CN108088823A - The detection method of calcium content in dairy products - Google Patents
The detection method of calcium content in dairy products Download PDFInfo
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- CN108088823A CN108088823A CN201611048129.5A CN201611048129A CN108088823A CN 108088823 A CN108088823 A CN 108088823A CN 201611048129 A CN201611048129 A CN 201611048129A CN 108088823 A CN108088823 A CN 108088823A
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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Abstract
The invention discloses a kind of detection methods of calcium content in dairy products.The described method includes：The dairy products are diluted processing with diluent, obtain dilution；The dilution and calcium reaction reagent are subjected to chromogenic reaction, to obtain reaction product；And the reaction product is measured using time-resolved fluorescence method, compared with the calcium calibration solution by equally handling, you can calcium in sample content is calculated, wherein, the diluent contains：The Triton X 100 of the hydrochloric acid of 10~30 mass % and 0.5 mass %.The detection method of calcium content can quickly and accurately detect calcium content in dairy products in the dairy products of the present invention.
Description
Technical field
The present invention relates to field of food.In particular it relates in dairy products calcium content detection method.
Background technology
Calcium is a kind of in vivo trace element of people, it is although very little in the in vivo content of people, but its effect is
Huge.Calcium can maintain to adjust in body without more physiological and biochemical procedures, adjust tramsmitter release, increase the secretion of incretory, tie up
Completion and the permeability of cell are held, promotes the regeneration of cell, increases Abwehrkraft des Koepers.Contain abundant calcium activated in milk,
It is the best source of human calcium, and calcium phosphorus ration is very appropriate, beneficial to the absorption of calcium.So edible milk be replenish the calcium it is most common
One of mode obtained, measuring various milk calcium contents contributes to the milk of calcium content needed for people's selection.
However, the detection method of calcium content still has much room for improvement in dairy products at present.
The content of the invention
It is contemplated that at least one technical problem in the prior art is solved at least to a certain extent.For this purpose,
The present invention proposes a kind of detection method of calcium content in dairy products.The present invention can quickly and accurately measure calcium in dairy products
Content.
It should be noted that the present invention is the following discovery based on inventor and completes：
The method of detection calcium content has complexometry (to titrate ox with disodium EDTA (EDTA) solution at present
Calcium in milk), calcium in GB 5413.21-2010-national food safety standard infant food and dairy products, iron, zinc, sodium, potassium,
Measure (the first method atomic absorption spectrophotometry) (hereinafter referred to as National Standard Method), ion selective electrode of magnesium, copper and manganese
Method (calcium ion meter method) etc., the accuracy of wherein National Standard Method testing result is higher.But time-consuming, method is cumbersome for detection,
Be not suitable for on-line quick detection, and existed during this method detection using the instrument and equipments such as acetylene gas, muffle furnace, detection process operation
Security risk.Since lactogenesis in purchase link needs to ensure freshness, the used time is examined to need to shorten, so that quick inspection can not be met
The demand of survey.
For this purpose, the present invention proposes a kind of detection method of calcium content in dairy products.According to an embodiment of the invention, it is described
Method includes：The dairy products are diluted processing with diluent, obtain dilution；By the dilution and calcium reaction reagent
Chromogenic reaction is carried out, to obtain reaction product；And measured using time-resolved fluorescence method, with the calcium school by equally handling
Quasi- liquid (standard items) is compared, you can calculates result.Wherein, the diluent contains：The hydrochloric acid of 10~30 mass % and
The Triton X-100 of 0.5 mass %.
