CN104945277B - A kind of cold labeling basic flavine O and its synthetic method - Google Patents
A kind of cold labeling basic flavine O and its synthetic method Download PDFInfo
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Abstract
The present invention relates to a kind of cold labeling basic flavine O and its synthetic method, described cold labeling basic flavine O for D or15N or13The basic flavine O of C flag, during preparation, with stable isotope D or15N or13C flag methyl alcohol, aniline be raw material, synthesis obtain stable isotope D or15N or13The N of C flag, N dimethyl benzene amine precursor, then through condensation, vulcanization, ammonification, be made stable isotope D or15N or13C flag basic flavine O.Compared with prior art, present invention process route is simple, it is readily synthesized, the easily separated purification of product, more than 99.0%, isotope abundance can fully meet the demand of field of food safety trace detection to product chemical purity in more than 98.0%atom, economy and use value are good, with preferable promotion prospect.
Description
Technical field
The invention belongs to isotope labelling techniques field, it is related to a kind of cold labeling basic flavine O and its synthesis
Method.
Background technology
Basic flavine O, yellow homogeneous powder.Cold water is dissolved in, hot water is soluble in glassy yellow, is boiled and is decomposed, be dissolved in second
Alcohol is in yellow, and its dye powder, in colourless, turns light yellow, in orange in concentrated nitric acid, in hydroxide in the concentrated sulfuric acid after dilution
It is that country forbids food additives into white precipitate in sodium solution.The chemical constitution of basic flavine O determines that it has carcinogenicity,
There is obvious toxicity to human body.At present, developed country and China forbid basic flavine O for food processing field.But part
Illegal businessman, is added into the edible products such as dried bean curd, and this can have a strong impact on the healthy of consumer, or even can cause greatly
Social repercussion.To prevent basic flavine O from being caused harm into field of food, health relevant departments need one kind can rapidly, it is accurate
The technology of true and smaller error detects it, and this has extremely important realistic meaning to the healthy of the people.
Stable isotope dilution mass spectrometry IDMS (Isotope Dilution Mass Spectrometry) be use with
Measured matter has the compound of the cold labeling of identical molecular structure as internal standard substance, with high-resolution liquid phase color
Spectrum-GC-MS (LC/MS) detected, measures the ratio of the ion of respective quality number by mass spectrograph, and with standard
Odds ratio is compared with and then reaching the purpose of accurate quantitative analysis.Sample can effectively be eliminated chemically and physically using Isotopic Internal Standard
Rate of recovery difference caused in pre-treatment step, so as to avoid what the loss because of sample handling processes was caused to testing result
Deviation.
This characteristic of stable isotope dilution mass spectrometry and the high sensitivity of LC/MS and the ability knot for the treatment of complex sample
Altogether so that chromatogram/isotope dilution mass spectrometry technology is acknowledged as a kind of micro and trace organic substance benchmark side of measurement
Method.And target is succeeded in developing in cold labeling basic flavine O, standard examination will be provided for more accurate quantitative analysis basic flavine O
Agent, improves China's food safety detection technical system, meets the demand of China's food security development.At present, both at home and abroad there is not yet
The document report of the synthetic method of cold labeling basic flavine O.
The content of the invention
The purpose of the present invention is exactly that a kind of process route is provided for the defect for overcoming above-mentioned prior art to exist simply,
The synthetic method of easily prepared cold labeling basic flavine O.
The purpose of the present invention can be achieved through the following technical solutions：
A kind of cold labeling basic flavine O, described cold labeling basic flavine O for D or15N or13C
The basic flavine O of mark, wherein,
The molecular structural formula of basic flavine O of D marks is：
Or
15The molecular structural formula of basic flavine O of N marks is：
13The molecular structural formula of the basic flavine O of C flag is：
Or
Wherein, * represents that carbon atom is at this13C atoms.
A kind of synthetic method of cold labeling basic flavine O, the method be with stable isotope D or15N or13C
Mark methyl alcohol, aniline be raw material, synthesis obtain stable isotope D or15N or13The DMA precursor of C flag, then pass through
Condensation, vulcanization, ammonification, be made stable isotope D or15N or13C flag basic flavine O.
In the present invention, the DMA of cold labeling is DMA-D6, N, N- dimethyl
Aniline-D4, N, accelerine-13C6, N, accelerine-15N, N, accelerine-13C2, synthetic method reference
Document carries out (the deuterium-labeled D of the stable isotopes such as Yang Weicheng6The synthesis of-crystal violet and sign [J] applied chemistries, 2013,31
(03)：262-267.).
