產品貨號：ZZK-K-ACETRM

產品品名：乙酸[乙酸激酶法]檢測試劑盒

英文品名：Acetic Acid (Acetate Kinase Manual Format) Assay Kit

規格型號：72 assays (manual) / 720 assays (microplate)

This rapid and reliable manual acetic acid kit is simple to perform(only two absorbance readings required), and because a true end-point ismeasured, does not involve complicated calculations like other kits.This product is very stable both during storage and use (> 2 years),has extended linearity (compared to ACS based kits), contains PVP toprevent tannin inhibition, and is performed at a relatively low pH(7.4), thus minimising ester hydrolysis related interference. Thismethod is suitable for the measurement of acetic acid/acetate in foods,beverages and other materials.

INTRODUCTION:

Given its importance and ubiquitous occurrence in foods, beverages andother materials, acetic acid ranks amongst the most measured analytes.However, unlike some other common analytes, traditional enzymaticmethods for this acid present significant limitations. For instance,manual acetyl-coenzyme A synthetase (ACS) based formats are typified byunstable reagents during use, slow reaction times,multiple cuvetteadditions / absorbance readings and complex

calculations. While the advanced acetic acid kit (K-ACET) recentlydeveloped by Megazyme overcomes the key stability issue relating to thereconstitution of ACS for manual use, the overall format of the assay isstill relatively inconvenient, requiring multiple cuvette additions andtime-consuming data recording / processing. Thus Megazyme developed thecurrent kit (K-ACETRM) based on acetate kinase (AK) andphosphotransacetylase (PTA) specifically to

overcome these limitations experienced by the analyst employing manualassay procedures. Unlike ACS based kits, this product is rapid (approx.4 min), does not present reagent stability issues,is simple to perform,has extended linearity and gives a change in absorbance stoichiometricwith the concentration of acetic acid (i.e. no complex equations arerequired and the raw absorbance data is processed like most otherenzymatic kits). In common with other Megazyme acetic acid kits,K-ACETRM also benefits from > 2 years stability during use, as thekey enzyme components are supplied as ready-to-use ammonium sulphatesuspensions. Polyvinylpyrrolidone (PVP) has also been incorporated intothe assay system to prevent interference from particular tannins foundin wines (especially red

wines). As the pH (7.4) of the buffer system employed is significantlylower than in traditional ACS based kits (8.4), interference fromspontaneous ester hydrolysis (causing creep reactions) is also minimised.

KITS:

Kits suitable for performing 72 assays in manual format (or 720 assaysin microplate format) are available from Megazyme. The kits contain thefull assay method plus:

Bottle 1: Buffer (24 mL, pH 7.4) and sodium azide (0.02% w/v) as apreservative.Stable for > 2 years at 4°C.

Bottle 2: Tablets (18) containing NADH plus ATP, PEP and PVP. Stablefor > 2 years at -20°C.

Bottle 3: (x2) CoA, lyophilised powder.Stable for > 2 years at -20°C.

Bottle 4: D-Lactate dehydrogenase, phosphotransacetylase and pyruvatekinase suspension (1.5 mL).Stable for >2 years at 4°C.

Bottle 5: Acetate kinase suspension (1.5 mL). Stable for > 2 yearsat 4°C.

Bottle 6: Acetic acid standard solution (5 mL, 0.10 mg/mL).Stable for> 2 years at 4°C.

EQUIPMENT (RECOMMENDED):

1. Glass test tubes (round bottomed; 16 x 100 mm).

2. Disposable plastic cuvettes (1 cm light path, 3.0 mL).

3. Micro-pipettors, e.g. Gilson Pipetman (20 μL and 100 μL).

4. Positive displacement pipettor, e.g. Eppendorf Multipette?

- with 5.0 mL Combitip [to dispense 0.5 mL aliquots of

NADH/ATP/PEP/PVP/buffer (solution 2)].

- with 25 mL Combitip (to dispense 2.0 mL aliquots of

distilled water).

5. Analytical balance.

6. Spectrophotometer set at 340 nm.

7. Vortex mixer (e.g. IKA Yellowline Test Tube Shaker TTS2).

8. Stop clock.

9. Whatman No.1 (9 cm) and GF/A glass fibre filter papers.

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