CN106248774A - A kind of method carrying out L leucine product analysis - Google Patents
A kind of method carrying out L leucine product analysis Download PDFInfo
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- CN106248774A CN106248774A CN201610781620.2A CN201610781620A CN106248774A CN 106248774 A CN106248774 A CN 106248774A CN 201610781620 A CN201610781620 A CN 201610781620A CN 106248774 A CN106248774 A CN 106248774A
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- leu
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/62—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating the ionisation of gases, e.g. aerosols; by investigating electric discharges, e.g. emission of cathode
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
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Abstract
The invention discloses a kind of method carrying out L leucine product analysis, utilize δ15L leucine contained by L leucine catalog is analyzed by N isotopic characteristic analytic process, determines that L leucine catalog is the L leucine prepared of plant material or L leucine prepared by animal material；Described δ15N isotopic characteristic analytic process includes: sample preparation and instrument regulation；Isotope analysis；Statistical analysis；Comprise the following steps: take body L leucine product approved sample that difference is prepared, obtain the sample of powder；The sample of powder is dried 8 hours at 40 ~ 50 DEG C further；Being placed in stannum capsule by every kind of sample and weigh, the weight of every kind of sample remains 1 10mg, must prepare sample；Stable Isotopic Analysis pattern is utilized to carry out L leucine source analysis, according to different δ15N value makes a distinction, so that it is determined that the leucine product Han L is to prepare in plant source or be prepared in animal sources.
Description
Technical field
The present invention relates to biological preparing technical field, specifically, be a kind of method carrying out L-Leu product analysis.
Background technology
The food of production safety high-quality is to ensure that consumer health and the prerequisite of trade the most both at home and abroad, and
And be the key of National agricultural resources sustainable development.Food and drug administration (U.S. FDA) carries on a shoulder pole the most always
The food (U.S. FDA, 2007) of heart Chinese exports.Although they think it can be seen that some of product quality of China are entered
Step, but still there are some Chinese companies to continue to the defective food of U.S. export.ANG (Ang, 2009) is in U.S.
State's food and agricultural import regulation and standard (FAIRS) report are pointed out: problem maximum in the sale of converted products and
Lasting is exactly food and medicine doping and the problem of personation.
L-Leu is that a kind of mankind cannot synthesize naturally, it is necessary to the essential amino acids obtained from diet or supplement
(Leuchtenberger et al., 2005; Parr, 2011).Viewpoint (2004) according to Mawatari et al., the bright ammonia of L-
Acid is one of composition most popular in dietary supplement.The most complete protein of the self-assembling formation that food contains is considered
Being a kind of important dietary protein source, L-Leu can serve as a kind of special diet and nourishing additive agent, it is intended to carries
The biological quality (U.S. FDA, 2009) of gross protein in high food.
L-Leu is as supplementary, dietary supplement, food supplement, sport nutrition, sports tonica, strong
Body tonic and medical material and be widely used (Mawatari et al., 2004; U.S. FDA, 2009; Renee,
2011).It can be obtained by plant source, such as corn, Semen sojae atricolor, Semen sojae atricolor, Seem Lablab Album and Semen arachidis hypogaeae, it is also possible to is obtained by animal sources, bag
Include but be not limited only to poultry, shrimp, Carnis caprae seu ovis and beef (Chew, 2009; Amerman, 2010; Shifko, 2010;
Marchini, 2012)。
Some fraudulent manufacturers use the raw material of separate sources in the production process of this very important product, lead
Causing doping or the product of personation occur, this will bring the risk in economy and health to consumer.It is known that some produces
Product consumer after point of purchase can be relatively easy determination they whether be spoofed to such an extent as to have purchased the product of inferior quality (Klein
and Leffler, 1981).But, for leucine product, consumer is difficult to by observing or making for distinguishing that L-is bright
Propylhomoserin is derived from plant or animal, but its source is to ensureing product safety, consumer health and successful domestic state
Border trade is particularly significant.
Stable Isotopic Analysis (SIA) is the most promising a kind of effective technology in terms of the test food true and false, especially
The field (Kelly, 2001) of definite result cannot be provided in traditional analysis method.Hobson and Wassenaar(1999) report
Road, in the last few years, in order to investigate several aspects of ecological Studies, is increasingly using stablizing of self-assembling formation in biomaterial
Isotopic measurement is as research tool.
Summary of the invention
It is an object of the invention to design a kind of method carrying out L-Leu product analysis, utilize stable isotope to divide
Analysis pattern carries out L-Leu source analysis, according to different δ15N value makes a distinction, so that it is determined that the product Han L-Leu be from
In plant source, preparation is still prepared in animal sources.
