CN1865925A - Method for detecting inorganic phosphorus in milk - Google Patents
Method for detecting inorganic phosphorus in milk Download PDFInfo
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- CN1865925A CN1865925A CN 200610012761 CN200610012761A CN1865925A CN 1865925 A CN1865925 A CN 1865925A CN 200610012761 CN200610012761 CN 200610012761 CN 200610012761 A CN200610012761 A CN 200610012761A CN 1865925 A CN1865925 A CN 1865925A
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- milk
- solution
- phos
- cuvette
- visible light
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- 239000008267 milk Substances 0.000 title claims abstract description 44
- 235000013336 milk Nutrition 0.000 title claims abstract description 42
- 210000004080 milk Anatomy 0.000 title claims abstract description 42
- 238000000034 method Methods 0.000 title claims abstract description 16
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 title abstract description 18
- 229910052698 phosphorus Inorganic materials 0.000 title abstract description 18
- 239000011574 phosphorus Substances 0.000 title abstract description 18
- 239000000243 solution Substances 0.000 claims abstract description 45
- 235000020256 human milk Nutrition 0.000 claims abstract description 21
- 210000004251 human milk Anatomy 0.000 claims abstract description 21
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 claims abstract description 9
- 239000007995 HEPES buffer Substances 0.000 claims abstract description 9
- 238000005259 measurement Methods 0.000 claims abstract description 4
- 239000007788 liquid Substances 0.000 claims description 18
- 238000010521 absorption reaction Methods 0.000 claims description 15
- HGPSVOAVAYJEIJ-XDHOZWIPSA-N 2-[(e)-(3,4-dihydroxyphenyl)-(3-hydroxy-4-oxoniumylidenecyclohexa-2,5-dien-1-ylidene)methyl]benzenesulfonate Chemical compound C1=CC(=O)C(O)=C\C1=C(C=1C(=CC=CC=1)S(O)(=O)=O)/C1=CC=C(O)C(O)=C1 HGPSVOAVAYJEIJ-XDHOZWIPSA-N 0.000 claims description 9
- 238000000108 ultra-filtration Methods 0.000 claims description 9
- 229910020366 ClO 4 Inorganic materials 0.000 claims description 5
- 235000020251 goat milk Nutrition 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 4
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 3
- 241001062009 Indigofera Species 0.000 claims description 3
- 229910001873 dinitrogen Inorganic materials 0.000 claims description 3
- 239000012528 membrane Substances 0.000 claims description 3
- 238000004321 preservation Methods 0.000 claims description 3
- 238000001514 detection method Methods 0.000 abstract description 6
- 229910052769 Ytterbium Inorganic materials 0.000 abstract description 2
- -1 ytterbium ions Chemical class 0.000 abstract description 2
- RRRCKIRSVQAAAS-UHFFFAOYSA-N 4-[3-(3,4-dihydroxyphenyl)-1,1-dioxo-2,1$l^{6}-benzoxathiol-3-yl]benzene-1,2-diol Chemical compound C1=C(O)C(O)=CC=C1C1(C=2C=C(O)C(O)=CC=2)C2=CC=CC=C2S(=O)(=O)O1 RRRCKIRSVQAAAS-UHFFFAOYSA-N 0.000 abstract 1
- 239000003795 chemical substances by application Substances 0.000 abstract 1
- 235000016709 nutrition Nutrition 0.000 abstract 1
- 238000000870 ultraviolet spectroscopy Methods 0.000 abstract 1
- 230000031700 light absorption Effects 0.000 description 9
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 6
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 6
- 239000011575 calcium Substances 0.000 description 5
- 229910052791 calcium Inorganic materials 0.000 description 5
- MWKXCSMICWVRGW-UHFFFAOYSA-N calcium;phosphane Chemical compound P.[Ca] MWKXCSMICWVRGW-UHFFFAOYSA-N 0.000 description 4
- 229910052742 iron Inorganic materials 0.000 description 3
- 208000007442 rickets Diseases 0.000 description 3
- 208000007502 anemia Diseases 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- BHEPBYXIRTUNPN-UHFFFAOYSA-N hydridophosphorus(.) (triplet) Chemical compound [PH] BHEPBYXIRTUNPN-UHFFFAOYSA-N 0.000 description 2
- 208000013038 Hypocalcemia Diseases 0.000 description 1
- 208000002720 Malnutrition Diseases 0.000 description 1
- 244000299461 Theobroma cacao Species 0.000 description 1
- 230000023555 blood coagulation Effects 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 235000019219 chocolate Nutrition 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 235000020247 cow milk Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000000705 hypocalcaemia Effects 0.000 description 1
