Glycated Albumin Assay Kit​

Glycated albumin (GA) assay kit (Peroxidase method)

Principle

1. Determination of glycated albumin In the sample (serum), specific proteases are added to decompose Glycated albumin to form glycosylated amino acids. Then glycosylated amino acids are converted into glucoaldosterone, amino acids and hydrogen peroxide by specific glycosylated amino acid oxidase. Under the action of peroxidase, hydrogen peroxide reacts with 4-amino-aminoantipyrine and TOOS to form red quinquinimine. The concentration of glycated albumin in the sample was obtained by measuring its absorbance.

2. Determination of albumin In pH 4.2, the albumin in the sample (serum) binds with the indicator bromocresol green to form a blue-green complex. The albumin concentration in the sample was obtained by measuring the change in absorbance.

Reagents:

ALB：

Sample Requirements

1. Serum, heparin or EDTA plasma, and urine are suitable for samples. Whole blood, hemolysis is not recommended for use as a sample. Freshly drawn serum is the preferred specimen.

2. Stability: Serum/plasma: 3 days at 2-8℃;

Calibrator Preparation

Carefully open the bottle, avoiding the loss of lyophilizate, and pipette in exactly 1.0 mL of distilled/deionized water. Carefully close the bottle and dissolve the contents completely by occasional gentle swirling within 30 minutes. Avoid the formation of foam. The dissolved calibrator can be used without any other Pretreatment.

Quality control Preparation

Carefully open the bottle, avoiding the loss of lyophilizate, and pipette in exactly 1.0 mL of distilled/deionized water. Carefully close the bottle and dissolve the contents completely by occasional gentle swirling within 30 minutes. Avoid the formation of foam. The dissolved control can be used without any other Pretreatment.

Method

1. Reagent preparation: Liquid reagent can be used when opened

2. Measurement：

Calibration

It is recommended to use the Calibrator from Hzymes and distilled/deionized water for two-point calibration.

Calibration frequency:

After reagent lot changed. As required following quality control procedures.

Quality control

At least two levels of control material should be analyzed with each batch of samples.Each laboratory should establish its own internal quality control scheme and procedures for corrective action if controls do not recover within the acceptable tolerances.

Reference Intervals

[3] Each laboratory should establish its own reference intervals based on its patient population. The reference intervals measured at 37 °C listed below were taken from the literature. Serum / Plasma: 10.8%~17.1%

Interferences/Specificity

The following substances were tested for interference with this methodology. Criterion: Recovery within ±10 % of the initial value.

Storage and stability

Up to expiration date indicated on the label, when stored unopened at 2-8℃ and protected from light. Once opened, the reagents are stable for 28 days when refrigerated on the analyzer or refrigerator. Contamination of the reagents must be avoided. Do not freeze the reagents. Once dissolved, the calibrator are stable for 7 days at 2– 8℃, the control are stable for 7 days at 2–8℃，do not freeze.

Reagent Blank Absorbance

The absorbance of reagent blank at 546 nm should be <0.020.

Sensitivity/Detection Limit

The lowest measurable GA concentration that can be distinguished from zero is 0.2g/dl with 99.7% confidence.

Precision

Within-run : CV≤5%

Between-run: CV≤5%

Linearity range

Conventional Units: GA: 0.2g/dl~4g/dl

ALB: 0.2g/dl~28g/dl

If the value of sample exceeds 4g/dl(GA) or 28g/dl (ALB), the sample should be diluted with 9 g/L NaCl solution (e.g. 1+3) and rerun; the result should be multiplied by 4.

Warnings and Precautions

1. For in vitro diagnostic use.

2. Take the necessary precautions for the use of laboratory reagents.

3. Preservative contained. Do not swallow. Avoid contact with skin and mucous membranes.

4. Disposal of all waste material should be in accordance with local guidelines.

5. Material safety data sheet is available on request for professional users.

References

[1] Guide for clinical application of blood glucose monitoring in China (2011 edition). Chinese Journal of Diabetes，2011, 3(1): 13 -21.

[2] Santiago Rodriguez-Segade et al. Progression of nephropathy in type 2 diabetes: The glycation gap is a significant predictor after adjustment for glycohemoglobin (HbA1c). Clinical Chemistry, 2011, 57(2):264-271.

[3] M. Koga, et al. Glycated albumin and glycated hemoglobin are influenced differently by endogenous insulin secretion in patients with type 2 diabetes. Diabetes care, 2010, 33(2): 270 – 272.

Used for the quantitative determination of glycated albumin (GA) and albumin (ALB) in human serum， the ratio of glycated albumin (%) is used glycated albumin concentration divided by the albumin concentration. Glycated albumin is a non-enzymatic reaction between glucose and albumin in the blood because the half-time of albumin in ivio is short (about 17-19 days）, so GA serves as a 2-3 week indicator of average blood glucose. Therefore， it has significant clinical application value When blood sugar needs to be evaluated for short-term control.