CN106404999A - Kit for detecting phosphor content and method for measuring phosphor content - Google Patents
Kit for detecting phosphor content and method for measuring phosphor content Download PDFInfo
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- CN106404999A CN106404999A CN201610773879.2A CN201610773879A CN106404999A CN 106404999 A CN106404999 A CN 106404999A CN 201610773879 A CN201610773879 A CN 201610773879A CN 106404999 A CN106404999 A CN 106404999A
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- Prior art keywords
- solution
- liquid
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- kit
- phosphorus
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N31/00—Investigating or analysing non-biological materials by the use of the chemical methods specified in the subgroup; Apparatus specially adapted for such methods
- 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
Abstract
The invention discloses a kit for detecting the phosphor content and a method using the same to measure the phosphor content. The kit comprises an indicator, which is a p-nitrophenol solution with a volume percentage of 0.2 vol.%; a color developing agent, which comprises ammonium molybdate, ammonium meta-vanadate, nitric acid, and water; and a mediating liquid, which comprises a nitric acid solution (0.2 mol/L), a first sodium hydroxide solution (6 mol/L), and a second sodium hydroxide solution (0.1 mol/L). Through the abovementioned technical scheme, the provided kit can accurately measure the phosphor content, and quick batch detection can be realized.
Description
Technical field
The present invention relates to field of food.In particular it relates to contain for the kit detecting phosphorus content and mensure phosphorus
The method of amount.
Background technology
The mensure Main Basiss national food safety standard GB 5413.22-2010 of phosphorus content in milk and milk productses《Baby children
Phosphorus yield in youngster's food and dairy products》, its principle is：Sample, through acid oxidase, makes phosphorus generate with vanadium ammonium molybdate in salpeter solution
Yellow complex.With spectrophotometer at wavelength 440nm mensuration absorbance, the depth of its color is directly proportional to the content of phosphorus.
However, the method measuring phosphorus content at present still has much room for improvement.
Content of the invention
It is contemplated that at least solving one of technical problem present in prior art.For this reason, one object of the present invention
It is to provide the kit for detecting phosphorus content and the method measuring phosphorus content, thereby, it is possible to measure phosphorus content exactly, and
It is capable of quick, batch detection.
In a first aspect of the present invention, the present invention proposes a kind of kit for detecting phosphorus content.According to the present invention
Embodiment, described kit includes：Indicator, described indicator is the paranitrophenol solution of 0.2 volume %；Developer, institute
State developer and include ammonium molybdate, ammonium metavanadate, nitric acid and water；And conciliation liquid, it is molten that described conciliation liquid includes 0.2mol/L nitric acid
Liquid, 6mol/L first sodium hydroxide solution and 0.1mol/L second sodium hydroxide solution.Thus, according to embodiments of the present invention
Kit for detecting phosphorus content can measure phosphorus content exactly, and is capable of quick, batch detection.
According to embodiments of the invention, it is special that the above-mentioned kit for detecting phosphorus content can also have following supplementary technology
Levy：
According to embodiments of the invention, described kit further includes：Phosphorus standard solution；And porous plate.Thus,
The kit for detecting phosphorus content according to embodiments of the present invention can measure phosphorus content exactly, and be capable of quick,
Batch detection.
According to embodiments of the invention, based on developer described in 20mL, described developer includes 0.5g ammonium molybdate, 0.025g
Ammonium metavanadate, 5mL nitric acid and 15mL distilled water.Thus, the kit for detecting phosphorus content according to embodiments of the present invention
Phosphorus content can be measured exactly, and be capable of quick, batch detection.
It should be noted that the offer form for developer does not make considered critical, both can be by every kind of composition with independence
Packaging provide, need use when according to said ratio configuration obtain it is also possible in the form of mixed liquor provide, need not voluntarily join
Put, directly use.
Also, it should be noted various solution are to be provided in the form of independent packaging respectively in conciliation liquid.
According to embodiments of the invention, described porous plate includes 6 orifice plates, 24 orifice plates or 96 orifice plates.Thus, according to the present invention
The kit for detecting phosphorus content of embodiment can measure phosphorus content exactly, and is capable of quick, batch detection.
Inventor finds, when GB GB5413.22-2010 measures phosphorus content, due to using spectrophotometric determination extinction
Degree, leads to every batch can only detect a small amount of sample it is impossible to complete the batch processing of sample, efficiency is low.Additionally, the volume of reaction system
Larger, required amount of reagent is more, causes to waste.Kit according to embodiments of the present invention includes porous plate, and then being capable of batch
Process sample, time-consuming and cost, improve efficiency.
