CN111380895A - Nuclear magnetic resonance hydrogen spectrometry for identifying whey protein powder added in formula powder - Google Patents
Nuclear magnetic resonance hydrogen spectrometry for identifying whey protein powder added in formula powder Download PDFInfo
- Publication number
- CN111380895A CN111380895A CN202010250441.2A CN202010250441A CN111380895A CN 111380895 A CN111380895 A CN 111380895A CN 202010250441 A CN202010250441 A CN 202010250441A CN 111380895 A CN111380895 A CN 111380895A
- Authority
- CN
- China
- Prior art keywords
- powder
- formula
- whey protein
- nuclear magnetic
- magnetic resonance
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000000843 powder Substances 0.000 title claims abstract description 132
- 102000007544 Whey Proteins Human genes 0.000 title claims abstract description 60
- 108010046377 Whey Proteins Proteins 0.000 title claims abstract description 60
- 235000021119 whey protein Nutrition 0.000 title claims abstract description 60
- 238000005481 NMR spectroscopy Methods 0.000 title claims abstract description 40
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 title claims abstract description 23
- 229910052739 hydrogen Inorganic materials 0.000 title claims abstract description 23
- 239000001257 hydrogen Substances 0.000 title claims abstract description 23
- 238000004611 spectroscopical analysis Methods 0.000 title claims abstract description 6
- 235000013350 formula milk Nutrition 0.000 claims abstract description 50
- XLYOFNOQVPJJNP-ZSJDYOACSA-N Heavy water Chemical compound [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 claims abstract description 24
- 238000001228 spectrum Methods 0.000 claims abstract description 22
- 241000283690 Bos taurus Species 0.000 claims abstract description 21
- 210000003022 colostrum Anatomy 0.000 claims abstract description 20
- 235000021277 colostrum Nutrition 0.000 claims abstract description 20
- 238000000034 method Methods 0.000 claims abstract description 19
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 19
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 19
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 claims abstract description 18
- 235000018102 proteins Nutrition 0.000 claims abstract description 18
- 239000007788 liquid Substances 0.000 claims abstract description 10
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims abstract description 9
- 229960001701 chloroform Drugs 0.000 claims abstract description 9
- 150000001875 compounds Chemical class 0.000 claims abstract description 8
- 238000005303 weighing Methods 0.000 claims abstract description 7
- 229910021642 ultra pure water Inorganic materials 0.000 claims abstract description 6
- 239000012498 ultrapure water Substances 0.000 claims abstract description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 11
- 238000012937 correction Methods 0.000 claims description 10
- 239000006228 supernatant Substances 0.000 claims description 9
- 238000003825 pressing Methods 0.000 claims description 8
- 239000000126 substance Substances 0.000 claims description 8
- 239000008186 active pharmaceutical agent Substances 0.000 claims description 5
- 238000000079 presaturation Methods 0.000 claims description 5
- 238000010438 heat treatment Methods 0.000 claims description 3
- 238000005259 measurement Methods 0.000 claims description 3
- 239000002253 acid Substances 0.000 claims description 2
- 235000008476 powdered milk Nutrition 0.000 claims description 2
- 238000006073 displacement reaction Methods 0.000 abstract description 12
- ISMLZRAEFVWAKT-UHFFFAOYSA-N C[SiH](C)C.C(C=C)(=O)O Chemical class C[SiH](C)C.C(C=C)(=O)O ISMLZRAEFVWAKT-UHFFFAOYSA-N 0.000 abstract description 4
- 238000005119 centrifugation Methods 0.000 abstract 1
- 239000000523 sample Substances 0.000 description 17
- 239000003153 chemical reaction reagent Substances 0.000 description 14
- 235000013336 milk Nutrition 0.000 description 11
- 239000008267 milk Substances 0.000 description 11
- 210000004080 milk Anatomy 0.000 description 11
- 238000004458 analytical method Methods 0.000 description 10
- 238000005160 1H NMR spectroscopy Methods 0.000 description 9
- 238000001514 detection method Methods 0.000 description 7
