KR101757175B1 - Method for detecting or quantifying starch in processed meat foods - Google Patents

Method for detecting or quantifying starch in processed meat foods Download PDF

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KR101757175B1
KR101757175B1 KR1020150155994A KR20150155994A KR101757175B1 KR 101757175 B1 KR101757175 B1 KR 101757175B1 KR 1020150155994 A KR1020150155994 A KR 1020150155994A KR 20150155994 A KR20150155994 A KR 20150155994A KR 101757175 B1 KR101757175 B1 KR 101757175B1
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starch
solution
processed meat
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phenol
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한상현
오홍식
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제주대학교 산학협력단
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    • G01N31/22—Investigating or analysing non-biological materials by the use of the chemical methods specified in the subgroup; Apparatus specially adapted for such methods using chemical indicators
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    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
    • G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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    • G01N33/02—Food
    • G01N33/12—Meat; Fish
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2400/00—Assays, e.g. immunoassays or enzyme assays, involving carbohydrates
    • G01N2400/10—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • G01N2400/12—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar
    • G01N2400/14—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar alpha-D-Glucans, i.e. having alpha 1,n (n=3,4,6) linkages between saccharide units, e.g. pullulan
    • G01N2400/16—Starch, amylose, amylopectin

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Abstract

The present invention relates to a method for detecting starch in processed meat products, a method for quantification, and an assay kit for the detection or quantification of starch in processed meat products. Using the starch detection or quantification method of the present invention, starch present in processed meat products can be specifically detected by distinguishing it from animal carbohydrates of meat components, and quantitative analysis with high accuracy is possible. Therefore, the present invention can be usefully used for the inspection of the amount and content of the extender of processed meat products.

Description

TECHNICAL FIELD The present invention relates to a method for detecting starch in processed meat products,

The present invention relates to a method for detecting starch in processed meat products, a method for quantification, and an assay kit for the detection or quantification of starch in processed meat products.

Food processing refers to a processing process that changes its shape, physical, chemical, sensory and nutritional characteristics without changing the essence of the food material. Advantages of processing food include increased processability, increased diversity, increased complexity, increased sensory value, increased product value, and increased shelf life. Disadvantages include reduced natural characteristics and processing costs. The types of food processing are classified into animal processed food and vegetable processed food depending on the sorting method, and they are roughly divided into agricultural products processing, livestock product processing, and aquatic product processing depending on the kinds of materials. Sausage, milk, ham, cheese , Butter, and ice cream, and processed marine products include soft products, dried products, smoked products, and canned foods. Among them, processed meat products are processed foods made from meat raw materials, and soybean protein and starch are used as an increasing agent in addition to animal main ingredients such as pork, chicken meat, and fish meat.

Extender means a food additive that does not change its properties but only increases in quantity. Processed meat products are required to show not only the content and content of pork, chicken meat, fish meat, but also the kind and content of soybean protein and starch. However, since it is difficult to distinguish starch from animal carbohydrates such as glycogen contained in meat ingredients in processed meat products, a technique capable of selectively detecting only starch in processed meat products has not been known so far.

Conventionally, techniques for distinguishing starch origin from a food or a starch mixture have been developed (Patent Document 1). However, in the past, starch-based processed foods such as sweet potato starch, tapioca starch, corn starch This method is used to distinguish starch-derived species using a mass spectrometer.

Accordingly, the inventors of the present invention have developed a method for selectively detecting and quantifying starch only in processed meat products, thereby completing the present invention.

Korean Registered Patent No. 1343981

It is an object of the present invention to provide a method of detecting starches in processed meat products.

It is another object of the present invention to provide a method for the determination of starch in processed meat products.

It is another object of the present invention to provide an assay kit for detecting or quantifying starch in processed meat products.

One embodiment of the present invention provides a method of detecting starches in processed meat products.

More specifically, the starch detection method in the processed meat product may comprise the following steps:

1) adding a first solution containing sucrose, saline sodium citrate (SSC), ethylenediamine-tetraacetic acid (EDTA) and sodium dodecyl sulfate (SDS) to a sample;

2) adding and mixing a second solution containing Proteinase K to the mixed solution prepared in step 1);

3) diluting the precipitate obtained by centrifuging the mixed solution prepared in the step 2) by adding distilled water;

4) adding a third solution containing phenol to the mixed solution diluted in step 3); And

5) adding phenol-sulfuric acid to the mixed solution prepared in step 4) by adding sulfuric acid.

