CN108732297A - The detection method and device of gluconic acid sodium content in a kind of zymotic fluid - Google Patents
The detection method and device of gluconic acid sodium content in a kind of zymotic fluid Download PDFInfo
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- CN108732297A CN108732297A CN201810811377.3A CN201810811377A CN108732297A CN 108732297 A CN108732297 A CN 108732297A CN 201810811377 A CN201810811377 A CN 201810811377A CN 108732297 A CN108732297 A CN 108732297A
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- bottle
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- glacial acetic
- acetic acid
- titration
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- UPMFZISCCZSDND-JJKGCWMISA-M sodium gluconate Chemical compound [Na+].OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C([O-])=O UPMFZISCCZSDND-JJKGCWMISA-M 0.000 title claims abstract description 91
- 239000012530 fluid Substances 0.000 title claims abstract description 80
- 238000001514 detection method Methods 0.000 title claims abstract description 64
- 239000000176 sodium gluconate Substances 0.000 claims abstract description 66
- 235000012207 sodium gluconate Nutrition 0.000 claims abstract description 64
- 229940005574 sodium gluconate Drugs 0.000 claims abstract description 64
- AEQDJSLRWYMAQI-UHFFFAOYSA-N 2,3,9,10-tetramethoxy-6,8,13,13a-tetrahydro-5H-isoquinolino[2,1-b]isoquinoline Chemical compound C1CN2CC(C(=C(OC)C=C3)OC)=C3CC2C2=C1C=C(OC)C(OC)=C2 AEQDJSLRWYMAQI-UHFFFAOYSA-N 0.000 claims abstract description 63
- 239000000243 solution Substances 0.000 claims description 133
- 238000004448 titration Methods 0.000 claims description 131
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 103
- 229960000583 acetic acid Drugs 0.000 claims description 97
- 239000012362 glacial acetic acid Substances 0.000 claims description 97
- SMUQFGGVLNAIOZ-UHFFFAOYSA-N quinaldine Chemical compound C1=CC=CC2=NC(C)=CC=C21 SMUQFGGVLNAIOZ-UHFFFAOYSA-N 0.000 claims description 87
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 77
- OKKJLVBELUTLKV-UHFFFAOYSA-N methanol Substances OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 55
- 239000007788 liquid Substances 0.000 claims description 44
- 239000012086 standard solution Substances 0.000 claims description 44
- 230000002572 peristaltic effect Effects 0.000 claims description 35
- 239000002699 waste material Substances 0.000 claims description 28
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 20
- 238000006243 chemical reaction Methods 0.000 claims description 16
- VLTRZXGMWDSKGL-UHFFFAOYSA-N perchloric acid Chemical compound OCl(=O)(=O)=O VLTRZXGMWDSKGL-UHFFFAOYSA-N 0.000 claims description 10
- 239000012488 sample solution Substances 0.000 claims description 9
- 241001536374 Indicator indicator Species 0.000 claims description 8
- 239000007864 aqueous solution Substances 0.000 claims description 6
- 238000004891 communication Methods 0.000 claims description 6
- 238000002360 preparation method Methods 0.000 claims description 4
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 claims 4
- SMWDFEZZVXVKRB-UHFFFAOYSA-N Quinoline Chemical compound N1=CC=CC2=CC=CC=C21 SMWDFEZZVXVKRB-UHFFFAOYSA-N 0.000 claims 4
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 claims 2
- 238000000034 method Methods 0.000 abstract description 77
- 238000005259 measurement Methods 0.000 abstract description 31
- 230000008569 process Effects 0.000 abstract description 27
- 238000000855 fermentation Methods 0.000 abstract description 23
- 230000004151 fermentation Effects 0.000 abstract description 23
- 238000002474 experimental method Methods 0.000 abstract description 17
- 241000228245 Aspergillus niger Species 0.000 abstract description 16
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 abstract description 12
- 239000008103 glucose Substances 0.000 abstract description 12
- 238000004519 manufacturing process Methods 0.000 abstract description 10
- 239000000203 mixture Substances 0.000 abstract description 6
- 238000012544 monitoring process Methods 0.000 abstract description 6
- YRKCREAYFQTBPV-UHFFFAOYSA-N acetylacetone Chemical compound CC(=O)CC(C)=O YRKCREAYFQTBPV-UHFFFAOYSA-N 0.000 description 26
- 239000000523 sample Substances 0.000 description 25
- 239000011734 sodium Substances 0.000 description 23
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 20
- 229910052708 sodium Inorganic materials 0.000 description 20
- 238000011161 development Methods 0.000 description 17
- RGHNJXZEOKUKBD-UHFFFAOYSA-N D-gluconic acid Natural products OCC(O)C(O)C(O)C(O)C(O)=O RGHNJXZEOKUKBD-UHFFFAOYSA-N 0.000 description 13
- RGHNJXZEOKUKBD-SQOUGZDYSA-N Gluconic acid Natural products OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C(O)=O RGHNJXZEOKUKBD-SQOUGZDYSA-N 0.000 description 13
- 239000000174 gluconic acid Substances 0.000 description 13
- 235000012208 gluconic acid Nutrition 0.000 description 13
- 238000004140 cleaning Methods 0.000 description 11
- 238000011084 recovery Methods 0.000 description 9
- 238000007789 sealing Methods 0.000 description 9
- 241000219095 Vitis Species 0.000 description 8
- 235000009754 Vitis X bourquina Nutrition 0.000 description 8
- 235000012333 Vitis X labruscana Nutrition 0.000 description 8
- 235000014787 Vitis vinifera Nutrition 0.000 description 8
- 239000003153 chemical reaction reagent Substances 0.000 description 8
- 238000006386 neutralization reaction Methods 0.000 description 8
- 239000006228 supernatant Substances 0.000 description 8
- 150000001875 compounds Chemical class 0.000 description 7
- 238000003760 magnetic stirring Methods 0.000 description 7
- 239000002253 acid Substances 0.000 description 6
- 239000012295 chemical reaction liquid Substances 0.000 description 6
- NAOLWIGVYRIGTP-UHFFFAOYSA-N 1,3,5-trihydroxyanthracene-9,10-dione Chemical compound C1=CC(O)=C2C(=O)C3=CC(O)=CC(O)=C3C(=O)C2=C1 NAOLWIGVYRIGTP-UHFFFAOYSA-N 0.000 description 5
- 238000003556 assay Methods 0.000 description 5
- 239000008367 deionised water Substances 0.000 description 5
- 229910021641 deionized water Inorganic materials 0.000 description 5
- 238000004128 high performance liquid chromatography Methods 0.000 description 5
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 4
- 238000002835 absorbance Methods 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- AVXURJPOCDRRFD-UHFFFAOYSA-N Hydroxylamine Chemical compound ON AVXURJPOCDRRFD-UHFFFAOYSA-N 0.000 description 3
- 239000002585 base Substances 0.000 description 3
- 238000009835 boiling Methods 0.000 description 3
- IDAGXRIGDWCIET-SDFKWCIISA-L disodium;(2s,3s,4s,5r)-2,3,4,5-tetrahydroxyhexanedioate Chemical compound [Na+].[Na+].[O-]C(=O)[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C([O-])=O IDAGXRIGDWCIET-SDFKWCIISA-L 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 description 3
- 239000012071 phase Substances 0.000 description 3
- 230000010076 replication Effects 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- NEAQRZUHTPSBBM-UHFFFAOYSA-N 2-hydroxy-3,3-dimethyl-7-nitro-4h-isoquinolin-1-one Chemical compound C1=C([N+]([O-])=O)C=C2C(=O)N(O)C(C)(C)CC2=C1 NEAQRZUHTPSBBM-UHFFFAOYSA-N 0.000 description 2
- 239000007836 KH2PO4 Substances 0.000 description 2
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 230000001276 controlling effect Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 230000005611 electricity Effects 0.000 description 2
- 229910052564 epsomite Inorganic materials 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 150000002596 lactones Chemical class 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 2
- 238000005457 optimization Methods 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 150000003839 salts Chemical group 0.000 description 2
- 239000003381 stabilizer Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000012795 verification Methods 0.000 description 2
- 241000228212 Aspergillus Species 0.000 description 1
- 101000765308 Aspergillus niger N-(5'-phosphoribosyl)anthranilate isomerase Proteins 0.000 description 1
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- VTLYFUHAOXGGBS-UHFFFAOYSA-N Fe3+ Chemical compound [Fe+3] VTLYFUHAOXGGBS-UHFFFAOYSA-N 0.000 description 1
- 235000003239 Guizotia abyssinica Nutrition 0.000 description 1
- WTDHULULXKLSOZ-UHFFFAOYSA-N Hydroxylamine hydrochloride Chemical class Cl.ON WTDHULULXKLSOZ-UHFFFAOYSA-N 0.000 description 1
