CN104132939A - Method for rapidly detecting amylase value in honey - Google Patents
Method for rapidly detecting amylase value in honey Download PDFInfo
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- CN104132939A CN104132939A CN201410330025.8A CN201410330025A CN104132939A CN 104132939 A CN104132939 A CN 104132939A CN 201410330025 A CN201410330025 A CN 201410330025A CN 104132939 A CN104132939 A CN 104132939A
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Abstract
The invention discloses a method for rapidly detecting the amylase value in honey. The method comprises the following steps: 1, sample preparation: accurately weighing 5.0g of a honey sample, putting the honey sample in a beaker, adding 7.5mL of an enzymatic hydrolysis buffer solution and 10mL of distilled water, stirring until the sample is completely dissolved, transferring the above obtained solution to a 25mL volumetric flask, adding distilled water to a constant volume, and shaking up; 2, enzymatic hydrolysis: accurately sucking 2.5mL of the prepared sample solution to a test tube with 12.5mL of a starch solution, uniformly mixing, and carrying out enzymatic hydrolysis at normal temperature for 10min or 20min; 3, coloration: sucking the obtained enzymatic hydrolysis liquid to a colorimetric pipe with a coloration solution and distilled water, and coloring at normal temperature for 5min; and 4, detection: comparing the color of the coloration solution with a multicolor colourimetric card, and determining the amylase value in the honey. The method is characterized in that amylase reacts with starch at normal temperature, and a colourimetric card technique is cooperated to detect. The method has the advantages of simple operation, simple conditions, rapidness and the like.
Description
Technical field
The present invention is specifically related to the method for quick of amylase value in a kind of honey, belongs to Safety of Food Quality detection technique field.
Background technology
The height of amylase value is one of important quality index of weighing honey biologically active, degree of ripeness and freshness.The method of measuring at present the amylase value in honey mainly contains spectrophotometer method, test tube ocular estimate, canonical reference color board method and kit method etc.First method is international method, and this method is accurate, quick, but needs the equipment such as spectrophotometer, higher to testing staff's technical requirement, is unsuitable for field quick detection; Second method detecting step complexity, and consuming time; The third method is the colorimetric card finding out on the basis of first method, more for convenience, but only has a kind of color, for the low sample of enzyme value, needs to detect more than one hour, and the 4th kind of method is in fact also spectrophotometer method.In addition, above all methods also all can not detect (need under 40 DEG C of water-baths enzymolysis) at normal temperatures.The present invention forefathers to honey on the basis of amylase value research, by the relation of enzymolysis under normal temperature and enzyme value, produce the colorimetric card of multiple color and corresponding enzyme value, set up a kind of detection method that can carry out at normal temperatures Fast Measurement to the amylase value in honey.
Summary of the invention
The object of the invention is to be to provide the method for quick of amylase value in a kind of honey, can on-the-spot complete the Fast Measurement to amylase value in honey, have easy and simple to handle, condition is simple, the advantage such as quick.
For achieving the above object, technical scheme of the present invention is:
(1) preparation of sample: accurately take 5.0 g honey sample liquids in beaker, add the distilled water of 7.5 mL enzymolysis damping fluids and 10 mL, stir sample is dissolved completely, then move in 25 mL volumetric flasks, add distilled water to be settled to scale, shake up;
(2) enzymolysis: accurately draw the sample solution that 2.5 mL prepare and mix in the test tube that fills 12.5 mL starch solutions; Enzymolysis 10 min or 20 min at normal temperatures;
(3) colour developing: draw above-mentioned enzymolysis liquid 5.0 mL in filling the color comparison tube of 2.5 mL nitrite ions and 6.6 mL distilled water, 5 min develop the color under normal temperature;
(4) detect: by the color of solution after developing the color and the color contrast of colorimetric card, determine the amylase value in honey.
Enzymolysis damping fluid described in above-mentioned steps (1), it consists of sodium acetate (0.43 moL/L), sodium chloride (0.1 moL/L) and glacial acetic acid (0.7wt.%); Starch solution concentration described in above-mentioned steps (2) is 0.04wt.%; Nitrite ion described in above-mentioned steps (3), it consists of potassium iodide (4.04wt.%) and iodine (0.018wt.%); The described color comparison tube of above-mentioned steps (3) is 25mL color-comparison tube (caliber 2cm, pipe range 15cm); Making and the scope of the colorimetric card described in above-mentioned (4) are as follows: at normal temperatures, the honey of a series of known enzyme values is pressed to step preparation described in 1, enzymolysis and colour developing, according to obtained color, make multi-colorimetric card, corresponding amylase value is respectively:
(1) 0-24: 0,6,12,18,24 (developing times 10 minutes);
(2) 0-14.5: 0,2,4,7.5,14.5(developing time 20 minutes).