It has furthermore been found that reaction system for different volumes, the interaction between each reactant is not complete for inventor
Exactly the same, and then, can not directly keep each reagent concentration in national standard method, only after volume scaled down, will be directly used in this
Application, otherwise will lead to testing result inaccurate.Inventor obtains concentration and the proportionate relationship of reagent through many experiments optimization,
So that the accuracy of testing result is higher.
It should be noted that the present invention does not make considered critical to the offer form of phosphorus standard solution, both can provide many
The phosphorus standard solution of individual variable concentrations, is directly applied to detection, to draw concentration-absorbance standard curve.Specifically,
The phosphorus standard solution of variable concentrations is provided with independent packaging form respectively.The phosphorus standard items of one high concentration can also be only provided
Solution, then according to user need it is diluted, in order to draw calibration curve.
According to embodiments of the invention, the described kit for detecting phosphorus content further includes：Perchloric acid and nitric acid.
Perchloric acid and nitric acid are used for being cleared up sample.
In a second aspect of the present invention, the present invention propose a kind of using kit measurement phosphorus content described above
Method.According to embodiments of the invention, methods described includes：Using known method, testing sample is carried out clearing up process, and
Obtained is cleared up product cooling, obtains predicting liquid；Described prediction liquid is transferred in volumetric flask, adds in described volumetric flask
Enter described indicator, be adjusted to solution being yellow with described first sodium hydroxide solution, then adjusted with described salpeter solution
It is colourless to solution, it is slightly yellow for finally being adjusted to solution with described second sodium hydroxide solution, and constant volume, obtains prepare liquid；
Respectively described phosphorus standard solution and prepare liquid are placed in described orifice plate, add described developer in described orifice plate, mix
After stand, and described orifice plate be placed in ELIASA detected；And phosphorus in described testing sample is determined based on testing result
Content.Thus, method according to embodiments of the present invention can measure phosphorus content exactly, and is capable of quick, batch detection.
It should be noted that considered critical is not made in the species for testing sample and source, can be according to actual conditions
Fixed, both can be solid sample or liquid sample.Specifically, can be foodstuff material.Concrete according to the present invention
Embodiment, testing sample can be dairy products.
According to embodiments of the invention, based on developer described in 1 μ L, the addition of described phosphorus standard solution and prepare liquid
It is 0.5 μ L.Thus, method according to embodiments of the present invention can measure phosphorus content exactly, and is capable of quick, in batches
Detection.
Inventor finds, the present invention is carried out in the orifice plate, and reaction volume is far smaller than the reaction volume of national standard method.
For the reaction system of different volumes, the interaction between each reactant is simultaneously incomplete same, and then, can not be directly by state
The reaction volume ratio (i.e. the volume ratio of prepare liquid/standard items and developer) of mark method is applied to the application, otherwise will lead to examine
The accuracy surveying result is low.Thus, inventor through many experiments optimization obtain above-mentioned optimum phosphorus standard solution, prepare liquid and
The addition of developer.Adding too much or too low all will impact testing result accuracy.
Additionally, the product of clearing up of cooling is carried out constant volume in national standard method, then the solution after 10mL constant volume is taken to hold in 50mL
Adjust pH value in measuring bottle, and directly the product of clearing up of cooling is transferred in volumetric flask in the method for the present invention, be adjusted pH
Value, decreases constant volume step, further shorten detection time.
According to embodiments of the invention, methods described includes：
Using known method, testing sample is carried out clearing up with process, and obtained is cleared up product cooling, obtain pre-
Survey liquid；
Described prediction liquid is transferred in volumetric flask, in described volumetric flask, adds indicator described in 0.10～0.15mL,
It is adjusted to solution being yellow with described first sodium hydroxide solution, then to be adjusted to solution with described salpeter solution be colourless,
It is slightly yellow for finally being adjusted to solution with described second sodium hydroxide solution, and is settled to 100mL with water, obtains prepare liquid；
Respectively phosphorus standard solution described in 100 μ L and 100 μ L prepare liquids are placed in described 96 orifice plates, to described 96 orifice plates
Middle addition developer described in 200 μ L, stands 10～15min after mixing, described 96 orifice plates is placed in ELIASA, in 440nm ripple
Long lower mensuration absorbance；And
Concentration according to described phosphorus standard solution and absorbance draw calibration curve, and the absorbance by described prepare liquid
Substitute in described calibration curve, be calculated the phosphorus content in described testing sample.