As used herein, the term "processed meat product" refers to a food which has been modified to be more delicious and easier to eat by taking advantage of the characteristics of livestock products and aquatic products, which are raw materials of foods, The meat can be processed foods mainly made of meat (meat), for example, sausage, ham, canned meat, fish smoked products, and the like, but the kind thereof is not limited. Soybean protein and starch may be used as an extender as well as raw materials such as meat for manufacturing processed meat products.

As used herein, the term "starch", also referred to as starch, is a polysaccharide composed of glucose (glucose), which is synthesized by a plant and exists as starch particles in cells. Potatoes, corn, sweet potatoes, It means dry powder that has settled sediment.

As used herein, the term "detection" may mean confirming the presence or absence of a specific component in a sample.

Therefore, the term " starch detection method in processed meat product "means a method of discriminating the presence of starch by distinguishing it from an animal carbohydrate such as glycogen contained in the meat component of the product in the processed meat product.

In step 1) above, the sample may be taken from the processed meat product, and may be collected from one or more portions of the processed meat product, but is not limited to this and is expected to contain both meat and starch Any sample can be used.

The sucrose contained in the first solution can be decomposed and fractionated by processed sucrose as a buffer solution, and the cell organelles can be fractionated when the cellular sample is dissolved, It is possible to store biomolecules in an aqueous solution. In addition, the sucrose may absorb glucose present in the cell, causing the cell to collapse and collapse by osmotic pressure.

SSC (Saline Sodium Citrate) contained in the first solution is a buffering solution and is a kind of cation chelating agent. SSC can bind to and hydrate cations in processed meat products. SSC includes sodium chloride and sodium citrate. In one embodiment of the invention, SSCs include, but are not necessarily limited to, 150 mM sodium chloride and 15 mM sodium citrate. The concentration of sodium chloride (NaCl) contained in the SSC can adjust the osmotic pressure in solution, containing sodium citrate, which (Sodium citrate) in the SSC can function sequestering agent for Ca + 2. In the above specific example, SSC may be used instead of SSC if it is a buffer capable of acting as a cationic sequestrant in a hypotonic solution state. For example, Tris-HCl, EDTA and the like may be used.

EDTA (Ethylene-Diamine-Tetraacetic acid) contained in the first solution is a chelating agent that reacts with cations. EDTA can block the activity inhibition of enzymes such as proteinase K by binding to cations in processed meat products during sample decomposition. In the above-mentioned embodiments, components which can perform a similar function to EDTA can be used in place. Examples thereof include ethylene glycol tetraacetic acid (EGTA), ethylenediamine-N, N'-disuccinic acid (EDDS), polyaspartic acid acid, IDS (Iminodisuccinic acid), and the like.

Sodium Dodecyl Sulfate (SDS) contained in the first solution is a kind of detergent, which can denature protein components in processed meat products. SDS is known to exhibit the strongest binding force with proteins. When about one SDS is bonded to two amino acids constituting a protein, the amino acid has a high charge (-) charge, and a high-order structure Tertiary or quaternary structure) can be completely collapsed and denatured into linear molecules. And thus can be easily hydrolyzed by a protein hydrolyzing enzyme such as proteinase K or the like.

The first solution may comprise 20 to 30% (w / v) sucrose, 1 x SSC, 1 to 10 mM EDTA and 0.5 to 2% (w / v) SDS, / v) sucrose, 1 x SSC, 2 mM EDTA, and 1% (w / v) SDS. The concentration range of the components contained in the first solution is such that the intracellular protein and the water-soluble carbohydrate derived from the animal raw material are easily dissolved. If the concentration is outside the range, the dissolution of the sample may take more time or may not be completely dissolved.

As used herein, the term "% (w / v) " means the number of solutes dissolved in 100 ml of solution. For example, 26% (w / v) sucrose means that 26 g of sucrose is dissolved in 100 ml of the first solution.

The first solution may be added at one time and divided into several portions. For example, a small amount of the first solution may be added to the sample to crush the sample, and then the first solution may be added again.

The step 1) may be carried out to dissolve intracellular proteins and water-soluble carbohydrates derived from an animal raw material.

Proteinase K contained in the second solution is a kind of serine endopeptidase among protein hydrolyzing enzymes and can decompose proteins of meat components in processed meat products. Proteinase K maintains its activity over a very wide range of conditions, generally maintained at a pH of 4 to 12, particularly when SDS is present. Therefore, it can be used simultaneously with SDS in the process of hydrolyzing cells or tissues.