Inventor has found that the dairy products handled by that will pass through dilution dilution agent carry out chromogenic reaction with calcium reaction reagent,
And chromogenic reaction product is detected using time-resolved fluorescence method, so as to realize the inspection to calcium content in dairy products
It surveys.Further, inventor has found, processing is diluted with the diluent containing hydrochloric acid and Triton X-100 so that dilution
Calcium content meets subsequent detection requirement in sample liquid afterwards.In addition, hydrochloric acid will cause the combination calcium in dairy products to become free calcium, just
In subsequently carrying out chromogenic reaction with calcium reaction reagent, Triton X-100 are able to maintain that the stabilization of dairy products system, prevent
Testing result wild effect.Inventor is it was unexpectedly observed that concentration of hydrochloric acid and Triton X-100 concentration significantly affect detection knot
The accuracy of fruit.For this purpose, inventor has found by many experiments, when hydrochloric acid and Triton X-100 concentration are respectively 10~30 matter
When measuring % and 0.5 mass %, the accuracy of testing result is stronger.If the excessively high sample liquid of content of hydrochloric acid precipitates, if Triton X-
100 is excessively high, can change the property of sample liquid system in itself；If the combination calcium in the too low sample liquid of content of hydrochloric acid cannot dissociate completely, if
Triton X-100 are too low, and testing result is relatively low, deviate the normal value of lactogenesis calcium content.It is according to embodiments of the present invention as a result,
The detection method of calcium content can quickly and accurately detect calcium content in dairy products in dairy products.
According to an embodiment of the invention, the detection method of calcium content can also have following supplementary technology in above-mentioned dairy products
Feature：
According to an embodiment of the invention, the diluent contains：The Trion X- of the hydrochloric acid of 20 mass % and 0.5 mass %
100.Inventor has found, can further detect calcium content in dairy products exactly with this condition.
According to an embodiment of the invention, the dairy products are raw milk.As a result, in dairy products according to embodiments of the present invention
The detection method of calcium content can quickly and accurately detect calcium content in raw milk.
According to an embodiment of the invention, based on dairy products described in 100 μ L, the extension rate of the dilution processing is 5~15
Times, the volume of the dilution of chromogenic reaction is carried out as 10 μ L with the calcium reaction reagent, the volume of the calcium reaction reagent
For 800~1500 μ L.Inventor has found that the adding proportion of dilution and calcium reaction reagent can influence follow-up chromogenic reaction and light splitting
The accuracy of photometry testing result.Inventor has found by many experiments, based on dairy products, the body of dilution described in 100 μ L
When product is 10 μ L, the accuracy of testing result is stronger.According to a preferred embodiment of the invention, based on dairy products, institute described in 100 μ L
The extension rate for stating dilution processing is 10 times.As a result, in dairy products according to embodiments of the present invention calcium content detection method energy
It is enough quickly and accurately to detect calcium content in dairy products.
According to an embodiment of the invention, the calcium reaction reagent is the arsenazo Ⅲ solution of 0.1~0.3g/L.Invention human hair
Existing, the calcium in dairy products can be reacted with arsenazo Ⅲ, generated blue complex, measured by time-resolved fluorescence method, with warp
It crosses the calcium calibration solution (standard items) equally handled to be compared, you can calculate calcium in sample content.It is real according to the present invention as a result,
Calcium content in dairy products can quickly and accurately be detected by applying the detection method of calcium content in the dairy products of example.
According to an embodiment of the invention, the chromogenic reaction includes：The dilution and calcium reaction reagent are shaken 30 seconds,
And in 30~40 DEG C of water-baths 1~6 minute, preferably 5 minutes.Thereby, it is possible to make the calcium in dairy products and the idol in calcium reaction reagent
Chromogenic reaction fully occurs for nitrogen arsine III, generates blue complex, is measured by time-resolved fluorescence method, with passing through what is equally handled
Calcium calibration solution (standard items) is compared, you can calculates calcium in sample content.Dairy products according to embodiments of the present invention as a result,
The detection method of middle calcium content can quickly and accurately detect calcium content in dairy products.
According to an embodiment of the invention, the Detection wavelength of the spectrophotometry is 650nm.It is real according to the present invention as a result,
Calcium content in dairy products can quickly and accurately be detected by applying the detection method of calcium content in the dairy products of example.
