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淀粉总量检测试剂盒 K-TSTA-100A
淀粉总量检测试剂盒 K-TSTA-100A

英文名:Total Starch (AA/AMG) Assay Kit

货号:K-TSTA-100A

规格:100 assays per kit

市场价: 5349
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K-TSTA-100中文版说明书equl.pdf

 

 

 

  爱尔兰Megazyme 淀粉总量检测试剂盒 

英文名:Total Starch (AA/AMG) Assay Kit

货号:K-TSTA-100A

规格:100 assays per kit

分析物意义:主要的食品组分

Content: 50 assays / 100 assays

Shipping Temperature: Ambient

Storage Temperature: Short term stability: 2-8oC,

Long term stability: See individual component labels

Stability: > 2 years under recommended storage conditions

Analyte: Total Starch

Assay Format: Spectrophotometer

Detection Method: Absorbance

Wavelength (nm): 510

Signal Response: Increase

Linear Range: 4 to 100 μg of D-glucose per assay

Limit of Detection: 0.18 g/100 g total starch “as is”

Total Assay Time: ~ 90 min

Application examples: Cereal flours, food products and other materials.

Method recognition: AACC Method 76-13.01, AOAC Method 996.11, ICC Standard Method No. 168 and RACI Standard Method

The Total Starch (AA/AMG) Assay Kit is used for the determination of total starch in cereal flours and food products.  AOAC Method 996.11, AACC Method 76-13.01.

 

This kit now contains an improved α-amylase that allows the amylase incubations to be performed at pH 5.0 (as well as pH 7.0). 

 

含量:50次分析/100次分析

运输温度:环境温度

储存温度:短期稳定性:2-8oC,

长期稳定性:参见单个组件标签

稳定性:在推荐的储存条件下超过2年

分析物:总淀粉

测定形式:分光光度计

检测方法:吸光度

波长(nm):510

信号响应:增加

线性范围:每次测定4至100μg D-葡萄糖

检测限:0.18g/100g总淀粉“原样”

总测定时间:~90分钟

应用示例:谷物粉、食品和其他材料。

方法识别:AACC方法76-13.01、AOAC方法996.11、ICC标准方法168和RACI标准方法

总淀粉(AA/AMG)测定试剂盒用于测定谷物粉和食品中的总淀粉。AOAC方法996.11,AACC方法76-13.01。

 

该试剂盒现在含有一种改进的α-淀粉酶,可以在pH 5.0(以及pH 7.0)下进行淀粉酶孵育。

 

淀粉大致包含两种成分,直链淀粉主要是通过α-1,4-糖苷键连接的 D-葡聚糖,含有极少的 α-1,6 支链;而支链淀粉中葡萄糖分子之间除了以 α-1,4-糖苷键相连外,还有以 α-1,6-糖苷键相连的分支结构。淀粉的酶解法主要是将淀粉置于高温下,通过耐热α-淀粉酶进行糊化,以产生一系列直链和支链糊精,随后用淀粉葡糖苷酶(AMG)定量水解为葡萄糖。在 ATP 和 NADP+ 存在下,使用葡萄糖氧化酶/过氧化物酶 (GOPOD) 试剂或己糖激酶/磷酸葡萄糖脱氢酶检测释放的葡萄糖。在一系列成功研究的基础上,该方法被采用为 AOAC 方法 996.11、AACC国际方法 76-13.01 和 ICC 方法 168。
本试剂盒在 AOAC 方法 996.11 的基础上进行了适当的优化,从而提供了一个更加方便用户的淀粉检测方法,即快速总淀粉检测法
 

The Total Starch (AA/AMG) test kit is used for the measurement and analysis of total starch in cereal flours and food products. This kit now contains an improved α-amylase that allows the amylase incubations to be performed at pH 5.0 (as well as pH 7.0).

