CN1056304A - 3,5,3 ', 5 '-preparation method of tetramethyl benzidine and hydrochloride thereof - Google Patents
3,5,3 ', 5 '-preparation method of tetramethyl benzidine and hydrochloride thereof Download PDFInfo
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- CN1056304A CN1056304A CN 90105657 CN90105657A CN1056304A CN 1056304 A CN1056304 A CN 1056304A CN 90105657 CN90105657 CN 90105657 CN 90105657 A CN90105657 A CN 90105657A CN 1056304 A CN1056304 A CN 1056304A
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Abstract
3,5,3 ', 5 '-preparation method of tetramethyl benzidine and hydrochloride thereof, with 2, the 6-xylidine is a raw material, through redox and molecular rearrangement reaction preparing product, reduce fluorizating agent high ferro potassium application rate, during the intermediate silica gel column chromatography with sherwood oil as eluent, obtain thick product after, without the free alkali stage, directly use recrystallizing methanol, make the product hydrochloride, product content is measured to adopt and is exchanged the extremely general volumetry of oscillography.The inventive method consumption raw material is few, shortened process, and the product efficiency height adapts to suitability for industrialized production.
Description
The invention relates to a kind of aromatic amine organic compound 3,5,3 that can be used as biochemical reagents ', 5 '-preparation method's of tetramethyl benzidine (TMB) and hydrochloride thereof improvement.
3,5,3 ', 5 '-tetramethyl benzidine is a kind of chromogen reagent, people such as Holland once reported it and synthesized, and saw " tetrahedron ", 3299 pages of (Tetrahedron of 1974 30 volumes, 30,3299,1974), and with it be made for the secure replacement thing of p-diaminodiphenyl, be used for blood stain and fecaluria occult blood examination, plasma hemoglobin quantitatively, enzyme immunoassay, cell dyeing and enzymatic analysis etc., existingly be worth certainly.People such as Pinkus introduced its concrete preparation method, saw 380 pages (J.Chemical Education, 54,380,1977) of " chemical education magazine " 1979 the 54th volumes.With 2, the 6-tolidine is a raw material, through redox reaction and molecular rearrangement reaction preparing product.Be oxygenant with the high-potassium ferricyanide wherein, oxidation in basic solution generates intermediate 2,6,2 ', 6 '-the tetramethyl-nitrogen benzide, in this intermediate silica gel column chromatography, with 1: 3 chloroform-hexane mixed solvent wash-out, with the zinc powder is reductive agent, and neutral reduction generates 2,6,2 ', 6 '-the tetramethyl-hydrazobenzene, in ether, carry out molecular rearrangement reaction again, obtain product, the large usage quantity of oxygenant high-potassium ferricyanide in the reaction, raw material weight ratio with it are about 1: 15, during silica gel column chromatography, use contains the mixed solvent of chloroform as eluent, toxicity is bigger, and difficult solvent recovery, the preparation of product hydrochloride, be to generate free alkali, again free alkali added hydrochloric acid and make hydrochloride with the sodium hydroxide neutralized product.The mensuration of product content is used nonaqueous titrations at present usually.The limitation of this method is, 1, can only test the product of free alkali state, can not survey hydrochloride; because molecular crystal water is arranged in the hydrochloride, 2, product is aromatic amine compounds, often needs to add aceticanhydride in the nonaqueous titration; make the aromatic amine acetylize, can make measurement result on the low side.3, this product also should not be surveyed its content with aryl primary amine heavy ammoniation process commonly used, because after amino ortho position occupied methyl, sterically hindered influence weight aminating reaction carried out.
In a word, prior art consumption raw material is more, and preparation process is longer, and the product yield is on the low side, the incompatibility suitability for industrialized production.
Purpose of the present invention is to improve at above-mentioned the deficiencies in the prior art part, provide a kind of preparation 3,5,3 ', 5 '-processing method of Gong the production of tetramethyl benzidine and hydrochloride thereof, provide a kind of method of measuring product content simultaneously.
