CN1181344C - Oxidized haematine solution and its preparing process and application - Google Patents
Oxidized haematine solution and its preparing process and application Download PDFInfo
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- CN1181344C CN1181344C CNB00109145XA CN00109145A CN1181344C CN 1181344 C CN1181344 C CN 1181344C CN B00109145X A CNB00109145X A CN B00109145XA CN 00109145 A CN00109145 A CN 00109145A CN 1181344 C CN1181344 C CN 1181344C
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- haematine
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- iodate
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Abstract
The present invention relates to an oxidized haematine solution, a preparing process thereof and uses thereof. The oxidized haematine solution is transparent dark red brown liquid; the preparing process of the oxidized haematine solution comprises: iodate is added to a haematine ethanol solution, a raw material; the haematine ethanol solution with the iodate is heated to the boil for reflux reaction, and filtered to obtain a filtrate; the oxidized haematine solution is obtained by adding glycerol to the filtrate. The present invention has the advantage of simple processing method, a haematine solution which is prepared by the oxidized haematine solution and a mordanting solution, and contains the mordanting solution has the advantage of strong affinity specificity of cell nucleuses, enables the cell nucleuses to show fine, clear and well-bedded blue black; collagenous fibre between tissues is colourless; besides, the haematine solution has the advantages of high stability and easy preservation.
Description
Technical field
The present invention relates to a kind of chemical reagent, particularly a kind of oxidized haematine solution and its production and use.
Background technology
Haematine solution is a kind of histocytology staining reagent, be widely used in biomedical clinical pathological diagnosis, biological morphology teaching and research field, prepared the required oxidation HAEMATOXYLIN of haematine solution is solid product in the past, and its processing technology complexity is prepared when needing temporarily, use inconvenience, the reagent dyeing selectivity is relatively poor, and a nuclear and a matter composition differentiation are unclear, and its stability is undesirable, storage life is shorter, therefore is necessary to provide a kind of oxidized haematine solution of good stability.
Summary of the invention
The purpose of this invention is to provide a kind of oxidized haematine solution and its production and use, oxidized haematine solution mixes with mordanting bath that to obtain haematine solution be the histocytology staining reagent, the method that the present invention prepares oxidized haematine solution is easy, can realize suitability for industrialized production, product stability is good, makes HAEMATOXYLIN reagent and has the affine specificity of stronger nucleus.
For achieving the above object, the present invention takes following design proposal: this oxidation Soviet Union apricot seminal fluid is the transparency liquid of dark brown red, be single dark brown red spot when checking with conventional thin layer chromatography, the chromatographic solution prescription is 82 milliliters of 95% ethanol, 18 milliliters in glacial acetic acid, chromatographic silica gel are tlc silica gel G (60 type).
The technology of oxidized haematine solution comprises: add iodate in the ethanolic solution of HAEMATOXYLIN, the boiling reflux of heating reaction, the concentration of described HAEMATOXYLIN ethanolic solution is the 10-25% weight percent, the concentration of alcohol of preparing above-mentioned solution is between 80-97%, the consumption of iodate is 1-4 with the ratio of raw material HAEMATOXYLIN weight: 10, described back flow reaction to the reactant liquor of heating is the dark brown red transparency liquid, available conventional thin layer chromatography is checked the extent of reaction, when being single dark brown red spot, cessation reaction, filter the back and in filtrate, add glycerine, glycerine: ethanol=1: 1-2 (volume ratio).
With oxidized haematine solution preparation haematine solution, in the described haematine solution mordanting bath is arranged, mordanting bath is inorganic salt solution or organic slat solution or phosphomolybdic acid or phosphotungstic acid.
The haematine solution of a kind of oxidized haematine solution preparation contains 1-5 part oxidized haematine solution in this haematine solution, 6-10 part mordanting bath (volume ratio), and the consisting of of described mordanting bath:
Distilled water 2000-4000 milliliter
Glycerine 100-500 milliliter
95% ethanol 1000-3000 milliliter
Glacial acetic acid 200-500 milliliter
The consumption of 12 water aluminium potassium sulfates or 12 water alums is controlled between the 0.01-0.21M that accounts for above-mentioned several mixed liquid concentration.
The molecular weight of the oxidation HAEMATOXYLIN in the haematine solution provided by the invention (containing mordanting bath) is 300.26, and its structural formula is
Haematine solution is represented with the HYJ1999 haematine solution among the present invention.
Preserving type: room temperature, storage life 2 years is kept in Dark Place at shady and cool place.