Colourimetric method for the determination of Total Starch in
cereal products, feeds, foodstuffs and other materials

 

Principle:
(α-amylase, 100°C ± DMSO)
(1) Starch granules + H2O → maltodextrins

(amyloglucosidase)
(2) Maltodextrins + H2O → D-glucose

(glucose oxidase)
(3) D-Glucose + H2O + O2 → D-gluconate + H2O2

(peroxidase)
(4) 2H2O2 + p-hydroxybenzoic acid + 4-aminoantipyrine →
quinoneimine + 4H2O

Kit size: 100 assays
Method: Spectrophotometric at 510 nm
Total assay time: ~ 90 min
Detection limit: 1-100% of sample weight
Application examples:
Cereal flours, food products and other materials
Method recognition:
AOAC (Method 996.11), AACC (Method 76-13.01), ICC (Standard Method
No. 168), and RACI (Standard Method)

Advantages

  • Very competitive price (cost per test)
  • All reagents stable for > 12 months after preparation
  • Simple format
  • Mega-Calc™ software tool is available from our website for hassle-free raw data processing
  • Standard included

 

 

 Q1. Should the pH of the sample be adjusted even for samples in acidic media?

The pH of the assay solution after the sample is added should be the same as that of the assay buffer that is supplied with the kit.
Low sample volumes (e.g. 0.1 mL) are not likely to affect the pH of the assay solution and therefore may not require pH adjustment.
Samples above 0.1 mL are more likely to affect the pH of the assay solution and therefore the pH of these samples should be adjusted as described in the data booklet, prior to addition to the assay.

Q2. Why does the quadruplicate glucose control in your Total Starch Assay Kit have to be incubated?

We feel more comfortable with quadruplicate glucose controls.  If the control is incorrect, or questionable, then all the results are in doubt.

Q3. Why do duplicate samples have to be measured?

Duplicate samples do not have to be measured.  We just suggest this for laboratories starting up. 

Q4. Does the Regular Maize Starch need to be analysed with pre-treating by DMSO? How do you store this Enclosed Control?

The Regular Maize Starch does not require DMSO pre-treatment.  The value should be about 84% with a moisture content of about 12%, the final dry weight value is about 96-97%.  Store the sample at room temperature, dry.

Q5. There is an issue with the performance of the kit; the results are not as expected.

If you suspect that the Megazyme test kit is not performing as expected such that expected results are not obtained please do the following:

  1. Ensure that you have tested the standard sample that is supplied with the Megazyme test kit.
  2. Send the results of the kit standard, blank samples and the results obtained for your sample, in the relevant MegaCalc spreadsheet (if available) to Megazyme (cs@megazyme.com). Where available the relevant MegaCalc spreadsheet can be downloaded from where the product appears on the Megazyme website.
  3. State the kit lot number being used (this is found on the outside of the kit box).
  4. State which assay format was used (refer to the relevant page in the kit booklet if necessary).
  5. State exact details of any modifications to the standard procedure that is provided by Megazyme.
  6. State the sample type and describe the sample preparation steps if applicable.

Q6. Does your kit with DMSO solubilise starch that has been vitrified due to malting/kilning?

Yes.  We believe that the DMSO step will solubilise vitrified starch in malt.  Make sure that the malt is milled to pass a 0.5 mm screen.  You could vary the time of cooking with DMSO to check solubilisation (i.e. 5 minutes, 10 minutes, or even up to 1 hour).

Q8. Is it possible to use the Total Starch Kit to measure starch levels in plasterboard and related products?

There should be no problem in measuring the starch in plasterboard.  I suggest that you grind about 100 g in a kitchen blender and then fine mill to pass 0.5 mm screen. Run a standard assay, but adjust volume to 10 mL after alpha-amylase treatment. Keep a close check on the pH.  Plasterboard may push the pH value up (pH up to about 8 should be fine).  You may be advised to run a DMSO format concurrently just to be sure.  When you treat with amyloglucosidase, I would advise that you take 0.2 and 0.4 mL aliquots of digest (to get the colour up), also, be careful about checking the pH.

Q9. Can the Total Starch Kit be used for samples containing 20% fat or higher?

A 20% fat content could cause a problem for the method.  We suggest that the sample be defatted before analysis for starch.

Q10. Do you have any kit or procedures for the determination of extractable starch in corn? This is of particular concern in the corn wet milling. I presume that extractable starch in corn is not the same as total starch?

Our Total Starch Assay Kit could measure starch left in a residue, or starch extracted. No method could measure potential extractable starch, as this will depend on numerous factors, including processing equipment, conditions etc.

Q12. I wish to measure Total Starch in several products. These products contain 10-20% starch + maltodextrins at similar levels. Is it possible to remove the maltodextrins from the sample? Will ethanol work?