The present invention prepares 3,5,3 ', 5 '-method of tetramethyl benzidine, still with 2, the 6-xylidine is a raw material, through redox reaction and molecular rearrangement reaction preparation, wherein, be oxygenant with the high-potassium ferricyanide, oxidation in basic solution generates intermediate 2,6,2 ', 6 '-the tetramethyl-nitrogen benzide, this intermediate is carried out silica gel column chromatography, with the zinc powder is reductive agent, and neutral reduction generates 2,6,2 ', 6 '-the tetramethyl hydrazobenzene, in ether, carry out molecular rearrangement reaction again, obtain product 3,5,3 ', 5 '-tetramethyl benzidine.It is characterized in that: 1, reduce the consumption of oxygenant high-potassium ferricyanide, raw material 2, the weight ratio of 6-xylidine and high-potassium ferricyanide is 1: 10～14,2, intermediate 2,6,2 ', 6 '-tetramethyl-nitrogen benzide silica gel column chromatography in, with the single solvent sherwood oil as eluent.The method of preparing product hydrochloride of the present invention, it is characterized in that obtaining thick product after, without the free alkali stage, use recrystallizing methanol, make 3,5,3 ', 5 '-the tetramethyl benzidine dihydrochloride.
Below in conjunction with embodiment preparation method of the present invention is described in detail:
One, 3,5,3 ', 5 '-preparation of tetramethyl benzidine
1, intermediate 2,6,2 ', 6 '-the preparation feedback formula of tetramethyl-nitrogen benzide:
Gradation adds B liquid (2 in A liquid (high-potassium ferricyanide, sodium hydroxide, water), 6-xylidine, concentrated hydrochloric acid, water), 93～96 ℃ of temperature stirred 10 minutes, use Petroleum ether extraction, silica gel column chromatography, the sherwood oil wash-out gets red component 2,6,2 ', 6 '-the tetramethyl-nitrogen benzide, 46～47 ℃ of fusing points.
Prepare four batches 2,6,2 ', 6 '-raw material and the product of tetramethyl-nitrogen benzide is as follows:
| Raw material and name of product specification | Lot number | ||||
| 1 | 2 | 3 | 4 | ||
| 2,6-xylidine (gram) | CP | 4.0 | 12 | 24 | 24 |
| High-potassium ferricyanide (gram) | CP | 40 | 120 | 300 | 330 |
| Hydrochloric acid (milliliter) | CP | 6.0 | 18 | 36 | 36 |
| Sodium hydroxide (gram) | CP | 8.4 | 25.2 | 50.4 | 50.4 |
| Water (milliliter) | 340 | 720 | 800 | 800 | |
| Chromatographic silica gel (gram) | 150 | 400 | 310 | 150 | |
| Sherwood oil (milliliter) | In right amount | In right amount | In right amount | In right amount | |
| 2,6,2, ' 6 '-tetramethyl-nitrogen benzide (gram) | 0.59 | 1.83 | 3.7 | 3.7 | |
| Yield (%) | 14.75 | 15.51 | 15.6 | 15.68 | |
2,2,6,2 ', 6 '-preparation of tetramethyl-hydrazobenzene
Reaction formula:
With 2,6,2 ', 6 '-tetramethyl-nitrogen benzide (ether solvent) adds in the mixture C (ammonium chloride, zinc powder, ethanol, water), and 20～25 ℃ of temperature are stirred to red the disappearance, tell the ether layer, extract water layer at twice with ether, combined ether liquid is 2 in the ether liquid, 6,2 ', 6 '-the tetramethyl-hydrazobenzene, can directly carry out rearrangement reaction.
3,3,5,3 ', 5 '-preparation of tetramethyl benzidine
Reaction formula:
Reaction solution described in 2 is added D liquid (hydrochloric acid, water), stirred 20 minutes, leaching grey mashed prod, water-soluble, to transfer more than the pH10, precipitation is with the benzene recrystallization, must 3,5,3 ', 5 '-tetramethyl benzidine, 166～168 ℃ of fusing points.
Prepare four batches 3,5,3 ', 5 '-raw material and the product of tetramethyl benzidine is as follows:
Two, 3,5,3 ', 5 '-preparation of tetramethyl biphenyl amine hydrochlorate
Reaction formula:
With aforementioned one, 2, the gained reaction solution adds D liquid (hydrochloric acid, water), stirred 20 minutes, after the leaching grey mashed prod, uses sherwood oil and washing filter cake instead, use recrystallizing methanol, make 3,5,3 ', 5 '-the tetramethyl benzidine dihydrochloride, fusing point is more than 300 ℃.