- 206010021037 Hyponatraemic syndrome Diseases 0.000 description 1
- 229910021578 Iron(III) chloride Inorganic materials 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 244000061456 Solanum tuberosum Species 0.000 description 1
- 235000002595 Solanum tuberosum Nutrition 0.000 description 1
- 229910000831 Steel Inorganic materials 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- 230000001476 alcoholic effect Effects 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- VTYYLEPIZMXCLO-UHFFFAOYSA-L calcium carbonate Substances [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000003638 chemical reducing agent Substances 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 239000012459 cleaning agent Substances 0.000 description 1
- 238000010835 comparative analysis Methods 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000002242 deionisation method Methods 0.000 description 1
- 238000009795 derivation Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005868 electrolysis reaction Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 230000003760 hair shine Effects 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-M hydroxide Chemical compound [OH-] XLYOFNOQVPJJNP-UHFFFAOYSA-M 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 210000001161 mammalian embryo Anatomy 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000011017 operating method Methods 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- KHIWWQKSHDUIBK-UHFFFAOYSA-N periodic acid Chemical compound OI(=O)(=O)=O KHIWWQKSHDUIBK-UHFFFAOYSA-N 0.000 description 1
- 230000005622 photoelectricity Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 239000012898 sample dilution Substances 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 238000011218 seed culture Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 239000010959 steel Substances 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
- G01N31/00—Investigating or analysing non-biological materials by the use of the chemical methods specified in the subgroup; Apparatus specially adapted for such methods
- G01N31/16—Investigating or analysing non-biological materials by the use of the chemical methods specified in the subgroup; Apparatus specially adapted for such methods using titration
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/38—Diluting, dispersing or mixing samples
-
- 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
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/38—Diluting, dispersing or mixing samples
- G01N2001/386—Other diluting or mixing processes
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Analytical Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Plasma & Fusion (AREA)
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
Abstract
The present invention provides a kind of detection method of gluconic acid sodium content in zymotic fluid, method provided by the invention can be with gluconic acid sodium content in Accurate Determining zymotic fluid.Methodology experiment shows, the method range of linearity provided by the invention is wide, precision, repeatability, stability and accuracy are good, other compositions do not influence measurement result in the zymotic fluids such as glucose, and method continuous mode provided by the invention is simple, is quick, the accurate method of gluconic acid sodium content during measuring fermentation of Aspergillus niger.In addition, the present invention provides a kind of detection devices of gluconic acid sodium content in zymotic fluid, use detection device provided by the invention can be with gluconic acid sodium content in Accurate Determining zymotic fluid, sample measurement can be completed in 3min, and device component easy to operate, simple in structure, without using costliness provided by the invention, it can realize the monitoring of fermentation of Aspergillus niger production sodium gluconate process.
Description
Technical field
The present invention relates to compound test technical fields, and in particular to the detection of gluconic acid sodium content in a kind of zymotic fluid
Method and apparatus.
Background technology
Sodium gluconate (C6H11O7Na) it is a kind of multi-hydroxy carboxy acid's sodium, appearance is white or light yellow crystalline powder.
Sodium gluconate is soluble easily in water, moderate notoxic, has saline taste, has excellent chelating ability, therefore be widely used in food
Product, building, chemical industry, medicine and other fields.In food service industry, sodium gluconate is a kind of important food additives, can conduct
PH regulating stabilizers, edible salt substituting agent, flavor improver etc..In building trade, it is gentle that sodium gluconate is used as water-reducing agent
Solidifying agent, can enhance the intensity of concrete and extend setting time.In chemical industry, sodium gluconate is added in cleaning agent
Very ideal cleaning performance can be reached, quilt should be in the cleaning of vial and steel embryo.In pharmaceuticals industry, sodium gluconate can be with
As water quality stabilizer and low sodium syndrome medicament.Therefore, the market demand of sodium gluconate is huge.
The production method of sodium gluconate mainly has fermentation method, electrolysis and catalytic oxidation.Wherein fermentation method is a kind of
Effective production method, mainly using aspergillus niger as production strain.It is monitored in real time in zymotic fluid during fermentation of Aspergillus niger
Gluconic acid na concn is critically important for predicting fermentation of Aspergillus niger process and fermentation process technique optimization being instructed to play the role of.Cause
This, find gluconic acid sodium content in effective zymotic fluid detection method value in ferment of sodium gluconate industry will produce it is positive
Facilitation.
The detection method of sodium gluconate have non-aqueous titration, acetylacetone,2,4-pentanedione development process, azanol-ferric trichloride development process,
High performance liquid chromatography etc..Wherein, the measurement result error of acetylacetone,2,4-pentanedione development process is larger, azanol-ferric trichloride development process
Determination step is excessively complicated, is not easy to realize the real-time monitoring of fermentation process.There are mobile phases to chromatography for high performance liquid chromatography
Column is affected, high performance liquid chromatograph costly the problems such as.Non-aqueous titration measurement result is accurate, easy to operate, is not required to
Expensive instrument is wanted, is sodium gluconate assay in People's Republic of China (PRC) light industry standard QB/T 4484-2013
Standard method.But non-aqueous titration is glacial acetic acid reaction system, can only measure the gluconic acid sodium content in solid powder, no
The content of sodium gluconate in aqueous solution can be directly measured, therefore may not apply to sodium gluconate yield in fermentation production process
Detection.
Invention content
The purpose of the present invention is to provide the detection methods and device of gluconic acid sodium content in a kind of zymotic fluid, using this
Inventing the method that provides can be and easy to operate with gluconic acid sodium content in Accurate Determining zymotic fluid.
In order to achieve the above-mentioned object of the invention, the present invention provides following technical scheme：
The present invention provides a kind of detection methods of gluconic acid sodium content in zymotic fluid, include the following steps：
Zymotic fluid is mixed with glacial acetic acid, obtains sample solution to be titrated；
Using quinaldine red-methanol solution as indicator, using perchloric acid-glacial acetic acid solution to the sample solution to be titrated
It is titrated, until terminating to titrate when indicator Indicator Reaction terminal, according to scheduled regression equation and required titration volumes, is obtained
The content of sodium gluconate in zymotic fluid, the regression equation are gluconic acid in titration volumes and gluconic acid sodium standard solution
Regression equation between sodium content, the gluconic acid sodium standard solution are the aqueous solution of sodium gluconate.
Preferably, the volume ratio of the zymotic fluid and glacial acetic acid is (0.001~1)：(5~15).
