Guaiacol Detection Kit - Cosmo Bio Co.,Ltd.

Antibodies

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Guaiacol Detection Kit

Catalog No.: KYO-08921
Size: 100TEST
Price: ¥25000
$334
catalog info: Catalog 2012-p155
Storage: 4C
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Purpose: Detection of guaiacole
component: Reagent 1: 50 mM Potassium hydrogen phthalate Buffer 60 mL x 2 bottles
Reagent 2: 1.3 % hydrogen peroxide solution 2.5 mL x 1 tube
Reagent 3: Peroxidase-Phosphoric Acid Buffer 2.5 mL x 1 tube
Positive Control: Guaiachole (1050 ppm) 2.5 mL x 1 tube
[Weight] 300g
[Size] 8.5 x 14.5 x 10cm

Other:

 Background 
Alicyclobacillus acidoterrestris is a gram-positive, spore-forming bacillus that occurs widely in nature. This species have been studied substantially after the incident of turbidity of transparent apple juice in 1984. The Alicyclobacillus genus bacteria, which peculiarly grow under relatively high temperature and acid conditions, are called Thermo-acidophilic Bacilli (TAB) (1, 2, 3).
TAB are not destroyed by pasteurization and may remain in final products of fabricated food and beverages. TAB are not known to be harmful to health but are known to degrade the quality of juices and other products by producing guaiacol, causing off-flavor. Even without the off-flavor problem, product values may be degraded by slight deterioration during distribution.
It is very difficult to avoid contamination of TAB, which are common in soil. Fortunately, however, A. acidocaldarius and other main species that cause contamination are known to be harmless, and damage caused by TAB can be minimized by monitoring the contamination of A. acidoterrestris, which is known to be harmful. This species, however, cannot be detected by usual isolation culture methods because it grows only under acid and relatively high temperature conditions.
Because of frequent occurrence of fruit juice deterioration incidents in Europe and other regions, Japan Fruit Juice Association developed and publicized a unified test method for thermo-acidophilic bacilli in March 2003(4). To alloweasy performance of this test method, we developed a test kit to differentiate A. acidoterrestris in terms of productivity of guaiacol, which causes off-flavor.
[Scheme of the unified test method for TAB]
TAB test:
TAB are detected selectivelybyculturing in an acid medium (pH 3.7}1) at temperatures best suited for their growth.
A. acidoterrestris differentiation:
After the TAB test, A. acidoterrestris is differentiated from the other Alicyclobacillus species by using either of thefollowing methods: 1) The differential growth temperature method, using differences in breeding ability at best suited temperatures, and 2)The peroxidase method, using tetraguaiacol production.
*This product adopts the peroxidase method to differentiate A.acidoterrestris from TAB. The method allows quick measurement of productivity of guaiacol, which causes off-flavor.
*Direct testing after the preliminary culture is also under consideration.
[Method]
1. Take with single inoculation loop,10 ul ,of young colony that has been isolated on YSG agar plate and that has not formed spores. Suspend the colony in a tube of ''Va-YSG medium (2 ml ), '' (†1 x 108 cfu/ ml ).
2. Incubate at 45 } 1‹C for 3 hours.
3. Add 1.0 ml of Reagent1of ''GuaiacoleDetection Kit''.
4. Add 20 ul of Reagent 2 of ''Guaiacole Detection Kit''.
5. Add 20 ul of Reagent 3 of ''Guaiacole Detection Kit''.
6. Mix well.
7. Let it stand for 5 - 10 minutes.
8. Check the color change of the solution.
*Readthe resultwithin 10 minutes
*Blank (Negative Control)
Use ''Va-YSG medium (2 ml )'' that has not been inoculated and follow steps 2 - 8.
*Positive Control
Mix well 2 ml of distilled water and 100 ul of Positive Control. Follow steps 3 - 8.
[Interpretationof Results]
Darker brown than blank = Positive *If the color change is not significant and difficult to determine, retest with 24 hours incubation instead of 3 hours, step 2.
Same color as blank = Negative
Verify the test results bycheckingthereading of positive control; positive control result should have significantly darker brown than blank.
[Handling instructions]
Please dispose used medium after high pressure sterilization(121‹C for more than 30 min)with appropriatecare.Pleasestrictly keep the storage condition (2 - 10‹C Celsiusand and dark place).
Expiration date : 12 month from date of manufactre.




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Scheme of TAB Test - Japan Fruit Juice Association -

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A.acidoterrestris Kindly provided by Mr. K. Goto. (MITSUI NORIN CO.,LTD.)

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Brown color indicates contamination of AAT 1.Negative Control 2. A. acidoterrestris B2067 3. A. acidoterrestris ATCC49025T 4. A. acidocaldarius ATCC27009T 5. A. acidocaldarius HP2 6. A. genomic sp. P2 7. A. genomic sp. DSM11983

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Experimental Procedure

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Reference:

•  G. Deinhard, et al.,_ Bacillus acidoterrestris sp. nov., Syst. Appl. Microbiol. 10, 47-53, 1987
•  GOTO, M., Bokin Bobai, 28 (8), 499-508, 2000 (in Japanese)
•  Niwa, M., Lecture on ''Classification and identification of microbes'' January 2001 issue of Soft Drink TechnicalData, 9-26, 2001 (in Japanese)
•  Niwa, M., et al., Fruit processing 13 (2) 102-107, 2003
•  Japan Fruit Juice Association, On theunifiedtest method for thermo-acidophilic bacilli, Report of Japan Fruit Juce Association, 535, 4-12, 2003 (in Japanese)
•  Niwa. M., et al., A. acidoterretris rapid detection kit, Fruit processing 13 (5) 328-331, 2003
•  Niwa, M., Control of deleteriousbacteria in acidic beverages by using a guaiacol detection kit (peroxidase method), Japan Food Science, 2004-2, 23-28, 2004 (in Japanese)
•  Japan Laid-Open P2003-259A ''Detection of guaiacol forming bacteria and/or its identification method''
◊  Keiichi Goto, Kaoru Mochida, Yuko Kato, Mika Asahara, Rieko Fujita, Sun-Young An, Hiroaki Kasai, and Akira Yokota Int J Syst Evol Microbiol, Jun 2007; 57: 1276 - 1285.
 
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