CN105974022B - A method of 5 hydroxymethyl furfural and its derivative are measured based on positive and reversed-phase high performance liquid chromatography combination - Google Patents
A method of 5 hydroxymethyl furfural and its derivative are measured based on positive and reversed-phase high performance liquid chromatography combination Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 24
- 238000004007 reversed phase HPLC Methods 0.000 title claims abstract description 19
- NOEGNKMFWQHSLB-UHFFFAOYSA-N 5-hydroxymethylfurfural Chemical compound OCC1=CC=C(C=O)O1 NOEGNKMFWQHSLB-UHFFFAOYSA-N 0.000 title claims description 40
- RJGBSYZFOCAGQY-UHFFFAOYSA-N hydroxymethylfurfural Natural products COC1=CC=C(C=O)O1 RJGBSYZFOCAGQY-UHFFFAOYSA-N 0.000 title claims description 40
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims abstract description 30
- CHTHALBTIRVDBM-UHFFFAOYSA-N furan-2,5-dicarboxylic acid Chemical class OC(=O)C1=CC=C(C(O)=O)O1 CHTHALBTIRVDBM-UHFFFAOYSA-N 0.000 claims abstract description 27
- 239000000126 substance Substances 0.000 claims abstract description 21
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims abstract description 18
- 238000001514 detection method Methods 0.000 claims abstract description 12
- PKAUICCNAWQPAU-UHFFFAOYSA-N 2-(4-chloro-2-methylphenoxy)acetic acid;n-methylmethanamine Chemical compound CNC.CC1=CC(Cl)=CC=C1OCC(O)=O PKAUICCNAWQPAU-UHFFFAOYSA-N 0.000 claims abstract description 5
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 5
- 239000012071 phase Substances 0.000 claims description 24
- PCSKKIUURRTAEM-UHFFFAOYSA-N 5-hydroxymethyl-2-furoic acid Chemical class OCC1=CC=C(C(O)=O)O1 PCSKKIUURRTAEM-UHFFFAOYSA-N 0.000 claims description 14
- 238000004587 chromatography analysis Methods 0.000 claims description 11
- 239000007788 liquid Substances 0.000 claims description 8
- 238000005259 measurement Methods 0.000 claims description 7
- 230000010354 integration Effects 0.000 claims description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N methanol Natural products OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 6
- 238000004366 reverse phase liquid chromatography Methods 0.000 claims description 6
- 238000005070 sampling Methods 0.000 claims description 6
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 claims description 4
- 239000007791 liquid phase Substances 0.000 claims description 4
- 238000000926 separation method Methods 0.000 claims description 4
- 239000012086 standard solution Substances 0.000 claims description 4
- 150000001299 aldehydes Chemical class 0.000 claims description 2
- -1 furyl dimethyl carbinols Chemical class 0.000 abstract description 7
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 abstract description 6
- 150000002240 furans Chemical class 0.000 abstract description 5
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 abstract description 3
- DNXDYHALMANNEJ-UHFFFAOYSA-N furan-2,3-dicarboxylic acid Chemical class OC(=O)C=1C=COC=1C(O)=O DNXDYHALMANNEJ-UHFFFAOYSA-N 0.000 abstract description 2
- 238000010829 isocratic elution Methods 0.000 abstract description 2
- SMNDYUVBFMFKNZ-UHFFFAOYSA-N 2-furoic acid Chemical compound OC(=O)C1=CC=CO1 SMNDYUVBFMFKNZ-UHFFFAOYSA-N 0.000 abstract 4
- CETXOEGRUBXUAL-UHFFFAOYSA-N 3-(hydroxymethyl)furan-2-carbaldehyde Chemical class OCC=1C=COC=1C=O CETXOEGRUBXUAL-UHFFFAOYSA-N 0.000 abstract 3
- TYNYLABSLJYTTH-UHFFFAOYSA-N 3-(hydroxymethyl)furan-2-carboxylic acid Chemical compound OCC=1C=COC=1C(O)=O TYNYLABSLJYTTH-UHFFFAOYSA-N 0.000 abstract 2
- 238000004458 analytical method Methods 0.000 description 7
- 239000000758 substrate Substances 0.000 description 6
- 238000006243 chemical reaction Methods 0.000 description 5
- 238000004128 high performance liquid chromatography Methods 0.000 description 5
- 238000004305 normal phase HPLC Methods 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 238000011002 quantification Methods 0.000 description 5
- 230000014759 maintenance of location Effects 0.000 description 4
- 239000002028 Biomass Substances 0.000 description 3