A kind of synthetic method of cold labeling basic flavine O, the method specifically includes following steps：
(1) condensation step：With stable isotope D or15N or13The DMA of C flag is raw material, with condensation
After agent, catalyst mixing, controlling reaction temperature is 30-150 DEG C, and controls reaction pressure for 0-5MPa, reacts 1-10h, preferably
1-6h, synthesizing stable isotope D or15N or13The methane bass of C flag；
(2) vulcanisation step：By step (1) synthesize stable isotope D or15N or13The methane bass of C flag, in liquid phase
Mix with vulcanizing agent in environment, controlling reaction temperature is 30-200 DEG C, and control reaction pressure for 0-5MPa, react 1-24h, it is excellent
Select 1-20h, synthesizing stable isotope D or15N or13The intermediate of C flag；
(3) ammonification is into salinization step：By step (2) synthesize stable isotope D or15N or13The intermediate of C flag,
Mix with aminating agent in liquid phase environment, controlling reaction temperature is 30-200 DEG C, and control reaction pressure for 0-5MPa, react 1-
24h, preferably 1-20h, that is, synthesize described cold labeling basic flavine O.
Condensing agent described in step (1) is one or two in metaformaldehyde, paraformaldehyde or urea, preferably trimerization first
Aldehyde and urea, described catalyst are the one kind in hydrochloric acid, NaOH, acetic acid or sodium carbonate, preferably NaOH, sodium carbonate
Or acetic acid.
Stable isotope D described in step (1) or15N or13The mol ratio of the N of C flag, accelerine and condensing agent
It is (2-6)：1, and described stable isotope D or15N or13The mol ratio of the N of C flag, accelerine and catalyst
It is 1：(0.1-0.3).
Vulcanizing agent described in step (2) is sulphur powder, and described stable isotope D or15N or13The methane shellfish of C flag
This is 1 with the mol ratio of sulphur powder：(1-6).
Described stable isotope D or15N or13The methane bass of C flag is 1 with the mol ratio of sulphur powder：(2-4).
Aminating agent described in step (3) is ammonium chloride, and described stable isotope D or15N or13The centre of C flag
Body is 1 with the mol ratio of ammonium chloride：(1-6).
Described stable isotope D or15N or13The intermediate of C flag is 1 with the mol ratio of ammonium chloride：(1-3).
Described liquid phase be methyl alcohol, ethanol, ether, acetonitrile, acetic anhydride, ethylene glycol or chloroform in one or more.
Stable isotope D of the present invention12Or15N or13The overall synthetic method of C flag basic flavine O compound can be briefly general
State for：
Wherein, * represents that carbon atom is at this13C atoms.
Specifically, in the present invention, cold labeling basic flavine O-D12The synthetic route of compound is：Stabilization is same
Position element mark methyl alcohol is raw material, after methylating, obtains the deuterium-labeled DMA of stable isotope, then through condensation, sulphur
Change, ammonification obtains cold labeling basic flavine O compound into salt, concrete technology route is：
In the present invention, isotope marks basic flavine O-D is determined8The synthetic route of compound is：Stable isotope deuterium first
Disjunction mark note aniline is raw material, obtains the deuterium-labeled DMA of stable isotope after methylating, then through condensation, vulcanization,
Ammonification obtains cold labeling basic flavine O compound into salt, and concrete technology route is：
In the present invention, cold labeling basic flavine O-13C12The synthetic route of compound is：Stable isotope13C phenyl ring mark aniline is raw material, and stable isotope is obtained after methylating13C flag DMA, then through being condensed,
Vulcanization, ammonification obtain cold labeling basic flavine O compound into salt, and concrete technology route is：
Wherein, * represents that carbon atom is at this13C。
In the present invention, cold labeling basic flavine O-13C4The synthetic route of compound is：Stable isotope13C
Methyl mark aniline is raw material, and stable isotope is obtained after methylating13C flag DMA, then through condensation, sulphur
Change, ammonification obtains cold labeling basic flavine O into salt, concrete technology route is：
Wherein, * represents that carbon atom is at this13C。
In the present invention, cold labeling basic flavine O-15N2The synthetic route of compound is：Stable isotope15N
Amino labeled aniline is raw material, and stable isotope is obtained after methylating15N marks DMA, then through condensation, sulphur
Change, ammonification obtains cold labeling basic flavine O compound into salt, concrete technology route is：
Compared with prior art, the invention has the characteristics that：
(1) present invention firstly discloses a kind of synthetic method of cold labeling basic flavine O；
(2) present invention is simple using process route, it is easy to synthesize；
(3) the easily separated purification of product of the present invention, more than 99.0%, isotope abundance is 98.0% for product chemical purity
More than atom, can fully meet the demand of field of food safety trace detection；
(4) economy of the present invention and use value are good, with preferable promotion prospect.
Specific embodiment
With reference to specific embodiment, the present invention is described in detail.
Embodiment 1
A, cold labeling methane bass-D12Synthesis
In flask, by DMA-D6It is 2 with the mol ratio of metaformaldehyde (mass concentration 37%)：1, and
With N, accelerine-D6It is 1 with the mol ratio of urea, acetic acid：1：0.1, urea, acetic acid are sequentially added, add reaction
The distilled water of volume 20%, stirring is heated to 60 DEG C, reacts 2h.After reaction terminates, washing stands suction filtration, and filter cake, washing is done
It is dry.Thick yield is 98.5%, recrystallization, yield 94.0%, HPLC detection purity 99.4%, abundance 99.2%atom.