Beneficial effect of the present invention: the method for amylase value in the employing multi-colorimetric block-regulations fast detecting honey that the present invention sets up.Its distinguishing feature is to measure at normal temperatures, does not need the instrument and equipments such as spectrophotometer, portable instrument and heating water bath, have easy and simple to handle, condition is simple, the advantage such as quick.Therefore the method is applicable to being applied to the amylase value in the field quick detection honey such as industry and commerce, quality inspection, supermarket and family, and the quality level of honey product is made to quick judgement, to safeguard people's economic interests and guarantee people's edible safety.
embodiment:
Embodiment 1:
The method for making of colorimetric card
(1) preparation of sample: get known enzyme value and be greater than 24 honey and be diluted with distilled water into enzyme value and be respectively 6,12,18 and 24 honey sample; Accurately take the above-mentioned honey sample liquid of 5.0g in beaker, add the distilled water of 7.5 mL enzymolysis damping fluids and approximately 10 mL, stir sample is dissolved completely, then move in 25mL volumetric flask, add distilled water be settled to scale and shake up; Replace honey, the sample that is 0 as enzyme value with distilled water.
(2) enzymolysis: accurately draw the above-mentioned solution of 2.5mL and mix in the test tube that fills 12.5mL starch solution, enzymolysis 10min at normal temperatures.
(3) colour developing: draw above-mentioned enzymolysis liquid 5.0mL and develop the color in the color comparison tube that fills 2.5mL nitrite ion and 6.6mL distilled water, according to obtained color, make colorimetric card, the enzyme value scope that colorimetric card is corresponding is 0-24.
Actual sample is measured
(1) preparation of sample: accurately take 5.0g honey sample liquid in beaker, add the distilled water of 7.5 mL enzymolysis damping fluids and approximately 10 mL, stir sample is dissolved completely.Then move in 25mL volumetric flask, add distilled water be settled to scale and shake up;
(2) enzymolysis: accurately draw the sample solution that 2.5mL prepares and mix in the test tube that fills 12.5mL starch solution, enzymolysis 10 min at normal temperatures.
(3) colour developing: draw above-mentioned enzymolysis liquid 5.0mL in filling the color comparison tube of 2.5mL nitrite ion and 6.6mL distilled water, 5 min develop the color under normal temperature.
(4) detect: by the color contrast of nitrite ion and colorimetric card, the color and luster that its color and enzyme value are 24 is the most approaching, therefore determine that the amylase value in honey sample is 24.
Embodiment 2:
The method for making of colorimetric card
(1) preparation of sample: get known enzyme value and be greater than 15 honey and be diluted with distilled water into enzyme value and be respectively 2,4,7.5 and 14.5 honey sample, accurately take the above-mentioned honey sample liquid of 5.0g in beaker, add the distilled water of 7.5 mL enzymolysis damping fluids and approximately 10 mL, stirring is dissolved sample completely, then moves in 25mL volumetric flask, adds distilled water be settled to scale and shake up; Replace honey, the sample that is 0 as enzyme value with distilled water.
(2) enzymolysis: accurately draw the above-mentioned solution of 2.5mL and mix in the test tube that fills 12.5mL starch solution, at normal temperatures enzymolysis 20 min.
(3) colour developing: draw above-mentioned enzymolysis liquid 5.0mL and develop the color in the color comparison tube that fills 2.5mL nitrite ion and 6.6mL distilled water, according to obtained color, make colorimetric card, the enzyme value scope that colorimetric card is corresponding is 0-14.5.
Actual sample is measured
(1) preparation of sample: accurately take 5.0g honey sample liquid in beaker, add the distilled water of 7.5 mL enzymolysis damping fluids and approximately 10 mL, stir sample is dissolved completely.Then move in 25mL volumetric flask, add distilled water be settled to scale and shake up;
(2) enzymolysis: accurately draw the sample solution that 2.5mL prepares and mix in the test tube that fills 12.5mL starch solution, enzymolysis 20 min at normal temperatures.
(3) colour developing: draw above-mentioned enzymolysis liquid 5.0mL in filling the color comparison tube of 2.5mL nitrite ion and 6.6mL distilled water, 5 min develop the color under normal temperature.
(4) detect: by the color contrast of nitrite ion and colorimetric card, its color is between 2 and 4 color and luster between enzyme value, therefore estimate that the amylase value in honey sample is 3.
Remarks: above the unit of amylase value is mL/ (gh).