- 238000006555 catalytic reaction Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000010828 elution Methods 0.000 description 3
- HYBBIBNJHNGZAN-UHFFFAOYSA-N furfural Chemical compound O=CC1=CC=CO1 HYBBIBNJHNGZAN-UHFFFAOYSA-N 0.000 description 3
- 230000005526 G1 to G0 transition Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 235000011149 sulphuric acid Nutrition 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 238000009849 vacuum degassing Methods 0.000 description 2
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 1
- MQIUGAXCHLFZKX-UHFFFAOYSA-N Di-n-octyl phthalate Natural products CCCCCCCCOC(=O)C1=CC=CC=C1C(=O)OCCCCCCCC MQIUGAXCHLFZKX-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 229960000074 biopharmaceutical Drugs 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 238000003889 chemical engineering Methods 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000001212 derivatisation Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 238000010812 external standard method Methods 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 239000000446 fuel Substances 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 230000026030 halogenation Effects 0.000 description 1
- 238000005658 halogenation reaction Methods 0.000 description 1
- KFZMGEQAYNKOFK-UHFFFAOYSA-N isopropyl alcohol Natural products CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 238000005374 membrane filtration Methods 0.000 description 1
- 238000005839 oxidative dehydrogenation reaction Methods 0.000 description 1
- 238000005192 partition Methods 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 238000007670 refining Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
Classifications
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- 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
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
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- 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)
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Abstract
The invention discloses a kind of methods measuring 5 hydroxymethylfurfurals and its derivative based on positive and reversed-phase high performance liquid chromatography combination.The present invention is to measure 5 hydroxymethylfurfurals, 2,5 furyl dimethyl carbinols, 2,5 furans dicarbaldehydes, 5 methylol, 2 furancarboxylic acid, 5 formoxyl, 2 furancarboxylic acid and 2, the method for 5 furandicarboxylic acids based on positive and reversed-phase high performance liquid chromatography combination.Differential refraction detector is used by sulfuric acid isocratic elution using Bio Rad Aminex HPX 87H (300mm × 7.8mm) forward chromatographic column, temperature is 35 DEG C, 2,5 furyl dimethyl carbinols of quantitative determination, 2,5 furandicarboxylic acids, 5 hydroxymethylfurfurals and 2,5 furans dicarbaldehydes；Using Zorbax Eclipse XDS C18 (300mm × 4.6mm) reverse-phase chromatographic column; pass through acetonitrile and acetic acid binary gradient elutes; using variable-wavelenght detector, ultraviolet wavelength 275nm quantitative determines 5 formoxyl, 2 furancarboxylic acid and 5 methylol, 2 furancarboxylic acid.It is combined by positive and reversed-phase high performance liquid chromatography, quantitative determines 6 kinds of substances, significantly improve the separating degree and accuracy in detection to above-mentioned furfuran compound.
Description
Technical field
The invention belongs to bioengineering, bio-pharmaceuticals and field of chemical engineering, disclose a kind of high based on positive and reverse phase
Effect liquid phase chromatogram is combined the method for being precisely separating and quantitative determining a variety of furan compound components, is related to containing 2,5- furans two
Methanol, 5 hydroxymethyl furfural, 2,5-furandaldehyde, 5- methylol -2- furancarboxylic acids, 5- formoxyl -2- furancarboxylic acids and 2,5- furans two
Positive and the reversed-phase high performance liquid chromatography combination of formic acid are precisely separating to be measured with accurate quantification.
