A continuous-flow (AutoAnalyzer II) procedure for measuring erythrocyte transketolase activity

Area of Science:

• Biochemistry
• Enzymology
• Clinical Chemistry

Background:

• Erythrocyte transketolase (EC 2.2.1.1) is a crucial enzyme in red blood cells.
• Accurate measurement of transketolase activity is important for clinical diagnostics.
• Existing manual methods can be time-consuming and prone to interference.

Purpose of the Study:

• To develop and validate a semiautomated method for assaying erythrocyte transketolase activity.
• To improve the sensitivity and reduce interference in transketolase assays.
• To establish a reliable and precise method for quantifying transketolase activity in hemolysates.

Main Methods:

• Utilized AutoAnalyzer II for semiautomated measurement of glyceraldehyde 3-phosphate production.
• Separated hemolysate and indicator reactions via dialysis to eliminate hemoglobin interference.
• Employed internal standards of glyceraldehyde 3-phosphate for accurate calibration and quantification.

Main Results:

• The semiautomated method demonstrated good precision and accuracy in measuring transketolase activity.
• Dialysis effectively removed hemoglobin interference, enhancing assay sensitivity.
• Results obtained using the AutoAnalyzer correlated well with established manual procedures.

Conclusions:

• The AutoAnalyzer II provides a reliable and efficient semiautomated method for erythrocyte transketolase activity assay.
• This method offers advantages in terms of sensitivity, accuracy, and reduced interference.
• The validated method can be valuable for clinical and research applications requiring transketolase activity measurements.