Inventor finds through many experiments, is reacted using 96 orifice plates, can detect substantial amounts of test substance simultaneously,
Drastically increase detection efficiency.Further, inventor finds, under the testing conditions of 96 orifice plates, required phosphorus standard items
Solution, prepare liquid and developer addition are respectively 100 μ L, 100 μ L, 200 μ L.Phosphorus can be measured exactly with this understanding to contain
Amount.Addition excessive or very few all will impact testing result accuracy.
Additionally, inventor finds, constant volume is carried out using the volumetric flask for 100mL for the range, be more beneficial for detecting.Specifically,
Due to directly carrying out constant volume so that the conciliation liquid measure required for subsequent adjustment pH value is on the high side by clearing up product.According to 50mL amount
The volumetric flask of journey is it is not easy to mix, and needs to add more bulk solution, easy plussage with sodium hydroxide solution when being adjusted
Journey.And then, inventor finds through many experiments, using the volumetric flask of 100mL range, can not only rapidly mix conciliation liquid
With clear up product, be easy to titrate, and without departing from range so that detection process is easy and simple to handle, quick.
Thus, method according to embodiments of the present invention can measure phosphorus content exactly, and is capable of quick, batch inspection
Survey.
According to a particular embodiment of the invention, the concrete step " using known method, testing sample being carried out clearing up process "
Suddenly as follows：
Weigh 0.5g solid sample or 2.5g liquor sample (being accurate to 0.1mg) in 125mL triangular flask, put into several glass
Glass ball, plus 10mL nitric acid, are then placed on heating on electric hot plate.Take off after vigorous reaction terminates, slightly cool down, add 10mL high
Chloric acid, is put in heating on electric hot plate again.If digestive juice blackening, need to take off and add the continuation digestion of 5mL nitric acid, until digestive juice
Become colourless or faint yellow, and white cigarette of emerging, stop when digestive juice is left 3mL～5mL clearing up process.
It should be noted that considered critical is not made in the source for perchloric acid and nitric acid, both can by commercially available acquisition,
Can be provided by kit described above.
The additional aspect of the present invention and advantage will be set forth in part in the description, and partly will become from the following description
Obtain substantially, or recognized by the practice of the present invention.
Specific embodiment
Below in conjunction with embodiment, the solution of the present invention is explained.It will be understood to those of skill in the art that it is following
Embodiment is merely to illustrate the present invention, and should not be taken as limiting the scope of the invention.Unreceipted particular technique or bar in embodiment
Part, carry out according to the technology described by document in the art or condition or according to product description.Agents useful for same or instrument
The unreceipted production firm person of device, be can by city available from conventional products.
Embodiment 1
In this embodiment, phosphorus content in determination sample in following manner.
1.1 sample determination
1.1.1 the drafting of calibration curve
Accurately draw 100 μ L phosphorus standard solutions in normal orifice, add developer 200 μ L, mix, 25～30 DEG C
Lower colour developing 15 minutes, measures its light absorption value on ELIASA under 440nm wavelength, and with light absorption value as ordinate, phosphorus content is horizontal stroke
Coordinate draws curve.
Table 1：100 μ L standard items+200 μ L developer detection data
Remarks：Each concentration point Parallel testing, takes absorbance values；Net light absorption value deducts zero for each concentration point detected value
The detected value of point light absorption value.
Result is as shown in table 1.With net light absorption value as ordinate, phosphorus standard concentration draws curve, curve side for abscissa
Journey is y=0.008x+0.002 (R2=0.997), curve linear is good, meets the R of method requirement2≥0.99.
1.1.2 sample determination
(1) weigh milk 2.5g (being accurate to 0.1mg), in 125mL triangular flask, put into several glass marbles, plus 10mL nitre
Acid, is then placed on electric hot plate, heats under 200～300 degrees Celsius.Take off after vigorous reaction terminates, slightly cool down, add
10mL perchloric acid, is put on electric hot plate again, heats under 200～300 degrees Celsius.Take when digestive juice is left 3mL～5mL
Under, cooling, proceed in 100mL volumetric flask；
(2) add 50mL water in volumetric flask, add 0.10～0.15mL dinitrophenol dinitrophenolate indicator, adjusted with 6mol/LNaOH
To yellow, use 0.2mol/LHNO3It is adjusted to colourless, is adjusted to 0.1mol/LNaOH slightly yellow, be settled to 100mL as prepare liquid,
Each sample arranges two parallel tests.Do blank test, as reference substance simultaneously；
(3) draw 100 μ L prepare liquids respectively and reference substance is placed in normal orifice, add developer 200 μ L, 25～30
Develop the color 15 minutes at DEG C, on ELIASA, measure its light absorption value under 440nm wavelength, and calculate phosphorus content and (participate in curve to calculate
Sample light absorption value be that blank absorbency is deducted by actually measured sample light absorption value).
