CN108872131A - A method of identifying shell-fish Glucosamine and microbial method Glucosamine - Google Patents
A method of identifying shell-fish Glucosamine and microbial method Glucosamine Download PDFInfo
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- CN108872131A CN108872131A CN201810933757.4A CN201810933757A CN108872131A CN 108872131 A CN108872131 A CN 108872131A CN 201810933757 A CN201810933757 A CN 201810933757A CN 108872131 A CN108872131 A CN 108872131A
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- MSWZFWKMSRAUBD-UHFFFAOYSA-N beta-D-galactosamine Natural products NC1C(O)OC(CO)C(O)C1O MSWZFWKMSRAUBD-UHFFFAOYSA-N 0.000 title claims abstract description 62
- MSWZFWKMSRAUBD-IVMDWMLBSA-N 2-amino-2-deoxy-D-glucopyranose Chemical compound N[C@H]1C(O)O[C@H](CO)[C@@H](O)[C@@H]1O MSWZFWKMSRAUBD-IVMDWMLBSA-N 0.000 title claims abstract description 61
- 229960002442 glucosamine Drugs 0.000 title claims abstract description 61
- 238000000034 method Methods 0.000 title claims abstract description 48
- 235000015170 shellfish Nutrition 0.000 title claims abstract description 25
- 230000000813 microbial effect Effects 0.000 title claims abstract description 23
- 230000000155 isotopic effect Effects 0.000 claims abstract description 24
- 239000000126 substance Substances 0.000 claims description 20
- IOLCXVTUBQKXJR-UHFFFAOYSA-M potassium bromide Chemical compound [K+].[Br-] IOLCXVTUBQKXJR-UHFFFAOYSA-M 0.000 claims description 12
- 238000004566 IR spectroscopy Methods 0.000 claims description 7
- 238000000862 absorption spectrum Methods 0.000 claims description 6
- 238000012360 testing method Methods 0.000 claims description 6
- 239000007789 gas Substances 0.000 claims description 5
- -1 aminoglucose saccharide Chemical class 0.000 claims description 4
- 238000004949 mass spectrometry Methods 0.000 claims description 4
- 238000005259 measurement Methods 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 4
- ATJFFYVFTNAWJD-UHFFFAOYSA-N Tin Chemical compound [Sn] ATJFFYVFTNAWJD-UHFFFAOYSA-N 0.000 claims description 3
- 239000012159 carrier gas Substances 0.000 claims description 3
- 239000001307 helium Substances 0.000 claims description 3
- 229910052734 helium Inorganic materials 0.000 claims description 3
- SWQJXJOGLNCZEY-UHFFFAOYSA-N helium atom Chemical compound [He] SWQJXJOGLNCZEY-UHFFFAOYSA-N 0.000 claims description 3
- 229910052757 nitrogen Inorganic materials 0.000 claims description 3
- 230000003647 oxidation Effects 0.000 claims description 3
- 238000007254 oxidation reaction Methods 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 2
- 238000000227 grinding Methods 0.000 claims description 2
- 230000009467 reduction Effects 0.000 claims description 2
- 238000005303 weighing Methods 0.000 claims description 2
- 239000003814 drug Substances 0.000 abstract description 10
- 229940079593 drug Drugs 0.000 abstract description 8
- 238000004519 manufacturing process Methods 0.000 abstract description 8
- 238000005516 engineering process Methods 0.000 abstract description 5
- 238000011156 evaluation Methods 0.000 abstract description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 21
- 238000002474 experimental method Methods 0.000 description 7
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 6
- 238000000855 fermentation Methods 0.000 description 5
- 230000004151 fermentation Effects 0.000 description 5
- CHVZQMAANSUXJU-JJKGCWMISA-N (2r,3s,4r,5r)-2,3,4,5,6-pentahydroxyhexanamide;hydrochloride Chemical compound Cl.NC(=O)[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO CHVZQMAANSUXJU-JJKGCWMISA-N 0.000 description 4
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 229920002101 Chitin Polymers 0.000 description 4
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 4
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 4
- 230000006196 deacetylation Effects 0.000 description 4
- 238000003381 deacetylation reaction Methods 0.000 description 4
