CN105987840A - Citric acid antigen repairing solution - Google Patents
Citric acid antigen repairing solution Download PDFInfo
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- CN105987840A CN105987840A CN201610385942.5A CN201610385942A CN105987840A CN 105987840 A CN105987840 A CN 105987840A CN 201610385942 A CN201610385942 A CN 201610385942A CN 105987840 A CN105987840 A CN 105987840A
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- citric acid
- repair liquid
- monohydrate
- antigen repair
- tween
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- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 title claims abstract description 160
- 239000000427 antigen Substances 0.000 title claims abstract description 81
- 102000036639 antigens Human genes 0.000 title claims abstract description 81
- 108091007433 antigens Proteins 0.000 title claims abstract description 81
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims abstract description 57
- 229960004106 citric acid Drugs 0.000 claims abstract description 53
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 48
- 229920001213 Polysorbate 20 Polymers 0.000 claims abstract description 31
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 claims abstract description 31
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 claims abstract description 31
- GSEJCLTVZPLZKY-UHFFFAOYSA-N Triethanolamine Chemical compound OCCN(CCO)CCO GSEJCLTVZPLZKY-UHFFFAOYSA-N 0.000 claims abstract description 28
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 19
- 230000008439 repair process Effects 0.000 claims description 59
- 239000007788 liquid Substances 0.000 claims description 57
- OAGSFHDUINSAMQ-UHFFFAOYSA-N 2-hydroxypropane-1,2,3-tricarboxylic acid;sodium;hydrate Chemical compound O.[Na].OC(=O)CC(O)(C(O)=O)CC(O)=O OAGSFHDUINSAMQ-UHFFFAOYSA-N 0.000 claims description 38
- 241000195474 Sargassum Species 0.000 claims description 38
- LTUDISCZKZHRMJ-UHFFFAOYSA-N potassium;hydrate Chemical compound O.[K] LTUDISCZKZHRMJ-UHFFFAOYSA-N 0.000 claims description 38
- 241001474374 Blennius Species 0.000 claims description 36
- 238000002360 preparation method Methods 0.000 claims description 14
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 12
- 239000011268 mixed slurry Substances 0.000 claims description 12
- 230000005070 ripening Effects 0.000 claims description 12
- 239000002002 slurry Substances 0.000 claims description 12
- 230000001954 sterilising effect Effects 0.000 claims description 12
- 238000001914 filtration Methods 0.000 claims description 11
- 238000000605 extraction Methods 0.000 claims description 10
- 238000004090 dissolution Methods 0.000 claims description 8
- 230000005855 radiation Effects 0.000 claims description 8
- 238000010025 steaming Methods 0.000 claims description 8
- 238000004140 cleaning Methods 0.000 claims description 7
- 235000019750 Crude protein Nutrition 0.000 claims description 6
- 238000004108 freeze drying Methods 0.000 claims description 6
- 230000007935 neutral effect Effects 0.000 claims description 6
- 239000007787 solid Substances 0.000 claims description 6
- 230000000694 effects Effects 0.000 abstract description 11
- 238000003364 immunohistochemistry Methods 0.000 abstract description 9
- 238000010186 staining Methods 0.000 abstract description 2
- YASYEJJMZJALEJ-UHFFFAOYSA-N Citric acid monohydrate Chemical compound O.OC(=O)CC(O)(C(O)=O)CC(O)=O YASYEJJMZJALEJ-UHFFFAOYSA-N 0.000 abstract 1
- 229960002303 citric acid monohydrate Drugs 0.000 abstract 1
- 239000012153 distilled water Substances 0.000 abstract 1
- 230000002055 immunohistochemical effect Effects 0.000 abstract 1
- 230000008595 infiltration Effects 0.000 abstract 1
- 238000001764 infiltration Methods 0.000 abstract 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 abstract 1
- 229960000999 sodium citrate dihydrate Drugs 0.000 abstract 1
- 239000000872 buffer Substances 0.000 description 18
- 210000001519 tissue Anatomy 0.000 description 18
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 12
- 238000000034 method Methods 0.000 description 8
- 239000000243 solution Substances 0.000 description 7
- 238000005516 engineering process Methods 0.000 description 4
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 3
