estimation of the glomerular filtration rate (GFR) for patients as an indicator of kidney function [ 20 ]. In this case, a Bland–Altman % difference plot confirms a good mutual agreement for serum creatinine determination by both methods with a mean bias of −5.6% ( Figure 2 F).…”

“…Measurement of serum creatinine was performed at St. Michael’s Hospital using a modified Jaffé colorimetric assay on a Beckman Coulter AU system/analyzer (Beckman Coulter, Inc., Brea, CA, USA). Briefly, creatinine standard reagent (OSR6678) was added to an aliquot of serum, where the serum creatinine reacted with picric acid under alkaline conditions to form a yellow-orange complex and the rate of change in absorbance at 520/800 nm was proportional to the serum creatinine concentration to minimize other reacting/absorbing interferences [ 20 ]. Serum creatinine measurements by the Jaffé colorimetric assay were compared with multisegment injection–capillary electrophoresis–mass spectrometry (MSI–CE–MS) using independent serum aliquots collected from the same participants.…”

A New Method for the Determination of Creatinine in Urine Samples Based on Disposable Pipette Extraction

“…Song and co-workers 30 developed a method for creatinine analysis using liquid chromatography-mass spectrometry (LC-MS) with mobile phase methanol:acetonitrile 55:45 (v/v) and a flow-rate of 0.3 mL min -1 to result in a retention time of approximately 1.5 and 3.0 min. Even using an higher flow-rate, which could make the developed method less attractive, the mobile phase consisted of ultrapure water acidified to pH 4 and acetonitrile (70:30 v/v) with a spectrophotometric detector, which has a lower cost comparing to MS detector.…”

BALSAM—An Interactive Online Platform for Breath Analysis, Visualization and Classification

“…Such an associated VOC might not only reveal a biomarker for disease detection, but also highlight pathways and potential drug targets involved in the disease. Several technologies for VOC analysis have emerged in recent decades, allowing for potential non-invasive and rapid discovery of VOC patterns: electronic nose [ 7 , 8 ], gas chromatography-mass spectrometry (GC-MS) [ 9 , 10 ], liquid chromatography-mass spectrometry (LC-MS) [ 11 , 12 ] and multi-capillary-column ion-mobility-spectrometry [ 13 , 14 , 15 ] based devices are used for a majority of trials. MCC-IMS devices have seen an increase in popularity—e.g., in the detection of anesthetics in the workplace [ 16 ], accurate tracking and quantification of gaseous propofol concentration during surgery [ 17 ], identification of VOCs in idiopathic pulmonary fibrosis [ 18 ] and for breast cancer detection [ 19 ].…”