Preferably, the mass concentration of the quinaldine red-methanol solution is 0.01~1%；Quinaldine red-the methanol is molten
The volume ratio of liquid and the zymotic fluid is (0.01~0.3)：(0.001~1).
Preferably, in the perchloric acid-glacial acetic acid solution perchloric acid a concentration of 0.01~0.5mol/L.
Preferably, the preparation method of the regression equation, includes the following steps：
Sodium gluconate is mixed with water, obtains the gluconic acid sodium standard solution of series concentration；
The gluconic acid sodium standard solution is mixed with glacial acetic acid, obtains gluconic acid sodium standard solution to be titrated；
Using quinaldine red-methanol solution as indicator, using perchloric acid-glacial acetic acid solution to the gluconic acid to be titrated
Sodium standard solution is titrated, until terminating to titrate when indicator Indicator Reaction terminal, titration volumes is recorded, with sodium gluconate mark
Quasi- glucose in solutions acid sodium content is abscissa, and titration volumes are ordinate, draws the standard curve of gluconic acid sodium content,
The regression equation of gluconic acid sodium content is calculated.
Preferably, a concentration of 0~2.4mol/L of the gluconic acid sodium standard solution.
The present invention provides a kind of detection devices of gluconic acid sodium content in zymotic fluid, including control-display system, light
Electricity detecting system, quinaldine red-methanol solution bottle, glacial acetic acid bottle and perchloric acid-glacial acetic acid solution bottle, wherein the control-is aobvious
Show and be provided with control switch, external display screen and control system in system, the control system respectively with the display screen, light
Electricity detecting system, quinaldine red-methanol solution bottle, glacial acetic acid bottle and perchloric acid-glacial acetic acid solution bottle communication connection, the control
System is to the display screen, photodetector system, quinaldine red-methanol solution bottle, glacial acetic acid bottle and perchloric acid-glacial acetic acid solution
The working condition of bottle is controlled and is adjusted；The photodetector system includes light source, photoelectric detection system and titration cell, described
Titration cell is connected to the quinaldine red-methanol solution bottle, glacial acetic acid bottle and perchloric acid-glacial acetic acid solution bottle respectively, the drop
Determine pond to be arranged between the light source and photoelectric detection system, the photoelectric detection system is used to detect the change of color in titration cell
Change.
Preferably, the titration cell and the quinaldine red-methanol solution bottle, glacial acetic acid bottle and perchloric acid-glacial acetic acid are molten
Peristaltic pump is respectively arranged between liquid bottle, the control system is communicated to connect with the peristaltic pump, and it is right that the control system passes through
The working condition of the peristaltic pump is controlled and is adjusted realization to the quinaldine red-methanol solution bottle, glacial acetic acid bottle and height
The control and adjusting of chloric acid-glacial acetic acid solution bottle working condition.
Preferably, the detection device further includes waste liquid bottle and sodium hydroxide solution bottle, the waste liquid bottle respectively with it is described
Titration cell is connected to sodium hydroxide solution bottle, the control system respectively with the waste liquid bottle and sodium hydroxide solution bottle communication link
It connects, the control system is controlled and adjusted to the working condition of the waste liquid bottle and sodium hydroxide solution bottle.
Preferably, it is arranged between the waste liquid bottle and the titration cell, between the waste liquid bottle and sodium hydroxide solution bottle
There are peristaltic pump, the control system to be communicated to connect with the peristaltic pump, the control system passes through the work to the peristaltic pump
State is controlled and is adjusted the control and adjusting realized to the titration cell and sodium hydroxide solution bottle working condition.
The present invention provides a kind of detection method of gluconic acid sodium content in zymotic fluid, detection methods provided by the invention
It has the advantages that：
1. method provided by the invention can directly to zymotic fluid, this Aquo System carries out aqueous titration, and then realizes hair
The measurement of gluconic acid sodium content in zymotic fluid.
2. being highly soluble in the characteristic of water based on sodium gluconate, the Portugal that aqueous titration provided by the invention not only avoids
Grape sodium saccharate difficulty is dissolved in the operating difficulties of glacial acetic acid, and range of linearity when greatly having widened gluconic acid sodium determination,
For 0~2.4mol/L (i.e. 0~523.5g/L, saturated concentration 529.1g/L when close to 15 DEG C of sodium gluconate, including entire black
Sodium gluconate concentration range in aspergillus fermentation process zymotic fluid), make the more non-water droplet of highest detection concentration of sodium gluconate
Determine method and improve 200 times or more (maximum linearity range of the non-aqueous titration of document report is 0~1.8g/L), compared with other glucose
Sour sodium content assay method at least improves 100~400 times.
3. zymotic fluid is as sample to be tested during fermentation of Aspergillus niger, without being diluted, after only filtering or centrifuge
It is directly measured, greatly improves the simplicity of method, avoid error caused by sample dilution.
4. method operating process provided by the invention is simple, expensive instrument and equipment is not needed, sample can be completed in 3min
It measures.Methodology experiment shows the method rate of recovery provided by the invention up to 99% or more, precision, repeatability and stability
Well, other compositions do not influence measurement result in the zymotic fluids such as glucose.Therefore, method provided by the invention can be fast
Gluconic acid sodium content in speed, Accurate Determining zymotic fluid, may be implemented using method provided by the invention to fermentation of Aspergillus niger mistake
The real-time monitoring of gluconic acid na concn in journey, and then instruct and optimize fermentation process.
The present invention provides a kind of detection device of gluconic acid sodium content in zymotic fluid, detection devices provided by the invention
It has the advantages that：
1. device provided by the invention can realize titration process automatically, and be titrated according to photocurrent variations rate automatic decision
Terminal, avoids titration by hand and naked eyes judge the error of titration end-point.
2. methodology experiment shows the method for measuring gluconic acid sodium content in zymotic fluid using device provided by the invention
The range of linearity is wide (0~2.4mol/L), and for the rate of recovery up to 99% or more, precision, stability and accuracy are good, can in 3min
To complete sample measurement, and device provided by the invention is easy to operate, simple in structure, without using expensive component.Using this
Invent the detection device that provides can easily and fast, gluconic acid sodium content in Accurate Determining zymotic fluid, utilize the present invention to provide
Device may be implemented the real-time monitoring to gluconic acid na concn during fermentation of Aspergillus niger, and then instruct and optimization was fermented
Journey.
Description of the drawings
Fig. 1 be zymotic fluid provided by the invention in gluconic acid sodium content detection device structural schematic diagram, 1,2,3,
4,5- peristaltic pumps；6- waste liquid bottles；7- quinaldine reds-methanol solution bottle；8- glacial acetic acid bottles；9- perchloric acid-glacial acetic acid solution bottle；
10-NaOH solution bottles；11- sealing-plugs；12- light sources；13- photoelectric detection systems；14- rotors；15- magnetic stirring apparatus；16- drops
Determine pond；17- display screens；18- is switched；19- control systems；20,21,22,23- sealing rings；24,25,26,27- check valves；
Fig. 2 is the standard curve of non-aqueous titration in embodiment 1；
Fig. 3 is the standard curve of acetylacetone,2,4-pentanedione development process in embodiment 1；
Fig. 4 is the standard curve of azanol-ferric trichloride development process in embodiment 1；
Fig. 5 is the standard curve of high performance liquid chromatography in embodiment 1；
Fig. 6 is the standard curve of the gluconic acid sodium content obtained using assay method provided by the invention in embodiment 2；
Fig. 7 is the linear relationship of the gluconic acid sodium content obtained using detection device provided by the invention in embodiment 3
Curve；
Fig. 8 is to produce sodium gluconate mistake using detection means measure fermentation of Aspergillus niger provided by the invention in embodiment 3
The variation diagram of gluconic acid sodium content in journey.