- 239000002131 composite material Substances 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 4
- OIYDLRPLPUQXCT-UHFFFAOYSA-M C(CC)(=O)[O-].[Na+].C[SiH](C)C Chemical class C(CC)(=O)[O-].[Na+].C[SiH](C)C OIYDLRPLPUQXCT-UHFFFAOYSA-M 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 239000011259 mixed solution Substances 0.000 description 3
- 235000016709 nutrition Nutrition 0.000 description 3
- 102000004407 Lactalbumin Human genes 0.000 description 2
- 108090000942 Lactalbumin Proteins 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 235000012907 honey Nutrition 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 108060003951 Immunoglobulin Proteins 0.000 description 1
- 102000008192 Lactoglobulins Human genes 0.000 description 1
- 108010060630 Lactoglobulins Proteins 0.000 description 1
- 102000007562 Serum Albumin Human genes 0.000 description 1
- 108010071390 Serum Albumin Proteins 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 230000008094 contradictory effect Effects 0.000 description 1
- 235000013365 dairy product Nutrition 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 235000015872 dietary supplement Nutrition 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 230000006806 disease prevention Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 235000013402 health food Nutrition 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 102000018358 immunoglobulin Human genes 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000001195 ultra high performance liquid chromatography Methods 0.000 description 1
- 235000021241 α-lactalbumin Nutrition 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N24/00—Investigating or analyzing materials by the use of nuclear magnetic resonance, electron paramagnetic resonance or other spin effects
- G01N24/08—Investigating or analyzing materials by the use of nuclear magnetic resonance, electron paramagnetic resonance or other spin effects by using nuclear magnetic resonance
Landscapes
- Physics & Mathematics (AREA)
- High Energy & Nuclear Physics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention relates to a nuclear magnetic resonance hydrogen spectrometry for identifying formula powder added with whey protein powder, which comprises the steps of weighing formula powder to be identified, dissolving the formula powder with ultrapure water, adding a proper amount of trichloromethane for vortex centrifugation, weighing a small amount of processed upper layer sample, adding a heavy water solution containing 2,2,3, 3-tetradeuterated trimethyl silane acrylic acid and sodium azide, uniformly mixing, and analyzing by using a liquid nuclear magnetic resonance instrument. The nuclear magnetic resonance hydrogen spectrum fingerprint of the obtained sample has the following characteristics: and if a convex peak exists within the displacement of 0.80-1.0 ppm and is connected with a saw-tooth-shaped multi-peak, a cluster of multi-peaks exists within the displacement of 1.37-1.43 ppm, and a peak exists within the displacement of 3.011-3.025 ppm, the whey protein powder is added into the identified formula powder. The method provided by the invention is simple and efficient, and can accurately identify the whey protein powder in the formula powder such as the bovine colostrum powder, the formula milk powder, the compound protein powder and the like.
Description
Technical Field
The invention belongs to the technical field of health food detection, and particularly relates to a method for identifying whey protein powder added in formula powder by adopting a nuclear magnetic resonance hydrogen spectrum fingerprint.
Background
Whey protein is a protein extracted from milk, accounts for 20% of total protein of milk, and mainly comprises β -lactoglobulin, α -lactalbumin, serum albumin, immunoglobulin and the like, has the advantages of high nutritional value, easy digestion and absorption, rich various bioactive components and the like, has important effects on human body metabolic balance, immunity, disease prevention and treatment and the like, is called 'the king of protein', and is an important raw material for producing various formula powder, sports nutritional supplements and other health-care foods.
