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Improved method for preparation of inoculum suspension for ino-inositol test

A technology of bacterial suspension and inositol, which is applied in biochemical equipment and methods, microorganism-based methods, and microbial determination/inspection, etc., can solve problems such as inability to complete, poor correlation, and low absorbance, and achieve good linear correlation. , the effect of high accuracy and large correlation coefficient

Inactive Publication Date: 2016-03-23
GUANGZHOU INST FOR FOOD INSPECTION(GUANGZHOU INSPECTION CENT FOR WINE & SPIRITS)
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  • Description
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Problems solved by technology

[0003] In the existing method for assaying myo-inositol, the quality of the bacteria is very important, because a certain amount of myo-inositol is accumulated in the ino-inositol inoculated bacteria, which interferes with the normal growth of microorganisms and makes the stability of the measurement results poor, especially when the ino-inositol is inoculated. When the inositol content in the test object is low, the gradient of the standard curve is not obvious, and the absorbance of the high-concentration inositol standard working solution is lower than the absorbance of the low-concentration inositol standard working solution. The correlation is poor or the standard curve cannot be completed, which directly affects the accuracy of the test results

Method used

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  • Improved method for preparation of inoculum suspension for ino-inositol test
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  • Improved method for preparation of inoculum suspension for ino-inositol test

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Embodiment 1

[0024] refer to figure 1 A method for detecting the content of inositol in food by microorganisms, comprising the following steps.

[0025] Step A, resuscitating the myo-inositol inoculum to obtain the resuscitated strain.

[0026] The inositol inoculum is grape juice yeast, which is activated and inoculated on the slant medium of malt extract powder agar, cultured at 30° C. for 16 hours, and then transferred for 2-3 generations to obtain the revived strain.

[0027] Step B, preparing the inoculum base solution.

[0028] Inoculate the resuscitated strain of step A into the inositol assay medium and continue to cultivate. After culturing at 30°C for 16 hours, the inoculum base solution is obtained. The inositol assay culture medium is 5 mL of inositol test medium and 5 mL of water. For the composition and preparation method of the inositol test medium, refer to the national food safety standard GB5413.25-2010 "Determination of inositol in food and dairy products for infants a...

Embodiment 2

[0065] The detection method of the present embodiment is different from embodiment 1 in that:

[0066] The absorbance of the inoculum base solution detected in step C is 0.2, and the inoculum base solution with an absorbance of 0.2 is taken to proceed to subsequent steps D and E.

[0067] Step E1, the standard curve made see image 3 .

[0068] Step E2. Weigh 2 samples, sample 1 has a mass of 2.0563 g, and sample 2 has a mass of 2.0095 g, and measure the absorbance of the liquid to be tested. The results are shown in Table 7.

[0069] Step E3, according to the absorbance of the liquid to be tested in Table 7, from image 3 The concentration of myo-inositol in the test solution was found in the shown standard curve, and the results are shown in Table 7. The calculated content of myo-inositol in the sample was 38.7mg / 100g.

[0070] Sample 1: m=2.0563g, Sample 2: 2.0095g,

[0071]

[0072] In order to test the reliability of the detection, the accuracy of the standard curv...

Embodiment 3

[0088] When the absorbance of the inoculum base liquid obtained after step A to step C of Example 180%, which is far from meeting the range of McFarland turbidity of 0.55-0.65, and the light transmittance required by the national standard In the range of 60%-80%, the concentration of the inoculated strain cannot meet the requirements of the test, and the inositol in the sample cannot be fully utilized, and accurate quantification cannot be performed. Therefore, the inoculum base solution with absorbance < 0.2 was discarded.

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Abstract

The present invention proposes an improved method for preparing an ino-inositol test inoculum suspension, comprising the steps of: A, resuscitating the inoculum inoculum to obtain the resuscitated strain; B, inoculating the resuscitated strain into the culture medium for inositol determination, and obtaining the inoculum Bacteria-based solution; C. The absorbance of the inoculated bacteria-based solution is detected, and the inoculated bacteria-based solution with an absorbance of 0.2 to 0.4 is taken for subsequent steps; D. The inoculated bacteria-based solution is placed in a sterile centrifuge tube, and after centrifugation and cleaning, Add 0.9% sterile saline, oscillate and mix to prepare a concentrated bacterial suspension; select an appropriate amount of concentrated bacterial suspension and add it to 0.9% sterile saline, and control the concentration within the range of 0.55-0.65 McFarland turbidity to obtain the inoculum Suspension; E. Take the inoculum suspension for testing the inositol content. In step C of the present invention, the inoculum base liquid whose absorbance is greater than 0.4 is replaced by the recovered strain in step B, and steps B to E are carried out again. The invention has the characteristics of easy operation and high accuracy.

Description

technical field [0001] The invention relates to a method for determining the content of inositol, in particular to a microbial determination method for the content of inositol in foods and dairy products. Background technique [0002] The existing method for utilizing microorganisms to determine the content of inositol mainly includes the following steps: 1. resuscitating the inoculated bacteria with inositol to obtain the resuscitated strain; 2. transferring the resuscitated strain to newly prepared broth or slant medium, Cultivate at the specified temperature for 16h-24h; 3. Take the broth culture directly into a sterile centrifuge tube, or take the slant culture into a centrifuge tube filled with 0.9% sterile saline for centrifugation, washing, and adding 1mL of 0.9% Shake and mix to prepare a concentrated bacterial suspension; select an appropriate amount of concentrated bacterial suspension and add it to 10mL of 0.9% sterile saline to prepare an inoculum suspension, and...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/04C12R1/645
Inventor 肖剑刘冬豪戚平梁智安易云婷
Owner GUANGZHOU INST FOR FOOD INSPECTION(GUANGZHOU INSPECTION CENT FOR WINE & SPIRITS)
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