Specific implementation mode
The present invention provides a kind of detection methods of gluconic acid sodium content in zymotic fluid, include the following steps：
Zymotic fluid is mixed with glacial acetic acid, obtains sample solution to be titrated；
Using quinaldine red-methanol solution as indicator, using perchloric acid-glacial acetic acid solution to the sample solution to be titrated
It is titrated, until terminating to titrate when indicator Indicator Reaction terminal, according to scheduled regression equation and required titration volumes, is obtained
The content of sodium gluconate in zymotic fluid, the regression equation are gluconic acid in titration volumes and gluconic acid sodium standard solution
Regression equation between sodium content, the gluconic acid sodium standard solution are the aqueous solution of sodium gluconate.
In the present invention, the volume ratio of the zymotic fluid and glacial acetic acid is preferably (0.001~1)：(5~15).The present invention
There is no special restriction for the source of the zymotic fluid, using source well known to those skilled in the art；In the present invention
Embodiment in, it be production strain that the zymotic fluid, which is specifically with aspergillus niger, prepares using fermentation method the reality of sodium gluconate
The zymotic fluid prepared when experiment in the zymotic fluid generated in production process either laboratory scope.It is detected to zymotic fluid
Before, zymotic fluid is preferably filtered or is centrifuged by the present invention, is then taken filtrate or supernatant to be mixed with glacial acetic acid and is subsequently dripped
It is fixed；In an embodiment of the present invention, the volume of the zymotic fluid is specifically with the filtrate or the stereometer of supernatant.In this hair
In bright, the filter cloth specification of the filtering is preferably 40~300 mesh, more preferably 80~150 mesh；The rate of the centrifugation is preferably
3000~15000r/min, more preferably 5000~10000r/min；Time is preferably 3~10min, more preferably 5~6min.
Using quinaldine red-methanol solution as indicator, using perchloric acid-glacial acetic acid solution to the sample solution to be titrated
It is titrated, until terminating to titrate when indicator Indicator Reaction terminal, according to scheduled regression equation and required titration volumes, is obtained
The content of sodium gluconate in zymotic fluid, the regression equation are gluconic acid in titration volumes and gluconic acid sodium standard solution
Regression equation between sodium content, the gluconic acid sodium standard solution are the aqueous solution of sodium gluconate.In the present invention, institute
The mass concentration for stating quinaldine red-methanol solution is preferably 0.01~1%, and more preferably 0.1~0.5%；The quinaldine red-
The volume ratio of methanol solution and the zymotic fluid is preferably (0.01~0.3)：(0.001~1).In the present invention, the high chlorine
The concentration of perchloric acid is preferably 0.01~0.5mol/L in acid-glacial acetic acid solution, more preferably 0.1~0.2mol/L.The present invention
Using quinaldine red-methanol solution as indicator, the sample solution to be titrated is dripped using perchloric acid-glacial acetic acid solution
Fixed, in titration process, with the increase of perchloric acid-glacial acetic acid solution addition, reaction system is become colorless by red, at this time
The addition of as reaction end, the perchloric acid-glacial acetic acid solution is titration volumes.
In the present invention, the preparation method of the regression equation, preferably includes following steps：
Sodium gluconate is mixed with water, obtains gluconic acid sodium standard solution；
The gluconic acid sodium standard solution is mixed with glacial acetic acid, obtains gluconic acid sodium standard solution to be titrated；
Using quinaldine red-methanol solution as indicator, using perchloric acid-glacial acetic acid solution to the gluconic acid to be titrated
Sodium standard solution is titrated, until indicator Indicator Reaction terminal, records titration volumes, with Portugal in gluconic acid sodium standard solution
Grape saccharic acid sodium content is abscissa, and titration volumes are ordinate, draws the standard curve of gluconic acid sodium content, Portugal is calculated
The regression equation of grape saccharic acid sodium content.
The present invention preferably mixes sodium gluconate with water, obtains the gluconic acid sodium standard solution of series concentration.At this
In invention, the concentration of the gluconic acid sodium standard solution is preferably 0~2.4mol/L.
After obtaining gluconic acid sodium standard solution, the present invention preferably mixes the gluconic acid sodium standard solution with glacial acetic acid
It closes, obtains gluconic acid sodium standard solution to be titrated.In the present invention, the body of the gluconic acid sodium standard solution and glacial acetic acid
Product is than being preferably (0.001~1)：(5~15).
After obtaining gluconic acid sodium standard solution to be titrated, the present invention preferably using quinaldine red-methanol solution as indicator,
The gluconic acid sodium standard solution to be titrated is titrated using perchloric acid-glacial acetic acid solution, until indicator Indicator Reaction
Terminal records titration volumes, and using gluconic acid sodium content in gluconic acid sodium standard solution as abscissa, titration volumes are vertical sit
Mark draws the standard curve of gluconic acid sodium content, the regression equation of gluconic acid sodium content is calculated.In the present invention,
The mass concentration of the quinaldine red-methanol solution is preferably 0.01~1%, and more preferably 0.1~0.5%；The quinaldine
The volume ratio of red-methanol solution and the gluconic acid sodium standard solution is preferably (0.01~0.3)：(0.001~1).At this
In invention, the concentration of the perchloric acid-glacial acetic acid solution is preferably 0.01~0.5mol/L, more preferably 0.1~0.2mol/L.
The present invention is using quinaldine red-methanol solution as indicator, using perchloric acid-glacial acetic acid solution to the sodium gluconate to be titrated
Standard titration solution is titrated, in titration process, with the increase of perchloric acid-glacial acetic acid solution addition, reaction system
It is become colorless by red, at this time as reaction end, the addition of the perchloric acid-glacial acetic acid solution is titration volumes.?
In the present invention, operation that the gluconic acid sodium standard solution to be titrated is titrated preferably with it is molten to the sample to be titrated
The operation that liquid is titrated is consistent.
In an embodiment of the present invention, specifically sodium gluconate is mixed with deionized water, compound concentration is respectively
0mol/L、0.1mol/L、0.2mol/L、0.3mol/L、0.4mol/L、0.5mol/L、0.6mol/L、0.7mol/L、0.8mol/
L、0.9mol/L、1.0mol/L、1.1mol/L、1.2mol/L、1.3mol/L、1.4mol/L、1.5mol/L、1.6mol/L、
The Portugal of 1.7mol/L, 1.8mol/L, 1.9mol/L, 2.0mol/L, 2.1mol/L, 2.2mol/L, 2.3mol/L and 2.4mol/L
Grape saccharic acid sodium standard solution.Take 10mL glacial acetic acid in conical flask, the sodium gluconate standard that 0.1mL various concentrations are added is molten
Liquid is uniformly mixed, and quinaldine red-methanol solution of 0.05mL a concentration of 0.1% is then added, using a concentration of 0.1mol/L's
Perchloric acid-glacial acetic acid solution is titrated, until being titration end-point when system is colourless, titration volumes is recorded, with sodium gluconate mark
Quasi- glucose in solutions acid sodium content is abscissa, and titration volumes are ordinate, draws the standard curve of gluconic acid sodium content,
The regression equation y=1.0903x+0.0532, R of gluconic acid sodium content is calculated2=0.9997.