B, cold labeling basic flavine O-D12Synthesis
In flask, the methane bass-D of cold labeling is added12, then by methane bass-D12With sulphur powder mole
Than being 1：2, sulphur powder is added, vacuum tightness is reacted, ammonia is passed through, reaction temperature is controlled at 120 DEG C, after reaction 4h, adds chlorination
Ammonium, is further continued for reacting 16h, stops reaction.Reactant is stood, question response thing cooling, crystallization, suction filtration is obtained stable isotope
Mark basic flavine O-D12.Yield is 85.5%, recrystallization, yield 80.0%.HPLC detects purity 99.2%, abundance
99.0atom%.
Embodiment 2
A, cold labeling methane bass-D12Synthesis
In flask, by DMA-D6It is 3 with the mol ratio of metaformaldehyde (mass concentration 37%)：1, and
With N, accelerine-D6It is 1 with the mol ratio of urea, sodium carbonate：1.5：0.15, urea, sodium carbonate are sequentially added, then add
Enter the distilled water of reaction volume 20%, stir, be heated to 100 DEG C, react 3h.After reaction terminates, washing, standing suction filtration, filter cake,
Washing, dries.Thick yield is 95.5%, recrystallization, yield 91.0%, HPLC detection purity 99.3%, abundance 98.9%atom.
B, cold labeling basic flavine O-D12Synthesis
Methane bass-the D of cold labeling is added in flask12, then by methane bass-D12With sulphur powder mole
Than being 1：2.5, sulphur powder is added, vacuum tightness is reacted, ammonia is passed through, reaction temperature is controlled at 160 DEG C, after reaction 5h, adds chlorine
Change ammonium, be further continued for reacting 14h, stop reaction.Reactant is stood, question response thing cooling, crystallization, suction filtration is obtained and stablizes same position
Element mark basic flavine O-D12.Yield is 86.5%, recrystallization, yield 81.4%.HPLC detects purity 99.1%, abundance
98.3%atom.
Embodiment 3
A, cold labeling methane bass-D12Synthesis
In flask, by DMA-D6It is 4 with the mol ratio of metaformaldehyde (mass concentration 37%)：1, and
With N, accelerine-D6It is 1 with the mol ratio of urea, acetic acid：2：0.15, urea, acetic acid are sequentially added, add reaction
The distilled water of volume 20%, stirring is heated to 140 DEG C, reacts 5h.After reaction terminates, washing stands suction filtration, and filter cake is washed,
Dry.Thick yield is 92.5%, recrystallization, yield 89.0%, HPLC detection purity 99.6%, abundance 99.2%atom.
B, cold labeling basic flavine O-D12Synthesis
Methane bass-the D of cold labeling is added in flask12, then by methane bass-D12With sulphur powder mole
Than being 1：3, sulphur powder is added, vacuum tightness is reacted, ammonia is passed through, reaction temperature is controlled at 180 DEG C, after reaction 7h, adds chlorination
Ammonium, is further continued for reacting 15h, stops reaction.Reactant is stood, question response thing cooling, crystallization, suction filtration is obtained stable isotope
Mark basic flavine O-D12.Yield is 88.5%, recrystallization, yield 82.4%.HPLC detects purity 99.0%, abundance 99.0%
atom。
Embodiment 4
A, cold labeling methane bass-D8Synthesis
In flask, by DMA-D4It is 3 with the mol ratio of metaformaldehyde (mass concentration 37%)：1, and
With N, accelerine-D4It is 1 with the mol ratio of urea, NaOH：2.5：0.15, urea, NaOH are sequentially added,
The distilled water of reaction volume 20% is added, is stirred, be heated to 90 DEG C, react 4h.After reaction terminates, washing stands suction filtration, filter
Cake, washing is dried.Thick yield is 97.5%, recrystallization, yield 93.0%, HPLC detection purity 99.1%, abundance 99.0%
atom。
B, cold labeling basic flavine O-D8Synthesis
Methane bass-the D of cold labeling is added in flask8, then by methane bass-D8With the mol ratio of sulphur powder
It is 1：3.5, sulphur powder is added, vacuum tightness is reacted, ammonia is passed through, reaction temperature is controlled at 100 DEG C, after reaction 8h, adds chlorination
Ammonium, is further continued for reacting 16h, stops reaction.Reactant is stood, question response thing cooling, crystallization, suction filtration is obtained stable isotope
Mark basic flavine O-D8.Yield is 82.5%, recrystallization, yield 76.2%.HPLC detects purity 99.0%, abundance 98.8%
atom。
Embodiment 5
A, cold labeling methane bass-D8Synthesis
In flask, by DMA-D4It is 4 with the mol ratio of metaformaldehyde (mass concentration 37%)：1, and
With N, accelerine-D4It is 1 with the mol ratio of urea, acetic acid：1.5：0.2, urea, acetic acid are sequentially added, add anti-
The distilled water of volume 20% is answered, is stirred, be heated to 125 DEG C, react 6h.After reaction terminates, washing stands suction filtration, and filter cake is washed
Wash, dry.Thick yield is 96.0%, recrystallization, yield 91.8%, HPLC detection purity 99.5%, abundance 99.0%atom.