The present invention is achieved through the following technical solutions: a kind of method carrying out L-Leu product analysis, utilizes δ15N coordination
L-Leu contained by L-Leu catalog is analyzed by element method for feature analysis, determines that L-Leu catalog is plant
L-Leu prepared by raw material or L-Leu prepared by animal material.
The most to better implement the present invention, following set-up mode is used especially: described δ15N isotopic characteristic is analyzed
Method includes:
1) sample preparation and instrument regulation；
2) isotope analysis；
3) statistical analysis.
The most to better implement the present invention, following set-up mode is used especially: described step 1) includes following step
Rapid:
1-1) take body L-Leu product approved sample that difference is prepared, obtain the sample of powder；
1-2) through step 1-1) after, the sample of powder is dried 8 hours at 40 ~ 50 DEG C further；
1-3) through step 1-2) after, every kind of sample is placed in stannum capsule and weighs, the weight of every kind of sample remains 1-10mg,
Prepare sample；
Protein standards 1-4) is used to carry out the test instrunment linearity as chemical standard and define instrument to different elemental compositions
The instrument response of assay.
The most to better implement the present invention, following set-up mode is used especially: described step 1) is particularly as follows: take not
With the body L-Leu product prepared approved sample, obtain the sample of powder；By the sample of powder at 45 DEG C further
Dry 8 hours；Being placed in stannum capsule by every kind of sample and weigh, the weight of every kind of sample is 5mg, must prepare sample；Use albumen
Matter standard is carried out the test instrunment linearity as chemical standard and defines the instrument instrument sound to the assay of different elemental compositions
Should.
The most to better implement the present invention, following set-up mode is used especially: described step 2) isotope analysis
Particularly as follows: utilize mass spectrograph to use stream isotopic ratio mass spectroscopy continuously to prepare δ in sample15The natural abundance of N.
The most to better implement the present invention, following set-up mode is used especially: described mass spectrograph uses Integra
2 CN special carbon nitrogen isotope mass spectrograph.
The most to better implement the present invention, following set-up mode is used especially: described step 3) includes following tool
Body step:
With the descriptive statistical analysis in the analytical tool case of Excel and one-dimensional variance analysis, the data obtained are analyzed；
Result of study being presented in graphical form, clearly illustrate the difference between sample, i.e. obtaining various sample is to use plant material
Prepared by preparation or animal material；
Using the confidence level of 95% in described descriptive statistical analysis, in one-dimensional variance analysis, alpha value is 0.05.
The most to better implement the present invention, following set-up mode is used especially: L-prepared by described plant material
Leucic δ15N isotopic composition scope is 0.8774 ‰ to 3.1245 ‰；The δ of L-Leu prepared by described animal material15N value scope is 3.6400 ‰ to 8.3400 ‰, and the δ of L-Leu prepared by animal material15The means standard deviation value of N value
Minima is > 5.0000 ± 0.0000 ‰.
The present invention compared with prior art, has the following advantages and beneficial effect:
The present invention utilizes Stable Isotopic Analysis pattern to carry out L-Leu source analysis, according to different δ15N value carries out district
Point, so that it is determined that the product Han L-Leu is to prepare in plant source or be prepared in animal sources.
Detailed description of the invention
Below in conjunction with embodiment, the present invention is described in further detail, but embodiments of the present invention are not limited to this.
Embodiment 1:
A kind of method carrying out L-Leu product analysis, utilizes Stable Isotopic Analysis pattern to carry out L-Leu source point
Analysis, according to different δ15N value makes a distinction, so that it is determined that the product Han L-Leu is to prepare or from animal sources in plant source
Inside it is prepared, utilizes δ15L-Leu contained by L-Leu catalog is analyzed by N isotopic characteristic analytic process, determines
The L-Leu that L-Leu catalog is the L-Leu prepared of plant material or prepared by animal material.
Embodiment 2:
The present embodiment is to optimize further on the basis of above-described embodiment, the most to better implement the present invention, especially
Use following set-up mode: described δ15N isotopic characteristic analytic process includes:
1) sample preparation and instrument regulation；
2) isotope analysis；
3) statistical analysis.
Embodiment 3:
The present embodiment is to optimize further on the basis of any of the above-described embodiment, the most to better implement the present invention,
Use following set-up mode especially: described step 1) comprises the following steps:
1-1) take body L-Leu product approved sample that difference is prepared, obtain the sample of powder；
1-2) through step 1-1) after, the sample of powder is dried 8 hours at 40 ~ 50 DEG C further；
1-3) through step 1-2) after, every kind of sample is placed in stannum capsule and weighs, the weight of every kind of sample remains 1-10mg,
Prepare sample；
Protein standards 1-4) is used to carry out the test instrunment linearity as chemical standard and define instrument to different elemental compositions
The instrument response of assay.