The second solution may contain 1 to 10% (w / v) of Proteinase K, specifically 1 to 5% (w / v) of Proteinase K, And 2% (w / v) of Proteinase K, but not always limited thereto.

The addition of the second solution in the step 2) can be carried out at a volume ratio of the mixed solution: the second solution = 1000: 0.5 to 2 prepared in the step 1), specifically, the addition of the second solution is carried out in the step 1) The second solution = 1000: 1, for example, 40ul of the second solution may be added to 40 ml of the mixed solution prepared in step 1), but the present invention is not limited thereto. Wherein the concentration of SDS in the first solution is within 0.5-2% (w / v), and the concentration of proteinase K in the second solution is 1-2% (w / v) If the concentration range is out of the range, the protein hydrolysis ability may be deteriorated.

The mixing of the step 2) may be performed by mixing the mixture in a thermostatic chamber at a specific temperature of 55 to 65 ° C for 6 to 20 hours, and specifically, culturing the mixture at a temperature of 60 to 65 ° C for 7 to 10 hours For example, by mixing in vortexing at intervals of 1 to 2 hours while culturing in a 65 ° C thermostatic chamber for 8 hours, but the present invention is not limited thereto. The incubation time can be controlled by judging the dissolution state of the sample. However, the incubation temperature is 55 to 65 ° C, which is the optimum activation temperature of the proteinase K. If the temperature is outside the above range, the activity of proteinase K may be decreased have.

The step 2) is performed to decompose the sample and hydrolyze the protein.

In step 3), the centrifugation may be performed at 1,000 to 1,500 rpm at 1 to 10 ° C for 5 to 20 minutes, for example, at 1,200 rpm at 4 ° C for 10 minutes, but is not limited thereto.

The method may further include the step of adding PBS (Phosphate buffer saline) to the precipitate and centrifuging the distillate before adding the distilled water in the step 3). The centrifugation can be performed at 3 to 3,500 rpm at 1 to 10 ° C for 5 to 20 minutes, for example, at 3,200 rpm at 4 ° C for 10 minutes, but is not limited thereto.

And adding PBS to the precipitate before adding the distilled water in the step 3).

The step 3) may be carried out to recover undissolved starch components.

When the reducing sugar dehydrated by the sulfuric acid treatment becomes furfural (C 5 H 4 O 2 , Furan-2-carbaldehyde) and a derivative, phenol contained in the third solution can be induced. Lt; RTI ID = 0.0 > color. ≪ / RTI >

The third solution of step 4) may contain 4 to 6% (w / v) of phenol, specifically 5 to 6% (w / v) of phenol, Phenol may be contained in an amount of 5.5% (w / v), but the present invention is not limited thereto. When the concentration of phenol is less than 4% (w / v), sufficient coloring reaction may not be induced when the amount of starch is high. When the concentration of starch is higher than 6% (w / v) The calculation of the total content may be wrong.

The addition of the third solution in the step 4) may be carried out at a volume ratio of the mixed solution: the third solution = 1 to 10: 10-1 diluted in the step 3), specifically, the addition of the third solution is performed in the step 3 1: 1 in the mixed solution diluted in the first solution: 1: 1 in the diluted mixed solution: the third solution = 1: 1, and in the calculation of the absorbance measurement of the appropriate reaction of the starch and the phenol in the mixed solution, For example, 100 제 of the third solution may be added to 100 혼합 of the mixed solution diluted in step 3), but it is not necessarily limited thereto.

In step 5), sulfuric acid (H 2 SO 4 ) may be added in an appropriate amount to perform a phenol-sulfuric acid reaction.

If the reaction solution after the phenol-sulfuric acid reaction in step 5) is brown or yellow, the sample contains starch. If the reaction solution is blue, it can be judged that the sample does not contain starch.

Another embodiment of the present invention provides a method for the determination of starch in processed meat products.

More specifically, the starch determination method in the processed meat product may comprise the following steps:

1) adding a first solution containing sucrose, SSC, Ethylene-Diamine-Tetraacetic acid (EDTA) and sodium dodecyl sulfate (SDS) to a sample;

2) adding and mixing a second solution containing Proteinase K to the mixed solution prepared in step 1);

3) diluting the precipitate obtained by centrifuging the mixed solution prepared in the step 2) by adding distilled water;

4) adding a third solution containing phenol to the mixed solution diluted in step 3);

5) adding sulfuric acid to the mixed solution prepared in step 4) to react with phenol-sulfuric acid; And

6) Measure the absorbance at 490 ~ 495 nm and calculate the starch content against the standard calibration equation.