B, cold labeling basic flavine O-D8Synthesis
Methane bass-the D of cold labeling is added in flask8, then by methane bass-D8With the mol ratio of sulphur powder
It is 1：2.5, sulphur powder is added, vacuum tightness is reacted, ammonia is passed through, reaction temperature is controlled at 120 DEG C, after reaction 4h, adds chlorination
Ammonium, is further continued for reacting 16h, stops reaction.Reactant is stood, question response thing cooling, crystallization, suction filtration is obtained stable isotope
Mark basic flavine O-D8.Yield is 86.5%, recrystallization, yield 82.0%.HPLC detects purity 99.3%, abundance 98.8%
atom。
Embodiment 6
A, cold labeling methane bass-D8Synthesis
In flask, by DMA-D4It is 2 with the mol ratio of metaformaldehyde (mass concentration 37%)：1, and
With N, accelerine-D4It is 1 with the mol ratio of urea, sodium carbonate：3.5：0.2, urea, sodium carbonate are sequentially added, then add
Enter the distilled water of reaction volume 20%, stir, be heated to 120 DEG C, react 7h.After reaction terminates, washing, standing suction filtration, filter cake,
Washing, dries.Thick yield is 96.0%, recrystallization, yield 91.8%, HPLC detection purity 99.7%, abundance 99.0%atom.
B, cold labeling basic flavine O-D8Synthesis
Methane bass-the D of cold labeling is added in flask8, then by methane bass-D8With the mol ratio of sulphur powder
It is 1：1.5, sulphur powder is added, vacuum tightness is reacted, ammonia is passed through, reaction temperature is controlled at 145 DEG C, after reaction 4h, adds chlorination
Ammonium, is further continued for reacting 16h, stops reaction.Reactant is stood, question response thing cooling, crystallization, suction filtration is obtained stable isotope
Mark basic flavine O-D8.Yield is 86.5%, recrystallization, yield 82.0%.HPLC detects purity 99.3%, abundance 98.8%
atom。
Embodiment 7
A, cold labeling methane bass-13C12Synthesis
In flask, by DMA-13C6It is 2.5 with the mol ratio of metaformaldehyde (mass concentration 37%)：1,
And with N, accelerine-13C6It is 1 with the mol ratio of urea, acetic acid：3：0.2, urea, acetic acid are sequentially added, add anti-
The distilled water of volume 20% is answered, is stirred, be heated to 60 DEG C, react 6h.After reaction terminates, washing stands suction filtration, and filter cake is washed,
Dry.Thick yield is 97.5%, recrystallization, yield 92.0%, HPLC detection purity 99.8%, abundance 99.0%atom.
B, cold labeling basic flavine O-13C12Synthesis
In flask add cold labeling methane bass-13C12, then by methane bass-13C12With rubbing for sulphur powder
You are than being 1：3, sulphur powder is added, vacuum tightness is reacted, ammonia is passed through, reaction temperature is controlled at 120 DEG C, after reaction 4h, adds chlorine
Change ammonium, be further continued for reacting 16h, stop reaction.Reactant is stood, question response thing cooling, crystallization, suction filtration is obtained and stablizes same position
Element mark basic flavine O-13C12.Yield is 88.5%, recrystallization, yield 85.8%.HPLC detects purity 99.5%, abundance
98.7%atom.
Embodiment 8
A, cold labeling methane bass-13C12Synthesis
In flask, by DMA-13C6It is 3.5 with the mol ratio of metaformaldehyde (mass concentration 37%)：1,
And with N, accelerine-13C6It is 1 with the mol ratio of urea, NaOH：2.5：0.25, sequentially add urea, hydroxide
Sodium, adds the distilled water of reaction volume 20%, stirring, is heated to 110 DEG C, reacts 2h.After reaction terminates, washing, standing is taken out
Filter, filter cake, washing is dried.Thick yield is 96.4%, recrystallization, yield 92.2%, HPLC detection purity 99.6%, abundance
99.2%atom.
B, cold labeling basic flavine O-13C12Synthesis
In flask add cold labeling methane bass-13C12, then by methane bass-13C12With rubbing for sulphur powder
You are than being 1：3.5, sulphur powder is added, vacuum tightness is reacted, ammonia is passed through, reaction temperature is controlled at 120 DEG C, after reaction 4h, is added
Ammonium chloride, is further continued for reacting 16h, stops reaction.Reactant is stood, question response thing cooling, crystallization, suction filtration is obtained stabilization same
The plain mark basic flavine O in position-13C12.Yield is 87.5%, recrystallization, yield 84.8%.HPLC detects purity 99.5%, abundance
98.9%atom.