- 238000004043 dyeing Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 230000008901 benefit Effects 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- 239000000975 dye Substances 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- JAHNSTQSQJOJLO-UHFFFAOYSA-N 2-(3-fluorophenyl)-1h-imidazole Chemical compound FC1=CC=CC(C=2NC=CN=2)=C1 JAHNSTQSQJOJLO-UHFFFAOYSA-N 0.000 description 1
- OFNISBHGPNMTMS-UHFFFAOYSA-N 3-methylideneoxolane-2,5-dione Chemical compound C=C1CC(=O)OC1=O OFNISBHGPNMTMS-UHFFFAOYSA-N 0.000 description 1
- FHVDTGUDJYJELY-UHFFFAOYSA-N 6-{[2-carboxy-4,5-dihydroxy-6-(phosphanyloxy)oxan-3-yl]oxy}-4,5-dihydroxy-3-phosphanyloxane-2-carboxylic acid Chemical compound O1C(C(O)=O)C(P)C(O)C(O)C1OC1C(C(O)=O)OC(OP)C(O)C1O FHVDTGUDJYJELY-UHFFFAOYSA-N 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 229940072056 alginate Drugs 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- HNEGQIOMVPPMNR-IHWYPQMZSA-N citraconic acid Chemical compound OC(=O)C(/C)=C\C(O)=O HNEGQIOMVPPMNR-IHWYPQMZSA-N 0.000 description 1
- 229940018557 citraconic acid Drugs 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000000093 cytochemical effect Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000004945 emulsification Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000011532 immunohistochemical staining Methods 0.000 description 1
- 238000005470 impregnation Methods 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- LVHBHZANLOWSRM-UHFFFAOYSA-N methylenebutanedioic acid Natural products OC(=O)CC(=C)C(O)=O LVHBHZANLOWSRM-UHFFFAOYSA-N 0.000 description 1
- 239000012046 mixed solvent Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 210000004877 mucosa Anatomy 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 238000010827 pathological analysis Methods 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
Classifications
-
- 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
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Chemical & Material Sciences (AREA)
- General Physics & Mathematics (AREA)
- Urology & Nephrology (AREA)
- General Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Pathology (AREA)
- Biochemistry (AREA)
- Hematology (AREA)
- Cell Biology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Medicinal Preparation (AREA)
- Cosmetics (AREA)
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Abstract
The invention relates to the technical field of immunohistochemistry and discloses a citric acid antigen repairing solution. Each 1000mL of the citric acid antigen repairing solution contains 0.35g-0.40g of citric acid monohydrate, 2.40g-2.42g of sodium citrate dihydrate, 400-600 microliters of Tween 20, 50-500mL of triethanolamine or ethylene glycol or trihydric alcohol and the balance of single-stage distilled water. The citric acid antigen repairing solution has good infiltration and repairing effects and can be used for repairing tissue slices with relatively less use amount; and the repaired tissue slices are easy to stain and the staining effect is good. The repairing solution is not easy to volatilize and can be matched with a full-automatic immunohistochemical instrument for utilization.
Description
Technical field
The present invention relates to immunohistochemistry technology field, particularly relate to a kind of citric acid antigen repair liquid.
Background technology
Immunohistochemistry technology is the detection technique that pathological diagnosis in recent years develops rapidly, as a kind of important pathology aided diagnosis method, is increasingly subject to pay attention in clinical application.Along with the application of immunohistochemistry autostainer, the requirement for dyeing quality improves the most day by day.Wherein, antigen retrieval has extremely important effect during whole immunohistochemical staining, and the quality of repairing condition directly decides the quality of final coloration result.Full-automatic dyeing instrument is because of the construction features of himself, it is less for the container volume holding repair liquid, the condition as repaired by hand, tissue slice can not being fully immersed in repair liquid, and immunohistochemistry autostainer needs when antigen retrieval to carry out at higher temperature (about 100 DEG C), therefore repair liquid is readily volatilized, further reduces the volume of repair liquid.Thus, how completing normal antigen retrieval on immunohistochemistry autostainer is the difficult problem that immunohistochemistry autostainer must solve.