The above steps 1) to 5) may be carried out in the same manner as described in the method for detecting starches in processed meat products according to one embodiment of the present invention, and may include all of the above limitations.

The phenol-sulfuric acid reaction of step 5) may be by total carbohydrate assay. The phenol sulfuric acid method for quantifying total carbohydrates is described in Dubios, M. et al. (Dubios, M., Gilles, KA, Hamilton, JK, Rebers, PA, and Smith, F. (1956) Colorimetry Method for Determination of Sugars and Related Substances. "Analytical Chemistry 28, 350-356).

The absorbance of step 6) is preferably measured at 492 nm, but is not limited thereto.

The absorbance of the step 6) can be measured by the absorbance method.

The absorbance method used in the present invention means a conventional method of determining the concentration of the target component by measuring the degree of light absorption at a specific wavelength by changing the target component in the sample solution to a light absorbing substance with an appropriate reagent.

In the absorbance method of the present invention, the Lambert-Beer law is applied to determine the concentration of the target component. A typical absorption spectrophotometric apparatus usually comprises a light source section, a wavelength selector, a sample section, and a photometric section. Methods for measuring the concentration in the absorbance method include a colorimetric method using the degree of color development and a turbidimetric method using the degree of suspension of the sample solution. The absorbance analyzer used for absorbance measurement in the present invention is preferably a microplate, but is not limited thereto.

In the absorbance method, the reference solution corresponding to the sample solution is prepared, and the absorbance is measured. After the calibration curve is prepared, the absorbance of the sample solution is measured, and the concentration is measured by comparing with the calibration curve.

The calculation of the starch content in step 6) above can be carried out by substituting the following formula 1:

[Formula 1]

Figure 112015108404535-pat00001

(In the above formula 1, the starch content means the weight percentage of the starch contained in the sample).

Another embodiment of the present invention provides an assay kit for detecting or quantifying starch in processed meat products.

More specifically, the assay kit for detecting or quantifying the starch in the processed meat product may comprise the following solutions: 20 to 30% (w / v) sucrose, 1 x SSC, 1 to 10 mM EDTA, 0.5 to 2% (w / v) SDS; A second solution comprising 1-10% (w / v) Proteinase K; And 4-6% (w / v) phenol.

The kit may include, for example, a first solution comprising 26% (w / v) sucrose, 1 x SSC, 2 mM EDTA, 1% (w / v) SDS; A second solution comprising 2% (w / v) Proteinase K; And a third solution containing 5 to 6% (w / v) phenol, but the present invention is not limited thereto.

The kit may further comprise a fourth solution that is a sulfuric acid solution.

Using the starch detection or quantification method of the present invention, starch present in processed meat products can be specifically detected by distinguishing it from animal carbohydrates of meat components, and quantitative analysis with high accuracy is possible. Therefore, the present invention can be usefully used for the inspection of the amount and content of the extender of processed meat products.

FIG. 1 is a graph showing the results of detection of starch in processed meat products according to an embodiment of the present invention: samples containing starch showed brown or yellow color and samples without starch showed blue color.
2 is a calibration curve prepared by measuring the absorbance (492 nm) according to the concentration of the starch standard solution (1-100 mg / ml): X-axis, concentration of starch in the mixed solution (ppm / ml); And Y-axis, absorbance at 492 nm wavelength obtained from phenol-sulfuric acid reaction.
3 is a diagram showing starch recovered after pretreatment of processed meat products: the end of the arrows in A, B and C is a precipitate precipitated by centrifugation of the recovered starch; Dashed arrows A and B indicate recovered starch; A solid arrow in C indicates that the recovered starch is dyed with a solution of iodine-potassium iodide.

Hereinafter, the present invention will be described in more detail in the following Examples. It should be noted, however, that the following examples are illustrative only and do not limit or limit the scope of the present invention. It will be understood by those of ordinary skill in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.

< Example 1> Processing Mixed meat  Composition of Reaction Solution for Starch Analysis

A total of three kinds of reaction solutions used for detecting or quantifying starches in the processed mixed meat were prepared. The composition of each reaction solution was as follows.

