CN102586391B - Method for determining colony count of pathogenic microbes in dehydrated garlic product - Google Patents
Method for determining colony count of pathogenic microbes in dehydrated garlic product Download PDFInfo
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- CN102586391B CN102586391B CN 201210043206 CN201210043206A CN102586391B CN 102586391 B CN102586391 B CN 102586391B CN 201210043206 CN201210043206 CN 201210043206 CN 201210043206 A CN201210043206 A CN 201210043206A CN 102586391 B CN102586391 B CN 102586391B
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Abstract
The invention discloses a method for determining colony count of pathogenic microbes in dehydrated garlic product. The method comprises taking garlic powder sample, adding sterilized diluent, homogenizing, filtering with homogenizing bag equipped with filter membrane, sucking filtrate, transferring into two centrifuge tubes, adding polyacrylamide cationic resin as flocculant, centrifuging, removing supernatant, adding sterile distilled water into precipitate, mixing, and counting bacteria in the precipitate according to national standard GB 4789.2-2010. The invention has fast detection, simple procedure, and precisely detects colony count in bacteria inhibitive samples such as dehydrated garlic in 3 hr.
Description
Technical field
The invention belongs to the microorganism detection technical field, be specifically related to a kind of method that adopts supercentrifugal process to separate and measure colony count of pathogenic microbes in dehydrated garlic product.
Background technology
The food sanitation and safety problem is healthy closely bound up with everyone, always by countries in the world government and main flow medium, is paid close attention to.In recent years along with people are more and more higher to the attention rate of food safety, particularly at China's food safety Frequent Accidents in recent years, under the overall background that the Safety of Food Quality event relevant with the Chinese exports product is reported in media often, importer is more and more stricter to the food inspection from Chinese.China is maximum in the world garlic (Allium sativum L.) and production of articles and export State, has become China first place and has exported the vegetables of exchanging, and its outlet increases year by year.Wherein the dehydrated garlic goods are exactly the large agricultural exports of Anhui Province's tradition, product main exit Japan and Korea S, European & American Market, and export volume is sure to occupy national prostatitis always.Strengthen the supervision of outlet dehydrated garlic goods sanitary inspection and not only be related to the single product outlet, protect especially the trade of the agricultural-food whole industry normally to carry out.Yet the dehydrated garlic goods do not have the related microorganisms inspecting standard to put into effect so far, use for a long time the check of food microorganisms GB4789 series standard, but because the dehydrated garlic goods have unique product performance, in actual the detection, expose a large amount of problems, show significantly and be not suitable for.There is no suitable inspecting standard to guaranteeing quality product, consumer safety, the very large hidden danger of the guarantee smooth and easy formation of export of farm produce trade.Therefore carry out the research and development of safety and sanitation detection technique of dehydrated garlic goods for guaranteeing that Agricultural Products Trade safety and human consumer's edible safety all have great significance.
The dehydrated garlic goods are mainly garlic powder, garlic grain and garlic sheet, are first-class seasonings and healthy food materials, and this product is selected the high-quality garlic, through multiple working procedures such as peeling, cleaning, oven dry, selected, abrasive dust, packings, process.Moisture content, lower than 6%, is not added any foodstuff additive, does not add sanitas.Research shows as protective foods, for maldigestion, flu, respiratory tract infection, digestive tract ulcer, Hyperlipidemia, arteriosclerosis, cancered grownup and middle-aged and old heart trouble, cardiovascular and cerebrovascular diseases person; The immunocompromised person all has the effect of assisting therapy.The garlic powder has broad-spectrum antimicrobial and antibacterial, germicidal action, can effectively suppress growth, the breeding of harmful mushrooms such as intestinal bacteria, Salmonella, and livestock and poultry are infected to respiratory tract, digestive tract diseases and the complication significant curative effect thereof caused because of Gram-negative and positive bacteria.But produce the amino acid whose formation of fragrance in garlicin effective stimulus animal body, increase the flavour ingredient in the meat egg, make meat egg delicate flavour more beautiful.Add in feed and can improve livestock and poultry and fishes and shrimps resistance against diseases, accelerate the speed of growth.Pest clearly, removing toxic substances, function promoting blood circulation and removing blood stasis are arranged, can significantly reduce the toxicity of active substance in feed.