But owing to current immunohistochemistry autostainer is domestic not yet common, few people use, and therefore nobody notices above-mentioned technical problem.The most external method solving above-mentioned technical problem is typically to control the environment of antigen retrieval, as controlled atmospheric pressure, ambient temperature etc., and these solution complicated conditions, it is difficult to reality application.Also do not find to solve by improving antigen retrieval buffers the report of above-mentioned technical problem at present.
Current antigen retrieval buffers has a large amount in variety, as EDTA antigen retrieval buffers, citric acid antigen repair liquid and EDTA-citric acid compound antigen retrieval buffers etc..The Chinese patent of Application No. 201180024136.0 discloses a kind of antigen retrieval buffers and antigen retrieval method, the antigen retrieval in the cytochemical staining procedure of immuning tissue and cell.Described antigen retrieval buffers is itaconic acid, itaconic anhydride or citraconic acid aqueous solution.Described antigen retrieval method carries out high temperature or High Temperature High Pressure reparation for using above-mentioned antigen retrieval buffers to tissue slice.It is high that the antigen retrieval buffers provided and antigen retrieval method have dyeing quality, reproducibility, safety, the advantage of good stability.
The solvent of above-mentioned antigen retrieval buffers is water, if be applied on immunohistochemistry autostainer, on the one hand, this antigen retrieval buffers needs repairing effect in the case of consumption is big better, it is impossible to less consumption cannot by tissue slice thorough impregnation in the case of complete preferably to repair；On the other hand, its boiling point is low, the most readily volatilized, is easily caused tissue slice generation mummification situation, and mummification once occurs, and the antigen of tissue slice is changing to irreversible destruction and cannot repair.
Summary of the invention
In order to solve above-mentioned technical problem, the invention provides a kind of citric acid antigen repair liquid.The citric acid antigen repair liquid of the present invention infiltrates, repairing effect is good, it is possible to repairing tissue slice under less consumption, after reparation, tissue slice easily dyes, and Color is good.And repair liquid is the most volatile, it is possible to coordinate full-automatic SABC instrument to use.
The concrete technical scheme of the present invention is: a kind of citric acid antigen repair liquid, the citric acid antigen repair liquid of every 1000mL includes: monohydrate potassium 0.35-0.40g, two citric acid monohydrate sodium 2.40-2.42g, tween 20 400-600 microlitre, the list of triethanolamine or ethylene glycol or trihydroxylic alcohol 50-500mL and surplus steams water.
In the antigen retrieval buffers of the present invention, using monohydrate potassium and two citric acid monohydrate sodium as main repairing activity composition, and using triethanolamine or ethylene glycol or trihydroxylic alcohol with single water that steams as mixed solvent.Wherein, triethanolamine or ethylene glycol or the addition of trihydroxylic alcohol, it is possible to increase the boiling point of antigen retrieval buffers, prevents it from excessively volatilizing.And the solvent phase ratio of triethanolamine or ethylene glycol or trihydroxylic alcohol and other high point, its advantage is that the impact on the active component in antigen retrieval buffers is less, and it is preferable with the affinity of tissue slice, toxicity is low, the reparation of tissue slice will not cause too much impact, and wherein trihydroxylic alcohol, the effect of triethylamine alcohol are better than ethylene glycol.Tween 20 plays emulsification, it is possible to increase the compatibility of each material in antigen retrieval buffers.
As preferably, the citric acid antigen repair liquid of every 1000mL includes: monohydrate potassium 0.378252g, two citric acid monohydrate sodium 2.411784g, tween 20 500 microlitre, and the list of triethanolamine or ethylene glycol or trihydroxylic alcohol 150mL and surplus steams water.
As preferably, the citric acid antigen repair liquid of every 1000mL includes: monohydrate potassium 0.378252g, two citric acid monohydrate sodium 2.411784g, tween 20 500 microlitre, and the list of triethanolamine 150mL and surplus steams water.