First solution: 26% (w / v) sucrose, 1 x SSC (150 mM Sodium chloride and 150 mM Sodium citrate), 2 mM Ethylene-Diamine- , 1% (w / v) SDS (sodium dodecyl sulfate)

- Second solution: 20 mg / ml (i.e., equal to 2% (w / v)) Proteinase K

- Third solution: 5.5% (w / v) phenol

< Example  2> Processing Mixed meat  Detection and quantification of starch

<2-1> Pretreatment of sample

Samples for the detection of starch or for the determination of starch content were taken at various sites of the processed meat product, and as far as possible the meat portion was uniformly distributed. The processed meat products were prepared using the prototype manufactured by the National Livestock Academy and sausage purchased from general retailers. 2 g each of 3-5 sites were randomly selected according to the size or quantity of the sample and transferred to a conical tube. The first solution (26% sucrose, 1 x SSC, 2 mM EDTA (pH 8.0), 1% SDS). 36 ml of the first solution was added again to make the total volume to be 40 ml. Then, 40 μl of the second solution (20 mg / ml Proteinase K) prepared in Example 1 was added, and the mixture was vortex- Were mixed for 1 minute. The mixed solution was transferred to a constant temperature water bath at 65 ° C. and cultured for 8 hours or more. The mixed solution was mixed with a vortex-mixer at intervals of 1 to 2 hours. 10 ml of the mixed solution was transferred to two 15 ml conical tubes and centrifuged at 1,200 rpm and 4 ° C for 10 minutes. The supernatant was removed, 10 ml of 1 × PBS (Phosphate Buffer Saline) was added, mixed with a vortex-mixer, and centrifuged at 3,200 rpm and 4 ° C for 10 minutes. The supernatant was removed and filled to 2 ml with PBS and mixed with a vortex-mixer.

The prepared solution was diluted to 1/10, 1/100 and 1/1000 with distilled water and used as a starch detection or quantitative analysis sample.

<2-2> Detection and determination of starch

100 μl each of starch standard solutions mixed to 1 to 100 mg / ml were put into a glass test tube. Separately, 100 μl of the analytical sample diluted 1/10, 1/100, and 1/1000 prepared in Example 2-1 was placed in each test tube. When the mixed solution was in a floating state, 100 μl of the third solution (5.5% phenol) prepared in Example 1 was added and mixed with a vortex-mixer. 0.5 ml of concentrated sulfuric acid (H 2 SO 4 ) was added to each of the mixed solutions, and phenol-sulfuric acid reaction was carried out. As a result, starch-containing samples showed brown or yellow color and starch-free samples showed blue color (FIG. 1). Therefore, it was possible to visually confirm whether or not the starch contained in the mixed and mixed sample was contained.

Next, a calibration curve and a standard calibration equation for the starch standard solution were prepared for quantitative analysis of starch. 200 [mu] l of the phenol-sulfuric acid-treated solution of the starch standard solution was placed in a microplate and measured at 492 nm. The standard curve was prepared by measuring the absorbance of the standard solution, and it was confirmed that it conformed to the Lambert-Beer law. As a result of the standard calibration formula, the relationship between the content (wt%) of the starch in the sample and the absorbance was as shown in the following formula 1, and the crystal coefficient (R 2 ) value was as high as 0.9913 ). Therefore, it was confirmed that the absorbance is proportional to the concentration of starch, and thus it is suitable as a method of detecting the starch content.

[Formula 1]

Figure 112015108404535-pat00002

The phenol-sulfuric acid-assayed analytes were put into a microplate in duplicate (200 쨉 l each), and the microplate was placed in a Versa Max Tunable Microplate Reader (Molecular Devices, USA) and the absorbance at 492 nm was measured. The concentration of the starch in the analytical sample was calculated by substituting the measured results into the standard calibration equation (Table 1). Separately, a control test was conducted in the same manner using distilled water instead of the sample solution, and the average value of the two times was used as a correction value of the absorbance calculated in the sample analysis.

Figure 112015108404535-pat00003

As shown in Table 1, it was possible to detect and quantify starch contained in each sample. Therefore, it was confirmed that the method of the present invention can specifically detect and quantify starch in processed meat products.

< Experimental Example  1> Evaluation of the effect of starch recovery by pretreatment of sample

Experiments were conducted to test the effect of starch recovery by pretreatment of processed meat products using the solution prepared in Example 1. 14 kinds of processed meat products were subjected to pretreatment in the same manner as in Example <2-1> and visually confirmed whether or not the starch was recovered through a starch-iodine reaction.