- 241000238557 Decapoda Species 0.000 description 3
- 239000002775 capsule Substances 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 239000007858 starting material Substances 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 241000238424 Crustacea Species 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 210000001503 joint Anatomy 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 230000008520 organization Effects 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- CBOJBBMQJBVCMW-BTVCFUMJSA-N (2r,3r,4s,5r)-2-amino-3,4,5,6-tetrahydroxyhexanal;hydrochloride Chemical compound Cl.O=C[C@H](N)[C@@H](O)[C@H](O)[C@H](O)CO CBOJBBMQJBVCMW-BTVCFUMJSA-N 0.000 description 1
- JCZPMGDSEAFWDY-SQOUGZDYSA-N (2r,3s,4r,5r)-2,3,4,5,6-pentahydroxyhexanamide Chemical compound NC(=O)[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO JCZPMGDSEAFWDY-SQOUGZDYSA-N 0.000 description 1
- UBDZFAGVPPMTIT-UHFFFAOYSA-N 2-aminoguanidine;hydron;chloride Chemical compound [Cl-].NC(N)=N[NH3+] UBDZFAGVPPMTIT-UHFFFAOYSA-N 0.000 description 1
- 239000003390 Chinese drug Substances 0.000 description 1
- FZHXIRIBWMQPQF-UHFFFAOYSA-N Glc-NH2 Natural products O=CC(N)C(O)C(O)C(O)CO FZHXIRIBWMQPQF-UHFFFAOYSA-N 0.000 description 1
- OVRNDRQMDRJTHS-UHFFFAOYSA-N N-acelyl-D-glucosamine Natural products CC(=O)NC1C(O)OC(CO)C(O)C1O OVRNDRQMDRJTHS-UHFFFAOYSA-N 0.000 description 1
- OVRNDRQMDRJTHS-FMDGEEDCSA-N N-acetyl-beta-D-glucosamine Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-FMDGEEDCSA-N 0.000 description 1
- MBLBDJOUHNCFQT-LXGUWJNJSA-N N-acetylglucosamine Natural products CC(=O)N[C@@H](C=O)[C@@H](O)[C@H](O)[C@H](O)CO MBLBDJOUHNCFQT-LXGUWJNJSA-N 0.000 description 1
- 241000219095 Vitis Species 0.000 description 1
- 235000009754 Vitis X bourquina Nutrition 0.000 description 1
- 235000012333 Vitis X labruscana Nutrition 0.000 description 1
- 235000014787 Vitis vinifera Nutrition 0.000 description 1
- 239000008186 active pharmaceutical agent Substances 0.000 description 1
- FZHXIRIBWMQPQF-SLPGGIOYSA-N aldehydo-D-glucosamine Chemical compound O=C[C@H](N)[C@@H](O)[C@H](O)[C@H](O)CO FZHXIRIBWMQPQF-SLPGGIOYSA-N 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 210000000544 articulatio talocruralis Anatomy 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000003962 counterfeit drug Substances 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 239000002019 doping agent Substances 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 229960001911 glucosamine hydrochloride Drugs 0.000 description 1
- 150000002302 glucosamines Chemical class 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 210000001624 hip Anatomy 0.000 description 1
- 210000000629 knee joint Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000003801 milling Methods 0.000 description 1
- 229950006780 n-acetylglucosamine Drugs 0.000 description 1
- 239000000820 nonprescription drug Substances 0.000 description 1
- 201000008482 osteoarthritis Diseases 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 210000000323 shoulder joint Anatomy 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 210000003857 wrist joint Anatomy 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/35—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light
- G01N21/3563—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light for analysing solids; Preparation of samples therefor
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
Abstract
The present invention provides a kind of methods for identifying shell-fish Glucosamine and microbial method Glucosamine, with isotopic ratio δ13C≤- 17 ‰ and △ δ ∣ C-N ∣ >=15 two parameter effectively improve the correctness of identification as evaluation index；The method of the present invention can effectively identify the source of drug Glucosamine, and the shell-fish Glucosamine that production technology is registered to meet national drug provides judgment basis.