Computing formula：
P (mg/100g)=(C × V) × 100/ (M × 1000)
In formula：
Phosphorus content in P--- sample, mg/100g；
The concentration of phosphorus in the prepare liquid that C--- checks in from calibration curve, μ g/mL；
V--- prepare liquid volume, mL, 100；
The quality of M--- sample, g.
Comparative example 1
In this comparative example, draw calibration curve according to the method in above-described embodiment 1, its difference is, standard items are molten
Liquid addition is 50 μ L, and developer addition is 150 μ L, and concrete detected value is as follows：
Table 2：50 μ L standard items+150 μ L developer detection data
According to table 2 data, with net light absorption value as ordinate, phosphorus standard concentration draws curve, curvilinear equation for abscissa
For y=0.004x+0.003 (R2=0.969) curve linear does not meet the R of method requirement2≥0.99.Thus standard items are worked as in explanation
When the addition of solution and developer is 50 μ L and 150 μ L, calibration curve is inaccurate, and then will affect phosphorus content in prepare liquid
Determine, lead to the accuracy of testing result low.
Comparative example 2
In this comparative example, draw calibration curve according to the method in above-described embodiment 1, its difference is, standard items are molten
Liquid addition is 200 μ L, and developer addition is 100 μ L, and concrete detected value is as follows：
Table 3：200 μ L standard items+100 μ L developer detection data
According to data above, with net light absorption value as ordinate, phosphorus standard concentration draws curve, curvilinear equation for abscissa
For y=0.011x+0.004 (R2=0.985) curve linear does not meet the R of method requirement2≥0.99.Thus standard items are worked as in explanation
When the addition of solution and developer is 200 μ L and 100 μ L, calibration curve is inaccurate, and then will affect phosphorus content in prepare liquid
Determination, lead to the accuracy of testing result low.
In summary, the addition of standard solution and developer significantly affects the accuracy of calibration curve, and then affects
The accuracy of final detection result.When the addition of standard solution and developer is respectively 100 μ L and 200 μ L, standard is bent
The accuracy of line is higher.Further, when prepare liquid is reacted with developer, when the addition of prepare liquid is similarly 100 μ L, can
Determine phosphorus content in sample exactly.
Embodiment 2
The method of the method and embodiment 1 that measure phosphorus content according to GB GB5413.22-2010 respectively measures milk for children
And modulate phosphorus content in breast, concrete detection data is as follows：
Table 4：Comparison between detecting methods result
According to data above, can meet with national standard method detection data relative deviation with embodiment 1 detection data
The tolerance that method in GB5413.22-2010 requires.Thus showing that the method for the present invention can measure phosphorus content exactly.
In the description of this specification, reference term " embodiment ", " some embodiments ", " example ", " specifically show
The description of example " or " some examples " etc. means specific features, structure, material or the spy describing with reference to this embodiment or example
Point is contained at least one embodiment or the example of the present invention.In this manual, to the schematic representation of above-mentioned term not
Identical embodiment or example must be directed to.And, the specific features of description, structure, material or feature can be in office
Combine in an appropriate manner in one or more embodiments or example.Additionally, in the case of not conflicting, the skill of this area
The feature of the different embodiments described in this specification or example and different embodiment or example can be tied by art personnel
Close and combine.
Although embodiments of the invention have been shown and described above it is to be understood that above-described embodiment is example
Property it is impossible to be interpreted as limitation of the present invention, 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 (7)
1. a kind of kit for detecting phosphorus content is it is characterised in that include：
Indicator, described indicator is the paranitrophenol solution of 0.2 volume %；
Developer, described developer includes ammonium molybdate, ammonium metavanadate, nitric acid and water；And
Reconcile liquid, described conciliation liquid includes 0.2mol/L salpeter solution, 6mol/L first sodium hydroxide solution and 0.1mol/L
Second sodium hydroxide solution.