The present invention provides a kind of detection devices of gluconic acid sodium content in zymotic fluid, as shown in Figure 1, including control-
Display system, photodetector system, quinaldine red-methanol solution bottle 7, glacial acetic acid bottle 8 and perchloric acid-glacial acetic acid solution bottle 9,
In, control switch 18, external display screen 17 and control system 19, the control system are provided in the control-display system
System 19 respectively with the display screen 17, photodetector system, quinaldine red-methanol solution bottle 7, glacial acetic acid bottle 8 and perchloric acid-ice
Acetum bottle 9 communicates to connect, and the control system 19 is molten to the display screen 17, photodetector system, quinaldine red-methanol
The working condition of liquid bottle 7, glacial acetic acid bottle 8 and perchloric acid-glacial acetic acid solution bottle 9 is controlled and is adjusted；The Photoelectric Detection system
System includes light source 12, photoelectric detection system 13 and titration cell 16, the titration cell 16 respectively with the quinaldine red-methanol solution
Bottle 7, glacial acetic acid bottle 8 are connected to perchloric acid-glacial acetic acid solution bottle 9, and the setting of the titration cell 16 is examined in the light source 12 and photoelectricity
It surveys between device 13, the photoelectric detection system 13 is used to detect the variation of color in titration cell 16.
In an embodiment of the present invention, the titration cell 16 and the quinaldine red-methanol solution bottle 7, glacial acetic acid bottle 8 and
It is respectively arranged with peristaltic pump between perchloric acid-glacial acetic acid solution bottle 9；Specifically, the titration cell 16 and the quinaldine red-first
It is provided with peristaltic pump 3 between alcoholic solution bottle 7, peristaltic pump 4 is provided between the titration cell 16 and the glacial acetic acid bottle 8, it is described
It is provided with peristaltic pump 5 between titration cell 16 and the perchloric acid-glacial acetic acid solution bottle 9.In an embodiment of the present invention, the control
System 19 processed is communicated to connect with the peristaltic pump 3, peristaltic pump 4 and peristaltic pump 5 respectively, and the control system 19 passes through to described compacted
The working condition of dynamic pump 3, peristaltic pump 4 and peristaltic pump 5 controlled and adjusted realization to the quinaldine red-methanol solution bottle 7,
The control and adjusting of glacial acetic acid bottle 8 and perchloric acid -9 working condition of glacial acetic acid solution bottle.
In an embodiment of the present invention, the detection device further includes waste liquid bottle 6 and sodium hydroxide solution bottle 10, described useless
Liquid bottle 6 is connected to the titration cell 16 and sodium hydroxide solution bottle 10 respectively, the control system 19 respectively with the waste liquid bottle 6
It is communicated to connect with sodium hydroxide solution bottle 10, work of the control system 19 to the waste liquid bottle 6 and sodium hydroxide solution bottle 10
It is controlled and is adjusted as state.The present invention titrates after being realized to titration by waste liquid bottle 6 and sodium hydroxide solution bottle 10
System in pond 16 is neutralized.
In an embodiment of the present invention, between the waste liquid bottle 6 and the titration cell 16, the waste liquid bottle 6 and hydroxide
It is provided with peristaltic pump between sodium solution bottle 10；Specifically, peristaltic pump 2 is provided between the waste liquid bottle 6 and the titration cell 16,
It is provided with peristaltic pump 1 between the waste liquid bottle 6 and the sodium hydroxide solution bottle 10.In an embodiment of the present invention, the control
System 9 processed is communicated to connect with the peristaltic pump 2 and peristaltic pump 1 respectively, and the control system 9 passes through to the peristaltic pump 2 and compacted
The working condition of dynamic pump 1 is controlled and is adjusted the control realized to 10 working condition of the titration cell 16 and sodium hydroxide solution bottle
System and adjusting.
In an embodiment of the present invention, the bottom of the titration cell 16 is provided with magnetic stirring apparatus 15, the titration cell
16 are internally provided with rotor 14；The present invention by magnetic stirring apparatus 15 and rotor 14 make titration process under agitation into
Row.
In an embodiment of the present invention, the top of the titration cell 16 is provided with sealing-plug 11；The present invention passes through sealing-plug
11 make titration process be carried out under air-proof condition.
In an embodiment of the present invention, the top of the waste liquid bottle 6 is provided with sealing ring 20, quinaldine red-methanol solution
The top of bottle 7 is provided with sealing ring 21, the top of glacial acetic acid bottle 8 is provided with sealing ring 22, perchloric acid-glacial acetic acid solution bottle 9
Top is provided with sealing ring 23；The present invention is made by sealing ring 20 between waste liquid bottle 6 and NaOH solution bottle 10 and titration cell 16
Liquid communication carries out under air-proof condition, makes quinaldine red-methanol solution bottle by sealing ring 21, sealing ring 22 and sealing ring 23
7, the liquid communication of glacial acetic acid bottle 8 and perchloric acid-between glacial acetic acid solution bottle 9 and titration cell 16 carries out under air-proof condition.
In an embodiment of the present invention, the top of the waste liquid bottle 6 is provided with check valve 24, quinaldine red-methanol solution
The top of bottle 7 is provided with check valve 25, the top of glacial acetic acid bottle 8 is provided with check valve 26, perchloric acid-glacial acetic acid solution bottle 9
Top is provided with check valve 27；The present invention is sucked or is discharged air by check valve, avoids occurring positive/negative pressure in bottle.
In the present invention, gluconic acid sodium content in zymotic fluid is examined using detection device described in above-mentioned technical proposal
When survey, due to titration time and titration volumes direct proportionality, with a concentration of abscissa of sodium gluconate, with titration time
For ordinate, titrimetric standard curve can be obtained.Two, which are detected, by two-point calibration defines concentration (M1And M2) calibration point (T1
And T2), wherein titration deionized water (a concentration of M of sodium gluconate1=0) required titration time is T when1, titration mark
Quasi- sodium gluconate solution (a concentration of M of sodium gluconate2) when required titration time T2, control in the detection device
System is according to T1、T2Data fit effective titrimetric standard curve equation：
AM+BT+C=0--------------------------------------- (1)
By (M1,T1)、(M2,T2) bring into formula (1) derivation can obtain,
A=T2-T1；B=M1-M2；C=M2T1-M1T2
Due to M1=0,
Then A=T2-T1；B=-M2；C=M2T1
(T2-T1)M-M2T+M2T1=0
M=M2(T-T1)/(T2-T1)---------------------------------------(2)
It is T to titrate required titration time when sample to be testedx, by TxBring titrimetric standard curve equation (2), the inspection into
Control system is according to T in survey devicexValue calculates the gluconic acid na concn (M in sample to be tested automaticallyx), Mx=M2(Tx-T1)/
(T2-T1)。
In the present invention, gluconic acid sodium content in zymotic fluid is examined using detection device described in above-mentioned technical proposal
The method of survey, preferably includes following steps：
(1) T is demarcated1：18 are turned on the switch, control system 19 controls peristaltic pump 3 will be in quinaldine red-methanol solution bottle 7
Quinaldine red-methanol solution (mass concentration be 0.01~1%, volume is 0.01~0.3mL) be pumped into (flow velocity is 0.01~
0.1mL/s) in titration cell 16, the glacial acetic acid (volume is 1~20mL) in glacial acetic acid bottle 8 is pumped into that (flow velocity is by control peristaltic pump 4
2~4mL/s) in titration cell 16, while the operating of magnetic stirring apparatus 15 is controlled, making the rotation of rotor 14, (rotating speed is 50~200r/
Min), with mixing reaction system；19 control display screen 17 of control system display " calibration T1", sealing-plug 11 is opened, by deionization