At present, the detection of related whey protein in China is limited to detecting the content of whey protein rather than identifying the components of whey protein, and the standard 'determination of whey protein in infant formula food and milk powder' (with the standard number of GB/T5413.21997) and the patent 'a method for determining the content of whey protein in milk-based infant formula powder' (with the application number of 201410722848.5) analyze the protein band based on the SDS-PAGE technology, so that the analysis steps are complex, and the test result is easy to generate errors; the patent 'method for measuring the content of bovine whey protein in milk powder and milk by ultra-high performance liquid chromatography' (application number is 201110283264.9) needs to use a specific chromatographic column, and has the defects of high consumption of organic reagents and high detection cost. Therefore, a technology for rapidly identifying whether whey protein powder is added to formula powder such as bovine colostrum powder, formula milk powder, compound protein powder and the like is urgently needed. The NMR (nuclear magnetic resonance) hydrogen spectrum fingerprint technology is used as a novel detection means, is used for the research of the sources and metabolites of dairy products by many scholars, but no research report for identifying whey protein powder added in formula powder exists at present.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a method for qualitatively detecting whey protein powder added in formula powder such as bovine colostrum powder, formula milk powder or compound protein powder, which is based on a liquid nuclear magnetic resonance hydrogen spectrometry fingerprint spectrum technology, has the advantages of simple sample pretreatment, high detection sensitivity, high detection efficiency and the like, can realize quick identification of the whey protein powder in the formula powder, and is suitable for daily analysis of the quality of the formula powder.
In order to achieve the purpose, the invention is realized by the following technical means:
the invention relates to a nuclear magnetic resonance hydrogen spectrometry for identifying whey protein powder added in formula powder, which comprises the following steps:
step 1: weighing 0.8-1.5 g of formula powder to be identified, and dissolving the formula powder in 10-15 mL of ultrapure water;
step 2: adding 10 mL of trichloromethane into the step 1, fully whirling, and centrifuging at a high speed for 10 min;
and step 3: taking 600-900-microliter of the sample supernatant treated in the step 2, adding a heavy water solution containing 0.1-0.2% of 2,2,3, 3-tetradeuterotrimethylsilylacrylic acid and 0.01-0.02% of sodium azide, fixing the volume to 1 mL, uniformly swirling, and measuring 500-600-microliter of the sample supernatant and putting the 500-600-microliter of the sample supernatant into a nuclear magnetic tube;
and 4, step 4: the sample in the nuclear magnetic resonance tube is measured by adopting a liquid nuclear magnetic resonance apparatus, and the measuring conditions are as follows: pressing water peak by adopting a pre-saturation method; the pulse program is as follows: noesygppr1 d; o1: 4.7-4.8; p1=10.4 mus; TD = 64K; NS = 32; DS = 4; SWH =8023 Hz; d1=4 s; d8=0.01 s;
and 5, obtaining a nuclear magnetic resonance hydrogen spectrum fingerprint, setting the chemical shift of the internal standard TSP as 0, then carrying out baseline correction and phase correction, wherein a convex enveloped peak is connected with a saw-tooth multi-peak in the spectrum within 0.80-1.0 ppm shift, a cluster of multi-peaks exist within 1.37-1.43 ppm shift, and a peak exists within 3.011-3.025 ppm shift, thus proving that whey protein powder is added into the identified formula powder.
The invention is further improved in that: the formula powder in the step 1 is prepared bovine colostrum powder or formula milk powder or compound protein powder.
The invention is further improved in that: the dissolving in the step 1 is ultrasonic at room temperature for 10min or water bath heating at 40 ℃.
The invention is further improved in that: the measurement temperature of the liquid nuclear magnetic resonance instrument in the step 4 is 20-30 ℃.
The invention has the beneficial effects that: the method provided by the invention has the advantages that the processing mode of the formula powder to be identified is simple and flexible, the consumed deuterated reagent is less, the cost is low, the detection result is accurate and reliable, whether the whey protein powder is added into the formula powder can be intuitively and accurately judged by setting the relevant measurement parameters of the liquid nuclear magnetic resonance spectrometer and observing and analyzing the obtained map, the method can be applied to identifying various nutritional health-care bovine colostrum powder, formula milk powder and composite protein powder which are declared to be added with the whey protein powder in the market, and the method has reference value for analyzing other nutritional health-care products added with the whey protein powder.
The method provided by the invention is simple and efficient, and can accurately identify the whey protein powder in the formula powder such as the bovine colostrum powder, the formula milk powder, the compound protein powder and the like.
Drawings
FIG. 1 is a nuclear magnetic spectrum of the pure whey protein powder, pure bovine colostrum powder, formulated milk powder without whey protein, bovine colostrum powder with whey protein, formulated milk powder with whey protein and composite protein powder with whey protein in examples 1-3 within 0.65-1.66 ppm.