Embodiment 9
A, cold labeling methane bass-13C12Synthesis
In flask, by DMA-13C6It is 1 with the mol ratio of metaformaldehyde (mass concentration 37%)：1, and
With N, accelerine-13C6It is 1 with the mol ratio of urea, acetic acid：4：0.3, urea, acetic acid are sequentially added, add reaction
The distilled water of volume 20%, stirring is heated to 130 DEG C, reacts 5h.After reaction terminates, washing stands suction filtration, and filter cake is washed,
Dry.Thick yield is 95.8%, recrystallization, yield 91.2%, HPLC detection purity 99.6%, abundance 98.7%atom.
B, cold labeling basic flavine O-13C12Synthesis
In flask add cold labeling methane bass-13C12, then by methane bass-13C12With rubbing for sulphur powder
You are than being 1：3.5, sulphur powder is added, vacuum tightness is reacted, ammonia is passed through, reaction temperature is controlled at 150 DEG C, after reaction 3h, is added
Ammonium chloride, is further continued for reacting 18h, stops reaction.Reactant is stood, question response thing cooling, crystallization, suction filtration is obtained stabilization same
The plain mark basic flavine O in position-13C12.Yield is 88.5%, recrystallization, yield 84.8%.HPLC detects purity 99.4%, abundance
98.0%atom.
Embodiment 10
A, cold labeling methane bass-13C4Synthesis
In flask, by DMA-13C2It is 4 with the mol ratio of metaformaldehyde (mass concentration 37%)：1, and
With N, accelerine-13C2It is 1 with the mol ratio of urea, sodium carbonate：3.5：0.25, urea, sodium carbonate are sequentially added, then
The distilled water of reaction volume 20% is added, stirring is heated to 70 DEG C, reacts 3.5h.After reaction terminates, washing stands suction filtration, filter
Cake, washing is dried.Thick yield is 97.3%, recrystallization, yield 93.0%, HPLC detection purity 99.5%, abundance 99.3%
atom。
B, cold labeling basic flavine O-13C4Synthesis
In flask add cold labeling methane bass-13C4, then by methane bass-13C4With sulphur powder mole
Than being 1：2.5, sulphur powder is added, vacuum tightness is reacted, ammonia is passed through, reaction temperature is controlled at 120 DEG C, after reaction 4h, adds chlorine
Change ammonium, be further continued for reacting 16h, stop reaction.Reactant is stood, question response thing cooling, crystallization, suction filtration is obtained and stablizes same position
Element mark basic flavine O-13C4.Yield is 86.5%, recrystallization, yield 83.0%.HPLC detects purity 99.4%, abundance
99.1%atom.
Embodiment 11
A, cold labeling methane bass-13C4Synthesis
In flask, by DMA-13C2It is 2.5 with the mol ratio of metaformaldehyde (mass concentration 37%)：1,
And with N, accelerine-13C2It is 1 with the mol ratio of urea, acetic acid：5.5：0.2, urea, acetic acid are sequentially added, add
The distilled water of reaction volume 20%, stirring is heated to 60 DEG C, reacts 4h.After reaction terminates, washing stands suction filtration, and filter cake is washed
Wash, dry.Thick yield is 97.3%, recrystallization, yield 93.0%, HPLC detection purity 99.5%, abundance 98.5%atom.
B, cold labeling basic flavine O-13C4Synthesis
In flask add cold labeling methane bass-13C4, then by methane bass-13C4With sulphur powder mole
Than being 1：2, sulphur powder is added, vacuum tightness is reacted, ammonia is passed through, reaction temperature is controlled at 130 DEG C, after reaction 6h, adds chlorination
Ammonium, is further continued for reacting 18h, stops reaction.Reactant is stood, question response thing cooling, crystallization, suction filtration is obtained stable isotope
Mark basic flavine O-13C4.Yield is 84.5%, recrystallization, yield 81.0%.HPLC detects purity 99.4%, abundance
98.1%atom.
Embodiment 12
A, cold labeling methane bass-13C4Synthesis
In flask, by DMA-13C2It is 3.5 with the mol ratio of metaformaldehyde (mass concentration 37%)：1,
And with N, accelerine-13C2It is 1 with the mol ratio of urea, NaOH：4.5：0.2, sequentially add urea, hydroxide
Sodium, adds the distilled water of reaction volume 20%, stirring, is heated to 145 DEG C, reacts 4h.After reaction terminates, washing, standing is taken out
Filter, filter cake, washing is dried.Thick yield is 96.3%, recrystallization, yield 92.0%, HPLC detection purity 99.3%, abundance
98.8%atom.
B, cold labeling basic flavine O-13C4Synthesis
In flask add cold labeling methane bass-13C4, then by methane bass-13C4With sulphur powder mole
Than being 1：2.5, sulphur powder is added, vacuum tightness is reacted, ammonia is passed through, reaction temperature is controlled at 130 DEG C, after reaction 6h, adds chlorine
Change ammonium, be further continued for reacting 18h, stop reaction.Reactant is stood, question response thing cooling, crystallization, suction filtration is obtained and stablizes same position
Element mark basic flavine O-13C4.Yield is 87.5%, recrystallization, yield 84.0%.HPLC detects purity 99.2%, abundance
98.3%atom.