Background technology
5 hydroxymethyl furfural, can hydrogenated, oxidative dehydrogenation, living things catalysis as a kind of important biology base platform chemicals
Even microbe conversion generates 2,5-FDM, 5 hydroxymethyl furfural, 2,5-furandaldehyde, 5- methylol -2- furancarboxylic acids, 5-
A series of derivatives such as formoxyl -2- furancarboxylic acids and 2,5-furandicarboxylic acid, they pass through ester again as important furans raw material
The reactions such as change, halogenation, polymerization can produce numerous high value added products, be widely used in fuel, medicine, engineering material and essence
Refine the various fields such as work, synthesis and the important content that application is biomass refining industry development.
Separating, analyzing and quantitatively detecting for current above-mentioned furfuran compound mainly uses high performance liquid chromatography and makings
Combination method.Due to the furan nucleus carbon skeleton of 5 hydroxymethyl furfural of the present invention and its five kinds of derivatives, the position of branch, official
It can roll into a ball that molecular size, physical parameter are all very close, there is mutual between substance in single high performance liquid chromatography detection
Interference causes to be difficult to efficiently separate identification or quantitative determination, is generally only capable of measuring 3 to 4 kinds of substances therein, and makings is used to join
Used time usually requires to perform the derivatization processing to high boiling substances such as 2,5-furandicarboxylic acid therein, cumbersome and be easy to draw
Measurement error is played, as a result poor reproducibility and testing cost is higher.
Based on the above, the present invention is combined using positive and reversed-phase high performance liquid chromatography, and it is a kind of quick, accurate to provide
The method of Synchronization Analysis and quantitative determination 5 hydroxymethyl furfural and its derivative is particularly suitable in the synthesis of biomass biorefinery
State the analysis and detection of furans platform chemicals.
Invention content
Goal of the invention：For deficiency in the prior art, it is high based on positive and reverse phase that the object of the present invention is to provide one kind
Effect liquid phase chromatogram is combined the method that accurate quantification measures 5 hydroxymethyl furfural and its derivative, to realize to 5 hydroxymethyl furfural
The accurate quantification detection of each component during the catalytic conversion reaction carried out for substrate.
Technical solution：In order to achieve the above-mentioned object of the invention, the technical solution adopted by the present invention is：
1, a kind of method that 5 hydroxymethyl furfural and its derivative are measured based on positive and reversed-phase high performance liquid chromatography combination,
It is characterized by comprising the following steps：
(1) highly effective liquid phase chromatographic system and chromatographic condition：
Normal-phase chromatography：U.S. Agilent1260 type high performance liquid chromatographs, using Bio-Rad Aminex HPX-87H
(300mm × 7.8mm) forward chromatographic column band guard column (30mm × 4.6mm), 55 DEG C of column temperature, automatic loading, 10.0 μ of sampling volume
L, using 5mmol/L H2SO4 as mobile phase, flow velocity 0.6mL/min；Detector is differential refraction detector, and detector temperature is
35 DEG C, the content of each component is measured using chromatographic peak area integration method.
Reverse-phase chromatography：U.S. Agilent1100 type high performance liquid chromatographs, using Zorbax Eclipse XDS-C18
(300mm × 4.6mm) reverse-phase chromatographic column band guard column (3.0mm × 4.6mm), 30 DEG C of column temperature, automatic loading, sampling volume 10.0
μL；Using 100% acetonitrile and 1.5% acetic acid as mobile phase, flow velocity 0.6mL/min, in 0min~20min acetonitrile concentration from
5% rises to 50%, and acetate concentration is down to 50% from 95%；Detector is variable-wavelenght detector (VWD), and Detection wavelength is
275nm measures the content of each component using chromatographic peak area integration method.
(2) chromatograph joint used measurement 5 hydroxymethyl furfural and its derivative：
By 2,5-FDM, 5 hydroxymethyl furfural, 2,5-furandaldehyde, 5- methylol -2- furancarboxylic acids, 5- formyls
Base -2- furancarboxylic acids, 2,5-furandicarboxylic acid standard items are configured to the standard solution of 0.4-1.0g/L.