Description
(1) technical field
The present invention relates to a kind of methods for identifying shell-fish Glucosamine and microbial method Glucosamine.
(2) background technique
Glucosamine (Glucosamine) and its hydrochloride (Glucosamine Hydrochloride), chemistry are entitled
2-amino-2-deoxy-D-Glucose or its hydrochloride are the active pharmaceutical ingredients of Chinese pharmaceuticals administration mechanism approval,
Preparation is OTC drug.It can be used for treating and preventing the osteoarthritis of whole body all sites, including knee joint, shoulder joint, hip close
Section, wrist joint, neck and spinal joint such as ankle-joint etc. can be relieved and eliminate the symptoms such as pain, the swelling of Bones and joints, improves and closes
Save movable function.
It is former that the production technology of shell-fish Glucosamine and its hydrochloride, which is with the shell of the crustaceans such as shrimp, crab,
Material through decalcification, de- albumen, goes the processes such as heavy metal to obtain chitin, then with hydrochloric acid deacetylation be refined into Glucosamine and its
Hydrochloride.This technique is the Registration regulations technique of Chinese pharmaceuticals administration mechanism approval.
The Glucosamine produced with microbial method and its hydrochloric acid product salt occurred currently on the market is not Chinese drug
The drug of supervision and administration organization approval, production technology (is contained in such as citric acid fermented residue with the thallus of microbial fermentation
Some black-koji mould filaments) in chitin extraction；Or N-acetylglucosamine is obtained by engineering bacterium fermentation method of Escherichia coli,
Glucosamine and its hydrochloride is made with sour deacetylation again.Since the starting material source of microbial method Glucosamine is numerous
More, the hidden dopant species deposited, structure, property and quantity are indefinite, it is difficult to it is controlled with existing target level of product quality, product
Safety not can guarantee.
Once there is Japanese Patent Publication [JP5687568] report：As isotopic ratio δ13C < -15 ‰ or δ15N > -3.5 ‰
When product be shell-fish Glucosamine, method isotopic ratio δ13C index is excessively wide in range, δ15N index cannot correctly identify not
With the Glucosamine in source.
(3) summary of the invention
For the deficiencies in the prior art, the present invention provides a kind of identification shell-fish Glucosamine and microorganisms
The method of method Glucosamine, with isotopic ratio δ13C≤- 17 ‰ and △ δ ∣ C-N ∣ >=15 two parameter as evaluation index,
Effectively improve the correctness of identification.
Technical scheme is as follows：
A method of identifying shell-fish Glucosamine and microbial method Glucosamine, the method are：
(1) sample to be tested is qualitative
Sample to be tested is taken, with determination of infrared spectroscopy infrared absorption spectrum, and the infrared light with aminoglucose saccharide
Spectrogram is compareed, and determines that sample to be tested is Glucosamine；
(2) sample to be tested identifies
It takes by step (1) qualitative sample to be tested, the same position of Elements C in sample to be tested, N is measured with isotopic mass spectrometry
Plain ratio delta13C、δ15N, and by formula △ δ ∣ C-N ∣=∣ δ13C—δ15The value of N ∣ calculating △ δ ∣ C-N ∣；
As the δ of sample to be tested13C≤- 17 ‰, and ∣ >=15 △ δ ∣ C-N, then it is shell-fish Glucosamine；
As the δ of sample to be tested13C >=-13 ‰, and ∣≤6 △ δ ∣ C-N, then it is microbial method Glucosamine；
The isotopic ratio δ that Elements C in sample to be tested, N are measured with isotopic mass spectrometry13C、δ15The method of N is：
The sample to be tested is taken to be ground to 100 mesh hereinafter, weighing 0.1~0.5mg after freeze-dried or 50 DEG C of drying
Sample after grinding is placed in tin can, and tight is placed in C/N elemental analyser combination isotope mass spectrometer, on-line determination
The isotopic ratio δ of Elements C in sample, N13C、δ15N, testing conditions are：