Prepare four batches 3,5,3 ', 5 '-raw material and the product of tetramethyl biphenyl amine hydrochlorate is as follows:
The present invention is an ac oscillopolarographic titration to product and hydrochloride content assaying method thereof, utilize AC oscillographic pole spectrum instrument and titration apparatus to carry out titration, concrete steps are: with 3,5,3 ', 5 '-the tetramethyl benzidine sample is dissolved in earlier in the water that contains an amount of dilute hydrochloric acid, add an amount of acetate buffer, add quantitatively excessive sodium tetraphenylborate (Na-TPB) reference liquid again, TMB and Na-TPB such as can form at the precipitation of mol ratio in this damping fluid, do filter after, divide and get quantitative filtrate, transfer to alkalescence, disappear directing terminal with thallous sulfate reference liquid titration otch of Tetraphenyl sodium borate to the oscillogram.Its reaction formula is as follows:
(white)
(white)
In this measuring method, the precipitation that tetramethyl benzidine and sodium tetraphenylborate are produced such as consists of at mol ratio, and acetate buffer pH value should be in 3.0～4.5 scopes, and optimal pH is 3.6～3.8.
The test of product hydrochloride, sample can be directly water-soluble after with damping fluid and reference liquid, all the other steps are the same.
3,5,3 ', 5 '-precipitation that tetramethyl benzidine and sodium tetraphenylborate generate has certain solubleness, makes rate of recovery result on the low side about 1%, and this measuring method adopts the blank correction method of adding, to improve the rate of recovery, promptly in working sample, all add a certain amount of test liquid in sample liquid and the blank solution, like this, all generate 3,5,3 in sample liquid and the blank solution ', 5 '-sodium tetraphenylborate precipitation of tetramethyl benzidine, thus the influence of offsetting a large portion solubleness.
The cubage formula is as follows:
(1) TMB cubage formula is as follows:
W＝M
T1+×（V
1-V
2）×2×2×240.35
W: TMB content (mg) in the sample
M
T1+: the volumetric molar concentration of thallous sulfate reference liquid
V
1: the milliliter number of surveying the thallous sulfate reference liquid that consumes when producing blank
V
2: the milliliter number that consumes the thallous sulfate reference liquid during working sample
240.35:TMB molecular weight
(2) TMB2HCl2H
2O cubage formula:
W＝M
T1+×（V
1-V
3）×2×2×349.30
W: TMB2HCl2H in the sample
2O content (mg)
M
T1+: the volumetric molar concentration of thallous sulfate reference liquid
V
1: the milliliter number of the thallous sulfate reference liquid that consumes when measuring blank
V
3: the milliliter number that consumes the thallous sulfate reference liquid during working sample
349.30:TMB2HCl2H
2The O molecular weight
Embodiment: 3,5,3 ', 5 '-tetramethyl benzidine free alkali sample test liquid 20.00ml, with blank amount test liquid 2.00ml together, add slow liquid (the pH value is 3.6～3.8) 10.00ml of 2mol/L acetate, 0.03mol1
-1Sodium tetraphenylborate solution 10.00ml mixes, and the water constant volume after dried the filtration, disperses 25.00ml filtrate to 50ml, is transferred to alkalescence with the 40%NaOH test solution, inserts electrode, uses 0.005mol1
-1The titration of thallous sulfate reference liquid, the sodium tetraphenylborate otch disappears to the oscillogram.Parallel work is blank, and test liquid adds test liquid 2.00ml for water.Every 1ml reference liquid is equivalent to 2.404mgC
6H
20ON
2Or 3.493mg C
6H
10ON
22HCl2H
1O.Measurement result, the inventive method makes 3,5,3 ', 5 '-the tetramethyl benzidine product purity is 98.0～99.6%.
The present invention prepares 3,5,3 ', 5 '-method of tetramethyl benzidine reduced the consumption of oxygenant high-potassium ferricyanide, improved intermediate product 2,6,2 ', 6 '-yield of tetramethyl-nitrogen benzide, reduce cost, during the intermediate product silica gel column chromatography, adopted the single solvent sherwood oil to make eluent, eliminated the murder by poisoning of chloroform, and made convenient solvent reclaiming.Preparation 3,5,3 ', 5 '-the tetramethyl biphenyl amine hydrochlorate, without the free alkali stage, shortened technical process.Adopt ac oscillopolarographic titration product and hydrochloride content thereof, improved measuring value accuracy, working method is also easier.Therefore, the inventive method can be applied in the suitability for industrialized production and go.
