Supplementary material: State of the art in currently available amylase detection technique Assay Design Detection Limits of detection Specificity Ref. Sequential Process Technique Flow system using a peroxide electrode and enzyme membrane with glucose oxidase, and optionally mutarotase cross-linked by α-glucosidase converts maltose to α-D-glucose. Mutarotase converts to which is determined via glucose oxidase AMP LOD: 2nkat/mL (0.117.64 units/mL) when reaction time 5 min 0.5 nkat/mL units/mL) when reaction time 30 min LR: 0.1-3 mmol/L Any 141 Screen-printed electrodes with immobilized glucose oxidase and mutarotase modified with Prussian Blue α-glucosidase converts maltose to α-D-glucose. Mutarotase converts to which is determined via glucose oxidase AMP LOD: 5 units/mL LR: 5-250 units/mL Any 142 Flow-injection device maltopentaose as substrate. α-glucosidase immobilised on membrane. Glucose oxidase immobilized on electrode α-glucosidase converts maltose to 5-d-glucose. Glucose oxidase coverts 5-d-glucose to gluconic acid and hydrogen peroxide which is measured AMP LOD: LR: 0-30 units/mL Any 143 Spectrophotometric flow injection measuring brick red complex formation at 540nm Amylose incubated with sample to produce maltose. 3,5 and maltose boiled SIA/FIA LOD: 0.0048 FAU LR: 0.005-0.06 FAU Any 144 Use of portable personal glucose meter Sample, and maltopentaose incubated 15 min at 370C. LOD: 20 U/L LR: 2.2-27.8 mM Any 145 Flat-chip micro analytical sensor used as part of a Systems. Pre-column and electrode incorporated into a flow cell where maltose Maltose phosphorylase phosphorylates maltose. Glucose oxidase converts phosphorylated maltose to gluconic acid and and Lateral flow LR: 0-190 kU/L Any 143
phosphorylase, glucose oxidase and peroxidase immobilised hydrogen peroxide which is measured Colorimetric assay biosensor system using Gal-G2-CNP, chromogenic substrate for α-amylase. CNP is a yellow product once hydrolysed which can be measured photometrically at 430nm. Disposable test strip placed under tongue (25 µl). Once strip inserted into reader and saliva moved onto the reagent paper. The entire test takes roughly 30 sec. AMP LR: 10-230 U/mL Any 146 Flow injection spectrophotometric analysis based on complexes Sample degradation of complexes measured in flow channel FIA LOD: 60 NU/mL LR: 0.25-5.0 KNU/mL Any 147 Immobilized layer of starch gel on thick-film magneto elastic sensor and presence of alters the resonance frequency Sample placed on starch gel LR: 75-125 U/mL Any 148 Spectrofluorimetric using the quenching of luminescence intensity (634nm) of nano CdS doped in sol-gel of different concentrations of maltose Sample incubated with starch in flow channels FIA LOD: 5.7 x 10-11 mol/L LR: 4.8 x 10 -10 – 1.2 x 10 -5 mol/L Any 149 Glycogen/amylopectin spin-coated on gold coated quartz crystals (case frequency of 10 MHz). Films cross-linked with hexamethylene diisocyanate. Film degradation measured with quartz crystal microbalance Sample incubated with film EIS Any 150 Degradation of measured using platinum redox sensor for direct potentiometric determination Sample incubated with LOD: 1.944 mU LR: 0-0.54 U Any 151 Glucose oxidase-based biosensor measuring the decrease in dissolved oxygen concentration related to starch concentration. LOD: LR: 0.66-9.83 U/mL Any 152