Embodiment 13
A, cold labeling methane bass-15N2Synthesis
In flask, by DMA-15N is 4 with the mol ratio of metaformaldehyde (mass concentration 37%)：1, and
With N, accelerine-15N is 1 with the mol ratio of urea, acetic acid：6：0.25, urea, acetic acid are sequentially added, add reaction
The distilled water of volume 20%, stirring is heated to 80 DEG C, reacts 5h.After reaction terminates, washing stands suction filtration, and filter cake, washing is done
It is dry.Thick yield is 95.3%, recrystallization, yield 93.3%, HPLC detection purity 99.5%, abundance 99.4%atom.
B, cold labeling basic flavine O-15N2Synthesis
In flask add cold labeling methane bass-15N2, then by methane bass-15N2With sulphur powder mole
Than being 1：4, sulphur powder is added, vacuum tightness is reacted, ammonia is passed through, reaction temperature is controlled at 100 DEG C, after reaction 4h, adds chlorination
Ammonium, is further continued for reacting 16h, stops reaction.Reactant is stood, question response thing cooling, crystallization, suction filtration is obtained stable isotope
Mark basic flavine O-15N2.Yield is 85.5%, recrystallization, yield 83.3%.HPLC detects purity 99.3%, abundance
99.2%atom.
Embodiment 14
A, cold labeling methane bass-15N2Synthesis
In flask, by DMA-15N is 3 with the mol ratio of metaformaldehyde (mass concentration 37%)：1, and
With N, accelerine-15N is 1 with the mol ratio of urea, acetic acid：5.5：0.3, urea, acetic acid are sequentially added, add anti-
The distilled water of volume 20% is answered, is stirred, be heated to 80 DEG C, react 6h.After reaction terminates, washing stands suction filtration, and filter cake is washed,
Dry.Thick yield is 92.3%, recrystallization, yield 90.3%, HPLC detection purity 99.7%, abundance 99.3%atom.
B, cold labeling basic flavine O-15N2Synthesis
In flask add cold labeling methane bass-15N2, then by methane bass-15N2With sulphur powder mole
Than being 1：2, sulphur powder is added, vacuum tightness is reacted, ammonia is passed through, reaction temperature is controlled at 120 DEG C, after reaction 8h, adds chlorination
Ammonium, is further continued for reacting 18h, stops reaction.Reactant is stood, question response thing cooling, crystallization, suction filtration is obtained stable isotope
Mark basic flavine O-15N2.Yield is 88.5%, recrystallization, yield 85.3%.HPLC detects purity 99.2%, abundance
99.0%atom.
Embodiment 15
A, cold labeling methane bass-15N2Synthesis
In flask, by DMA-15N is 2.5 with the mol ratio of metaformaldehyde (mass concentration 37%)：1,
And with N, accelerine-15N is 1 with the mol ratio of urea, sodium carbonate：4.5：0.3, urea, sodium carbonate are sequentially added, then
The distilled water of reaction volume 20% is added, stirring is heated to 80 DEG C, reacts 4.5h.After reaction terminates, washing stands suction filtration, filter
Cake, washing is dried.Thick yield is 94.3%, recrystallization, yield 92.3%, HPLC detection purity 99.4%, abundance 99.2%
atom。
B, cold labeling basic flavine O-15N2Synthesis
In flask add cold labeling methane bass-15N2, then by methane bass-15N2With sulphur powder mole
Than being 1：4, sulphur powder is added, vacuum tightness is reacted, ammonia is passed through, reaction temperature is controlled at 120 DEG C, after reaction 6h, adds chlorination
Ammonium, is further continued for reacting 17h, stops reaction.Reactant is stood, question response thing cooling, crystallization, suction filtration is obtained stable isotope
Mark basic flavine O-15N2.Yield is 83.5%, recrystallization, yield 81.3%.HPLC detects purity 99.0%, abundance
98.7%atom.
Embodiment 16
The present embodiment cold labeling basic flavine O-D8, molecular structural formula is
Specific preparation method is as follows：
(1) condensation step：With stable isotope D4The DMA of mark is raw material, with condensing agent, catalyst
After mixing, controlling reaction temperature is 150 DEG C, and controls reaction pressure for 5MPa, reacts 1h, synthesizing stable isotope D4Mark
Methane bass-D8；
(2) vulcanisation step：The stable isotope D that step (1) is synthesized4Methane bass-the D of mark8, in liquid phase environment
Mix with vulcanizing agent, controlling reaction temperature is 200 DEG C, and control reaction pressure for 5MPa, react 1h, synthesizing stable isotope D8
The intermediate of mark；
(3) ammonification is into salinization step：The stable isotope D that step (2) is synthesized4Intermediate-the D of mark8, in liquid phase ring
Mix with aminating agent in border, controlling reaction temperature is 200 DEG C, and control reaction pressure for 5MPa, reaction 1h, i.e. synthesizing stable is same
Position element marks basic flavine O-D8。
Wherein, the condensing agent in step (1) is metaformaldehyde, and catalyst is hydrochloric acid, and stable isotope D4Mark
N, accelerine is 6 with the mol ratio of metaformaldehyde：1, it is 1 with the mol ratio of hydrochloric acid：0.3.