FIG. 2 is a nuclear magnetic spectrum of the pure whey protein powder, the pure bovine colostrum powder, the formulated milk powder without whey protein, the bovine colostrum powder with whey protein, the formulated milk powder with whey protein and the composite protein powder with whey protein in examples 1-3 within 2.10-3.28 ppm.
Detailed Description
In the following description, for purposes of explanation, numerous implementation details are set forth in order to provide a thorough understanding of the various embodiments of the present invention. It should be understood, however, that these implementation details are not to be interpreted as limiting the invention. That is, in some embodiments of the invention, such implementation details are not necessary.
In addition, technical solutions between various embodiments may be combined with each other, but must be realized by a person skilled in the art, and when the technical solutions are contradictory or cannot be realized, such a combination should not be considered to exist, and is not within the protection scope of the present invention.
Example 1
1. Instruments, reagents and samples:
1.1 Main instruments:
a 400 MHz liquid nuclear magnetic resonance spectrometer, BBI probe, from brueck, switzerland; an IT0079 electronic balance of mettler-toledo instruments (shanghai) ltd; 5mm nuclear magnetic tubes from Wilmad-labglass, USA; kunshan Seama ultrasonic Instrument Co., Ltd is KH-500B model ultrasonic cleaner; centrifuge of Shanghai' an pavilion scientific instrument factory.
1.2 reagent:
sodium azide (NaN)3) The purity specification is analytical purity, purchased from Tianjin Fuchen chemical reagent factory; heavy water (D)2O), a deuteration rate of 99.9%, available from shanghai' an jie chemical co; the deuteration rate of 2,2,3, 3-deuterated Trimethylsilane Sodium Propionate (TSP) is 98 percent; trichloromethane (CHCl)3) The purity specification is analytical purity, and is purchased from Shanghai national drug group chemical reagent company Limited; TSP with mass fraction of 0.15 percent and NaN with mass fraction of 0.015 percent3The mixed solution of (2) is prepared with heavy water.
1.3 sample Collection
Whey protein powder, pure bovine colostrum powder without whey protein powder and bovine colostrum powder with whey protein powder are provided with sample raw materials by corresponding production enterprises, and the number of each sample is not less than 5.
A method for identifying whey protein powder added in formula powder by adopting nuclear magnetic resonance hydrogen spectrum fingerprint comprises the following steps:
step 1: weighing whey protein powder, pure bovine colostrum powder and 1.0 g of bovine colostrum powder added with the whey protein powder, and ultrasonically dissolving for 10min by using 12 mL of ultrapure water;
step 2: adding 10 mL of trichloromethane into the step 1, fully whirling, and centrifuging at a high speed for 10 min;
and step 3: measuring 900 muL from the supernatant of the layered sample, adding 100 muL of a heavy water solution containing 0.15 mass percent of 2,2,3, 3-tetradeuterato trimethyl silane acrylic acid and 0.015 mass percent of sodium azide, uniformly swirling, measuring 600 muL, and loading into a nuclear magnetic tube;
and 4, step 4: nuclear magnetic resonance hydrogen spectrum (1H NMR) analysisComprises the following steps: measuring temperature: 25 ℃; pressing water peak by adopting a pre-saturation method; the pulse program is as follows: noesygppr1 d; o1: 4.7-4.8, depending on the water peak pressing conditions; p1=10.4 mus; TD = 64K; NS = 32; DS = 4; SWH =8023 Hz; d1=4 s; d8=0.01 s;
and 5: nuclear magnetic resonance hydrogen spectrum (1H NMR) analysis of fingerprint: analysis of the resulting samples Using the software Topspin3.51H NMR data, setting the chemical shift of the internal standard TSP as 0, and then carrying out baseline correction and phase correction;
determining the characteristic mark of whey protein powder added into the bovine colostrum powder: if the obtained nuclear magnetic resonance hydrogen spectrum fingerprint has a convex enveloped peak connected with jagged multiple peaks within 0.80-1.0 ppm displacement, see the attached figure 1, and a cluster of multiple peaks within 1.37-1.43 ppm displacement, see the attached figure 1, and a peak within 3.011-3.025 ppm displacement, see the attached figure 2, the fact that the identified bovine colostrum powder is added with whey protein powder, see the attached figure 1-2, or the bovine colostrum powder is pure bovine colostrum powder without whey protein powder, see the attached figure 1-2 is proved.