- 229910052816 inorganic phosphate Inorganic materials 0.000 description 1
- 230000006651 lactation Effects 0.000 description 1
- 230000001071 malnutrition Effects 0.000 description 1
- 235000000824 malnutrition Nutrition 0.000 description 1
- 235000020124 milk-based beverage Nutrition 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000002232 neuromuscular Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 208000015380 nutritional deficiency disease Diseases 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 230000005195 poor health Effects 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 230000001228 trophic effect Effects 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
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Abstract
本发明使用镱离子、以邻苯二酚紫为显色剂在pH6.5－7.5的HEPES缓冲溶液在紫外可见分光光度仪上定性、定量的去测定奶中无机磷的含量，测定过程简单、方便、无干扰，测量结果准确；用检测体系去检测母乳和商业牛奶中的无机磷，通过测定母乳中无机磷的含量，可以了解新生婴儿无机磷的供应状况；通过测定商业牛奶中无机磷的指标，可以提高牛奶加工的质量，从而增加牛奶的营养价值；该方法可适用于临床和商业。The present invention uses ytterbium ions, catechol violet as a chromogenic agent, and HEPES buffer solution at pH 6.5-7.5 to qualitatively and quantitatively measure the content of inorganic phosphorus in milk on an ultraviolet-visible spectrophotometer, and the determination process is simple and convenient. Convenient, no interference, and accurate measurement results; use the detection system to detect inorganic phosphorus in breast milk and commercial milk, and by measuring the content of inorganic phosphorus in breast milk, you can understand the supply status of inorganic phosphorus in newborn babies; Indicators that can improve the quality of milk processing, thereby increasing the nutritional value of milk; the method can be applied clinically and commercially.
Description
Technical field:
The present invention relates to a kind of detection method of Phos, specifically belong to a kind of method that detects Phos in the milk, promptly detect the method for Phos in breast milk, milk or the goat milk.
Background technology:
For new life's baby, breast milk or milk or goat milk are the topmost food sources that they grow up and grow.And this period exactly is the fastest period of infant growth, is in the not perfect stage of skeletonization animated period and brain and intelligence, needs to take in an amount of nutrient, could guarantee to grow normally, and prevent malnutrition, rickets, anaemia etc.Ionization element such as calcium, phosphorus are that normal blood coagulation and neuromuscular function are necessary, because they are important component parts of bone, so claim big element again.If but the calcium phosphorus ration that absorbs imbalance can cause hypocalcemia, causes rickets, and the iron in the milk be vulnerable to suckle in the influence of high phosphorus, high calcium form insoluble iron containing compounds, the utilization that can not be absorbed by the body, can cause children's in poor health, easily be stranded easily tiredly, hypoferric anemia takes place.The ratio of calcium phosphorus is 2: 1 in the normal breast milk, is easy to absorb, and to preventing and treating rickets certain effect is arranged; And milk is 1.2: 1, is difficult for absorbing.Therefore, the index of calcium phosphorus is very important in mensuration breast milk and the milk.By the content of calcium phosphorus in the mensuration consumption by infants milk, thereby in time rationally add diatery supplement.Certainly along with the raising of people's standard of living, commercial milk becomes the basic nutriment of people.But the milk after the commercial processing of milk and process, the index of quite a lot of important component is not enough, and calcic is few, phosphorous many in the cow's milk, forms the calcium phosphate of slightly solubility easily, influences enteron aisle calcium, phosphorus absorption.If eat the child of milk, add many, the phosphorous few ternips of calcic, correct calcium, phosphorus ratio, can improve the calcium absorption amount.So, need reasonably to add and regulate just to be unlikely to owing to milk drink causes trophic disturbance.For a long time, people just pay close attention to the mensuration of Phos in urine and the serum both at home and abroad, and the method for measuring Phos in the milk is seldom mentioned.