Water (volume is 0.01~0.5mL) is added in titration cell 16, and control system 19 controls peristaltic pump 5 by perchloric acid-glacial acetic acid solution
Perchloric acid-glacial acetic acid solution (a concentration of 0.01~0.5mol/L) in bottle 9 is pumped into (flow velocity is 0.01~0.2mL/s) titration cell
In 16, while the light source 12 for controlling 16 one end of titration cell shines, and the photoelectric detection system 13 of 16 other end of titration cell automatically records
Photocurrent variations, according to photocurrent variations rate automatic decision titration end-point, record titration time T1, demarcate T1Terminate, display screen 7
Show T1Time；
Empty titration cell, neutralization reaction liquid：After titration, control system 19 controls light source 12, photoelectric detection system 13
It is closed with magnetic stirring apparatus 15, while controlling peristaltic pump 2 and being pumped into the reaction solution in titration cell 16 (flow velocity is 2~20mL/s)
In waste liquid bottle 6, control peristaltic pump 1 by NaOH solution bottle 10 NaOH solution (a concentration of 1~5mol/L, volume be 2~
It 60mL) is pumped into (flow velocity is 2~20mL/s) waste liquid pool 6, neutralization reaction liquid；
It cleans titration cell, neutralize cleaning solution：Control system 19 controls peristaltic pump 4 by the glacial acetic acid (volume in glacial acetic acid bottle 8
For 1~20mL) it is pumped into (flow velocity is 2~4mL/s) titration cell 16, while the operating of magnetic stirring apparatus 15 is controlled, make rotor 14
Rotation (rotating speed is 50~200r/min, and the time is 10~15s), cleans titration cell 16；After cleaning, control system
19 control magnetic stirring apparatus 15 are closed, at the same control peristaltic pump 2 glacial acetic acid in titration cell 16 is pumped into (flow velocity is 2~
20mL/s) in waste liquid bottle 6, by the NaOH solution in NaOH solution bottle 10, (a concentration of 1~5mol/L, volume are control peristaltic pump 1
2~60mL) it is pumped into (flow velocity is 2~20mL/s) waste liquid pool 6, neutralize glacial acetic acid；
(2) T is demarcated2：T is demarcated according to the method in step (1)2, the difference is that, 19 control display screen of control system
17 displays " calibration T2", sealing-plug 11 is opened, (a concentration of 0.1~2.4mol/L, volume are by standard glucose acid sodium solution
0.01~1mL) it is added in titration cell 16, after reaching titration end-point, record titration time T2, demarcate T2Terminate, display screen 7 is shown
T2Time；
Empty titration cell, neutralization reaction liquid：According to method emptying titration cell, the neutralization reaction liquid in step (1)；
It cleans titration cell, neutralize cleaning solution：According to method cleaning titration cell, the neutralization cleaning solution in step (1)；
(3) sample measures：Sample measurement is carried out according to the method in step (1), the difference is that, control system 19 is controlled
Display screen 17 processed shows " sample measurement ", opens sealing-plug 11, and sample to be tested (zymotic fluid, volume are 0.01~0.5mL) is added
Enter in titration cell 16, after reaching titration end-point, the titration time T used in record titration sample to be testedx, sample, which measures, to be terminated；Control
System processed is according to TxValue calculates the gluconic acid na concn in sample to be tested automatically, and display screen 7 shows TxTime and gluconic acid
Na concn；
Empty titration cell, neutralization reaction liquid：According to method emptying titration cell, the neutralization reaction liquid in step (1)；
It cleans titration cell, neutralize cleaning solution：According to method cleaning titration cell, the neutralization cleaning solution in step (1)；
Start sample measurement next time after cleaning titration cell under the control of control system 19.
Below in conjunction with the embodiment in the present invention, the technical solution in the present invention is clearly and completely described.It is aobvious
So, described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.Based on the reality in the present invention
Example is applied, every other embodiment obtained by those of ordinary skill in the art without making creative efforts all belongs to
In the scope of protection of the invention.
Embodiment 1
The comparative analysis of sodium gluconate assay method
(1) non-aqueous titration
Accurate compound concentration is the sodium gluconate-of 0.3g/L, 0.6g/L, 0.9g/L, 1.2g/L, 1.5g/L and 1.8g/L
Quinaldine red-methanol solution that 0.05mL mass concentrations are 0.1% is added, with a concentration of 0.1mol/L in glacial acetic acid standard solution
Perchloric acid-glacial acetic acid solution be titration end-point when being titrated to colourless.With a concentration of abscissa of sodium gluconate, with perchloric acid-
The titration volumes of glacial acetic acid solution are that ordinate draws standard curve, see Fig. 2.
The measuring principle of non-aqueous titration is that sodium gluconate is alkaline compound, and glacial acetic acid system can enhance its alkali
Property, apparent titration jump is observed that when with perchloric acid titration.
As shown in Fig. 2, the regression equation of non-aqueous titration is y=4.6024x-0.0279, R2=0.9996.Nonaquepous tration
Although method linear relationship is good, operating method is easy, does not need expensive instrument, and this method is only used for analysis glucose
Sour sodium dry product, so non-aqueous titration may not apply to the measurement of gluconic acid sodium content in zymotic fluid.
(2) acetylacetone,2,4-pentanedione development process
Take respectively volume be 0mL, 0.1mL, 0.2mL, 0.3mL, 0.4mL, 0.5mL, 0.6mL, 0.7mL, 0.8mL,
0.9mL, 1.0mL gluconic acid sodium standard solution (5mmol/L) add 1mL 0.02mol/L periodic acid solution in test tube,
It is placed at room temperature for 20min, NaHSO is added dropwise3It has just been moved back to brown, 2mL acetylacetone,2,4-pentanedione reagents is added, deionized water is used in combination to be settled to
25mL；3min in boiling water bath is placed it in, it is rapid cooling, with blank tube zeroising, absorbance value is measured at 420nm.With Portugal
A concentration of abscissa of grape sodium saccharate draws standard curve by ordinate of absorbance value, sees Fig. 3.
As shown in figure 3, the regression equation of acetylacetone,2,4-pentanedione development process is y=0.2761x-0.2208, R2=0.9342.Acetyl
Acetone development process linear relationship is undesirable, and the glucose in zymotic fluid influences measurement result very big, and institute is in this way not
It can apply to the measurement of gluconic acid sodium content in zymotic fluid.
(3) azanol-ferric trichloride development process
1. preparation of reagents：
Azanol base reagent：4mol/L hydroxylamine hydrochlorides and 4mol/LNaOH are mixed in equal volume, and adjust pH value to 8, current existing
Match.
Ferric trichloride reagent：Weigh 10g FeCl3·6H2O is settled to 100mL with 0.1mol/LHCl, as a concentration of
The FeCl of 0.37mol/L3Reagent.
2. Specification Curve of Increasing：The sodium gluconate mark of a concentration of 0.1mol/L of 1mL, 3mL, 5mL, 7mL and 9mL is taken respectively
Quasi- solution is diluted to 100mL and adjusts pH value to 1.5 with 4mol/LHCl；The above-mentioned Portugals 1mL are added into 7 10mL test tubes respectively
Grape saccharic acid sodium standard solution, is placed in boiling water bath and heats 30min, be cooled to room temperature, and sequentially adds 2mL azanol base reagents, and 1mL is dense
Degree is the FeCl of the HCl and 1mL of 4mol/L3Reagent colour development measures absorbance at 500nm；Blank control is with 1mL pH value
1.5 distilled water replaces gluconic acid sodium standard solution, other reagents and dosage are constant；Measuring work need to complete in 10min.
With a concentration of abscissa of sodium gluconate, standard curve is drawn by ordinate of absorbance value, sees Fig. 4.
The measuring principle of azanol-ferric trichloride development process is that sodium gluconate generates lactone, lactone under strongly acidic conditions
Hydroxamic acid is generated with azanol reaction, hydroxamic acid can be complexed with iron (III) to develop the color.