Example 2
1. Instruments, reagents and samples:
1.1 Main instruments:
a 400 MHz liquid nuclear magnetic resonance spectrometer, BBI probe, from brueck, switzerland; an IT0079 electronic balance of mettler-toledo instruments (shanghai) ltd; 5mm nuclear magnetic tubes from Wilmad-labglass, USA; (ii) a HH-4 digital display water bath kettle of Changzhou Guohua electric appliance Co Ltd; centrifuge of Shanghai' an pavilion scientific instrument factory
1.2 reagent:
sodium azide (NaN)3) The purity specification is analytical purity, purchased from Tianjin Fuchen chemical reagent factory; heavy water (D)2O), a deuteration rate of 99.9%, available from shanghai' an jie chemical co; the deuteration rate of 2,2,3, 3-deuterated Trimethylsilane Sodium Propionate (TSP) is 98 percent; trichloromethane (CHCl)3) The purity specification is analytical purity, and is purchased from Shanghai national drug group chemical reagent company Limited; TSP with mass fraction of 0.2% and NaN with mass fraction of 0.015%3The mixed solution of (2) is prepared with heavy water.
1.3 sample Collection
The formula milk powder added with the whey protein and without the whey protein is collected from various supermarkets, and the number of each sample is not less than 5.
A method for identifying whey protein powder added in formula powder by adopting nuclear magnetic resonance hydrogen spectrum fingerprint comprises the following steps:
step 1: weighing 0.8 g of formula milk powder without whey protein and 0.8 g of formula milk powder with whey protein, and heating and dissolving the milk powder at 40 ℃ by using 10 mL of ultrapure water;
step 2: adding 10 mL of trichloromethane, fully whirling, and centrifuging at high speed for 10 min;
and step 3: measuring 800 muL from the supernatant of the layered sample, adding 200 muL of a heavy water solution containing 0.2% by mass of 2,2,3, 3-tetradeuterato trimethyl silane acrylic acid and 0.015% by mass of sodium azide, uniformly swirling, measuring 550 muL, and loading into a nuclear magnetic tube;
and 4, step 4: nuclear magnetic resonance hydrogen spectrum (1H NMR) analysis are: measuring temperature: 25 ℃; pressing water peak by adopting a pre-saturation method; the pulse program is as follows: noesygppr1 d; o1: 4.7-4.8, depending on the water peak pressing conditions; p1=10.4 mus; TD = 64K; NS = 32; DS = 4; SWH =8023 Hz; d1=4 s; d8=0.01 s;
and 5: nuclear magnetic resonance hydrogen spectrum (1H NMR) analysis of fingerprint: analysis of all raw Honey species Using the software Topspin3.51H NMR data, setting the chemical shift of the internal standard TSP as 0, and then carrying out baseline correction and phase correction;
determining a characteristic marker of whey protein powder added in the formula milk powder: if the obtained nuclear magnetic resonance hydrogen spectrum fingerprint has a convex peak packet connected with jagged multiple peaks within 0.80-1.0 ppm displacement, see the attached figure 1, and a cluster of multiple peaks within 1.37-1.43 ppm displacement, see the attached figure 1, and a peak within 3.011-3.025 ppm displacement, see the attached figure 2, the identified formula milk powder is proved to be added with whey protein powder, see the attached figure 1-2, otherwise, the formula milk powder is the formula milk powder without whey protein powder, see the attached figure 1-2.
Example 3
1. Instruments, reagents and samples:
1.1 Main instruments:
a 400 MHz liquid nuclear magnetic resonance spectrometer, BBI probe, from brueck, switzerland; an IT0079 electronic balance of mettler-toledo instruments (shanghai) ltd; 5mm nuclear magnetic tubes from Wilmad-labglass, USA; kunshan Seama ultrasonic Instrument Co., Ltd is KH-500B model ultrasonic cleaner; centrifuge of Shanghai' an pavilion scientific instrument factory.
