COMPARISON OF HbA1C TESTING METHODS USING ION EXCHANGE HIGH PERFORMANCE LIQUID CHROMATOGRAPHY WITH UV-VISIBLE REFLECTOMETRY AND PRELIMINARY STUDY FOR DETECTION OF HbA1C BASED ON PHENYLBORONIC ACID AFFINITY

• Main Author: Kafesa, Ally
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/45241
• Institution: Institut Teknologi Bandung

Background and objective: Glycated hemoglobin, measured as HbA1c for diabetes monitoring. There are several different testing methods for measuring glycated hemoglobin (HbA1c). The standard method used is an ion exchange High-Performance Liquid Chromatography (HPLC). One of development methods have been carried out is a reflectometry UV-Visible method. Many factors intruder in HbA1c measurement with this method. The purpose of this research is to determine the performance reflectometry UV-visible method against ion exchange HPLC and preliminary study for detection of HbA1c based on phenylboronic acid affinity using spectrophotometer UV-Visible. Methods: Performance testing reflectometry UV-Visible methods was screened using test Total Error (TE) of the Total Error allowable (TEa) in a way measure the performance comparison with ion exchange HPLC using patient samples. The decision of performance method is based on Westgard rules. Preliminary testing of HbA1c detection based on phenylboronic acid affinity is done by reacting the phenylboronic acid with HbA1c standard in polar solvents with alkaline conditions. Results: TE on reflectometry UV-Visibel method (6.4%) greater than TEa (6.0%). In the preliminary test detection of HbA1c, the visibel spectrum HbA1c was not different with Hb at ?415 nm, so the detection of HbA1c can not be observed at these wavelengths. Phenylboronic acid pattern on UV spectrum does not change at ?275 nm so it can not detect the presence or absence of reaction between phenylboronic acid with HbA1c. Proof of the reaction between the phenylboronic acid with glucose done with optimum conditions at pH 8.0 with r = 0.9871. Conclusions: Performance of reflectometry UV-Visibel are not comparable with ion exchange HPLC method. This method can be used in clinical laboratories but it is not satisfactory for the quality requirements. HbA1c detection using UV-Visible spectrophotometer by phenylboronic acid affinity can not be observed because of an increase in absorbance due to hemoglobin which gives the same absorption maximum at a wavelength of phenylboronic acid.