Advantage of the present invention: the method that the present invention prepares oxidized haematine solution is easy, can realize suitability for industrialized production, and product stability is good, makes HAEMATOXYLIN reagent and has the affine specificity of stronger nucleus.
Description of drawings
Fig. 1: the biological tissue section figure (human stomach tissue) that utilizes haematine solution provided by the invention to obtain
The invention will be further described below in conjunction with embodiment.
Embodiment
Embodiment 1:
One, take by weighing HAEMATOXYLIN pressed powder 20 grams, be dissolved in 100 milliliters of 80% ethanol, fully stir dissolving fully, add sodium iodate powder 4 grams, fully stir under the room temperature, reaction suspension is poured in the refluxing reactor, ebuillition of heated, return time 2 hours, at this moment reactant liquor is the transparent liquid of dark brown red, drawing reactant liquor with glass pipette is coated with on white quantitative filter paper, present dark brown, also available conventional thin layer chromatography is checked the extent of reaction, and (the chromatographic solution prescription is 95% ethanol g2ml to being single dark brown red spot, glacial acetic acid 18ml, chromatographic silica gel is tlc silica gel G (60 type), gets final product cessation reaction.Naturally cool to room temperature, filter, obtain filtrate, get glycerine and add in the filtrate for 80 milliliters, fully mix to such an extent that contain glycerine filtrate, i.e. oxidized haematine solution.Adopting ebuillition of heated backflow purpose in this technology is for the accelerated oxidation reaction rate, and removes excessive iodate.
Two, mordanting bath preparation
Get 4000 milliliters of distilled water, add 12 water aluminium potassium sulfates or 12 water alums 500 gram, fully dissolving adds 3000 milliliters of 95% ethanol successively, 500 milliliters of glycerine, 500 milliliters in glacial acetic acid, mixing and getting.
Three, haematine solution preparation:
Get 4 parts of oxidized haematine solutions and add 7 parts of mordanting baths, fully stirring and evenly mixing and getting.It is darker transparent salmon liquid, evenly, wall built-up is strong, pH value 2-4.
Oxidized haematine solution provided by the invention can also be used with other known inorganic salts or organic slat solution (mordanting bath), for example: change aluminium potassium sulfate among the embodiment 1 into ammonium ferric sulfate or aluminium sulphate or aluminum chloride.Also has other mordants, for example ferric trichloride or ferric acetate solution etc.Be used with phosphotungstic acid or phosphomolybdic acid in addition, be used for neurocyte dyeing etc.
Embodiment 2
One, takes by weighing HAEMATOXYLIN pressed powder 10 gram, be dissolved in 100 milliliters of 95% ethanol, fully stir dissolving fully, add Potassiumiodate powder 2 grams, fully stir under the room temperature, above-mentioned suspension is poured in the refluxing reactor, be heated to boiling, refluxed 3 hours, other operations are with embodiment 1.
Two, mordanting bath preparation
Get 2000 milliliters of distilled water, add 12 water aluminium potassium sulfates, 250 grams, fully dissolving adds 100 milliliters of glycerine, 1000 milliliters of 95% ethanol, 200 milliliters in glacial acetic acid, mixing successively.
Three, haematine solution preparation
Get 3 parts of oxidized haematine solutions and add 7 parts of mordanting baths, fully stirring and evenly mixing and getting.
Haematine solution provided by the invention (including mordanting bath) is used for cell smear dyeing, tissue section strain, histochemistry's nucleus and redyes.The related reagent of above-mentioned test has 0.5% eosin stain, and 0.011M ammonium chloride returns blue liquid, 0.01M citric acid differentiation liquid.
Hematoxylin eosin stain (HE) method pair cell smear staining routinely, the result is: nucleus presents clear well-bedded black-and-blue, and tenuigenin is beautiful Yihong look, mucus and is well-bedded pink; To section statining, the result is according to a conventional method: nucleus presents clear well-bedded black-and-blue, and tenuigenin is beautiful redness, and interstitial fibers is the pale red (see figure 1) of levels are rich.
According to a conventional method histochemistry's nucleus is redyed, the result is: present dark, shallow differ black-and-blue according to the different nucleus of dyeing time, nucleus clear-cut, clear and definite.
The dyeability test method of haematine solution of the present invention (containing mordanting bath)
1, paraffin wax is cut into slices and is taken off cured secondary into dimethylbenzene, each 10 minutes.
2, section is gone into absolute ethyl alcohol and is taken off the benzene secondary, each 5 minutes.
3, section is gone into 95% ethanol and is taken off the benzene secondary, 5 minutes.