1.2 reagent:
sodium azide (NaN)3) The purity specification is analytical purity, purchased from Tianjin Fuchen chemical reagent factory; heavy water (D)2O), a deuteration rate of 99.9%, available from shanghai' an jie chemical co; the deuteration rate of 2,2,3, 3-deuterated Trimethylsilane Sodium Propionate (TSP) is 98 percent; trichloromethane (CHCl)3) The purity specification is analytical purity, and is purchased from Shanghai national drug group chemical reagent company Limited; TSP with mass fraction of 0.12 percent and NaN with mass fraction of 0.015 percent3The mixed solution of (2) is prepared with heavy water.
1.3 sample Collection
The composite protein powder added with the lactalbumin is collected from a large supermarket, and the number of samples is not less than 5.
A method for identifying whey protein powder added in formula powder by adopting nuclear magnetic resonance hydrogen spectrum fingerprint comprises the following steps:
step 1: weighing 1.0 g of whey protein powder-added compound protein powder to be identified, and ultrasonically dissolving the protein powder by 10 mL of ultrapure water;
step 2: adding 10 mL of trichloromethane, fully whirling, and centrifuging at high speed for 10 min;
and step 3: measuring 800 muL from the supernatant of the layered sample, adding 200 muL of a heavy water solution containing 0.12% by mass of 2,2,3, 3-tetradeuterato trimethyl silane acrylic acid and 0.015% by mass of sodium azide, uniformly swirling, measuring 600 muL, and loading into a nuclear magnetic tube;
and 4, step 4: nuclear magnetic resonance hydrogen spectrum (1H NMR) analysis are: measuring temperature: 25 ℃; pressing water peak by adopting a pre-saturation method; the pulse program is as follows: noesygppr1 d;o1: 4.7-4.8, depending on the water peak pressing conditions; p1=10.4 mus; TD = 64K; NS = 32; DS = 4; SWH =8023 Hz; d1=4 s; d8=0.01 s;
and 5: nuclear magnetic resonance hydrogen spectrum (1H NMR) analysis of fingerprint: analysis of all raw Honey species Using the software Topspin3.51H NMR data, setting the chemical shift of the internal standard TSP as 0, and then carrying out baseline correction and phase correction;
verifying that whey protein powder is added into the composite protein powder: if a convex peak is connected with a jagged multi-peak in the obtained nuclear magnetic resonance hydrogen spectrum fingerprint within 0.80-1.0 ppm displacement, a cluster of multi-peaks exists in the displacement of 1.37-1.43 ppm shown in the attached figure 1, and a peak exists in the displacement of 3.011-3.025 ppm shown in the attached figure 2, the whey protein powder is added into the identified composite protein powder, which is shown in the attached figure 1-2.
The method provided by the invention is simple and efficient, and can accurately identify the whey protein powder in the formula powder such as the bovine colostrum powder, the formula milk powder, the compound protein powder and the like.
The above description is only an embodiment of the present invention, and is not intended to limit the present invention. Various modifications and alterations to this invention will become apparent to those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the scope of the claims of the present invention.
Claims (4)
1. A nuclear magnetic resonance hydrogen spectrometry for identifying whey protein powder added in formula powder is characterized in that: the method comprises the following steps:
step 1: weighing 0.8-1.5 g of formula powder to be identified, and dissolving the formula powder in 10-15 mL of ultrapure water;
step 2: adding 10 mL of trichloromethane into the step 1, fully whirling, and centrifuging at a high speed for 10 min;
and step 3: taking 600-900-microliter of the sample supernatant treated in the step 2, adding a heavy water solution containing 0.1-0.2% of 2,2,3, 3-tetradeuterotrimethylsilylacrylic acid and 0.01-0.02% of sodium azide, fixing the volume to 1 mL, uniformly swirling, and measuring 500-600-microliter of the sample supernatant and putting the 500-600-microliter of the sample supernatant into a nuclear magnetic tube;
and 4, step 4: the sample in the nuclear magnetic resonance tube is measured by adopting a liquid nuclear magnetic resonance apparatus, and the measuring conditions are as follows: pressing water peak by adopting a pre-saturation method; the pulse program is as follows: noesygppr1 d; o1: 4.7-4.8; p1=10.4 mus; TD = 64K; NS = 32; DS = 4; SWH =8023 Hz; d1=4 s; d8=0.01 s;
and 5, obtaining a nuclear magnetic resonance hydrogen spectrum fingerprint, setting the chemical shift of the internal standard TSP as 0, then carrying out baseline correction and phase correction, wherein a convex enveloped peak is connected with a saw-tooth multi-peak in the spectrum within 0.80-1.0 ppm shift, a cluster of multi-peaks exist within 1.37-1.43 ppm shift, and a peak exists within 3.011-3.025 ppm shift, thus proving that whey protein powder is added into the identified formula powder.