Summary of the invention:
The object of the present invention is to provide a kind of method that detects Phos in the milk, this method is applicable to clinical and commercial use.
The invention provides a kind of method that detects Phos in the milk, this method Phos that to be developer based on ytterbium ion, with the pyrocatechol violet detect in pH is the HEPES buffer solution of 6.5-7.5 quantitatively in the milk, it is as follows that it specifically detects step:
1, gets breast milk or milk or goat milk, the centrifugal upper strata liquid that gets on high speed tabletop centrifuge;
2, get above-mentioned upper strata liquid, change the ultrafiltration cup over to, in room temperature and 5 * 10
5Use the PM-10 ultra filtration membrane ultrafilter ultrafiltration of molecular cut off under the condition of nitrogen gas of Pa, collect the colourless limpid liquid that leaches ,-17 ℃ to-20 ℃ preservations, for measurement as 10kD;
3, the HEPES buffer solution of preparation pH6.5-7.5, and prepare 2 * 10
-3The Yb of M
3+Solution and 2 * 10
-3The pyrocatechol violet solution of M;
4, the HEPES buffer solution of 2ml is added in the clean ultraviolet cuvette,, draws 2 * 10 with microsyringe as blank
-3The pyrocatechol violet solution 10-100 μ l of M is added in this cuvette, and solution is detected on the UV, visible light spectrophotometer by colourless flavescence at this moment, and 443nm has absorption maximum;
5, in above-mentioned cuvette, add 2 times of molal quantitys to the Yb of pyrocatechol violet solution
3+Solution, solution is detected on the UV, visible light spectrophotometer by xanthochromia indigo plant at this moment, finds that maximum absorption band becomes 623nm by above-mentioned 443nm;
6, get above-mentioned colourless limpid liquid, be added in this cuvette with microsyringe gradually, application of sample limit, limit is detected on the UV, visible light spectrophotometer, adding along with limpid liquid, maximum absorption band is become again to 443nm gradually by 623nm again, and the color of solution is also returned yellow by blue stain gradually, when absorption peak reaches maximum at 443nm, during the complete flavescence of the color of solution, stop to add limpid liquid;
7, by [Yb
3+] * 2 * 10
3/ V
MilkCalculate the content of Phos in the milk;
Wherein, described Yb
3+Solution is YbCl
3Solution or Yb (ClO
4)
3Solution.
Described Yb
3+Can use Nd
3+, Sm
3+, Gd
3+, Er
3+Or Ho
3+Replace.
The present invention has following advantage: 1, do not need synthetic any reagent, detect on the UV, visible light spectrophotometer, testing process is simple, and can with the naked eye obviously observe this effect; 2, we to milk do super consider handle after, remaining other many compositions in the milk such as calcium, iron, candy part etc., to the detection of Phos without any interference; 3, detection of the present invention is quantitative, the very big range of linearity is arranged, and error is very little; 4, remove to detect Phos in breast milk and the commercial milk with detection architecture,, can understand the supply situation of ewborn infant Phos by measuring the content of Phos in the breast milk; By measuring the index of Phos in the commercial milk, can improve the quality of milk processing, thereby increase the nutritive value of milk; Therefore, our method can be used for clinical and commercial, will have very big social value.