As shown in figure 4, the regression equation of azanol-ferric trichloride development process is y=0.0932x-0.0031, R2=
0.9999.Azanol-ferric trichloride development process has good linear relationship, can be applied to gluconic acid sodium content in zymotic fluid
Detection, but the determination step of this method is very cumbersome, and the glucose of zymotic fluid middle and high concentration can generate measurement result
It influences.
(4) high performance liquid chromatography
Compound concentration is the gluconic acid sodium standard solution of 0~0.015mol/L, is analyzed on high performance liquid chromatograph；It is high
Effect liquid phase chromatogram condition：50mL methanol and 5mL phosphoric acid (are settled to by mobile phase for the aqueous solution of methanol and phosphoric acid with deionized water
1L), column temperature is 25 DEG C, flow velocity 1mL/min, Detection wavelength 210nm, and sample size is 20 μ L.It is a concentration of with sodium gluconate
Abscissa draws standard curve by ordinate of peak area, sees Fig. 5.
As shown in figure 5, the regression equation of high performance liquid chromatography is y=122829x+50.459, R2=0.9992.Efficiently
Liquid chromatography has good linear relationship, can quickly and accurately detect the content of sodium gluconate in zymotic fluid.But
It is that, since sodium gluconate belongs to salt, the pH value of mobile phase is less than the tolerance range of chromatographic column, therefore this method is to chromatographic column
Be affected；In addition, high performance liquid chromatograph device is costly, the application of this method is limited.
Embodiment 2
(1) sodium gluconate is mixed with water, compound concentration be respectively 0mol/L, 0.1mol/L, 0.2mol/L,
0.3mol/L、0.4mol/L、0.5mol/L、0.6mol/L、0.7mol/L、0.8mol/L、0.9mol/L、1.0mol/L、
1.1mol/L、1.2mol/L、1.3mol/L、1.4mol/L、1.5mol/L、1.6mol/L、1.7mol/L、1.8mol/L、
The gluconic acid sodium standard solution of 1.9mol/L, 2.0mol/L, 2.1mol/L, 2.2mol/L, 2.3mol/L and 2.4mol/L, point
Do not take the gluconic acid sodium standard solution of 0.1mL various concentrations in 10mL glacial acetic acid, it is 0.1% that 0.05mL mass concentrations, which are added,
Quinaldine red-methanol solution, be when being titrated to colourless with perchloric acid-glacial acetic acid solution of a concentration of 0.1mol/L titration eventually
Point.With a concentration of abscissa of sodium gluconate, it is bent to draw standard using the titration volumes of perchloric acid-glacial acetic acid solution as ordinate
Line is shown in Fig. 6.
It will be appreciated from fig. 6 that the regression equation y=1.0903x+0.0532, R of gluconic acid sodium content2=0.9997, it is linear to close
System is ideal.
(2) Precision Experiment
Zymotic fluid is centrifuged into (10000r/min, 10min), takes supernatant as sample to be tested, according to titration in step (1)
Method, replication 8 times calculate gluconic acid sodium content in zymotic fluid, and measurement result is shown in Table 1.
1 Precision Experiment measurement result of table
As shown in Table 1, the relative standard deviation (RSD) between each concentration is 0.0059, illustrates method tool provided by the invention
There is higher precision.
(3) repeated experiment
It takes 8 portions of zymotic fluids to centrifuge (10000r/min, 10min) respectively, takes supernatant as sample to be tested, according to step
(1) titration method in is measured 8 parts of samples to be tested respectively, calculates gluconic acid sodium content in zymotic fluid, and measurement result is shown in
Table 2.
2 repeated experiment measurement result of table
As shown in Table 2, the relative standard deviation (RSD) between each concentration is 0.016, shows method variation provided by the invention
Coefficient is small, reproducible.
(4) stability experiment
Zymotic fluid is centrifuged into (10000r/min, 10min), takes supernatant as sample to be tested, every 10min according to step
(1) titration method is measured sample to be tested in, and measurement result is shown in Table 3.
3 stability experiment measurement result of table
As shown in Table 3, the relative standard deviation (RSD) between each concentration is 0.0059, shows method tool provided by the invention
There is preferable stability.
(5) accuracy is tested
According to titration method in step (1), recovery of standard addition experiment is carried out to zymotic fluid, the accuracy of verification method is surveyed
Surely 4 be the results are shown in Table.
4 accuracy measuring result of table
As shown in Table 4, sample recovery rate is between 94.64~104.43%, and average recovery rate is 97.90~100.66%
Between, illustrate that method accuracy provided by the invention is good.
(6) influence of other disturbing factors
According to titration method in step (1), measures other compositions in aspergillus niger culture medium and perchloric acid-glacial acetic acid solution is dripped
The influence for determining volume, the results are shown in Table 5.
The measurement result that other disturbing factors of table 5 influence titration volumes
As shown in Table 5, required perchloric acid-when other compositions (concentration is identical with culture medium) in culture medium is titrated
Glacial acetic acid solution volume is identical as required perchloric acid-glacial acetic acid solution volume when titration deionized water.So aspergillus niger is trained
The other compositions supported in base do not cause error to method provided by the invention.
(7) it is compared with azanol-ferric trichloride determination of color result
Using glucose in aqueous titration provided by the invention and azanol-same zymotic fluid of ferric trichloride determination of color
Sour sodium content, replication 8 times, measurement result is shown in Table 6.Wherein, Portugal in the azanol-ferric trichloride determination of color zymotic fluid
The step of grape saccharic acid sodium content, is specific as follows：
The standard curve of azanol-ferric trichloride development process is drawn according to the method for (3) part in embodiment 1；
Sample measures：Zymotic fluid dilute 100 times after adjust pH value to 1.5, be placed in boiling water bath and heat 30min, then according to
The operation for drawing standard curve handles gained sample solution, and sodium gluconate in zymotic fluid is obtained according to standard curve
Concentration.
The aqueous titration provided by the invention of table 6 and azanol-ferric trichloride determination of color Comparative result
As shown in Table 6, Portugal in the zymotic fluid that aqueous titration provided by the invention and azanol-ferric trichloride development process measure
Not notable (the P of grape sodium saccharate content difference>0.05), i.e., aqueous titration provided by the invention and azanol-ferric trichloride development process
Measurement result it is consistent, the gluconic acid that the aqueous titration for further illustrating the present invention offer can be in Accurate Determining zymotic fluid
Sodium content.
Embodiment 3
(1) sodium gluconate is mixed with water, compound concentration be respectively 0mol/L, 0.2mol/L, 0.4mol/L,
0.6mol/L、0.8mol/L、1mol/L、1.2mol/L、1.4mol/L、1.6mol/L、1.8mol/L、2mol/L、2.2mol/L
With the gluconic acid sodium standard solution of 2.4mol/L, using detection device provided by the invention to the sodium gluconate in zymotic fluid
Content is measured, and using sodium gluconate concentration of standard solution as abscissa, is drawn by ordinate of detection means measure result
Linear relationship curve, is shown in Fig. 7.
As shown in Figure 7, detection means measure result is consistent with sodium gluconate concentration of standard solution, regression equation y=
0.9968x+0.0012, R2=0.9999.
(2) Precision Experiment
Zymotic fluid is centrifuged into (10000r/min, 10min), takes supernatant as sample to be tested, is measured and filled using the present invention
Set replication 8 times, measurement result is shown in Table 7.
7 Precision Experiment measurement result of table
As shown in Table 7, the fluctuation of concentration of sodium gluconate is little, and the relative standard deviation (RSD) between each concentration is
0.0023, illustrate the method coefficient of variation using gluconic acid sodium content in detection means measure zymotic fluid provided by the invention
It is small, there is higher precision.
(3) stability experiment
Zymotic fluid is centrifuged into (10000r/min, 10min), takes supernatant as sample to be tested, this hair is used every 10min
The detection device of bright offer is measured sample to be tested, and measurement result is shown in Table 8.
