Glucose Determination Procedure for Stanbio Glucose LiquiColor (PDF)

Summary

This document provides instructions for the quantitative determination of glucose in serum, plasma, or CSF using the Stanbio Glucose LiquiColor (Oxidase) method. The procedure outlines reagent preparation, storage, and material requirements. It also details the procedure itself, including manual and automated options.

Full Transcript

Non reactive ingredients and preservatives Interfering Substances: Excessive levels of Glucose Standard, 100 mg/dL, Ref. No. 1072 ascorbic acid can produce falsely low glucose...

Non reactive ingredients and preservatives Interfering Substances: Excessive levels of Glucose Standard, 100 mg/dL, Ref. No. 1072 ascorbic acid can produce falsely low glucose Contains 5.55 mmol/L Glucose in 0.5 mol/L values. For a more comprehensive review of benzoic acid. factors affecting glucose assays, refer to the Precautions: The reagent is for "In Vitro publication by Young.9 Stanbio Glucose LiquiColor® Diagnostic Use". Normal precautions exercised in handling laboratory reagents should be followed. Automated Procedure Consult your instrument manual for programming (Oxidase) Procedure No. 1070 The reagent contains sodium azide, which may be instructions. Specific programming applications for Intended Use: For the Quantitative toxic if ingested. Sodium azide may also react with most automated analyzers are available from Determination of Glucose in Serum, Plasma or lead and copper plumbing to form highly explosive Stanbio’s Technical Service Department. metal azides. Refer to Material Safety Data Sheet CSF for any updated risk, hazard or safety information. Manual Procedure (Linear to 400 mg/dL)* Summary and Principle 1. Remove the amount of reagent to be used for The accurate estimation of glucose is important in Reagent Preparation: The Glucose reagent is supplied ready-to-use. testing and allow to warm to ambient temperature. the diagnosis and management of hyperglycemia 2. Glucose reagent is supplied ready-to-use. and hypoglycemia. Hyperglycemia may occur as a Reagent Storage and Stability: The reagent is 3. Zero spectrophotometer at 500 nm with distilled result of diabetes mellitus, in patients receiving stable up to the end of its labeled expiration date, water. intravenous glucose fluids, during severe stress or if properly stored at 2-8°C, protected from light and 4. For each standard, sample and control, add 1.0 as a result of cerebrovascular accidents. contamination is avoided. Do not freeze the mL reagent to cuvettes/test tubes and warm to Hypoglycemia may be the result of an insulinoma, reagent! Measurement is not influenced by 37°C for 5 minutes. insulin administration, reagent color changes as long as absorbance of 5. Add 10 μL (0.010 mL) of each sample to its inborn error of carbohydrate metabolism or fasting.1 the reagent is < 0.80 at 500 nm in a 1 cm respective cuvette/test tube, mix gently and return Measurement of blood glucose levels was among lightpath. Discard if it is found to contain to 37°C incubation. the first chemical procedures employed in clinical particulate matter. The standard is stable up to the 6. After 5 minutes of incubation, read and record laboratory medicine.2 The glucose oxidase end of the labeled expiration date, if properly the absorbance of all samples. methodology was introduced by Keilin and Hartree3 stored at 2-8°C and contamination is avoided. * If linearity is desired to 500 mg/dL, increase in 1948. Keston4 later reported use of the combined Material Required But Not Provided reagent volume to 1.5 mL and proceed using 10 glucose oxidase-peroxidase reagent, followed by the Spectrophotometer capable of absorbance µL of sample. Teller5 addition of a chromogenic reagent to reading at 500 nm (492-520 nm) Constant Keston's procedure. The Stanbio single reagent Manual Procedure (Linear to 650 mg/dL) temperature block or bath, 37°C, or temperature glucose method is based on a technique described 1. Remove the amount of reagent to be used for controlled cuvette Accurate pipetting devices by Trinder et al.6 testing and allow to warm to ambient temperature. Test tubes Interval timer Glucose is oxidized in the presence of glucose 2. Glucose reagent is supplied ready-to-use. oxidase (GOD). The hydrogen peroxide formed Specimen Collection and Storage 3. Zero spectrophotometer at 500 nm with distilled reacts, under the influence of peroxidase (POD), Non-hemolyzed serum is the specimen of choice. water. with phenol and 4-aminoantipyrine to form a red- Remove from clot within 30 minutes of collection in 4. For each standard, sample and control, add 1.0 violet quinone complex. The intensity of the color is order to prevent glycolysis. An anticoagulant mL reagent to cuvettes/test tubes and warm to proportional to glucose concentration. containing fluoride is recommended, but any of the 37°C for 5 minutes. β-D-Glucose + H2O + O2 GOD H2O2 + D-Gluconic Acid common anticoagulants may be used if the 5. Add 5 μL (0.005 mL) of each sample to its plasma is separated from the cells promptly after respective cuvette/test