The dehydrated garlic goods garlicky strong, pure, do not develop immunity to drugs, noresidue in nontoxic, body.Exactly because the good moral character of dehydrated garlic goods, used more and more widely and consume, strengthen its hygienic safety research is just seemed to important all the more.The production of dehydrated garlic goods will be passed through a plurality of links, and wherein all there is the possibility of microbial contamination and breeding in a lot of links.Are so why, present GB 4789 series standards not suitable for the safety and sanitation detection of dehydrated garlic goods? because contain a kind of natural phant fungistat---garlicin in garlic, garlicin is the general name of sulfocompound in garlic bulblet and garlic essential oil, there is (pressing down) the bacterium effect of killing in various degree, be described as natural bactericidal agent, have and report that the ability of its sterilization is almost 100 times of penicillin.1/80000 garlicin can suppress or kill staphylococcus, suis, typhoid fever, dysentery bacterium etc.The garlic bulb extracting solution is to mould, head mold, aspergillus, the wooden mould obvious restraining effect that has.
Summary of the invention
In order to ensure Agricultural Products Trade safety and human consumer's edible safety, the invention provides a kind of method of separating quickly and efficiently and measuring colony count of pathogenic microbes in dehydrated garlic product.
Concrete operation step of the present invention is as follows:
(1) get garlic powder 25g, add 225mL sterilizing diluent to mix, obtain mixed solution;
The configuration of described sterilizing diluent: the potassium primary phosphate that takes 34.0g is dissolved in 500mL distilled water, and regulating the pH value with the 175mL sodium hydroxide solution of concentration 1mol/L is 7.2, with distilled water diluting, to the 1000mL constant volume, obtains mother liquor; Get the 1.25mL mother liquor, with distilled water diluting, to 1000mL, 121 ℃ of autoclavings of temperature 15 minutes, obtain the sterilizing diluent;
(2) homogenizing bag of mixed solution band filter membrane is filtered, described filter membrane aperture is 330 μ m, diameter 50mm, draws clear liquid, and the 1.5mL clear liquid is sub-packed in 2 2mL centrifuge tubes, and the 0.75mL clear liquid is housed in every centrifuge tube;
(3) add the coagulant aids of polyacrylamide resin cation (R.C.) in every centrifuge tube of clear liquid is housed, in clear liquid, the concentration of coagulant aids is 0.1ppm; Under 4 ℃ of temperature, 20000 rev/mins of conditions of rotating speed centrifugal 40 minutes, remove supernatant liquor, in throw out, add the 1.5mL sterile distilled water to mix, obtain test solution, by GB GB 4789.2-2010, test solution is carried out to the amount of bacteria counting.
Useful technique effect of the present invention: centrifugal separation has solved the impact of garlicin bacteriostatic action on the total number of bacterial colony counting in the dehydrated garlic goods well, and counting is compared more accurate and effective with standard method.The present invention detects fast, step is simple, can accurately detect dehydrated garlic one class containing the total number of bacterial colony in the antibacterial substance sample at 3 hours.
Embodiment
Below in conjunction with embodiment, the present invention is further described.
Embodiment 1
1 material and instrument
1.1 material
The Salmonellas bacterial classification (
salmonella) by Beijing Lan Borui company, provided, various substratum are provided by Beijing road and bridge company, Ciba series food grade polyacrylamide resin cation (R.C.), and sample keeps sample from Anhui Dong Qian Food Co., Ltd submitted sample;
1.2 instrument
Table-type high-speed refrigerated centrifuge (BECKMAN 64R), vertical and high-speed refrigerated centrifuge (BECKMAN J30I), micro sample adding appliance: 0.5 ul～10ul, 10 ul～100ul, high-pressure sterilizing pot.
2 degerming rate experiments
Take 25g dehydrated garlic product sample (irradiation sterilization), a Maxwell unit (3.5 * 10 of artificial quantitatively interpolation
8individual/g) Salmonellas bacterial classification, add 225mL sterilizing diluent homogeneous, and with the homogenizing bag filtration of filter membrane, the filter membrane aperture is 330 μ m, diameter 50mm; Draw clear liquid, the 1.5mL clear liquid is sub-packed in 2 2mL centrifuge tubes, the 0.75mL clear liquid is housed in every centrifuge tube; In every centrifuge tube of clear liquid is housed, add a small amount of polyacrylamide resin cation (R.C.) to make coagulant aids, in clear liquid, the concentration of coagulant aids is 0.1ppm; Centrifugation under 4 ℃ of temperature, 20000 rev/mins of conditions of rotating speed, draw supernatant liquor, by GB GB 4789.2-2010, measures residual bacterial count in supernatant liquor.