Embodiment 4:
The present embodiment is to optimize further on the basis of any of the above-described embodiment, the most to better implement the present invention,
Use following set-up mode especially: described step 1), particularly as follows: take body L-Leu product the approved sample that difference is prepared, obtains
The sample of powder；The sample of powder is dried 8 hours at 45 DEG C further；Every kind of sample is placed in stannum capsule and claims
Weight, the weight of every kind of sample is 5mg, must prepare sample；Protein standards is used to carry out the test instrunment linearity as chemical standard
And define the instrument instrument response to the assay of different elemental compositions.
Embodiment 5:
The present embodiment is to optimize further on the basis of any of the above-described embodiment, the most to better implement the present invention,
Using especially following set-up mode: described step 2) isotope analysis flows isotope ratio continuously particularly as follows: utilize mass spectrograph to use
Value mass spectroscopy prepares δ in sample15The natural abundance of N.
Embodiment 6:
The present embodiment is to optimize further on the basis of any of the above-described embodiment, the most to better implement the present invention,
Use following set-up mode especially: described mass spectrograph uses Integra 2 CN special carbon nitrogen isotope mass spectrograph.
Embodiment 7:
The present embodiment is to optimize further on the basis of any of the above-described embodiment, the most to better implement the present invention,
Use especially following set-up mode: described step 3) includes step in detail below:
With the descriptive statistical analysis in the analytical tool case of Excel and one-dimensional variance analysis, the data obtained are analyzed；
Result of study being presented in graphical form, clearly illustrate the difference between sample, i.e. obtaining various sample is to use plant material
Prepared by preparation or animal material；
Using the confidence level of 95% in described descriptive statistical analysis, in one-dimensional variance analysis, alpha value is 0.05.
Embodiment 8:
The present embodiment is to optimize further on the basis of any of the above-described embodiment, the most to better implement the present invention,
The following set-up mode of employing especially: the δ of L-Leu prepared by described plant material15N isotopic composition scope is 0.8774 ‰
To 3.1245 ‰；The δ of L-Leu prepared by described animal material15N value scope is 3.6400 ‰ to 8.3400 ‰, and animal is former
The δ of the L-Leu of material preparation15The means standard deviation value minima of N value is > 5.0000 ± 0.0000 ‰.
Owing to Stable Isotopic Analysis (SIA) can apply the measurement condition at the scene of the nitrogen stable isotope that nature generates
Lower estimation trophic level.Theory (2010) according to Kelly, the main application in animal ecology of the nitrogen-stable isotope ratio
It is the relation of it and trophic level.According to estimates, the trophic level from plant to herbivore or from herbivore to carnivore
Increase, the δ in consumer15N will have growth (the Vander Zanden and of 2.2 to 3.4 ‰ relative to its diet
Rasmussen 2001; McCutchan et al. 2003).DeNiro and Epstein(1981) test at first and consumed
Person is higher by 0 10 ‰ than δ 15N in their diet, and meansigma methods is 3 ‰.Therefore δ15N is often applied to food_chain lengths and nutrition
The estimation of position, because it has the δ in predictable nutrition accumulation, i.e. consumer and diet15Different (the Minagawa and of N
Wada 1984)。
It addition, McClelland and Montoya(2002) also report, some aminoacid roll up along with trophic transfer,
Including L-Trp, altheine, Pidolidone, ALANINE, ILE, L-Leu, Valine and L-dried meat
Propylhomoserin, meanwhile, (glycine, Serine, L-threonine, L-phenylalanine, TYR and L-rely ammonia to another group aminoacid
Acid) in δ15N value is basically unchanged along with each trophic transfer；The present invention makes δ15N isotopic content becomes differentiation L-Leu and produces
Employed in the unique and highly reliable instrument of raw material.
Present invention application δ15N isotopic characteristic is distinguished L-Leu and produced the raw material used is plant source raw material
Or animal sources raw material.
The above, be only presently preferred embodiments of the present invention, and the present invention not does any pro forma restriction, every depends on
Any simple modification of being made above example according to the technical spirit of the present invention, equivalent variations, each fall within the protection of the present invention
Within the scope of.
Claims (8)
1. the method carrying out L-Leu product analysis, it is characterised in that: utilize δ15N isotopic characteristic analytic process is bright to L-
L-Leu contained by propylhomoserin catalog is analyzed, and determines that L-Leu catalog is L-Leu prepared by plant material
Or L-Leu prepared by animal material.
A kind of method carrying out L-Leu product analysis the most according to claim 1, it is characterised in that: described δ15N is same
Position element method for feature analysis includes:
1) sample preparation and instrument regulation；
2) isotope analysis；
3) statistical analysis.