Correcting gas：CO2(purity >=99.995%), N2(purity >=99.999%)；
Internal standard substance：USGS24 and GBW4407 (are used for CO2Calibration), IAEA-N-I (be used for N2Calibration)；
Standard substance：δ13The standard substance of C is PDB, δ15The standard substance of N is air (Air-N2)；
Analyze parameter：1000~1250 DEG C of furnace temperature of oxidation restores 600~700 DEG C of furnace temperature, chromatographic column column temperature 30~50
DEG C, carrier gas (helium), 10~100mL/min of flow velocity；
Demarcate detection gas CO respectively with internal standard substance2And N2, in the initial phase, intermediate stage and knot of sample test
The tail stage is corrected with internal standard substance, controls the stability of experimentation；
Wherein,
δ13The calculation method of C is：δ13C=[(RCSample/RCStandard) -1] × 1000, the RCRefer to：The weight of carbon
The ratio between isotope atom abundance and light isotope atom abundance, i.e. RC=13C/12C, the standard substance of carbon isotope are PDB；
δ15The calculation method of N is：δ15N=[(RNSample/RNStandard) -1] × 1000, the RNRefer to：The weight of nitrogen
The ratio between isotope atom abundance and light isotope atom abundance, i.e. RN=15N/14N, the standard substance of nitrogen isotope are air
(Air-N2)。
In the present invention, the Glucosamine is compound Glucosamine or the form of its hydrochloride.
The shell-fish Glucosamine refers to the shell extraction chitin with crustaceans such as shrimp, crabs, then uses hydrochloric acid
The Glucosamine or its hydrochloride (being also possible to sulfate, generally hydrochloride) product that (or sulfuric acid) deacetylation obtains.
The microbial method Glucosamine refers to (such as to be contained in Citric Acid Fermentation Residue with the thallus of microbial fermentation
Black-koji mould filament) in chitin extraction, then the Glucosamine or its hydrochloride that are obtained with hydrochloric acid (or sulfuric acid) deacetylation
(being also possible to sulfate, generally hydrochloride) product；Or N- acetylamino Portugal is obtained by engineering bacterium fermentation method of Escherichia coli
Grape sugar, then deacetylated obtained Glucosamine or its esters product.
Specifically, the qualitative method of step (1) sample to be tested is：
Take sample to be tested appropriate (1~1.5mg), with potassium bromide powder in mass ratio 1：200 fine ground mixings, are subsequently placed in pressure
Transparence potassium bromide thin slice is pressed into piece machine, with the infrared absorption spectrum of determination of infrared spectroscopy thin slice；Take Glucosamine
Standard items operate in the same way, measure the infrared absorption spectrum of standard items；If the infrared spectroscopy of sample to be tested and standard items
Figure is consistent, it is determined that product to be measured is Glucosamine.
Compared with the existing technology, the beneficial effects of the present invention are：The method of the present invention can effectively identify drug amino Portugal
The source of grape sugar, the shell-fish Glucosamine that production technology is registered to meet national drug provide judgment basis.
(4) specific embodiment
Below by specific embodiment, the invention will be further described, but protection scope of the present invention is not limited in
This.
Embodiment 1
Experiment condition
Instrument：It is same that Thermofisher company EA1112 type C/N elemental analyser is combined Delta Plus Advantage
Position quality spectrometer
Correcting gas：CO2(purity >=99.995%), N2(purity >=99.999%)
Internal standard substance：USGS24 and GBW4407 (are used for CO2Calibration), IAEA-N-I (is used for N2Calibration)
Standard substance：δ13The standard substance of C be PDB (South Carolina, United States Cretaceous period skin Di organize quasi- belemnite fossil),
δ15The standard substance of N is air (Air-N2)
Analyze parameter：1020 DEG C of furnace temperature of oxidation, 650 DEG C of furnace temperature of reduction, 40 DEG C of polarity chromatographic column temperature, carrier gas (helium)
90~100ml/min of flow.