The foregoing is only preferred embodiment of the present invention, all equalizations of doing according to the present patent application the scope of the claims change and modify, and all should belong to covering scope of the present invention.
Claims (7)
1. a method for quick for amylase value in honey, is characterized in that comprising the following steps:
(1) preparation of sample: accurately take 5.0 g honey sample liquids in beaker, add the distilled water of 7.5 mL enzymolysis damping fluids and 10 mL, stir sample is dissolved completely, then move in 25 mL volumetric flasks, add distilled water to be settled to scale, shake up;
(2) enzymolysis: accurately draw the sample solution that 2.5 mL prepare and mix in the test tube that fills 12.5 mL starch solutions; Enzymolysis 10 min or 20 min;
(3) colour developing: draw above-mentioned enzymolysis liquid 5.0 mL in filling the color comparison tube of 2.5 mL nitrite ions and 6.6 mL distilled water, 5 min develop the color under normal temperature;
(4) detect: by the color of solution after developing the color and the color contrast of colorimetric card, determine the amylase value in honey.
2. the method for quick of amylase value in a kind of honey according to claim 1, is characterized in that: described enzymolysis damping fluid consist of 0.43 moL/L sodium acetate, 0.1 moL/L sodium chloride and 0.7wt.% glacial acetic acid.
3. the method for quick of amylase value in a kind of honey according to claim 1, is characterized in that: the concentration of step (2) starch solution is 0.04wt.%.
4. the method for quick of amylase value in a kind of honey according to claim 1, is characterized in that: step (2) enzymolysis at normal temperatures.
5. the method for quick of amylase value in a kind of honey according to claim 1, is characterized in that: nitrite ion consists of 4.04wt.% potassium iodide and 0.018wt.% iodine in step (3).
6. the method for quick of amylase value in a kind of honey according to claim 1, is characterized in that: color comparison tube is 25mL color-comparison tube, and caliber 2 cm, manage high 15 cm.
7. the method for quick of amylase value in a kind of honey according to claim 1, it is characterized in that: at normal temperatures, the honey of a series of known enzyme values is pressed to described step preparation, enzymolysis and colour developing, according to obtained color, make multi-colorimetric card, the enzyme value scope that colorimetric card is corresponding is respectively 0-24 and 0-14.5.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104359900A (en) * | 2014-11-07 | 2015-02-18 | 福建农林大学 | Kit for quickly detecting content of proline in honey |
| CN104359901A (en) * | 2014-11-07 | 2015-02-18 | 福建农林大学 | Rapid detecting method for hydroxymethyl furfural in honey |
| CN108226151A (en) * | 2018-01-31 | 2018-06-29 | 安徽泓顺源生物科技有限公司 | A kind of assay method of Fructose in Honey and glucose content |
| CN109001189A (en) * | 2017-06-06 | 2018-12-14 | 中粮营养健康研究院有限公司 | Detect the method and its application of the amylase content range in the sugar products of sugarcane |
| CN109536575A (en) * | 2018-12-10 | 2019-03-29 | 石家庄君乐宝乳业有限公司 | A kind of detection method of amylase |
| CN110609033A (en) * | 2019-10-31 | 2019-12-24 | 中国农业科学院蜜蜂研究所 | Method for detecting enzyme value of sucrose invertase in honey |
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| CN1067896A (en) * | 1991-06-26 | 1993-01-13 | 龟甲万株式会社 | The malto-oligosaccharide glycoside derivates is used to measure the reagent and the method for alpha-amylase activity |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104359900A (en) * | 2014-11-07 | 2015-02-18 | 福建农林大学 | Kit for quickly detecting content of proline in honey |
| CN104359901A (en) * | 2014-11-07 | 2015-02-18 | 福建农林大学 | Rapid detecting method for hydroxymethyl furfural in honey |
| CN109001189A (en) * | 2017-06-06 | 2018-12-14 | 中粮营养健康研究院有限公司 | Detect the method and its application of the amylase content range in the sugar products of sugarcane |
| CN108226151A (en) * | 2018-01-31 | 2018-06-29 | 安徽泓顺源生物科技有限公司 | A kind of assay method of Fructose in Honey and glucose content |
| CN109536575A (en) * | 2018-12-10 | 2019-03-29 | 石家庄君乐宝乳业有限公司 | A kind of detection method of amylase |
| CN109536575B (en) * | 2018-12-10 | 2022-07-05 | 君乐宝乳业集团有限公司 | Detection method of amylase |
| CN110609033A (en) * | 2019-10-31 | 2019-12-24 | 中国农业科学院蜜蜂研究所 | Method for detecting enzyme value of sucrose invertase in honey |
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Application publication date: 20141105 |