2. The NMR spectroscopy for identifying whey protein powder added to formula powder as claimed in claim 1, wherein the NMR spectroscopy is characterized in that: the formula powder in the step 1 is prepared bovine colostrum powder or formula milk powder or compound protein powder.
3. The NMR spectroscopy for identifying whey protein powder added to formula powder as claimed in claim 1, wherein the NMR spectroscopy is characterized in that: the dissolving in the step 1 is ultrasonic at room temperature for 10min or water bath heating at 40 ℃.
4. The NMR spectroscopy for identifying whey protein powder added to formula powder as claimed in claim 1, wherein the NMR spectroscopy is characterized in that: the measurement temperature of the liquid nuclear magnetic resonance instrument in the step 4 is 20-30 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010250441.2A CN111380895A (en) | 2020-04-01 | 2020-04-01 | Nuclear magnetic resonance hydrogen spectrometry for identifying whey protein powder added in formula powder |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010250441.2A CN111380895A (en) | 2020-04-01 | 2020-04-01 | Nuclear magnetic resonance hydrogen spectrometry for identifying whey protein powder added in formula powder |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111380895A true CN111380895A (en) | 2020-07-07 |
Family
ID=71220222
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010250441.2A Pending CN111380895A (en) | 2020-04-01 | 2020-04-01 | Nuclear magnetic resonance hydrogen spectrometry for identifying whey protein powder added in formula powder |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111380895A (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003069331A1 (en) * | 2002-02-11 | 2003-08-21 | Foss Analytical A/S | Method for casein determination in milk |
| CN103698353A (en) * | 2013-12-30 | 2014-04-02 | 厦门出入境检验检疫局检验检疫技术中心 | Bird's nest identification method |
| CN107300565A (en) * | 2017-08-25 | 2017-10-27 | 江苏出入境检验检疫局动植物与食品检测中心 | The NMR spectrum method of rice syrup is mixed in a kind of quick discriminating honey |
| CN107561110A (en) * | 2017-08-25 | 2018-01-09 | 江苏出入境检验检疫局动植物与食品检测中心 | A kind of discrimination method for being used for chaste honey, rape honey and acacia honey based on nuclear magnetic resonance technique |
| CN107860705A (en) * | 2017-11-06 | 2018-03-30 | 贝因美婴童食品股份有限公司 | A kind of detection method of baby milk powder oxidation stability |
| CN109444200A (en) * | 2018-12-21 | 2019-03-08 | 江苏中谱检测有限公司 | A method of northeast eucalyptus honey is identified using nuclear magnetic resonance spectroscopy finger-print |
-
2020
- 2020-04-01 CN CN202010250441.2A patent/CN111380895A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003069331A1 (en) * | 2002-02-11 | 2003-08-21 | Foss Analytical A/S | Method for casein determination in milk |
| CN103698353A (en) * | 2013-12-30 | 2014-04-02 | 厦门出入境检验检疫局检验检疫技术中心 | Bird's nest identification method |
| CN107300565A (en) * | 2017-08-25 | 2017-10-27 | 江苏出入境检验检疫局动植物与食品检测中心 | The NMR spectrum method of rice syrup is mixed in a kind of quick discriminating honey |
| CN107561110A (en) * | 2017-08-25 | 2018-01-09 | 江苏出入境检验检疫局动植物与食品检测中心 | A kind of discrimination method for being used for chaste honey, rape honey and acacia honey based on nuclear magnetic resonance technique |
| CN107860705A (en) * | 2017-11-06 | 2018-03-30 | 贝因美婴童食品股份有限公司 | A kind of detection method of baby milk powder oxidation stability |
| CN109444200A (en) * | 2018-12-21 | 2019-03-08 | 江苏中谱检测有限公司 | A method of northeast eucalyptus honey is identified using nuclear magnetic resonance spectroscopy finger-print |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103278574B (en) | Method for detecting hemoglobin by liquid chromatogram-triple tandem quadrupole mass spectrometer | |