In another aspect of this invention, the present invention proposes a kind of detection method of calcium content in raw milk.According to this hair
Bright embodiment, the described method includes：(1) 100 μ L raw milks is taken to be placed in centrifuge tube, 900 μ L are added in into the centrifuge tube
Diluent with whirlpool instrument mixing 30s, obtains 10 times of dilutions, wherein the diluent contains the hydrochloric acid and 0.5 matter of 20 mass %
Measure the Trion X-100 of %；(2) dilution, distilled water and each 10 μ L of standard calcium product solution are taken respectively, it is anti-with 1000 μ L calcium
Reagent is answered to shake 30 seconds after 37 DEG C of water-baths 5 minutes, sample prepare liquid, blank control liquid and standard items is respectively obtained and treats
Liquid is surveyed, wherein, the calcium reaction reagent is the arsenazo Ⅲ solution of 0.1~0.3g/L；(3) take respectively 200 μ L samples prepare liquids,
Blank control liquid and standard items prepare liquid are placed in micropore cup, then each micropore cup is positioned in time-resolved fluorescence instrument card slot
It is detected, Detection wavelength 650nm, records the extinction of the sample prepare liquid, blank control liquid and standard items prepare liquid
Degree；And (4) concentration and absorbance based on the standard items prepare liquid, the absorbance of the blank control liquid and the sample
Calcium content in the raw milk is calculated in the absorbance of product prepare liquid.Calcium in raw milk according to embodiments of the present invention as a result,
The detection method of content can quickly and accurately detect calcium content in raw milk.
The additional aspect and advantage of the present invention will be set forth in part in the description, and will partly become from the following description
It obtains substantially or is recognized by the practice of the present invention.
Description of the drawings
The above-mentioned and/or additional aspect and advantage of the present invention will become in the description from combination accompanying drawings below to embodiment
Substantially and it is readily appreciated that, wherein：
Fig. 1 shows influence of the bath temperature according to an embodiment of the invention to testing result；And
Fig. 2 shows influence of the water bath time according to an embodiment of the invention to testing result.
Specific embodiment
The embodiment of the present invention is described below in detail.The embodiments described below is exemplary, and is only used for explaining this hair
It is bright, and be not considered as limiting the invention.
The solution of the present invention is explained below in conjunction with embodiment.It will be understood to those of skill in the art that following
Embodiment is merely to illustrate the present invention, and should not be taken as limiting the scope of the invention.Particular technique or item are not specified in embodiment
Part, it is carried out according to the described technology of document in the art or condition or according to product description.Agents useful for same or instrument
Production firm person is not specified in device, and being can be with conventional products that are commercially available.
Embodiment 1
In this embodiment, calcium content in raw milk is measured in following manner：
1st, instrument and reagent
1.1 instruments and consumptive material
Time-resolved fluorescence instrument：Model SS-330, the calcium content detection range of linearity is 0-20mg/100mL；
Pipettor (0.5-10 μ L), (20-200 μ L), (100-1000 μ L)；
Whirlpool instrument；
Water-bath；
Sample introduction micropore cup.
1.2 reagent
Calcium reaction reagent：Arsenazo Ⅲ concentration is 0.1~0.3g/L；
Diluent：The Trion X-100 of the hydrochloric acid of 20 mass % and 0.5 mass %；
Calcium calibration object：Calcium content is 10mg/100ml.
2nd, step
Reach room temperature using all reagents, sample before surveying (at 20 ± 5 DEG C)
(1) 100 μ L raw milks is taken to be placed in centrifuge tube；
(2) 900 μ L diluents are added in centrifuge tube, with whirlpool instrument mixing 30s, obtain 10 times of dilutions；
(3) dilution, distilled water and each 10 μ L of standard calcium product solution are taken respectively, are shaken 30 seconds with 1000 μ L calcium reaction reagents
After 37 DEG C of water-baths 5 minutes, sample prepare liquid, blank control liquid and standard items prepare liquid are respectively obtained；
(4) 200 μ L samples prepare liquids, blank control liquid and standard items prepare liquid is taken to be placed in micropore cup, then will be each micro-
Hole cup is positioned in SS-330 instrument card slots to be measured with time-resolved fluorescence method, Detection wavelength 650nm, and record sample is to be measured
The absorbance of liquid, blank control liquid and standard items prepare liquid；
(5) concentration and absorbance, the absorbance of blank control liquid and sample prepare liquid based on standard items prepare liquid
Calcium content in raw milk is calculated in absorbance.