Specifically, 14 kinds of processed meat products were used for the prototype manufactured by National Livestock Academy and sausage purchased from general retail store. 2 g of each of the three sites was randomly selected and transferred to a conical tube. The first solution (26% sucrose, 1 x SSC, 2 mM EDTA (pH 8.0), 1% SDS ) Was added to 2 ml of the solution. 36 ml of the first solution was added again to make the total volume to be 40 ml. Then, 40 μl of the second solution (20 mg / ml Proteinase K) prepared in Example 1 was added, and the mixture was vortex- Were mixed for 1 minute. The mixed solution was transferred to a constant-temperature water bath at 65 ° C and cultured for 8 hours. The mixed solution was mixed at an interval of 1 hour using a vortex-mixer. 10 ml of the mixed solution was transferred to two 15 ml conical tubes and centrifuged at 1,200 rpm and 4 ° C for 10 minutes. The supernatant was removed, 10 ml of 1 × PBS was added, mixed with a vortex-mixer, and centrifuged at 3,200 rpm at 4 ° C for 10 minutes. The supernatant was removed and filled to 2 ml with PBS and mixed with a vortex-mixer. As a result, the precipitate, starch, was visually confirmed (Figs. 3A and 3B).

The starch-iodine reaction experiment was carried out to confirm that the precipitate was a starch. This was done by dropping the iodine-potassium iodide solution to the precipitate recovered after the pretreatment. As a result, it was confirmed that the precipitate was starch because the color of the precipitate was dyed with a deep blue color by iodine staining (Fig. 3C). Also, the higher the amount of starch in the processed meat products, the more the proportion of the recovered starch was.

Accordingly, it was confirmed that the starch can be recovered with excellent efficiency through the pretreatment of the processed meat product, and the recovered starch can be used for starch detection or quantitative analysis with high accuracy.

Claims (10)

1) To a processed meat product sample, 20 to 30% (w / v) sucrose, 1 × SSC (saline sodium citrate), 1 to 10 mM ethylenediamine-tetraacetic acid (EDTA) w / v) Sodium Dodecyl Sulfate (SDS);
2) A second solution containing 1 to 10% (w / v) of Proteinase K was added to the mixed solution prepared in the step 1) at a volume ratio of 1000: 0.5 to 2, In a thermostatic chamber at any specific temperature for 6 to 20 hours while mixing;
3) diluting the precipitate obtained by centrifuging the mixed solution prepared in the step 2) by adding distilled water;
4) adding a third solution containing 4-6% (w / v) phenol to the mixed solution diluted in step 3) at a volume ratio of 1: 1;
5) adding the sulfuric acid to the mixed solution prepared in the step 4) to react with phenol-sulfuric acid to visually detect starch in the processed meat product sample by a coloring reaction;
6) if the reaction solution is brown or yellow in step 5), the processed meat product sample contains starch, and if the reaction solution is blue, it is determined that the processed meat product sample does not contain starch;
7) measuring the absorbance of the starch standard solution in the wavelength range of 490 to 495 nm and preparing a calibration curve and a standard calibration equation of the following formula 1; And
8) The absorbance of the processed meat product sample subjected to the phenol-sulfuric acid reaction in the step 5) within the wavelength range of 490 to 495 nm was measured and substituted for the standard calibration equation of the formula 1 prepared in the step 7) Calculating the content of starch in the sample; and detecting the amount of starch in the sample.

[Formula 1]
Starch (%) = (a / 0.031) × 100 × b (a, absorbance of test sample; b, dilution factor)
The coefficient of determination (R 2 ) in Equation 1 is 0.9913.
delete delete delete delete delete delete delete delete A first solution comprising 20 to 30% (w / v) sucrose, 1 x SSC, 1-10 mM EDTA, 0.5-2% (w / v) SDS; A second solution comprising 1-10% (w / v) Proteinase K; A third solution comprising 4-6% (w / v) phenol; And a calibration curve for the starch standard solution and a standard calibration formula of the following formula 1:

[Formula 1]
Starch (%) = (a / 0.031) × 100 × b (a, absorbance of test sample; b, dilution factor)
The coefficient of determination (R 2 ) in Equation 1 is 0.9913.
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CN101526510A (en) * 2009-04-22 2009-09-09 南昌大学 Method for rapidly determining content of lactosucrose in food
KR101391091B1 (en) * 2011-06-01 2014-04-30 제이-오일 밀스, 인코포레이티드 Starch processed with oil or fat and method for producing same

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