A kind of method carrying out L-Leu product analysis the most according to claim 2, it is characterised in that: described step 1)
Comprise the following steps:
1-1) take body L-Leu product approved sample that difference is prepared, obtain the sample of powder；
1-2) through step 1-1) after, the sample of powder is dried 8 hours at 40 ~ 50 DEG C further；
1-3) through step 1-2) after, every kind of sample is placed in stannum capsule and weighs, the weight of every kind of sample remains 1-10mg,
Prepare sample；
Protein standards 1-4) is used to carry out the test instrunment linearity as chemical standard and define instrument to different elemental compositions
The instrument response of assay..
A kind of method carrying out L-Leu product analysis the most according to claim 3, it is characterised in that: described step 1)
Particularly as follows: take body L-Leu product the approved sample that difference is prepared, obtain the sample of powder；By the sample of powder 45
Dry 8 hours further at DEG C；Being placed in stannum capsule by every kind of sample and weigh, the weight of every kind of sample is 5mg, must prepare sample
This；Protein standards is used to carry out the test instrunment linearity as chemical standard and define instrument different elemental compositions are contained measurement
Fixed instrument response.
5. according to a kind of method carrying out L-Leu product analysis described in claim 3 or 4, it is characterised in that: described step
Rapid 2) isotope analysis is particularly as follows: utilize mass spectrograph to use stream isotopic ratio mass spectroscopy continuously to prepare δ in sample15N from
So abundance.
A kind of method carrying out L-Leu product analysis the most according to claim 5, it is characterised in that: described mass spectrograph
Use Integra 2 CN special carbon nitrogen isotope mass spectrograph.
7. according to a kind of method carrying out L-Leu product analysis described in claim 3 or 4, it is characterised in that: described step
Rapid 3) step in detail below is included:
With the descriptive statistical analysis in the analytical tool case of Excel and one-dimensional variance analysis, the data obtained are analyzed；
Result of study being presented in graphical form, clearly illustrate the difference between sample, i.e. obtaining various sample is to use plant material
Prepared by preparation or animal material；
Using the confidence level of 95% in described descriptive statistical analysis, in one-dimensional variance analysis, alpha value is 0.05.
8. according to a kind of method carrying out L-Leu product analysis described in claim 3 or 4, it is characterised in that plant described in:
The δ of L-Leu prepared by raw material15N isotopic composition scope is 0.8774 ‰ to 3.1245 ‰；Prepared by described animal material
The δ of L-Leu15N value scope is 3.6400 ‰ to 8.3400 ‰, and the δ of L-Leu prepared by animal material15N value
Means standard deviation value minima is > 5.0000 ± 0.0000 ‰.
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| CN101603944A (en) * | 2009-07-14 | 2009-12-16 | 中山大学 | A kind of analytical approach of tilapia mossambica food source |
| CN101738461A (en) * | 2008-11-14 | 2010-06-16 | 周京石 | 13C abundance method for distinguishing sources of glucosamine hydrochloride |
| EP1495320B1 (en) * | 2002-04-12 | 2013-01-09 | SNU R&DB Foundation | Method for discriminating organic agricultural products from conventional agricultural products by using nitrogen isotope index |
| CN103472124A (en) * | 2013-08-30 | 2013-12-25 | 大连海事大学 | Fingerprint identification method of marine organisms as well as products thereof |
-
2016
- 2016-08-31 CN CN201610781620.2A patent/CN106248774A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1495320B1 (en) * | 2002-04-12 | 2013-01-09 | SNU R&DB Foundation | Method for discriminating organic agricultural products from conventional agricultural products by using nitrogen isotope index |
| US20090042304A1 (en) * | 2006-04-21 | 2009-02-12 | Anderson Kim A | Method for analyzing foods |
| CN101738461A (en) * | 2008-11-14 | 2010-06-16 | 周京石 | 13C abundance method for distinguishing sources of glucosamine hydrochloride |
| CN101603944A (en) * | 2009-07-14 | 2009-12-16 | 中山大学 | A kind of analytical approach of tilapia mossambica food source |
| CN103472124A (en) * | 2013-08-30 | 2013-12-25 | 大连海事大学 | Fingerprint identification method of marine organisms as well as products thereof |
Non-Patent Citations (3)
| Title |
|---|
| JINGYU HUANG ET AL.: "Authentication of Pure L-leucine Products Manufactured in China by Discriminating between Plant and Animal Sources Using Nitrogen Stable Isotope Technique", 《JOURNAL OF FOOD SCIENCE》 * |
| JINGYU HUANG ET AL.: "The use of nitrogen-stable isotope technique to identify L-leucine extracted from human air", 《JOURNAL OF CONSUMER PROTECTION AND FOOD SAFETY》 * |
| 袁玉伟 等: "氮稳定同位素的印迹规律与有机食品鉴别", 《核农学报》 * |
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Application publication date: 20161221 |