Description of drawings:
Fig. 1 to Fig. 8 is the UV, visible light absorption figure of Phos in the detection breast milk of corresponding embodiment 1 to embodiment 8 and the milk.
Embodiment:
Embodiment 1:
Get breast milk from a certain mother who gives birth to 3 months and lactation, centrifugal upper strata liquid (separating a large amount of fat) on TGL-16B type high speed tabletop centrifuge; Get above-mentioned upper strata liquid 6ml, change 10ml ultrafiltration cup over to, in room temperature and 5 * 10
5Use the PM-10 ultra filtration membrane of molecular cut off under the condition of nitrogen gas of Pa,, collect the colourless limpid liquid that leaches (collect former milk volume 3/4) ,-17 ℃ to-20 ℃ preservations, for measurement with Amicon Model 8010 ultrafilter ultrafiltration as 10kD; HEPES (10mM) buffer solution of preparation pH7.0, and prepare 2 * 10
-3The YbCl of M
3Solution and 2 * 10
-3The pyrocatechol violet solution of M; The HEPES buffer solution of 2ml is added in the clean ultraviolet cuvette,, draws 2 * 10 with microsyringe as blank
-3The pyrocatechol violet solution 50 μ l of M are added in this cuvette, and solution is detected on the UV, visible light spectrophotometer by colourless flavescence at this moment, and 443nm has absorption maximum; In above-mentioned cuvette, add 2 * 10
-3The Yb of M
3+Solution 100 μ l, solution is detected on the UV, visible light spectrophotometer by xanthochromia indigo plant at this moment, finds that maximum absorption band becomes 623nm by above-mentioned 443nm; Get above-mentioned limpid liquid, be added in this cuvette with microsyringe gradually, application of sample limit, limit is detected on HP8453 UV, visible light spectrophotometer, adding along with limpid liquid, maximum absorption band is become again to 443nm gradually by 623nm again, and the color of solution is also returned yellow by blue stain gradually, when absorption peak reaches maximum at 443nm, during the complete flavescence of the color of solution, this moment, limpid liquid addition was 62 μ l; Calculate:
In this breast milk content of inorganic phosphorus be 2.42mmol/L, UV, visible light absorption figure sees Fig. 1.
Embodiment 2: HEPES (10mM) buffer solution of preparation pH6.5, all the other are with embodiment 1, in the breast milk content of inorganic phosphorus be 2.42mmol/L, UV, visible light absorption figure sees Fig. 2.
Embodiment 3: with Yb (ClO
4)
3Solution, get another people's breast milk, all the other are with embodiment 1,
In the breast milk content of inorganic phosphorus be 2.34mmol/L, UV, visible light absorption figure sees Fig. 3.
Embodiment 4: use ErCl
3Solution, all the other are with embodiment 1,
In the breast milk content of inorganic phosphorus be 2.42mmol/L, UV, visible light absorption figure sees Fig. 4.
Embodiment 5: with Nd (ClO
4)
3Solution, all the other are with embodiment 1,
In the breast milk content of inorganic phosphorus be 2.42mmol/L, UV, visible light absorption figure sees Fig. 5.
Embodiment 6: with Sm (ClO
4)
3Solution, get commercial packed in flexible pouchy Mongolia Ox brand milk,
In the milk content of inorganic phosphorus be 12.5mmol/L, UV, visible light absorption figure sees Fig. 6.
Embodiment 7: use GdCl
3Solution, get commercial packed in flexible pouchy Erie brand milk,
In the milk content of inorganic phosphorus be 12.5mmol/L, UV, visible light absorption figure sees Fig. 7.
Embodiment 8: use HoCl
3Solution, get the permanent health brand of commercial packed in flexible pouchy milk,
In the milk content of inorganic phosphorus be 11.54mmol/L, UV, visible light absorption figure sees Fig. 8.