On normal-phase chromatography, the appearance time weight of the appearance time and 5- formoxyl -2- furancarboxylic acids of 5- methylol -2- furancarboxylic acids
It is folded, cause two kinds of substances cannot achieve and is kept completely separate；On reverse-phase chromatography, appearance time and the 5- hydroxyl first of 2,5-FDM
The appearance time of base furfural is overlapped, and is caused two kinds of substances cannot achieve and is kept completely separate.Pass through positive and reversed-phase high performance liquid chromatography
Joint-detection finally realize to 5 hydroxymethyl furfural, 2,5-FDM, 2,5-furandaldehyde, 5- methylol -2- chaffs
The fast, accurately separation and quantitative determination of acid, 5- formoxyl -2- furancarboxylic acids and 2,5-furandicarboxylic acid.
2, it is according to claim 1 based on positive and reversed-phase high performance liquid chromatography combination measure 5 hydroxymethyl furfural and
The method of its derivative, which is characterized in that in step (2), the standard working strategy of separated substance is on normal-phase chromatography：
2,5-FDM A=806393.489 × c1-4722.254
2,5-furandicarboxylic acid A=186241.361 × c2-51.022341
5 hydroxymethyl furfural A=187580.332 × c3-42.131853
2,5-furandaldehyde A=199336.41 × c4-34.177678
In above-mentioned standard working strategy, A indicates that chromatographic peak area (nRIU × s), c indicate concentration of component (g/L)
The standard working strategy of separated substance is on reverse-phase chromatography：
5- formoxyl -2- furancarboxylic acids A=53901.787 × c5-1228.0761
5- methylol -2- furancarboxylic acids A=18727.7606 × c6-376.46279
In above-mentioned standard working strategy, A indicates that chromatographic peak area (mAu × s), c indicate concentration of component (g/L).
High performance liquid chromatography is a chromatographic important branch, and separation principle is being flowed according to the component detached
Mutually and stationary phase in different solubility and detach.Separation process is a partition equilibrium process.High performance liquid chromatography has analysis
Speed is fast, high resolution, high sensitivity, pillar can Reusability and sample size it is few, the advantages of being easy recycling.And according to stationary phase
It is different with the polarity of mobile phase to be divided into normal phase chromatography (NPC) and RP chromatography (RPC).Normal phase chromatography is usually used in detaching
Middle polarity and the stronger compound of polarity；RP chromatography is suitable for detaching the weaker compound of non-polar and polar.5- hydroxyls
The furan nucleus carbon skeleton of methyl furfural and its five kinds of derivatives, the position of branch, molecule of functional group size, material parameters are all very
It is close, in single high performance liquid chromatography detection between substance there is mutual interference cause to be difficult to efficiently separate identification or
Quantitative determination.
The side that 5 hydroxymethyl furfural and its derivative are measured based on positive and reversed-phase high performance liquid chromatography combination of the present invention
Method is exactly effectively combined according to above-mentioned positive and reversed-phase high performance liquid chromatography feature and the property of 6 kinds of substances itself, by 2,5-
Furyl dimethyl carbinol, 5 hydroxymethyl furfural, 2,5-furandaldehyde, 5- methylol -2- furancarboxylic acids, 5- formoxyl -2- furancarboxylic acids and 2,5-
After the standard items of furandicarboxylic acid are configured to standard solution, in given positive and rp-hplc system and chromatostrip
The standard working strategy of 6 kinds of substances is measured under part.Then it will be waited for using 5 hydroxymethyl furfural as what substrate progress catalyzed conversion obtained
Sample is measured under same positive and rp-hplc system and chromatographic condition, and according to standard work side
Journey conversion obtains the quantitative measure of sample to be tested.
Advantageous effect：Being combined based on positive and reversed-phase high performance liquid chromatography for the present invention is measured 5 hydroxymethyl furfural and its spread out
Biology method, compared with existing assay method, have the advantage that including：It is combined based on positive and reversed-phase high performance liquid chromatography
The method that accurate quantification measures 5 hydroxymethyl furfural and its derivative, using Bio-Rad Aminex HPX-87H (300mm ×
7.8mm) forward chromatographic column, by sulfuric acid isocratic elution, using differential refraction detector, temperature measures under conditions of being 35 DEG C inhales
Light value, quantitative determination 2,5-FDM, 2,5-furandicarboxylic acid, 5 hydroxymethyl furfural and 2,5-furandaldehyde；Using
Zorbax Eclipse XDS-C18 (300mm × 4.6mm) reverse-phase chromatographic column is made by acetonitrile and acetic acid binary gradient elutes
With variable-wavelenght detector, light absorption value is measured at ultraviolet wavelength 275nm, quantitative determines 5- methylol -2- furancarboxylic acids and 5- formyls
Base -2- furancarboxylic acids.A kind of quick, accurate synchronization analysis and the method for quantitatively detecting 5 hydroxymethyl furfural and its derivative are provided,
It is particularly suitable for analysis and detection that biomass biorefinery synthesizes above-mentioned furans platform chemicals.
