HPLC with enzymatic detection as a candidate reference method for serum creatinine.
نویسندگان
چکیده
We present a candidate Reference Method for determining the concentration of serum creatinine. The method is based on HPLC combined with enzymatic determination. Creatinine plus 14[C]creatinine is extracted by cation-exchange chromatography, subjected to reversed-phase HPLC, and finally quantified enzymatically. Enzymatic measurement ensures no interference from co-eluting compounds, which has been a problem for some reported HPLC assays relying on ultraviolet detection. The average corrected recovery was 100.1% (SEM = 1.1%; n = 15). The accuracy was verified by assaying five sera with target values determined by isotope dilution mass spectrometry. The total interassay CV was less than or equal to 2.5%. We applied the method to study the specificity of HPLC-ultraviolet detection, using 72 plasma samples from hospitalized patients; no interference was noted. Thus, HPLC-ultraviolet detection appears to be specific, provided that sample cleanup is based on cation-exchange chromatography. Our diode-array detector studies of peak homogeneity supported this conclusion. Still, combined HPLC-enzymatic determination ensures even greater accuracy, ranking close to that by isotope dilution mass spectrometry.
منابع مشابه
Candidate reference method for determining creatinine in serum: method development and interlaboratory validation.
We describe a "high-performance" liquid chromatographic (HPLC) method for accurately determining creatinine in serum. After prechromatographic precipitation of protein, we performed isocratic ion-exchange chromatography with ultraviolet detection (234 nm). Analytical results showed linearity up to 1770 mumol/L, a detection limit of 22 mumol/L, an average analytical recovery of 101%, and a CV ra...
متن کاملEvaluation of Accuracy, Precision and Agreement of Five Hba1c Measurement Methods with HPLC Reference Method
ABSTRACT Background and Objective: The current challenge of diabetes mellitus is to prevent its complications. These complications are directly associated with hyperglycemia in diabetics. The HbA1c measurement is essential for long-term glycemic control. Synchronization of HbA1c measurement is important in order to avoid discrepancies between resu...
متن کاملApplicability of common reference intervals for serum creatinine concentrations to the Croatian population.
BACKGROUND In accordance with an ongoing activity for worldwide harmonization based on traceability in laboratory methods, the goal of this study was to validate the applicability of recommended "common" reference intervals for serum creatinine concentrations using a specific enzymatic method to the Croatian population. METHODS The reference group consisted of 240 healthy subjects (120 males ...
متن کاملDeveloped Method Application for Nitrite Ion (NO2¯ ) Analysis of Tib -186 Macrophage Like Cell Lines by Rapid Isocratic HPLC System with High Sensitive Glassy Carbon Electrochemical Detector
A rapid isocratic method of high performance liquid chromatography system (HPLC) with a glassy carbon working electrode of electrochemical detector is set up for quantitative detection of trace amount of nitrite ion (NO2¯) in aqueous protein containing cell lysate, cell media, plasma, serum, urine and other body fluids. The solid extraction reversed phase cartridges ...
متن کاملDetermination of creatinine in serum and urine by a rapid liquid-chromatographic method.
We describe an HPLC ion-pair procedure for rapid and specific evaluation of creatinine in serum and urine. We used a 15 cm X 4.6 mm ODS column with a 50/50 (by vol) mixture of sodium decanesulfonic acid (10 mmol/L, pH 3.2) and methanol and measured absorbance at 236 nm. Serum (100 microL) or 30-fold-diluted urine (100 microL) was added to 400 microL of acetone. After centrifugation, the superna...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Clinical chemistry
دوره 37 10 Pt 1 شماره
صفحات -
تاریخ انتشار 1991