Vulcanizing agent in step (2) is sulphur powder, and stable isotope D4Methane bass-the D of mark8With sulphur powder mole
Than being 1：6.
Aminating agent in step (3) is ammonium chloride, and stable isotope D4Intermediate-the D of mark8With rubbing for ammonium chloride
You are than being 1：6.
In the present embodiment, liquid phase is that ether, ethylene glycol and chloroform are 1 in mass ratio：1：1 blended liquid phase.
Embodiment 17
The present embodiment cold labeling basic flavine O-D12, molecular structural formula is
Specific preparation method is as follows：
(1) condensation step：With stable isotope D6The DMA of mark is raw material, with condensing agent, catalyst
After mixing, controlling reaction temperature is 30 DEG C, and controls reaction pressure for 0MPa, reacts 10h, synthesizing stable isotope D6Mark
Methane bass-D12；
(2) vulcanisation step：The stable isotope D that step (1) is synthesized6Methane bass-the D of mark12, in liquid phase environment
Mix with vulcanizing agent, controlling reaction temperature is 30 DEG C, and control reaction pressure for 0MPa, react 24h, synthesizing stable isotope D12
The intermediate of mark；
(3) ammonification is into salinization step：The stable isotope D that step (2) is synthesized6Intermediate-the D of mark12, in liquid phase ring
Mix with aminating agent in border, controlling reaction temperature is 30 DEG C, and control reaction pressure for 0MPa, reaction 24h, i.e. synthesizing stable is same
Position element marks basic flavine O-D12。
Wherein, the condensing agent in step (1) is urea, and catalyst is sodium carbonate, and stable isotope D6The N of mark,
Accelerine is 2 with the mol ratio of urea：1, it is 1 with the mol ratio of sodium carbonate：0.1.
Vulcanizing agent in step (2) is sulphur powder, and stable isotope D6Methane bass-the D of mark12With sulphur powder mole
Than being 1：1.
Aminating agent in step (3) is ammonium chloride, and stable isotope D6Intermediate-the D of mark12With rubbing for ammonium chloride
You are than being 1：1.
In the present embodiment, liquid phase is methyl alcohol.
Embodiment 18
The present embodiment cold labeling basic flavine O-15N2, molecular structural formula is
Specific preparation method is as follows：
(1) condensation step：With stable isotope15The DMA of N marks is raw material, with condensing agent, catalyst
After mixing, controlling reaction temperature is 80 DEG C, and controls reaction pressure for 3MPa, reacts 6h, synthesizing stable isotope15N marks
Methane bass-15N2；
(2) vulcanisation step：The stable isotope that step (1) is synthesized15The methane bass of N marks-15N2, in liquid phase environment
In mix with vulcanizing agent, controlling reaction temperature is 120 DEG C, and controls reaction pressure for 3MPa, reacts 12h, the same position of synthesizing stable
Element15The intermediate of N marks-15N2；
(3) ammonification is into salinization step：The stable isotope that step (2) is synthesized15The intermediate of N marks-15N2, in liquid phase
Mix with aminating agent in environment, controlling reaction temperature is 120 DEG C, and control reaction pressure for 3MPa, react 12h, that is, synthesize steady
Determine isotope marks basic flavine O-15N2。
Wherein, the condensing agent in step (1) is paraformaldehyde, and catalyst is NaOH, and stable isotope15N is marked
The N of note, accelerine is 3 with the mol ratio of paraformaldehyde：1, it is 1 with the mol ratio of NaOH：0.2.
Vulcanizing agent in step (2) is sulphur powder, and stable isotope15The methane bass of N marks-15N2With rubbing for sulphur powder
You are than being 1：4.
Aminating agent in step (3) is ammonium chloride, and stable isotope15The intermediate of N marks-15N2With ammonium chloride
Mol ratio is 1：3.
In the present embodiment, liquid phase is that ethanol, acetonitrile and acetic anhydride are 1 in mass ratio：1：1 blended liquid phase.
Embodiment 19
The present embodiment cold labeling basic flavine O-13C4, molecular structural formula is
Wherein, * represents that carbon atom is at this13C atoms.