Description of the drawings
Fig. 1 is 2,5-FDM, 2,5-furandicarboxylic acid, 5 hydroxymethyl furfural and 2,5-furandaldehyde standard sample
The collection of illustrative plates that the Normal-phase HPLC of product measures.Abscissa indicates that the retention time RT (min) of component, ordinate are indicated in figure
The obtained signal of Composition distribution (nRIU).
Fig. 2 is the rp-hplc of 5- formoxyl -2- furancarboxylic acids and 5- methylol -2- furancarboxylic acid standard samples
Collection of illustrative plates.Abscissa indicates that the retention time RT (min) of various components, ordinate indicate that variable-wavelenght detector is obtained in figure
Signal (mAu).
Specific implementation mode
With reference to specific embodiment, the present invention will be further explained.
1 Normal-phase HPLC system of embodiment and chromatographic condition
Normal-phase HPLC system：1260 liquid chromatographic systems of German Agilent, configuration vacuum degassing machine show difference
B.01.01 the runs software of refraction detector (RID) and system autosampler, chromatographic system uses Agilent chem workstation
Version, chromatographic condition：Bio-Rad Aminex HPX-87H (300mm × 7.8mm) chromatographic column band guard column (30mm ×
4.6mm), 55 DEG C of column temperature, 10.0 μ L of sampling volume；
Mobile phase condition：Gradient elution, flow velocity 0.6ml/min are carried out by mobile phase of 5mmol/L H2SO4；
Signal detection：Detector is differential refraction detector, and detector temperature is 35 DEG C, using chromatographic peak area integration method
The content of measurement 2,5-FDM, 2,5-furandicarboxylic acid, 5 hydroxymethyl furfural and 2,5-furandaldehyde.
2 rp-hplc system of embodiment and chromatographic condition
Rp-hplc system：1100 liquid chromatographic systems of German Agilent, configuration vacuum degassing machine can be changed
A.10.XX the runs software of wavelength detecting (VWD) and system autosampler, chromatographic system uses Agilent chem workstation
Version, chromatographic condition：Zorbax Eclipse XDS-C18 (300mm × 4.6mm) reverse-phase chromatographic column band guard column (3.0mm ×
4.6mm), 30 DEG C of column temperature, 10.0 μ L of sampling volume；
Elution requirement：Variable Composition gradient elution is carried out as mobile phase using 1.5% acetic acid and 100% acetonitrile, flow velocity is
0.6mL/min.Acetonitrile concentration rises to 50% from 5% in 0min~20min, and acetate concentration is down to 50% from 95%；20min~
Acetonitrile concentration is rapidly decreased to 5% from 50% in 20.1min, and acetate concentration is rapidly increased to 95% from 50%；20.1min~25min
It is interior that pillar is regenerated and balanced with the acetonitrile of 5% concentration and the acetic acid of 95% concentration.
Signal detection：Variable-wavelenght detector (VWD), Detection wavelength 275nm are measured using chromatographic peak area integration method
The content of 5- formoxyl -2- furancarboxylic acids, 5- methylol -2- furancarboxylic acids.