Most of the maltodextrins can be removed with 50% ethanol washing.  If the starch is not gelatinised, it can be washed with cold water.  This will remove all of the soluble maltodextrins, but the starch will spin down.  If the starch has been gelatinised, then the best material which can be used for washing is 50% ethanol.

Q14. Is the accuracy of the Total Starch test affected by the presence of other inorganic chemicals and ground calcium carbonate in pulp?

We think that calcium carbonate etc. will not cause any problems.  However, this of course depends on the amount present and if it changes the pH of the incubation mixture.

Q15. Does the Megazyme Total Starch method work well on all the new chemically modified starches that are now appearing, e.g. highly crosslinked, dextrinised and highly propylene oxide substituted?

The method will work for some chemically modified starches (e.g. crosslinked) however, if the degree of chemical modification is high, there will be an underestimation as the modification will interfere with complete hydrolysis to glucose and subsequent measurement.

Q16. Is it necessary to pre-wash ground cereal samples prior to analysis for Total Starch?

You only need to wash samples which you feel may contain glucose and/or maltodextrins, e.g. breakfast cereals.  There is little glucose in ground cereals, so it is not necessary to pre-wash these materials.

Q17. What is the stability of the enzymes from the Total Starch Kit?

The enzymes from this kit are stable at room temperature for at least 6 months.  At 4˚C, they are stable for several years.

Q18. Can the Total Starch Kit determine the degree of gelatinisation? Sample : Corn Flour.

The Starch Damage Kit may be best for this.  If the starch is gelatinised and dried before analysis the correct results for gelatinisation will not be obtained.

Q19. Are there any limits to the sensitivity of the Total Starch Kit?

The Total Starch Kit can accurately measure starch levels as low as 1% w/w.

Q20. What is the sensitivity and how much is the absorbance of glucose standard (100 micrograms)?

The absorbance for 100 micrograms of glucose (in 3 mL of GOPOD Reagent) is about 0.97.

Q21. Is it possible to raise sensitivity by modifying dilution of GOPOD reagent?

Yes, you can reduce the volume of GOPOD to 1 mL and use micro cuvettes.  This will increase sensitivity by ~ 3-fold.

Q22. Does DMSO solubilise resistant starch, i.e. crystallised amylose and amylopectin?

DMSO does solubilise resistant starch (crystallise amylose and amylopectin).  The only starch material we have had problems in dissolving in DMSO is potato amylose.

Q23. When analysing samples containing sugars, an 80% v/v solution of ethanol is used to solubilise and remove the sugars. About how large are the smallest dextrins that are left in the starch (not solubilised) in this treatment?

We believe that for starch fragments, oligosaccharides of a DP up to 10 would be soluble in 80% alcohol.  The degree of solubility of other oligosaccharides would depend on the sugar type and linkage type.

Q24. What is the sensitivity of the Total Starch Method for measurement in liquids containing low levels of starch?

The Total Starch Kit can be used for liquids containing as little as 200 micrograms per mL with some adjustments of conditions, as below:
Mix 0.5 mL of sample with 0.5 mL of 100 mM sodium acetate buffer (pH 4.5). Incubate at 40˚C and add 0.1 mL of Amyloglucosidase and incubate for 30 minutes.  Add GOPOD reagent as usual.  You will need to run an AMG blank as this enzyme preparation contains a very small amount of glucose.

Q25. AMYLOGLUCOSIDASE. The activity is stated as being 3260 U/mL (Soluble Starch). How was this determined?

The AMG activity was determined with soluble starch as substrate (10 mg/mL) in 0.1 M sodium acetate buffer at pH 4.5 and 40˚C.  One Unit is the amount of enzyme required to hydrolyse one micromole of maltose per minute (i.e. to release 2 micromoles of glucose).  Glucose release is measured with Glucose Determination Reagent.

Equl爱尔兰Megazyme 淀粉总量检测试剂盒操作视频 中文版(K-TSTA) 高清版

Megazyme 溶解淀粉 操作视频

Megazyme 试剂盒样品前处理准备操作视频

 
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淀粉总量检测试剂盒 K-TSTA-100A
淀粉总量检测试剂盒 K-TSTA-100A

100 assays per kit

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