As preferably, the citric acid antigen repair liquid of every 1000mL includes: monohydrate potassium 0.35-0.40g, two citric acid monohydrate sodium 2.40-2.42g, tween 20 400-600 microlitre, triethanolamine or ethylene glycol or trihydroxylic alcohol 50-500mL, the list of Sargassum extract 0.5-1.5g and surplus steams water.
As preferably, the citric acid antigen repair liquid of every 1000mL includes: monohydrate potassium 0.378252g, two citric acid monohydrate sodium 2.411784g, tween 20 500 microlitre, triethanolamine or ethylene glycol or trihydroxylic alcohol 150mL, the list of Sargassum extract 1g and surplus steams water.
As preferably, the citric acid antigen repair liquid of every 1000mL includes: monohydrate potassium 0.378252g, two citric acid monohydrate sodium 2.411784g, tween 20 500 microlitre, triethanolamine 150mL, and the list of Sargassum extract 1g and surplus steams water.
As preferably, described citric acid antigen repair liquid preparation method is as follows: weigh monohydrate potassium and two citric acid monohydrate sodium by proportioning, after single steaming water dissolution monohydrate potassium and two citric acid monohydrate sodium, add the triethanolamine of formula ratio or ethylene glycol or trihydroxylic alcohol, add tween 20 after being completely dissolved, be finally settled to rated capacity with single water that steams.
As preferably, described citric acid antigen repair liquid preparation method also can be as follows: weighs monohydrate potassium and two citric acid monohydrate sodium by proportioning, after single steaming water dissolution monohydrate potassium and two citric acid monohydrate sodium, add the triethanolamine of formula ratio or ethylene glycol or trihydroxylic alcohol, add Sargassum extract and tween 20 after being completely dissolved, be finally settled to rated capacity with single water that steams.
As preferably, the preparation method of described Sargassum extract is: the Sargassum after cleaning mixes with water 2-4:10 in mass ratio, and makes seaweed slurry with beater；Seaweed slurry is mixed with the hydrochloric acid solution that pH value is 3-5 1:1-2 by volume, obtain mixed slurry, mixed slurry is carried out microwave radiation exaraction, it is filtered to remove solids after extraction and obtains seaweed extracted liquor, to seaweed extracted liquor at 100-110 DEG C, ripening sterilizing 0.5-1.5h in the environment of 4-8MPa, can make par-tial polysaccharide degrade in maturing process；Seaweed extracted liquor after ripening sterilizing is filtered the crude protein removing degeneration；Seaweed extracted liquor pH regulator after filtering, to neutral, prepares Sargassum extract after last lyophilization.
In order to further optimize, the antigen retrieval buffers of the present invention also adds Sargassum extract.The Sargassum extract prepared in the present invention contains the materials such as natural alginate and natural polysaccharide, and biological mildness is good, good with tissue slice affinity.A small amount of Sargassum extract viscosity that liquid can be made certain soluble in water, lock water is effective, it is possible to prevent solvent volatilization in antigen retrieval buffers further.In addition, Sargassum extract can physical bond to the repairing activity composition in antigen retrieval buffers, after Sargassum extract contacts with tissue slice, one layer of thin mucosa can be formed on tissue slice surface, so that repairing activity composition in connection can fully contact with tissue slice, in the case of less consumption antigen retrieval buffers, improve repairing effect.But the used in amounts of Sargassum extract if consumption is too much, can cause antigen retrieval fluid viscosity too high strictly control, and mobility is the poorest, and is combined excessively with tissue slice, is difficult to follow-up cleaning.
As preferably, the condition of described microwave radiation exaraction is: temperature 70-90 DEG C, time 5-10min, microwave power 800-1000W, microwave frequency 2450MHZ.
As preferably, described trihydroxylic alcohol can be glycerol.
It is compared with the prior art, the invention has the beneficial effects as follows:
The citric acid antigen repair liquid of the present invention infiltrates, repairing effect is good, it is possible to repair tissue slice under less consumption.Repair liquid needed for averagely every tissue slice only needs 100-150uL.
After reparation, tissue slice easily dyes, and Color is good.