2. kit according to claim 1 is it is characterised in that further include：
Phosphorus standard solution；And
Porous plate.
3. it is characterised in that being based on developer described in 20mL, described developer includes kit according to claim 1
0.5g ammonium molybdate, 0.025g ammonium metavanadate, 5mL nitric acid and 15mL distilled water.
4. kit according to claim 1 is it is characterised in that described porous plate includes 6 orifice plates, 24 orifice plates or 96 holes
Plate.
5. a kind of method of the kit measurement phosphorus content described in utilization any one of claim 1～4 is it is characterised in that include：
Using known method, testing sample is carried out clearing up with process, and obtained is cleared up product cooling, obtain predicting liquid；
Described prediction liquid is transferred in volumetric flask, adds described indicator in described volumetric flask, with described first hydroxide
It is yellow that sodium solution is adjusted to solution, then to be adjusted to solution with described salpeter solution be colourless, finally with described second hydrogen
It is slightly yellow that sodium hydroxide solution is adjusted to solution, and constant volume, obtains prepare liquid；
Respectively described phosphorus standard solution and prepare liquid are placed in described orifice plate, in described orifice plate, add described developer,
Stand after mixing, and described orifice plate is placed in ELIASA is detected；And
Phosphorus content in described testing sample is determined based on testing result.
6. method according to claim 5 it is characterised in that be based on developer described in 1 μ L, described phosphorus standard solution and
The addition of prepare liquid is 0.5 μ L.
7. method according to claim 5 is it is characterised in that include：
Using known method, testing sample is carried out clearing up with process, and obtained is cleared up product cooling, obtain predicting liquid；
Described prediction liquid is transferred in volumetric flask, adds indicator described in 0.10～0.15mL in described volumetric flask, use institute
State the first sodium hydroxide solution and be adjusted to solution being yellow, then to be adjusted to solution with described salpeter solution be colourless, finally
It is slightly yellow for being adjusted to solution with described second sodium hydroxide solution, and is settled to 100mL with water, obtains prepare liquid；
Respectively phosphorus standard solution described in 100 μ L and 100 μ L prepare liquids are placed in described 96 orifice plates, add in described 96 orifice plates
Enter developer described in 200 μ L, stand 10～15min after mixing, described 96 orifice plates are placed in ELIASA, under 440nm wavelength
Mensuration absorbance；And
Concentration according to described phosphorus standard solution and absorbance draw calibration curve, and the absorbance of described prepare liquid is substituted into
In described calibration curve, it is calculated the phosphorus content in described testing sample.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610773879.2A CN106404999A (en) | 2016-08-30 | 2016-08-30 | Kit for detecting phosphor content and method for measuring phosphor content |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610773879.2A CN106404999A (en) | 2016-08-30 | 2016-08-30 | Kit for detecting phosphor content and method for measuring phosphor content |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106404999A true CN106404999A (en) | 2017-02-15 |
Family
ID=58002287
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610773879.2A Pending CN106404999A (en) | 2016-08-30 | 2016-08-30 | Kit for detecting phosphor content and method for measuring phosphor content |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106404999A (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1687787A (en) * | 2005-04-08 | 2005-10-26 | 山西大学 | Method for visual detection of inorganic phosphorus in urine and kit thereof |
| CN101968452A (en) * | 2010-03-05 | 2011-02-09 | 浙江省疾病预防控制中心 | Kit for quickly detecting tripolycyanamide and use thereof |
| CN105567788A (en) * | 2016-01-18 | 2016-05-11 | 北京农业职业学院 | Kit for detecting urea nitrogen in milk |
-
2016
- 2016-08-30 CN CN201610773879.2A patent/CN106404999A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1687787A (en) * | 2005-04-08 | 2005-10-26 | 山西大学 | Method for visual detection of inorganic phosphorus in urine and kit thereof |
| CN101968452A (en) * | 2010-03-05 | 2011-02-09 | 浙江省疾病预防控制中心 | Kit for quickly detecting tripolycyanamide and use thereof |
| CN105567788A (en) * | 2016-01-18 | 2016-05-11 | 北京农业职业学院 | Kit for detecting urea nitrogen in milk |
Non-Patent Citations (1)
| Title |
|---|
| 无: "GB 5413.22-2010 婴幼儿配方食品和乳粉中磷的测定", 《中华人民共和国国家标准》 * |
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