tube, mix gently and return H2O2 + 4-Aminoantipyrine + Phenol POD Quinone Complex + H2O centrifugation. No special preparation is required to 37°C incubation. Reagents for CSF specimens. Glucose in serum/plasma 6. After 5 minutes of incubation, read and record processed in the manner described is stable for 48 the absorbance of all samples. Glucose Reagent, Ref. No. 1071 hours at 2-8oC. For long term storage, samples Phosphate Buffer 200 mmol/L should be placed in sealed containers and frozen Phenol 4 mmol/L at -20oC.7,8 CSF samples should be analyzed 4-Aminoantipyrine 0.2 mmol/L immediately because of possible bacterial Glucose Oxidase > 15 KU/L contamination. Peroxidase > 1.2 KU/L Expected Values10 Normal Range: Serum/Plasma: 70 -105 mg/dL, (3.89 – 5.83 mmol/L) CSF: 40 - 75 mg/dL, (2.22 – 4.17 mmol/L) These ranges should serve only as a guideline. It is ultimately the responsibility of the laboratory to Quality Control: Stanbio Ser-T-Fy® 1, Level 1 establish its own range of expected values, since Control Serum, Ref. No. G427-86, and Ser-T-Fy® 2, differences exist between instruments, Level 2 Control Serum, Ref. No. G428-86, are laboratories, and local populations. recommended for each run. Other commercially Performance Characteristics11 available controls with glucose values assayed by Precision: Using a serum containing glucose in this method are also suitable. Glucose values the normal range and another with an elevated determined in these materials, by this procedure, value, a series of 5 assays were performed on STANBIO LABORATORY DISCLAIMS ALL EXPRESS AND should fall within the ranges stated for the controls. each of 5 days. Coefficients of variation (CV) were IMPLIED WARRANTIES OF THE MERCHANTABILITY AND Two levels of controls should be analyzed with each within runs of 1.6% and 1.2 % and between runs FITNESS PERTAINING TO THIS PRODUCT WHICH ARE run. of 3.0% and 2.0 %, respectively. NOT EXPRESSLY DETAILED IN THIS PACKAGING INFORMATION OR A WRITTEN AGREEMENT BETWEEN Calibration: Calibration is required. Stanbio Correlation: Determination of glucose by the THE BUYER AND SELLER OF THIS PRODUCT. Laboratory recommends the use of the standard procedure described (y) and by the powder form STANBIO LABORATORY MAINTAINS THAT THIS provided in the kit when using a manual method (x) on 64 sera showed a correlation PRODUCT CONFORMS TO THE INFORMATION spectrophotometer. For automated analyzers, coefficient (r) of 0.995 and the regression equation CONTAINED IN THIS INSERT. PURCHASER MUST DETERMINE THE SUITABILITY OF THE PRODUCT FOR Stanbio Laboratory recommends the use of the of y = 0.98x - 1.99 mg/dL. THEIR PARTICULAR USE. USE ONLY IN ACCORDANCE Ser-T-Cal® Multi-Calibrator, Ref. No. 0550-605. For Linearity: When performed as directed, this WITH LABELING INSTRUCTIONS. more information about automated analyzer method is linear as listed for each test procedure. calibration consult the calibration guidelines of your For Technical Service call: 1-800-531-5535 analyzer. References (830) 249-0772 1. Zilva JF, Pannall PR. Carbohydrate Metabolism in Results "Clinical Chemistry in Diagnosis and Treatment". Lloyd- Index of Symbols Attention, see Tests per Manufactured Values are derived by comparing the absorbance of Luke London 1979, Chap 9:174-214. instructions for use kit by the unknown (u) with that of a standard (s) 2. Folin O., Wu H.: J Biol Chem 41:367, 1920 For in vitro Use by Do not reuse diagnostic use only identically treated. 3. Keilin D., Hartree E.F.: Biochem J 42:230, 1948 Store between Lot 4. Keston AS: Abstr 129th Meeting, Am Chem Soc, temperature indicated Number REF Reference No. Glucose (mg/dL) = Au x 100 1956, p 31c. As 5. Teller J.D.: Abstr 130th Meeting, Am Chem Soc, Where Au and As are the absorbances of unknown 1956, p 69c. Stanbio Laboratory 6. Trinder, P., "Determination of Blood Glucose Using 4- 1261 North Main Street BDS International and standard, respectively, and 100 the Aminophenazone." J. Clin. Path., 22:246 (1959). Boerne, Texas 78006, USA Diagnostics GmbH Anselm-Feuerbach-Str. 12, Toll Free: (800) 531-5535 concentration of standard (mg/ dL) 7. Pencock CA, et al. Clin Chem Acta 1973; 49:193. Phone: (830) 249-0772 68723 Schwetzingen, Germany Fax: (830) 249-0851 e-mail: info@bds- 8. Shepard MDS, Mazzachi RD. The Clin Biochem e-mail: [email protected] international.de Example: Au = 0.370, As = 0.280 1983;4:61-67. www.stanbio.com 9. Young DS, Effects of Drugs on Clinical Laboratory Glucose (mg/dL) = 0.370 x 100 = 132 Tests, 3rd Ed, 1990; 3:168-182. 0.280 10. Cooper G.R., McDaniel V: Manual of Methods for RBR.1070.CE.03 Last Revision: 4/11 the Determination of Glucose, CDC, USPHS, Atlanta. Limitations 11. Caraway W.T.: IN Fundamentals of Clinical If the glucose value exceeds the linearity as Chemistry, 2nd ed., N.W. Tietz, Ed. Saunders, described in the method run, the specimen should Philadelphia, 1976, p 242. be diluted 2-fold (1+1) with distilled water, the assay 12. Stanbio Laboratory Data repeated and results multiplied by the dilution factor of 2.

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