中文名称：乙酸[AF法]检测试剂盒
货号：K-ACETAF
英文名称：Acetic Acid Assay Kit (ACS Analyser Format)
规格：141.6 mL of prepared reagent (e.g. 456 assays of 0.31 mL)
Analyser format UV-method for the determination of Acetic Acid in foodstuffs, beverages and other materials
(乙酰-CoA合成酶)(1)(柠檬酸合成酶)(2)2(L-苹果酸脱氢酶)+170.5 mL 配好的试剂 (R1 + R2)分光光度计，340 nm~15min10mg/L(使用推荐方法)葡萄酒、啤酒、水果和果汁、软饮料、醋、蔬菜、泡菜乳制品（如奶酪）、肉、鱼、面包、焙烤食品（和发酵粉）、番茄酱、酱油、蛋黄酱、调味汁、纸（硬纸板）、茶、医药品（如注射液）、饲料和其他原料（生物培养基、样品等）该实验方法已通过EN、ISO、ICUMSA、德国和荷兰的认证
INTRODUCTION:
The most widely used method for enzymatic quantification of acetic acidis that based on the use of acetyl-coenzyme A synthetase (ACS),according to equations 1 – 3 below. For auto-analyser applications itbecomes necessary to prepare a “master mix” reagent (R1), containing allcomponents of the assay except the reaction initiation enzyme, ACS.However, reagent R1 when prepared from some kits has very limitedon-machine stability, owing to a rapidly increasing absorbance value. Toovercome this issue,
Megazyme developed this kit (K-ACETAF), thatdoesn’t exhibit this instability phenomenon. Additionally, theanti-inhibitory compound polyvinylpyrollidone (PVP) has also beenincorporated into the assay to prevent inhibition caused by certaintannins found in grape juice and wine.
试剂盒组成:
Kits suitable for the preparation of 141.6 mL of reagent (equivalent to456 reactions of 0.31 mL) are available from
Megazyme.
The kits contain the full assay method plus:
Bottle 1: Buffer (30 mL, pH 8.4) plus L-malic acid, PVP and sodiumazide (0.02 % w/v) as a preservative. Stable for > 2 years at 4°C.
Bottle 2: (x2) NAD+ plus ATP and CoA.Freeze dried powder.Stable for> 5 years at -20°C.
Bottle 3: L-Malate dehydrogenase plus citrate synthase suspension, 2.2mL. Stable for > 2 years at 4°C.
Bottle 4: Acetyl-coenzyme A synthetase suspension (1.1 mL). Stable for> 2 years at 4°C.
REAGENT PREPARATION:
Preparation of R1:
Component Volume
bottle 1 (buffer) 5.50 mL
bottle 2 (NAD+/ATP/CoA) 2.20 mL (after adding 5.50 mL of H2O to bottle 2)
bottle 3 (L-MDH/CS) 0.44 mL (swirl to mix before use)
H2O 18.35 mL
Total volume 26.49 mL
Preparation of R2:
Component Volume
bottle 4 (ACS) 0.22 mL (swirl to mix before use)
H2O 1.60 mL
Total volume 1.82 mL
EXAMPLE METHOD:
R1: 0.290 mL
Sample: ~ 0.005 mL
R2: 0.020 mL
Reaction time: 15 min at either 20-25°C or 37°C
Wavelength: 340 nm
Prepared reagent stability: > 3 days when refrigerated
Calculation: endpoint
Reaction direction: increase
Linearity: up to 30 μg/mL of acetic acid in final reaction solution
Principle原理:
(acetyl-CoA synthetase)
(1) Acetic acid + ATP + CoA → acetyl-CoA + AMP + pyrophosphate
(citrate synthase)
(2) Acetyl-CoA + oxaloacetate + H2O → citrate + CoA
(L-malate dehydrogenase)
(3) L-Malate + NAD+ ↔ oxaloacetate + NADH + H+Kit size: 141.6 mL of prepared reagent (R1 + R2)
Method: Spectrophotometric at 340 nm
Reaction time: ~ 15 min
Detection limit: 10 mg/L (recommended assay format)
Application examples:
Wine, beer, fruit and fruit juices, soft drinks, vinegar, vegetables,
pickles, dairy products (e.g. cheese), meat, fish, bread, bakery products
(and baking agents), ketchup, soy sauce, mayonnaise, dressings,
paper (and cardboard), tea, pharmaceuticals (e.g. infusion solutions),
feed and other materials (e.g. biological cultures, samples, etc.)
Method recognition:
Methods based on this principle have been accepted by EN, ISO,
ICUMSA, IFU and MEBAK
Advantages
- No wasted ACS solution (stable suspension supplied)
- PVP incorporated to prevent tannin inhibition
- Very stable reagent when prepared for auto-analyser applications (> 3 days at 4°C)
- Linear calibration up to 30 μg/mL of acetic acid in final reaction solution
- Validated by the University of Wine, Suze la Rousse, France
- Very competitive price (cost per mL of reagent)
- All reagents stable for > 2 years after preparation
优点：
不会浪费ACS溶液（提供的 为稳定的悬浮液）加入PVP防止丹宁酸的抑制自动分析检测时配置好的试剂非常稳定 (> 5天， 4摄氏度)
最终反应液中的乙酸线性高达已通过法国葡萄酒大学验证
价格低廉 (每ml试剂的成本)所有试剂配制后的稳定性 >2年