The measurement of 36 kinds of standard of physical working strategies of embodiment
Measure the standard working strategy of 6 kinds of substances：2,5-FDM is purchased from Shanghai Mike woods company；5- methylol chaffs
Aldehyde, 2,5-furandicarboxylic acid are purchased from Shanghai Aladdin company；5- methylol -2- furancarboxylic acids, 2,5-furandaldehyde, 5- formoxyls -
2- furancarboxylic acids are purchased from TCI companies of Japan, the standard solution of 0.4~1.0g/L are configured to, using above-mentioned positive and reversed phase high efficiency
Liquid chromatographic system and chromatographic condition measure the standard working strategy of 6 kinds of substances, and measurement result is as shown in Figure 1, Figure 2 and table 1,2 institute of table
Show.Each chromatographic peak is respectively in figure：1.2,5- furyl dimethyl carbinols；2.2,5- furandicarboxylic acids；3.5- hydroxymethylfurfural；4.2,5-
Furans dicarbaldehyde；5.5- formoxyl -2- furancarboxylic acids；6.5- methylol -2- furancarboxylic acids.
15 hydroxymethyl furfural of table and its derivative Normal-phase HPLC standard working strategy measure
Chromatographic peak retention time RT (min) on Normal-phase HPLC：2,5-FDM 13.703；2,5- furans
Dioctyl phthalate 16.151；5 hydroxymethyl furfural 31.995；2,5-furandaldehyde 39.114.
The standard working strategy of separated substance is on normal-phase chromatography：
2,5-FDM A=806393.489 × c1-4722.254, coefficient R 2=0.99807
2,5-furandicarboxylic acid A=186241.361 × c2-51.022341, coefficient R 2=1.00000
5 hydroxymethyl furfural A=187580.332 × c3-42.131853, coefficient R 2=0.99999
2,5-furandaldehyde A=199336.41 × c4-34.177678, coefficient R 2=1.00000
In above-mentioned standard working strategy, A indicates that chromatographic peak area (nRIU × s), c indicate concentration of component (g/L)
25 hydroxymethyl furfural of table and its derivative reversed-phase high performance liquid chromatography standard working strategy measure
Chromatographic peak retention time RT (min) in reversed-phase high performance liquid chromatography：5- formoxyl -2- furancarboxylic acids 6.505；5- hydroxyl first
Base -2- furancarboxylic acids 6.837；
Standard working strategy：
5- formoxyl -2- furancarboxylic acid A=53901.787 × c2-1228.0761, coefficient R 2=0.99644
5- methylol -2- furancarboxylic acid A=18727.7606 × c3-376.46279, coefficient R 2=0.99782
In standard working strategy, A indicates that chromatographic peak area (mAu × s), c indicate concentration of component (g/L)
The constituent analysis of 4 actual sample of embodiment and measurement
Practical converted product is obtained after microorganism catalysis reacts using 5 hydroxymethyl furfural as substrate, zymotic fluid is existed
Sample liquid obtained with 0.2 μm of micro-filtrate membrane filtration supernatant after centrifuging 5min under the conditions of 10000rpm, then be transferred to chromatograph it is automatic on
Sample bottle carries out chromatographic determination.
Analysis and quantitative determination 5 hydroxymethyl furfural and its derivative sample：Use external standard method with the mark of above-mentioned 6 kinds of components
The content of each component during quasi- working strategy accurate quantification is measured using 5 hydroxymethyl furfural as the converted product that substrate obtains.
Using embodiment 1, the method for embodiment 2 and embodiment 3 can realize 6 kinds of efficient quick separatings of component in sample and
Accurate quantitative analysis detects, and using 5 hydroxymethyl furfural as substrate progress microorganism catalysis conversion, after 12h, substrate 5 hydroxymethyl furfural turns
Rate is 80%, and each converted product yield is respectively：2,5-furandicarboxylic acid 10%, 5- formoxyl -2- furancarboxylic acids 5%, 5- hydroxyl first
Base -2- furancarboxylic acids 60%, 2,5-furandaldehyde 3%, 2,5-FDM 22%.