Specific preparation method is as follows：
(1) condensation step：With stable isotope13The DMA of C flag is raw material, with condensing agent, catalyst
After mixing, controlling reaction temperature is 100 DEG C, and controls reaction pressure for 4MPa, reacts 4h, synthesizing stable isotope13C flag
Methane bass-13C4；
(2) vulcanisation step：The stable isotope that step (1) is synthesized13The methane bass of C flag-13C4, in liquid phase environment
In mix with vulcanizing agent, controlling reaction temperature is 150 DEG C, and controls reaction pressure for 4MPa, reacts 20h, the same position of synthesizing stable
Element13The intermediate of C flag-13C4；
(3) ammonification is into salinization step：The stable isotope that step (2) is synthesized13The intermediate of C flag-13C4, in liquid phase
Mix with aminating agent in environment, controlling reaction temperature is 150 DEG C, and control reaction pressure for 4MPa, react 20h, that is, synthesize steady
Determine isotope marks basic flavine O-13C4。
Wherein, the condensing agent in step (1) is metaformaldehyde, and catalyst is acetic acid, and stable isotope13C flag
N, accelerine is 5 with the mol ratio of metaformaldehyde：1, it is 1 with the mol ratio of acetic acid：0.25.
Vulcanizing agent in step (2) is sulphur powder, and stable isotope13The methane bass of C flag-13C4With rubbing for sulphur powder
You are than being 1：3.
Aminating agent in step (3) is ammonium chloride, and stable isotope13The intermediate of C flag-13C4With ammonium chloride
Mol ratio is 1：2.
In the present embodiment, liquid phase is ether.
Claims (8)
1. a kind of synthetic method of cold labeling basic flavine O, it is characterised in that described cold labeling alkali
Property light yellow O be D or15N or13The basic flavine O of C flag, wherein,
The molecular structural formula of basic flavine O of D marks is：
15The molecular structural formula of basic flavine O of N marks is：
13The molecular structural formula of the basic flavine O of C flag is：
Wherein, * represents that carbon atom is at this13C atoms；
Described synthetic method be with stable isotope D or15N or13C flag methyl alcohol, aniline are raw material, and synthesis obtains stablizing same position
Plain D or15N or13The DMA precursor of C flag, then through condensation, vulcanization, ammonification, be made stable isotope D or15N
Or13C flag basic flavine O, specifically includes following steps：
(1) condensation step：With stable isotope D or15N or13The DMA of C flag is raw material, with condensing agent, catalysis
After agent mixing, controlling reaction temperature is 30-150 DEG C, and controls reaction pressure for 0-5MPa, reacts 1-10h, the same position of synthesizing stable
Plain D or15N or13The methane bass of C flag；
(2) vulcanisation step：By step (1) synthesize stable isotope D or15N or13The methane bass of C flag, in liquid phase environment
Mix with vulcanizing agent, controlling reaction temperature is 30-200 DEG C, and control reaction pressure for 0-5MPa, react 1-24h, synthesizing stable
Isotope D or15N or13The intermediate of C flag；
(3) ammonification is into salinization step：By step (2) synthesize stable isotope D or15N or13The intermediate of C flag, in liquid phase ring
Mix with aminating agent in border, controlling reaction temperature is 30-200 DEG C, and control reaction pressure for 0-5MPa, react 1-24h, that is, close
Into described cold labeling basic flavine O.
2. a kind of synthetic method of cold labeling basic flavine O according to claim 1, it is characterised in that step
Suddenly the condensing agent described in (1) is one or two in metaformaldehyde, paraformaldehyde or urea, described catalyst is hydrochloric acid,
One kind in NaOH, acetic acid or sodium carbonate.
3. a kind of synthetic method of cold labeling basic flavine O according to claim 1, it is characterised in that step
Suddenly stable isotope D described in (1) or15N or13The N of C flag, accelerine is (2-6) with the mol ratio of condensing agent：1,
And described stable isotope D or15N or13The N of C flag, accelerine is 1 with the mol ratio of catalyst：(0.1-
0.3)。
4. a kind of synthetic method of cold labeling basic flavine O according to claim 1, it is characterised in that step
Suddenly the vulcanizing agent described in (2) is sulphur powder, and described stable isotope D or15N or13The methane bass of C flag and sulphur powder
Mol ratio is 1：(1-6).
5. a kind of synthetic method of cold labeling basic flavine O according to claim 4, it is characterised in that institute
The stable isotope D that states or15N or13The methane bass of C flag is 1 with the mol ratio of sulphur powder：(2-4).
6. a kind of synthetic method of cold labeling basic flavine O according to claim 1, it is characterised in that step
Suddenly the aminating agent described in (3) is ammonium chloride, and described stable isotope D or15N or13The intermediate and ammonium chloride of C flag
Mol ratio be 1：(1-6).
7. a kind of synthetic method of cold labeling basic flavine O according to claim 6, it is characterised in that institute
The stable isotope D that states or15N or13The intermediate of C flag is 1 with the mol ratio of ammonium chloride：(1-3).
8. a kind of synthetic method of cold labeling basic flavine O according to claim 1, it is characterised in that institute
The liquid phase stated be methyl alcohol, ethanol, ether, acetonitrile, acetic anhydride, ethylene glycol or chloroform in one or more.
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