And repair liquid is the most volatile, it is possible to coordinate full-automatic SABC instrument to use.
Detailed description of the invention
Below in conjunction with embodiment, the invention will be further described.
Embodiment 1
A kind of citric acid antigen repair liquid, the citric acid antigen repair liquid of every 1000mL includes: monohydrate potassium 0.378252g, two citric acid monohydrate sodium 2.411784g, tween 20 500 microlitre, and the list of triethanolamine 150mL and surplus steams water.
Preparation method is as follows: weigh monohydrate potassium and two citric acid monohydrate sodium by proportioning, after single steaming water dissolution monohydrate potassium and two citric acid monohydrate sodium, add the triethanolamine of formula ratio, add tween 20 after being completely dissolved, be finally settled to rated capacity with single water that steams.
Embodiment 2
A kind of citric acid antigen repair liquid, the citric acid antigen repair liquid of every 1000mL includes: monohydrate potassium 0.378252g, two citric acid monohydrate sodium 2.411784g, tween 20 500 microlitre, and the list of ethylene glycol 325mL and surplus steams water.
Preparation method is as follows: weigh monohydrate potassium and two citric acid monohydrate sodium by proportioning, after single steaming water dissolution monohydrate potassium and two citric acid monohydrate sodium, add the ethylene glycol of formula ratio, add tween 20 after being completely dissolved, be finally settled to rated capacity with single water that steams.
Embodiment 3
A kind of citric acid antigen repair liquid, the citric acid antigen repair liquid of every 1000mL includes: monohydrate potassium 0.35g, two citric acid monohydrate sodium 2.40g, tween 20 400 microlitre, and the list of triethanolamine 50mL and surplus steams water.
Embodiment 4
A kind of citric acid antigen repair liquid, the citric acid antigen repair liquid of every 1000mL includes: monohydrate potassium 0.40g, two citric acid monohydrate sodium 2.42g, tween 20 600 microlitre, and the list of triethanolamine 500mL and surplus steams water.
Embodiment 5
A kind of citric acid antigen repair liquid, the citric acid antigen repair liquid of every 1000mL includes: monohydrate potassium 0.378252g, two citric acid monohydrate sodium 2.411784g, tween 20 500 microlitre, and the list of glycerol 300mL and surplus steams water.
Embodiment 6
A kind of citric acid antigen repair liquid, the citric acid antigen repair liquid of every 1000mL includes: monohydrate potassium 0.378252g, two citric acid monohydrate sodium 2.411784g, tween 20 500 microlitre, triethanolamine 150mL, and the list of Sargassum extract 1g and surplus steams water.
Preparation method is as follows: weigh monohydrate potassium and two citric acid monohydrate sodium by proportioning, after single steaming water dissolution monohydrate potassium and two citric acid monohydrate sodium, add the triethanolamine of formula ratio, add Sargassum extract and tween 20 after being completely dissolved, be finally settled to rated capacity with single water that steams.
The preparation method of described Sargassum extract is: the Sargassum after cleaning mixes with water 3:10 in mass ratio, and makes seaweed slurry with beater；Being mixed with the hydrochloric acid solution that pH value is 4 1:1.5 by volume by seaweed slurry, obtain mixed slurry, mixed slurry carries out microwave radiation exaraction, wherein extraction conditions is: temperature 80 DEG C, time 7.5min, microwave power 900W, microwave frequency 2450MHZ.It is filtered to remove solids after extraction and obtains seaweed extracted liquor, to seaweed extracted liquor at 105 DEG C, ripening sterilizing 1h in the environment of 6MPa；Carry out the seaweed extracted liquor after ripening sterilizing filtering and remove crude protein；Seaweed extracted liquor pH regulator after filtering, to neutral, prepares Sargassum extract after last lyophilization.
Embodiment 7
A kind of citric acid antigen repair liquid, the citric acid antigen repair liquid of every 1000mL includes: the citric acid antigen repair liquid of every 1000mL includes: monohydrate potassium 0.378252g, two citric acid monohydrate sodium 2.411784g, tween 20 500 microlitre, triethanolamine 150mL, the list of Sargassum extract 0.5g and surplus steams water.