Measure sample isotopic ratio δ13C、δ15N：
(1) determine that sample to be tested is Glucosamine：Product 1mg to be measured and potassium bromide powder are weighed (by about 1：200) it grinds
It is thin to mix, it sets press machine transparence potassium bromide thin slice is made and take aminoglucose with the infrared spectroscopy of determination of infrared spectroscopy thin slice
Sugar or its corresponding salt standard items are operated with method, if the infrared spectrogram of product to be measured is consistent with standard items spectrogram, to
Survey product is Glucosamine.
(2) sample preparation：Will have determined that test substance be Glucosamine sample, through 50 DEG C dry 8 hours after, it is fine ground extremely
100 mesh are set in tin can, tight hereinafter, weigh about 0.1~0.5mg sample.Three parts of each sample.
(3) isotopic ratio measures：By the δ of above-mentioned experiment condition on-line determination sample13C、δ15N isotopic ratio.
(4) it corrects：Calibration experiment room gas CO is distinguished with internal standard substance USGS24, GBW4407 and IAEA-N-12And N2。
It is carried out and is corrected with internal standard substance in the initial phase of sample test, intermediate stage and ending stage, control the stability of experiment.
(5) isotopic ratio δ is used13The starting material of C (PDB) and △ δ ∣ C-N two parameter decision products to be measured of ∣ is traced to the source,
The δ of shell-fish Glucosamine and its hydrochloride13∣ >=15 C (PDB)≤- 17 ‰, △ δ ∣ C-N；Microbial method Glucosamine
And its δ of hydrochloride13∣≤6 C (PDB) >=-13 ‰, △ δ ∣ C-N.
Embodiment 2
From the enterprise and 3 production microbial method Glucosamines for producing shell-fish aminoglucose hydrochloride known to 4
Sample (being shown in Table 1) is bought by hydrochloride enterprise, describes by " experiment condition ", " the measurement sample isotopic ratio " in embodiment 1
Program determination sample (is shown in Table 1) as the result is shown：The isotopic ratio δ of shell-fish aminoglucose hydrochloride13C≤- 17 ‰, △ δ
∣C— N∣≥15；The isotopic ratio δ of microbial method aminoglucose hydrochloride13∣≤6 C >=-13 ‰, △ δ ∣ C-N.
Table 1：The isotopic ratio (average value n=3) of the Glucosamine of separate sources
| Number | Type | The place of production | δ13C(‰) | δ15N(‰) | △δ∣C—N∣ |
| 1 | Shell-fish Glucosamine | Taizhou of Zhejiang | -23.29 | -5.88 | 17.31 |
| 2 | Shell-fish Glucosamine | Zhoushan Of Zhejiang Province | -17.55 | -2.29 | 15.26 |
| 3 | Shell-fish Glucosamine | Fujian | -23.81 | -6.38 | 17.43 |
| 4 | Shell-fish Glucosamine | Jiangsu | -25.56 | -4.68 | 20.88 |
| 5 | Microbial method Glucosamine | Shandong | -11.20 | -6.05 | 5.15 |
| 6 | Microbial method Glucosamine | Jiangsu | -12.00 | -7.58 | 4.42 |
| 7 | Microbial method Glucosamine | Zhejiang | -12.71 | -7.55 | 5.16 |
Embodiment 3
China National Drug supervision and administration organization, which is bought, from pharmacy ratifies the aminoguanidine hydrochloride Portugal that 3 pharmaceutical producing enterprises produce
Grape carbohydrate gum capsule is sample to be tested, takes capsule 's content fine ground to 100 mesh hereinafter, by " experiment condition ", " measurement in embodiment 1
The program of sample isotopic ratio " description, measures sample, (is shown in Table 2) as the result is shown：1~2 sample of number is shell-fish amino Portugal
Grape sugar starting material, meets legal manufacturing technique requirent.3 sample of number is microbial method Glucosamine, according to the Chinese people
Republic's Drugs Clause " not obtaining the production of authentication code " drug, should be punished by counterfeit drug.