2 analysis of the accuracy of embodiment
Calcium content in same raw milk is measured according to embodiment 1 and National Standard Method respectively, the results are shown in Table 1.As can be seen that
The result error of two methods is not higher than 10%, meets national standard, so as to show that 1 method of embodiment can accurately detect raw ox
Calcium content in breast.In addition, when the whole used time of National Standard Method is 16 small or so, the whole used time of 1 method of embodiment is 15 minutes left sides
The right side, in comparison, 1 method of embodiment are detected faster, and efficiency is higher.
The comparison of 1 method of 1 National Standard Method of table and embodiment
Embodiment 3
Calcium content in raw milk is measured according to the method for embodiment 1, difference lies in concentration of hydrochloric acid is 10 matter in diluent
Measure %.
The results are shown in Table 2.It is wanted as can be seen that the testing result that concentration of hydrochloric acid is 10 mass % in diluent meets national standard
10% asked, wherein having 1 group of data deviation between 5%~10%, deviation is higher for embodiment 1.
The comparison of 3 method of 2 National Standard Method of table and embodiment
Embodiment 4
Calcium content in raw milk is measured according to the method for embodiment 1, difference lies in concentration of hydrochloric acid is 30 matter in diluent
Measure %.
The results are shown in Table 3.It is wanted as can be seen that the testing result that concentration of hydrochloric acid is 30 mass % in diluent meets national standard
10% asked, wherein having 2 groups of data between 5%~10%, deviation is higher for embodiment 1.
The comparison of 4 method of 3 National Standard Method of table and embodiment
Embodiment 5
Calcium content in raw milk is measured according to the method for embodiment 1, difference lies in step (1), dilution dosage is 5 μ
L。
The results are shown in Table 4.As can be seen that testing result is higher, and National Standard Method testing result deviation is above 10%, standard
True property is poor.
The comparison of 5 method of 4 National Standard Method of table and embodiment
Embodiment 6
Calcium content in raw milk is measured according to the method for embodiment 1, difference lies in step (1), dilution dosage is 20
μL。
The results are shown in Table 5.As can be seen that testing result is relatively low, wherein 2 groups of results and National Standard Method testing result deviation are high
In 10%, accuracy is poor.
The comparison of 6 method of 5 National Standard Method of table and embodiment
Embodiment 7
Calcium content in raw milk is measured according to the method for embodiment 1, difference lies in, in step (1), the reaction of calcium reaction reagent
It measures as 1500 μ L.
The results are shown in Table 6.As can be seen that and National Standard Method testing result deviation between 5%-10%, compared to implementation
Deviation is higher for example 1.
The comparison of 7 method of 6 National Standard Method of table and embodiment
Embodiment 8
Calcium content in raw milk is measured according to the method for embodiment 1, difference lies in, in step (1), the reaction of calcium reaction reagent
It measures as 800 μ L.
The results are shown in Table 7.As can be seen that and National Standard Method testing result deviation between 5%-10%, compared to implementation
Deviation is higher for example 1.
The comparison of 8 method of 7 National Standard Method of table and embodiment
Embodiment 9
Calcium content in raw milk is measured according to the method for embodiment 1, difference lies in do not contain Triton X- in diluent
100。
The results are shown in Table 8.As can be seen that it is relatively low not contain Triton X-100 testing results, detects and tie with National Standard Method
Fruit deviation is higher than 10%, and accuracy is poor.
The comparison of 9 method of 8 National Standard Method of table and embodiment
Embodiment 10
Calcium content in raw milk is measured according to the method for embodiment 1, is 30~40 DEG C and water-bath difference lies in bath temperature
Reaction time is 1~6 minute, as a result as illustrated in fig. 1 and 2.As can be seen that 36-37 DEG C of bath temperature and water-bath time 5-6
Minute result is optimal.
In the description of this specification, reference term " one embodiment ", " some embodiments ", " example ", " specifically show
The description of example " or " some examples " etc. means specific features, structure, material or the spy for combining the embodiment or example description
Point is contained at least one embodiment of the present invention or example.In the present specification, schematic expression of the above terms is not
It must be directed to identical embodiment or example.Moreover, particular features, structures, materials, or characteristics described can be in office
It is combined in an appropriate manner in one or more embodiments or example.In addition, without conflicting with each other, the skill of this field
Art personnel can tie the different embodiments described in this specification or example and different embodiments or exemplary feature
It closes and combines.