The preparation method of described Sargassum extract is: the Sargassum after cleaning mixes with water 2:10 in mass ratio, and makes seaweed slurry with beater；Being mixed with the hydrochloric acid solution that pH value is 3-5 1:1 by volume by seaweed slurry, obtain mixed slurry, mixed slurry carries out microwave radiation exaraction, wherein extraction conditions is: temperature 70 C, time 10min, microwave power 800W, microwave frequency 2450MHZ.It is filtered to remove solids after extraction and obtains seaweed extracted liquor, to seaweed extracted liquor at 100 DEG C, ripening sterilizing 0.5h in the environment of 8MPa；Carry out the seaweed extracted liquor after ripening sterilizing filtering and remove crude protein；Seaweed extracted liquor pH regulator after filtering, to neutral, prepares Sargassum extract after last lyophilization.
Embodiment 8
The citric acid antigen repair liquid of every 1000mL includes: monohydrate potassium 0.378252g, two citric acid monohydrate sodium 2.411784g, tween 20 500 microlitre, triethanolamine 150mL, and the list of Sargassum extract 1.5g and surplus steams water.
The preparation method of described Sargassum extract is: the Sargassum after cleaning mixes with water 4:10 in mass ratio, and makes seaweed slurry with beater；Being mixed with the hydrochloric acid solution that pH value is 3-5 1:2 by volume by seaweed slurry, obtain mixed slurry, mixed slurry carries out microwave radiation exaraction, wherein extraction conditions is: temperature 90 DEG C, time 5min, microwave power 1000W, microwave frequency 2450MHZ.It is filtered to remove solids after extraction and obtains seaweed extracted liquor, to seaweed extracted liquor at 110 DEG C, ripening sterilizing 1.5h in the environment of 4MPa；Carry out the seaweed extracted liquor after ripening sterilizing filtering and remove crude protein；Seaweed extracted liquor pH regulator after filtering, to neutral, prepares Sargassum extract after last lyophilization.
Embodiment 9
A kind of citric acid antigen repair liquid, the citric acid antigen repair liquid of every 1000mL includes: monohydrate potassium 0.378252g, two citric acid monohydrate sodium 2.411784g, tween 20 500 microlitre, glycerol 150mL, and the list of Sargassum extract 1g and surplus steams water.
Preparation method is as follows: weigh monohydrate potassium and two citric acid monohydrate sodium by proportioning, after single steaming water dissolution monohydrate potassium and two citric acid monohydrate sodium, add the glycerol of formula ratio, add Sargassum extract and tween 20 after being completely dissolved, be finally settled to rated capacity with single water that steams.
The preparation method of described Sargassum extract is: the Sargassum after cleaning mixes with water 3:10 in mass ratio, and makes seaweed slurry with beater；Being mixed with the hydrochloric acid solution that pH value is 4 1:1.5 by volume by seaweed slurry, obtain mixed slurry, mixed slurry carries out microwave radiation exaraction, wherein extraction conditions is: temperature 80 DEG C, time 7.5min, microwave power 900W, microwave frequency 2450MHZ.It is filtered to remove solids after extraction and obtains seaweed extracted liquor, to seaweed extracted liquor at 105 DEG C, ripening sterilizing 1h in the environment of 6MPa；Carry out the seaweed extracted liquor after ripening sterilizing filtering and remove crude protein；Seaweed extracted liquor pH regulator after filtering, to neutral, prepares Sargassum extract after last lyophilization.Raw materials used, equipment in the present invention, unless otherwise noted, is the conventional raw material of this area, equipment；Method therefor in the present invention, unless otherwise noted, is the conventional method of this area.
The above, be only presently preferred embodiments of the present invention, not impose any restrictions the present invention, every any simple modification, change and equivalent transformation made above example according to the technology of the present invention essence, all still falls within the protection domain of technical solution of the present invention.
Claims (10)
1. a citric acid antigen repair liquid, it is characterized in that: the citric acid antigen repair liquid of every 1000mL includes: monohydrate potassium 0.35-0.40g, two citric acid monohydrate sodium 2.40-2.42g, tween 20 400-600 microlitre, the list of triethanolamine or ethylene glycol or trihydroxylic alcohol 50-500mL and surplus steams water.