| JPS6336143A (en) | Method and instrument for measuring stable type saccharogenic hemoglobin | |
| CN104711279B (en) | It is prepared by a kind of VEGF detection kit and its raw material | |
| CN107561110A (en) | A kind of discrimination method for being used for chaste honey, rape honey and acacia honey based on nuclear magnetic resonance technique | |
| CN101514991A (en) | Alpha-fetoprotein magnetic particle chemoluminescence immunoassay kit and method for preparing same | |
| Tea et al. | Simultaneous determination of natural‐abundance δ15N values and quantities of individual amino acids in proteins from milk of lactating women and from infant hair using gas chromatography/isotope ratio mass spectrometry | |
| CN103454433A (en) | Novel immunomic mass spectrometry kit for detecting endogenous and exogenous insulins | |
| Fan et al. | Determination of tropomyosin in shrimp and crab by liquid chromatography–tandem mass spectrometry based on immunoaffinity purification | |
| CN105445409B (en) | A kind of method that utilization isotope dilution mass spectrometry measures glycosylated hemoglobin | |
| CN111380895A (en) | Nuclear magnetic resonance hydrogen spectrometry for identifying whey protein powder added in formula powder | |
| CN101303360A (en) | Mass spectrogram antibody kit of human chorionic gonadotrophin isomer and preparation method thereof | |
| CN111650296A (en) | Method for improving matrix of calibrator and quality control material in blood detection type in-vitro diagnostic kit and product | |
| CN103163225A (en) | High-sensitivity quantitative detection kit for cyclosporine A in whole blood and preparation method thereof | |
| CN108008060A (en) | The assay method and reagent of hydroxyproline in a kind of feed | |
| CN106546729A (en) | A kind of new process for removing serum matrix effect in the detection of dry type immunofluorescence sizing technique | |
| CN104730231A (en) | Sample buffering solution used for fluorescence immunoassay quantitative determination and application of sample buffering solution | |
| Petzke et al. | Stable isotope ratio analysis in human hair | |
| CN106370843A (en) | Quick measurement method of lean meat powder on the basis of gold magnetic immuno-chromatography | |
| Spencer et al. | Thyroid testing for the new millenium | |
| Branch et al. | Atomic spectrometry update—clinical and biological materials, foods and beverages | |
| JP2000513438A (en) | In vitro proton MRS detection of frequency and amount of alcohol self-administration | |
| CN116990423A (en) | Vitamin B12 quantitative detection method and method for rapidly drawing standard curve | |
| Kouwenhoven et al. | Early‐Life Nutrition and Stable Isotope Techniques | |
| CN114563504B (en) | Method and kit for determining content of free aldosterone in blood plasma | |
| CN208888254U (en) | A kind of a variety of remaining colloid gold reagent bar assemblies of cephalosporins of detection |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB02 | Change of applicant information | ||
| CB02 | Change of applicant information |
Address after: 210000 building B1-3, building 16, Shuwu, No. 73, tanmi Road, Jiangbei new district, Nanjing City, Jiangsu Province Applicant after: Titanium and spectrum detection technology (Jiangsu) Co.,Ltd. Address before: 210000 building B1-3, building 16, Shuwu, No. 73, tanmi Road, Jiangbei new district, Nanjing City, Jiangsu Province Applicant before: JIANGSU SINOGRAPHY TESTING Co.,Ltd. |
|
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20200707 |