8 stability experiment measurement result of table
As shown in Table 8, the fluctuation of concentration of sodium gluconate is little, and the relative standard deviation (RSD) between each concentration is
0.0038, it is higher to illustrate that the method using gluconic acid sodium content in detection means measure zymotic fluid provided by the invention has
Stability.
(4) accuracy is tested
Using detection device provided by the invention to zymotic fluid carry out recovery of standard addition experiment, the accuracy of verification method,
Measurement result is shown in Table 9.
9 accuracy measuring result of table
As shown in Table 9, for sample recovery rate between 96.50~100.25%, average recovery rate is 98.63~100.03%
Between, illustrate to use the method rate of recovery of gluconic acid sodium content in detection means measure zymotic fluid provided by the invention good,
The needs of sodium gluconate assay in zymotic fluid can be met.
(5) fermentation of Aspergillus niger production sodium gluconate process is monitored using detection device provided by the invention, is had
Body is as follows：
Seed culture medium：Potato 200g/L, glucose 20g/L, MgSO4·7H2O 1g/L, KH2PO41.5g/L
CaCO34g/L；
Fermentation medium：Glucose 217.6g/L, MgSO4·7H2O 0.01g/L, KH2PO40.13g/L, urea 0.2g/
L；
Aspergillus niger seed liquor is inoculated into the 500mL shaking flasks equipped with 250mL fermentation mediums, shaking table culture (120r/
Min, 28 DEG C).PH value is adjusted to 5.5 with NaOH solution (4mol/L) every for 24 hours in superclean bench, and takes appropriate fermentation
Liquid centrifuges (10000r/min, 6min), supernatant is taken, using detection means measure provided by the invention gluconic acid therein
Na concn, measurement result are shown in Fig. 8.
As shown in Figure 8, detection device provided by the invention can realize fermentation of Aspergillus niger production sodium gluconate process
Monitoring.
As seen from the above embodiment, method provided by the invention can with gluconic acid sodium content in Accurate Determining zymotic fluid,
Methodology experiment shows that the method range of linearity provided by the invention, precision, repeatability, stability and accuracy are good, grape
Other compositions do not influence measurement result in the zymotic fluids such as sugar, and method provided by the invention is easy to operate, are as measurement
The ideal method of gluconic acid sodium content during fermentation of Aspergillus niger.In addition, can be accurate using detection device provided by the invention
Gluconic acid sodium content in zymotic fluid is really measured, and apparatus structure provided by the invention is simple, component without using costliness, energy
Enough realize the monitoring of fermentation of Aspergillus niger production sodium gluconate process.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (10)
1. the detection method of gluconic acid sodium content, includes the following steps in a kind of zymotic fluid：
Zymotic fluid is mixed with glacial acetic acid, obtains sample solution to be titrated；
Using quinaldine red-methanol solution as indicator, the sample solution to be titrated is carried out using perchloric acid-glacial acetic acid solution
Titration, until terminating to titrate when indicator Indicator Reaction terminal, according to scheduled regression equation and required titration volumes, is fermented
The content of sodium gluconate in liquid, the regression equation are that sodium gluconate contains in titration volumes and gluconic acid sodium standard solution
Regression equation between amount, the gluconic acid sodium standard solution are the aqueous solution of sodium gluconate.
2. detection method according to claim 1, which is characterized in that the volume ratio of the zymotic fluid and glacial acetic acid is
(0.001~1)：(5~15).
3. detection method according to claim 1, which is characterized in that the mass concentration of the quinaldine red-methanol solution
It is 0.01~1%；The volume ratio of the quinaldine red-methanol solution and the zymotic fluid is (0.01~0.3)：(0.001~
1)。
4. detection method according to claim 1, which is characterized in that perchloric acid in the perchloric acid-glacial acetic acid solution
A concentration of 0.01~0.5mol/L.
5. according to Claims 1 to 4 any one of them detection method, which is characterized in that the preparation method of the regression equation,
Include the following steps：
Sodium gluconate is mixed with water, obtains the gluconic acid sodium standard solution of series concentration；
The gluconic acid sodium standard solution is mixed with glacial acetic acid, obtains gluconic acid sodium standard solution to be titrated；
Using quinaldine red-methanol solution as indicator, using perchloric acid-glacial acetic acid solution to the sodium gluconate mark to be titrated
Quasi- solution is titrated, until terminating to titrate when indicator Indicator Reaction terminal, records titration volumes, molten with sodium gluconate standard
Gluconic acid sodium content is abscissa in liquid, and titration volumes are ordinate, draws the standard curve of gluconic acid sodium content, is calculated
Obtain the regression equation of gluconic acid sodium content.
6. detection method according to claim 5, which is characterized in that a concentration of the 0 of the gluconic acid sodium standard solution
~2.4mol/L.
7. the detection device of gluconic acid sodium content in a kind of zymotic fluid, including control-display system, photodetector system, quinoline
Which pyridine is red-methanol solution bottle, glacial acetic acid bottle and perchloric acid-glacial acetic acid solution bottle, wherein it is arranged in the control-display system
Have control switch, external display screen and a control system, the control system respectively with the display screen, photodetector system,
Quinaldine red-methanol solution bottle, glacial acetic acid bottle and perchloric acid-glacial acetic acid solution bottle communication connection, the control system is to described
The work shape of display screen, photodetector system, quinaldine red-methanol solution bottle, glacial acetic acid bottle and perchloric acid-glacial acetic acid solution bottle
State is controlled and is adjusted；The photodetector system includes light source, photoelectric detection system and titration cell, the titration cell difference
It is connected to the quinaldine red-methanol solution bottle, glacial acetic acid bottle and perchloric acid-glacial acetic acid solution bottle, the titration cell setting exists
Between the light source and photoelectric detection system, the photoelectric detection system is used to detect the variation of color in titration cell.
8. detection device according to claim 7, which is characterized in that the titration cell and the quinaldine red-methanol are molten
Peristaltic pump, the control system and the wriggling are respectively arranged between liquid bottle, glacial acetic acid bottle and perchloric acid-glacial acetic acid solution bottle
Pump communication connection, the control system controlled and adjusted by the working condition to the peristaltic pump realization to the quinoline which
The control and adjusting of red-methanol solution bottle, glacial acetic acid bottle and perchloric acid-the glacial acetic acid solution bottle working condition of pyridine.
9. detection device according to claim 7 or 8, which is characterized in that further include waste liquid bottle and sodium hydroxide solution bottle,
The waste liquid bottle is connected to the titration cell and sodium hydroxide solution bottle respectively, the control system respectively with the waste liquid bottle and
Sodium hydroxide solution bottle communicates to connect, and the control system carries out the working condition of the waste liquid bottle and sodium hydroxide solution bottle
Control and adjusting.
10. detection device according to claim 9, which is characterized in that between the waste liquid bottle and the titration cell, described
Peristaltic pump is provided between waste liquid bottle and sodium hydroxide solution bottle, the control system is communicated to connect with the peristaltic pump, described
It is molten to the titration cell and sodium hydroxide that control system is controlled and adjusted realization by the working condition to the peristaltic pump
The control and adjusting of liquid bottle working condition.
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| CN114849606A (en) * | 2022-03-31 | 2022-08-05 | 中国科学院兰州化学物理研究所 | Preparation of gel type liquid chromatography filler and method for detecting sodium gluconate, syrup and white sugar in water reducer compound liquid by using gel type liquid chromatography filler |
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| CN117607346A (en) * | 2024-01-17 | 2024-02-27 | 长春黄金研究院有限公司 | Method for measuring sodium gluconate in water quality |
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