Press the breast milk that embodiment 1 surveys 5 ursing mothers, in the breast milk inorganic phosphate content be 2.31-2.46mmol/L; Survey 5 kinds of commercial brand plain chocolates, in the commercially pure milk content of inorganic phosphorus be 11.54-12.5mmol/L.
Claims (2)
1, a kind of method that detects Phos in the milk is characterized in that comprising the steps:
(1) gets breast milk or milk or goat milk, the centrifugal upper strata liquid that gets on high speed tabletop centrifuge;
(2) get above-mentioned upper strata liquid, change the ultrafiltration cup over to, in room temperature and 5 * 10
5Use the PM-10 ultra filtration membrane ultrafilter ultrafiltration of molecular cut off under the condition of nitrogen gas of Pa, collect the colourless limpid liquid that leaches ,-17 ℃ to-20 ℃ preservations, for measurement as 10kD;
(3), the HEPES buffer solution of preparation pH6.5-7.5, and prepare 2 * 10
-3The Yb of M
3+Solution and 2 * 10
-3The pyrocatechol violet solution of M;
(4), the HEPES buffer solution of 2ml is added in the clean ultraviolet cuvette, as blank, with microsyringe absorption 2 * 10
-3The pyrocatechol violet solution 10-100 μ l of M is added in this cuvette, and solution is detected on the UV, visible light spectrophotometer by colourless flavescence at this moment, and 443nm has absorption maximum;
(5), in above-mentioned cuvette, add 2 times of molal quantitys to the Yb of pyrocatechol violet solution
3+Solution, solution is detected on the UV, visible light spectrophotometer by xanthochromia indigo plant at this moment, and maximum absorption band becomes 623nm by above-mentioned 443nm;
(6), get colourless limpid liquid, be added in this cuvette with microsyringe gradually, application of sample limit, limit is detected on the UV, visible light spectrophotometer, adding along with limpid liquid, maximum absorption band is become again to 443nm gradually by 623nm again, and the color of solution is also returned yellow by blue stain gradually, when absorption peak reaches maximum at 443nm, during the complete flavescence of the color of solution, stop to add colourless limpid liquid;
(7), by [Yb
3+] * 2/V
MilkCalculate the content of Phos in the milk;
Wherein, described Yb
3+Solution is YbCl
3Solution or Yb (ClO
4)
3Solution.
2, by the described a kind of method that detects Phos in breast milk and the milk of claim 1, it is characterized in that described Yb
3+Can use Nd
3+, Sm
3+, Gd
3+, Er
3+Or Ho
3+Replace.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200610012761 CN1865925A (en) | 2006-05-26 | 2006-05-26 | Method for detecting inorganic phosphorus in milk |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200610012761 CN1865925A (en) | 2006-05-26 | 2006-05-26 | Method for detecting inorganic phosphorus in milk |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1865925A true CN1865925A (en) | 2006-11-22 |
Family
ID=37425003
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200610012761 Pending CN1865925A (en) | 2006-05-26 | 2006-05-26 | Method for detecting inorganic phosphorus in milk |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1865925A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103257137A (en) * | 2012-02-17 | 2013-08-21 | 谱尼测试科技股份有限公司 | Method of determining content of methyl ethyl ketone peroxide in workplace air through UV spectrophotometer |
| CN104777119A (en) * | 2015-04-27 | 2015-07-15 | 泰安市康宇医疗器械有限公司 | Breast milk micro-measuring method |
-
2006
- 2006-05-26 CN CN 200610012761 patent/CN1865925A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103257137A (en) * | 2012-02-17 | 2013-08-21 | 谱尼测试科技股份有限公司 | Method of determining content of methyl ethyl ketone peroxide in workplace air through UV spectrophotometer |
| CN104777119A (en) * | 2015-04-27 | 2015-07-15 | 泰安市康宇医疗器械有限公司 | Breast milk micro-measuring method |
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