4, slightly control dried ethanol, went into haematine solution (containing mordanting bath) 20 minutes.
5, water is rinsed and is washed 2 seconds.
6, go into 0.01M aqueous citric acid solution differentiation 2 seconds.
7, water is rinsed and is washed.
8, go into 0.01M ammonium chloride and return blue liquid 10-30 second fast
9, water is rinsed and is washed.
10,95% ethanol dehydration is one time 2 minutes
11, absolute ethyl alcohol dehydration secondary, each 2 minutes
12, dimethylbenzene takes off the transparent secondary of ethanol.Each 2 minutes, glass cover thing sheet was added a cover in neutral optical resin gum sealing.
13, result: observation of cell nuclear is presented fine and smoothly and clear well-bedded black-and-blue by specific stain under 40-100 times of optical microscope, organizes the interstitial collagen fiber colourless.
Claims (5)
1, a kind of oxidized haematine solution is characterized in that it comprises following material: the HAEMATOXYLIN ethanolic solution, and the concentration of this solution is the 20-25% percentage by weight, the concentration of alcohol of preparing above-mentioned solution is between 80-97%; Also comprise iodate, the weight ratio of iodate and HAEMATOXYLIN ethanolic solution is 1-4: 10, and these materials are made by heating, boiling, and the terminal point of ebuillition of heated adopts conventional thin layer chromatography inspection, and when being single punctation, experiment stops.
2, a kind of technology for preparing the described oxidized haematine solution of claim 1, it is characterized in that: it comprises: add iodate in the ethanolic solution of HAEMATOXYLIN, the boiling reflux of heating reaction, the concentration of described HAEMATOXYLIN ethanolic solution is the 20-25% percentage by weight, the concentration of alcohol of preparing above-mentioned solution is between 80-97%, the consumption of iodate is 1-4 with the ratio of raw material HAEMATOXYLIN weight: 10, described back flow reaction to the reactant liquor of heating is the dark brown red transparency liquid, and when being single dark brown red spot when checking with conventional thin layer chromatography, cessation reaction, filter the back and add glycerine in filtrate, the volume ratio of glycerine and ethanol consumption is 1: 1-2.
3, the technology of the described oxidized haematine solution of preparation claim 1 according to claim 2, it is characterized in that: described iodate is sodium iodate or Potassiumiodate.
4, a kind of haematine solution with the described oxidized haematine solution preparation of claim 1, it is characterized in that: in the described haematine solution mordanting bath is arranged, mordanting bath is inorganic salt solution or organic slat solution or phosphomolybdic acid or phosphotungstic acid.
5, according to the haematine solution with the preparation of the described oxidized haematine solution of claim 4, it is characterized in that: contain 1-5 part oxidized haematine solution in this haematine solution, 6-10 part mordanting bath, both amount ratios are volume ratio, the consisting of of described mordanting bath:
Distilled water 2000-4000 milliliter
Glycerine 100-500 milliliter
95% ethanol 1000-3000 milliliter
Glacial acetic acid 200-500 milliliter
12H
2OKAl (SO
4)
2Or 12H
2ONH
4Al (SO
4)
2Consumption be controlled between the 0.01-0.21M that accounts for above-mentioned several mixed liquid concentration.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB00109145XA CN1181344C (en) | 2000-06-12 | 2000-06-12 | Oxidized haematine solution and its preparing process and application |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB00109145XA CN1181344C (en) | 2000-06-12 | 2000-06-12 | Oxidized haematine solution and its preparing process and application |
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| Publication Number | Publication Date |
|---|---|
| CN1328259A CN1328259A (en) | 2001-12-26 |
| CN1181344C true CN1181344C (en) | 2004-12-22 |
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| Application Number | Title | Priority Date | Filing Date |
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| CNB00109145XA Expired - Fee Related CN1181344C (en) | 2000-06-12 | 2000-06-12 | Oxidized haematine solution and its preparing process and application |
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Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2012205849A1 (en) * | 2011-01-10 | 2013-07-04 | Ventana Medical Systems, Inc. | Hematoxylin staining method |
| CN102167914A (en) * | 2011-01-21 | 2011-08-31 | 哈尔滨格林标本技术开发有限公司 | Mercury-free environment-friendly hematoxylin staining solution |
| CN104264454A (en) * | 2014-09-27 | 2015-01-07 | 无锡市东北塘宏良染色厂 | Mordant for textiles |
| CN116426142A (en) * | 2023-04-03 | 2023-07-14 | 安徽信灵检验医学科技股份有限公司 | Hematoxylin staining solution and preparation method and application thereof |
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