Claims (3)
1, preparation aromatic amine organic compound 3,5,3 ', 5 '-method of tetramethyl benzidine and hydrochloride thereof, with 2, the 6-xylidine is a raw material, through redox and molecular rearrangement reaction preparation 3,5,3 ', 5 '-tetramethyl benzidine.Wherein, be oxygenant with the high-potassium ferricyanide, oxidation in basic solution, generation intermediate 2,6,2 ', 6 '-the tetramethyl-nitrogen benzide.This intermediate is carried out silica gel column chromatography, under neutrallty condition, reduce with zinc powder, generation 2,6,2 ', 6 '-the tetramethyl-hydrazobenzene, in ether, carry out molecular rearrangement reaction again, it is characterized in that:
The weight ratio of (1) 2,6-xylidine and high-potassium ferricyanide is 1: 10～14,
(2) use the single solvent sherwood oil as eluent in the silica gel column chromatography,
(3) obtain thick product after, without the free alkali stage, use recrystallizing methanol, make 3,5,3,5-tetramethyl benzidine dihydrochloride.
2, measure 3,5,3 ', 5 '-method of tetramethyl benzidine and hydrochloride content thereof, respectively with 3,5,3 ', 5 '-tetramethyl benzidine free alkali and 3,5,3 ', 5 '-the tetramethyl benzidine dihydrochloride makes sample liquid, it is characterized in that sample liquid is carried out titration with ac oscillopolarographic titration, key step is: with 3,5,3 ', 5 '-the tetramethyl benzidine sample is dissolved in (hydrochloride is then directly water-soluble) in an amount of dilute hydrochloric acid, add an amount of acetate buffer, the pH value is 3.0～4.5, adds quantitatively excessive sodium tetraphenylborate reference liquid again, after dried the filtration, divide and get quantitative filtrate, transfer to alkalescence, disappear directing terminal with thallous sulfate reference liquid titration sodium tetraphenylborate otch to the oscillogram that exchanges oscillographic polarograph.
3, mensuration 3,5,3 according to claim 2 ', 5 '-method of tetramethyl benzidine and hydrochloride content thereof, it is characterized in that acetate buffer pH value is 3.6～3.8.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 90105657 CN1056304A (en) | 1990-05-04 | 1990-05-04 | 3,5,3 ', 5 '-preparation method of tetramethyl benzidine and hydrochloride thereof |
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|---|---|---|---|
| CN 90105657 CN1056304A (en) | 1990-05-04 | 1990-05-04 | 3,5,3 ', 5 '-preparation method of tetramethyl benzidine and hydrochloride thereof |
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|---|---|
| CN1056304A true CN1056304A (en) | 1991-11-20 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102875412A (en) * | 2012-10-22 | 2013-01-16 | 浙江万丰化工有限公司 | Method for oxidatively preparing azoic compound by amino compounds |
| CN106631825A (en) * | 2016-11-18 | 2017-05-10 | 河南师范大学 | Preparation method of 3,3,5,5-tetramethyl benzidine |
| CN107652187A (en) * | 2017-10-20 | 2018-02-02 | 荆楚理工学院 | A kind of biochemical preparation TMB synthetic method |
| CN108997139A (en) * | 2018-07-12 | 2018-12-14 | 南京艾普特生物医药有限公司 | A kind of synthetic method of 3,3`, 5,5`- tetramethyl benzidine and its hydrochloride |
-
1990
- 1990-05-04 CN CN 90105657 patent/CN1056304A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102875412A (en) * | 2012-10-22 | 2013-01-16 | 浙江万丰化工有限公司 | Method for oxidatively preparing azoic compound by amino compounds |
| CN106631825A (en) * | 2016-11-18 | 2017-05-10 | 河南师范大学 | Preparation method of 3,3,5,5-tetramethyl benzidine |
| CN106631825B (en) * | 2016-11-18 | 2019-02-01 | 山东凌凯药业有限公司 | A kind of preparation method of 3,3,5,5- tetramethyl benzidine |
| CN107652187A (en) * | 2017-10-20 | 2018-02-02 | 荆楚理工学院 | A kind of biochemical preparation TMB synthetic method |
| CN107652187B (en) * | 2017-10-20 | 2020-10-30 | 荆楚理工学院 | Synthesis method of biochemical preparation TMB |
| CN108997139A (en) * | 2018-07-12 | 2018-12-14 | 南京艾普特生物医药有限公司 | A kind of synthetic method of 3,3`, 5,5`- tetramethyl benzidine and its hydrochloride |
| CN108997139B (en) * | 2018-07-12 | 2020-11-10 | 南京艾普特生物医药有限公司 | Synthetic method of 3,3',5,5' -tetramethyl benzidine and hydrochloride thereof |
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| WD01 | Invention patent application deemed withdrawn after publication |