Date of Completion

2023

Document Type

Thesis

Degree Name

Bachelor of Science in Pharmacy

Keywords

Alpha-amylase

Abstract

Knowing drug activities is crucial in early drug discovery in identifying which molecules have a certain activity towards a particular target, and in pharmacology in terms of determining whether a drug is safe and effective. Drug activity confirmation requires superior specificity and sensitivity in order to prevent false positive or false negative results. Meanwhile, concentration of drugs and the body’s response have a direct relationship to one another.

Affinity Chromatography is widely used as a means of separation and purification. It is based on the high affinity binding between the target substances. In Weak Affinity Chromatography (WAC), separation is facilitated by pairing ligands with lower affinity to the target. WAC allows ligands to be retained and eluted according to their inherent affinity, resulting in distinct peaks viewed in the chromatogram (Ohlson et al., 2017). It has been determined that WAC may be used to measure the activity of commercially available herbal drug supplements. Hence this study aims to validate its simultaneous Quantification Method and alpha-Amylase Inhibition Bioassay using the ICH Guidelines.

In this study, the Amylase Affinity column assembled by Cosio, et. al. (2022) was prepared via a Schiff-based Reduction method and packed in a 2.5 mm x 50 mm stainless-steel column using the PerkinElmer HPLC system. A total of 322 nmol or 133 mg amylase/g silica was immobilized in the affinity column. Five concentrations of Quercetin samples were prepared in triplicates for three separate days.

The results for precision parameters only passed repeatability (intra-day), and linearity. On the other hand, intermediate precision did not pass the requirements of ICH while specificity was not conducted hence cannot specifically determine the accuracy and range of WAC. Overall, AMY-WAC does not conform to the ICH Bioanalytical M10 and Analytical Q2 (R1) Guidelines.

First Advisor

Timothy Jay L. Bengala

Download

Share

COinS