Claims (2)
1. a kind of method measuring 5 hydroxymethyl furfural and its derivative based on positive and reversed-phase high performance liquid chromatography combination, special
Sign is, includes the following steps：
(1) highly effective liquid phase chromatographic system and chromatographic condition：
Normal-phase chromatography：U.S. Agilent1260 type high performance liquid chromatographs, using Bio-Rad Aminex HPX-87H 300mm
× 7.8mm forward chromatographic column band guard column 30mm × 4.6mm, 55 DEG C of column temperature, automatic loading, 10.0 μ L of sampling volume, with
5mmol/L H2SO4For mobile phase, flow velocity 0.6mL/min；Detector is differential refraction detector, and detector temperature is 35 DEG C,
The content of each component is measured using chromatographic peak area integration method；
Reverse-phase chromatography：U.S. Agilent1100 type high performance liquid chromatographs, using Zorbax Eclipse XDS-C18 300mm
× 4.6mm reverse-phase chromatographic column band guard column 3.0mm × 4.6mm, 30 DEG C of column temperature, automatic loading, 10.0 μ L of sampling volume；With
100% acetonitrile and 1.5% acetic acid are mobile phase, and flow velocity 0.6mL/min, acetonitrile concentration rises to from 5% in 0min~20min
50%, acetate concentration is down to 50% from 95%；Detector is variable-wavelenght detector (VWD), and Detection wavelength 275nm is used
Chromatographic peak area integration method measures the content of each component；
(2) chromatograph joint used measurement 5 hydroxymethyl furfural and its derivative：
By 2,5- furyl dimethyl carbinols, 5 hydroxymethyl furfural, 2,5- furans dicarbaldehyde, 5- methylol -2- furancarboxylic acids, 5- formoxyls -2-
Furancarboxylic acid and 2,5- furandicarboxylic acid standard items are configured to the standard solution of 0.4-1.0g/L；
On normal-phase chromatography, the appearance time of 5- methylol -2- furancarboxylic acids is Chong Die with the appearance time of 5- formoxyl -2- furancarboxylic acids, leads
It causes two kinds of substances cannot achieve to be kept completely separate；On reverse-phase chromatography, appearance time and the 5- methylol chaffs of 2,5-FDM
The appearance time of aldehyde is overlapped, and is caused two kinds of substances cannot achieve and is kept completely separate；
It is final to realize to 5 hydroxymethyl furfural, 2,5-FDM, 2 by the combination of positive and reversed-phase high performance liquid chromatography,
The fast, accurately separation of 5- furans dicarbaldehyde, 5- methylol -2- furancarboxylic acid, 5- formoxyl -2- furancarboxylic acids and 2,5- furandicarboxylic acids
And quantitative determination.
2. according to claim 1 measure 5 hydroxymethyl furfural based on positive and reversed-phase high performance liquid chromatography combination and its spread out
The method of biology, which is characterized in that in step (2), the standard working strategy of separated substance is on normal-phase chromatography：
2,5- furyl dimethyl carbinols A=806393.489 × c1-4722.254
2,5- furandicarboxylic acids A=186241.361 × c2-51.022341
5 hydroxymethyl furfural A=187580.332 × c3-42.131853
2,5- furans dicarbaldehydes A=199336.41 × c4-34.177678
In above-mentioned standard working strategy, A indicates that chromatographic peak area nRIU × s, c indicate concentration of component g/L
The standard working strategy of separated substance is on reverse-phase chromatography：
5- formoxyl -2- furancarboxylic acids A=53901.787 × c5-1228.0761
5- methylol -2- furancarboxylic acids A=18727.7606 × c6-376.46279
In above-mentioned standard working strategy, A indicates that chromatographic peak area mAu × s, c indicate concentration of component g/L.
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Non-Patent Citations (4)
| Title |
|---|
| Enzyme cascade reactions: synthesis of furandicarboxylic acid (FDCA) and carboxylic acids using oxidases in tandem;Shane M. McKenna et al.;《Green Chemistry》;20150421;第17卷;第3271-3275页 * |
| Selective aerobic oxidation of the biomass-derived precursor 5-hydroxymethylfurfural to 2,5-furandicarboxylic acid under mild conditions over a magnetic palladium nanocatalyst;Zehui Zhang et al.;《Green Chemistry》;20141211;第17卷;第1308-1317页 * |
| Selective oxidation of 5-hydroxymethylfurfural with H O catalyzed by a molybdenum complex;Shuang Li et al.;《Green Chemistry》;20151209;第18卷;第2122-2128页 * |
| 高效液相色谱法测定变压器油中糠醛及相关呋喃类化合物;徐燕锋;《上海计量测试》;20121231;第18-21页 * |
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