The configuration of sterilizing diluent: the potassium primary phosphate that takes 34.0g is dissolved in 500mL distilled water, and regulating the pH value with the 175mL sodium hydroxide solution of concentration 1mol/L is 7.2, with distilled water diluting, to the 1000mL constant volume, obtains mother liquor; Get the 1.25mL mother liquor, with distilled water diluting, to 1000mL, 121 ℃ of autoclavings of temperature 15 minutes, obtain the sterilizing diluent.
Table 1 supercentrifugal process degerming rate experimental result
| Centrifugation time (min.) | The clear liquid amount of bacteria | The degerming rate |
| 0 | 3.5×10 7 | / |
| 20 | 4.3×10 5 | 98.77 |
| 40 | 3.6×10 5 | 98.97 |
| 60 | 2.1×10 5 | 99.4 |
| 80 | 8.9×10 4 | 99.75 |
| 100 | 3.8×10 4 | 99.89 |
By upper table 1 high speed centrifugation bacterium effect experimental result, can be found out, in sample liquid thalline through 20000 rev/mins, more than 40 minutes after high speed centrifugation, the thalline more than 99% can separate from sample liquid.The mensuration that can be used for total number of bacterial colony, can meet the demand of food hygiene detection fully.
Embodiment 2
1 material is identical with embodiment 1 with instrument
2 supercentrifugal process are measured and the traditional method contrast for total number of bacterial colony
Get garlic powder sample 25g, add 225mL sterilizing diluent, homogeneous filters by the homogenizing bag with filter membrane, draws clear liquid, and packing 1.5mL is in 2 2mL centrifuge tubes; In every centrifuge tube of clear liquid is housed, add a small amount of polyacrylamide resin cation (R.C.) as coagulant aids, in clear liquid, the concentration of coagulant aids is 0.1ppm; Under 4 ℃ of temperature, 20000 rev/mins of conditions of rotating speed centrifugal 40 minutes, remove supernatant liquor, add the 1.5mL sterile distilled water and mix in throw out, measure the amount of bacteria in throw out by GB GB 4789.2-2010, Application standard method contrast simultaneously; Other condition is with embodiment 1.
Table 2 supercentrifugal process is measured with the traditional method result and is compared for total number of bacterial colony
In upper table 2, from practice the method and standard method, compare, measuring the result of total number of bacterial colony in actual dehydrated garlic goods can find out simultaneously, at 1000 times below dilution, the obvious reliable use of having disturbed standard method of the antipathogenic composition in garlic powder sample liquid, national standard method is subject to severe jamming, is difficult to accurate judgement.While only being diluted to more than 1000 times, result is just credible, and supercentrifugal process has solved this problem well.
Claims (1)
1. measure the method for colony count of pathogenic microbes in dehydrated garlic product, it is characterized in that operation steps is as follows:
(1) get garlic powder 25g, add 225mL sterilizing diluent to mix, obtain mixed solution;
(2) homogenizing bag of mixed solution band filter membrane is filtered, described filter membrane aperture is 330 μ m, diameter 50mm, draws clear liquid, and the 1.5mL clear liquid is sub-packed in 2 2mL centrifuge tubes, and the 0.75mL clear liquid is housed in every centrifuge tube;
(3) add the coagulant aids of polyacrylamide resin cation (R.C.) in every centrifuge tube of clear liquid is housed, in clear liquid, the concentration of coagulant aids is 0.1ppm; Under 4 ℃ of temperature, 20000 rev/mins of conditions of rotating speed centrifugal 40 minutes, remove supernatant liquor, in the throw out of every centrifuge tube, add the 1.5mL sterile distilled water to mix, obtain test solution, by GB GB4789.2-2010, the test solution of two centrifuge tubes is carried out respectively to the amount of bacteria counting;
The configuration of described sterilizing diluent: the potassium primary phosphate that takes 34.0g is dissolved in 500mL distilled water, and regulating the pH value with the 175mL sodium hydroxide solution of concentration 1mol/L is 7.2, with distilled water diluting, to the 1000mL constant volume, obtains mother liquor; Get the 1.25mL mother liquor, with distilled water diluting, to 1000mL, 121 ℃ of autoclavings of temperature 15 minutes, obtain the sterilizing diluent.
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| CN 201210043206 CN102586391B (en) | 2011-06-29 | 2012-02-24 | Method for determining colony count of pathogenic microbes in dehydrated garlic product |
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| CN201110177983.2 | 2011-06-29 | ||
| CN201110177983 | 2011-06-29 | ||
| CN 201210043206 CN102586391B (en) | 2011-06-29 | 2012-02-24 | Method for determining colony count of pathogenic microbes in dehydrated garlic product |
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| CN102586391B true CN102586391B (en) | 2013-12-18 |
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