Table 2：The isotopic ratio (average value n=3) of the aminoglucose hydrochloride capsule of different manufacturers
| Number | Specification (mg/) | δ13C(‰) | δ15N(‰) | △δ∣C—N∣ |
| 1 | 240 | -24.02 | -5.20 | 18.82 |
| 2 | 240 | -17.07 | -1.68 | 15.39 |
| 3 | 750 | -11.90 | -6.50 | 5.40 |
Embodiment 4
It takes microbial method ammonia known to number 5 in shell-fish Glucosamine sample known to number 1 in table 1 and table 1
Base glucose sample presses (7：3) ratio mixes, and mixing of sufficiently milling obtains sample to be tested, by embodiment 1 " experiment condition ",
The program determination sample to be tested of " measurement sample isotopic ratio " description, obtains isotopic ratio δ13C (PDB) is -19.70 ‰, δ15N (Air-N2) it is -5.95 ‰, △ δ ∣ C-N ∣ is 13.75, though δ13C numerical value is less than -17 ‰, but △ δ ∣ C-N ∣ < 15, discloses
Test substance is the shell-fish Glucosamine sample for mixing certain amount microbial method Glucosamine.
Bibliography [1]：Japanese Patent Publication speciallys permit No. 5687568 (B2).
Claims (2)
1. a kind of method for identifying shell-fish Glucosamine and microbial method Glucosamine, which is characterized in that the method
For：
(1) sample to be tested is qualitative
Sample to be tested is taken, with determination of infrared spectroscopy infrared absorption spectrum, and the infrared spectrogram with aminoglucose saccharide
It is compareed, determines that sample to be tested is Glucosamine；
(2) sample to be tested identifies
It takes by step (1) qualitative sample to be tested, with Elements C, the isotope ratio of N in isotopic mass spectrometry measurement sample to be tested
Value δ13C、δ15N, and by formula △ δ ∣ C-N ∣=∣ δ13C—δ15The value of N ∣ calculating △ δ ∣ C-N ∣；
As the δ of sample to be tested13C≤- 17 ‰, and ∣ >=15 △ δ ∣ C-N, then it is shell-fish Glucosamine；
As the δ of sample to be tested13C >=-13 ‰, and ∣≤6 △ δ ∣ C-N, then it is microbial method Glucosamine；
The isotopic ratio δ that Elements C in sample to be tested, N are measured with isotopic mass spectrometry13C、δ15The method of N is：
The sample to be tested is taken, after freeze-dried or 50 DEG C of drying, is ground to 100 mesh hereinafter, weighing 0.1~0.5mg grinding
Sample afterwards is placed in tin can, and tight is placed in C/N elemental analyser combination isotope mass spectrometer, on-line determination sample
The isotopic ratio δ of middle Elements C, N13C、δ15N, testing conditions are：
Correcting gas：CO2、N2；
Internal standard substance：USGS24 and GBW4407, IAEA-N-I；
Standard substance：δ13The standard substance of C is PDB, δ15The standard substance of N is air；
Analyze parameter：1000~1250 DEG C of furnace temperature of oxidation, 600~700 DEG C of furnace temperature of reduction, 30~50 DEG C of chromatographic column column temperature,
Carrier gas helium, 10~100mL/min of flow velocity.
2. identifying the method for shell-fish Glucosamine and microbial method Glucosamine as described in claim 1, feature exists
In the qualitative method of step (1) sample to be tested is：
1~1.5mg of sample to be tested is taken, with potassium bromide powder in mass ratio 1：200 fine ground mixings, are subsequently placed in tablet press machine and suppress
At transparence potassium bromide thin slice, with the infrared absorption spectrum of determination of infrared spectroscopy thin slice；Take aminoglucose saccharide by same
The method of sample operates, and measures the infrared absorption spectrum of standard items；If sample to be tested is consistent with the infrared spectrogram of standard items, really
Fixed product to be measured is Glucosamine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810933757.4A CN108872131A (en) | 2018-08-16 | 2018-08-16 | A method of identifying shell-fish Glucosamine and microbial method Glucosamine |
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| CN111521730A (en) * | 2020-06-10 | 2020-08-11 | 江苏澳新生物工程有限公司 | Method for identifying production source of N-acetyl-D-glucosamine |
| CN112946052A (en) * | 2021-02-02 | 2021-06-11 | 江苏澳新生物工程有限公司 | Method for identifying production source of D-glucosamine derivative and application thereof |
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