Although the embodiment of the present invention has been shown and described above, it is to be understood that above-described embodiment is example
Property, it is impossible to limitation of the present invention is interpreted as, those of ordinary skill in the art within the scope of the invention can be to above-mentioned
Embodiment is changed, changes, replacing and modification.
Claims (9)
1. a kind of detection method of calcium content in raw milk, which is characterized in that including：
(1) 100 μ L raw milks is taken to be placed in centrifuge tube, 900 μ L diluents are added in into the centrifuge tube, with whirlpool instrument mixing
30s obtains 10 times of dilutions, wherein the diluent contains the hydrochloric acid of 20 mass % and the Trion X-100 of 0.5 mass %；
(2) dilution, distilled water and each 10 μ L of standard calcium product solution are taken respectively, are shaken 30 seconds with 1000 μ L calcium reaction reagents
After 37 DEG C of water-baths 5 minutes, sample prepare liquid, blank control liquid and standard items prepare liquid are respectively obtained, wherein, it is described
Calcium reaction reagent is the arsenazo Ⅲ solution of 0.1~0.3g/L；
(3) 200 μ L samples prepare liquids, blank control liquid and standard items prepare liquid is taken to be placed in micropore cup respectively, then will be each micro-
Hole cup, which is positioned in time-resolved fluorescence instrument card slot, to be detected, and Detection wavelength 650nm records the sample prepare liquid, sky
The absorbance of white comparison liquid and standard items prepare liquid；And
(4) concentration and absorbance, the absorbance of the blank control liquid and the sample based on the standard items prepare liquid
Calcium content in the raw milk is calculated in the absorbance of prepare liquid.
2. a kind of detection method of calcium content in dairy products, which is characterized in that including：
The dairy products are diluted processing with diluent, obtain dilution；
The dilution and calcium reaction reagent are subjected to chromogenic reaction, to obtain reaction product；And
The reaction product is detected using time-resolved fluorescence method, and calcium in the dairy products is determined based on testing result
Content, wherein, the diluent contains：The Triton X-100 of the hydrochloric acid of 10~30 mass % and 0.5 mass %.
3. according to the method described in claim 2, it is characterized in that, the diluent contains：The hydrochloric acid and 0.5 matter of 20 mass %
Measure the Trion X-100 of %.
4. according to the method described in claim 2, it is characterized in that, the dairy products are raw milk.
5. according to the method described in claim 2, it is characterized in that, based on dairy products described in 100 μ L,
The extension rate of the dilution processing is 5~15 times,
The volume of the dilution of chromogenic reaction is carried out as 10 μ L with the calcium reaction reagent,
The volume of the calcium reaction reagent is 800~1500 μ L.
6. according to the method described in claim 2, it is characterized in that, based on dairy products described in 100 μ L, the dilution handles dilute
Multiple is released as 10 times.
7. according to the method described in claim 2, it is characterized in that, the calcium reaction reagent is the arsenazo of 0.1~0.3g/L
III solution.
8. according to the method described in claim 2, it is characterized in that, the chromogenic reaction includes：
The dilution and calcium reaction reagent are shaken 30 seconds, and in 30~40 DEG C of water-baths 1~6 minute.
9. according to the method described in claim 2, it is characterized in that, the time-resolved fluorescence method Detection wavelength is 650nm.
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| CN101408515A (en) * | 2007-10-11 | 2009-04-15 | 上海电气电站设备有限公司 | Method for measuring zirconium in steel |
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2016
- 2016-11-23 CN CN201611048129.5A patent/CN108088823B/en active Active
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| CN101408515A (en) * | 2007-10-11 | 2009-04-15 | 上海电气电站设备有限公司 | Method for measuring zirconium in steel |
| CN101315387A (en) * | 2008-07-23 | 2008-12-03 | 内蒙古蒙牛乳业（集团）股份有限公司 | Detection method for micronutrient levels in liquid milk or milk powder |
| CN102466582A (en) * | 2010-11-10 | 2012-05-23 | 内蒙古蒙牛乳业（集团）股份有限公司 | Pretreatment method of sweetened bean paste for detection of sodium element therein and method for detecting sodium element in sweetened bean paste |
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