2. a kind of citric acid antigen repair liquid as claimed in claim 1, it is characterized in that, the citric acid antigen repair liquid of every 1000mL includes: monohydrate potassium 0.378252g, two citric acid monohydrate sodium 2.411784g, tween 20 500 microlitre, the list of triethanolamine or ethylene glycol or trihydroxylic alcohol 150mL and surplus steams water.
3. a kind of citric acid antigen repair liquid as claimed in claim 2, it is characterized in that, the citric acid antigen repair liquid of every 1000mL includes: monohydrate potassium 0.378252g, two citric acid monohydrate sodium 2.411784g, tween 20 500 microlitre, the list of triethanolamine 150mL and surplus steams water.
4. a kind of citric acid antigen repair liquid as claimed in claim 1, it is characterized in that, the citric acid antigen repair liquid of every 1000mL includes: monohydrate potassium 0.35-0.40g, two citric acid monohydrate sodium 2.40-2.42g, tween 20 400-600 microlitre, triethanolamine or ethylene glycol or trihydroxylic alcohol 50-500mL, the list of Sargassum extract 0.5-1.5g and surplus steams water.
5. a kind of citric acid antigen repair liquid as claimed in claim 4, it is characterized in that, the citric acid antigen repair liquid of every 1000mL includes: monohydrate potassium 0.378252g, two citric acid monohydrate sodium 2.411784g, tween 20 500 microlitre, triethanolamine or ethylene glycol or trihydroxylic alcohol 150mL, the list of Sargassum extract 1g and surplus steams water.
6. a kind of citric acid antigen repair liquid as claimed in claim 5, it is characterized in that, the citric acid antigen repair liquid of every 1000mL includes: monohydrate potassium 0.378252g, two citric acid monohydrate sodium 2.411784g, tween 20 500 microlitre, triethanolamine 150mL, the list of Sargassum extract 1g and surplus steams water.
7. a kind of citric acid antigen repair liquid as described in one of claim 1-3, it is characterized in that preparation method is as follows: weigh monohydrate potassium and two citric acid monohydrate sodium by proportioning, after single steaming water dissolution monohydrate potassium and two citric acid monohydrate sodium, add the triethanolamine of formula ratio or ethylene glycol or trihydroxylic alcohol, add tween 20 after being completely dissolved, be finally settled to rated capacity with single water that steams.
8. a kind of citric acid antigen repair liquid as described in one of claim 4-6, it is characterized in that preparation method is as follows: weigh monohydrate potassium and two citric acid monohydrate sodium by proportioning, after single steaming water dissolution monohydrate potassium and two citric acid monohydrate sodium, add the triethanolamine of formula ratio or ethylene glycol or trihydroxylic alcohol, add Sargassum extract and tween 20 after being completely dissolved, be finally settled to rated capacity with single water that steams.
9. a kind of citric acid antigen repair liquid as described in one of claim 4-6, it is characterised in that the preparation method of described Sargassum extract is: the Sargassum after cleaning mixes with water 2-4:10 in mass ratio, and makes seaweed slurry with beater；Seaweed slurry is mixed with the hydrochloric acid solution that pH value is 3-5 1:1-2 by volume, obtain mixed slurry, mixed slurry is carried out microwave radiation exaraction, be filtered to remove solids after extraction and obtain seaweed extracted liquor, to seaweed extracted liquor at 100-110 DEG C, ripening sterilizing 0.5-1.5h in the environment of 4-8MPa；Carry out the seaweed extracted liquor after ripening sterilizing filtering and remove crude protein；Seaweed extracted liquor pH regulator after filtering, to neutral, prepares Sargassum extract after last lyophilization.
10. a kind of citric acid antigen repair liquid as claimed in claim 9, it is characterised in that the condition of described microwave radiation exaraction is: temperature 70-90 DEG C, time 5-10min, microwave power 800-1000W, microwave frequency 2450MHZ.
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