CN105561346B - A kind of preparation method of acoustic contrast agent - Google Patents
A kind of preparation method of acoustic contrast agent Download PDFInfo
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- CN105561346B CN105561346B CN201511015312.0A CN201511015312A CN105561346B CN 105561346 B CN105561346 B CN 105561346B CN 201511015312 A CN201511015312 A CN 201511015312A CN 105561346 B CN105561346 B CN 105561346B
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- buffered saline
- saline solution
- phosphate buffered
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- 239000002872 contrast media Substances 0.000 title claims abstract description 33
- 238000002360 preparation method Methods 0.000 title claims abstract description 8
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims abstract description 46
- 241000287828 Gallus gallus Species 0.000 claims abstract description 38
- 239000006228 supernatant Substances 0.000 claims abstract description 33
- 239000013049 sediment Substances 0.000 claims abstract description 32
- 239000000243 solution Substances 0.000 claims abstract description 28
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 claims abstract description 26
- 239000002953 phosphate buffered saline Substances 0.000 claims abstract description 26
- 239000008280 blood Substances 0.000 claims abstract description 24
- 210000004369 blood Anatomy 0.000 claims abstract description 24
- 235000011187 glycerol Nutrition 0.000 claims abstract description 23
- 239000012267 brine Substances 0.000 claims abstract description 20
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical compound O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 claims abstract description 20
- 239000003918 blood extract Substances 0.000 claims abstract description 16
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 claims abstract description 14
- 238000005119 centrifugation Methods 0.000 claims abstract description 13
- 102000004882 Lipase Human genes 0.000 claims abstract description 8
- 108090001060 Lipase Proteins 0.000 claims abstract description 8
- 239000004367 Lipase Substances 0.000 claims abstract description 8
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims abstract description 8
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims abstract description 8
- 235000011130 ammonium sulphate Nutrition 0.000 claims abstract description 8
- 235000019421 lipase Nutrition 0.000 claims abstract description 8
- 238000002156 mixing Methods 0.000 claims abstract description 7
- 235000010344 sodium nitrate Nutrition 0.000 claims abstract description 7
- 239000004317 sodium nitrate Substances 0.000 claims abstract description 7
- 238000000034 method Methods 0.000 claims abstract description 6
- 238000003756 stirring Methods 0.000 claims description 24
- 238000007710 freezing Methods 0.000 claims description 6
- 230000008014 freezing Effects 0.000 claims description 6
- 239000000155 melt Substances 0.000 claims description 6
- 150000002500 ions Chemical class 0.000 claims description 2
- 238000004587 chromatography analysis Methods 0.000 claims 1
- 230000002708 enhancing effect Effects 0.000 abstract description 12
- 230000000694 effects Effects 0.000 abstract description 10
- 238000003384 imaging method Methods 0.000 abstract description 10
- 230000017531 blood circulation Effects 0.000 abstract description 9
- 230000004088 pulmonary circulation Effects 0.000 abstract description 5
- 239000011248 coating agent Substances 0.000 abstract description 4
- 238000000576 coating method Methods 0.000 abstract description 4
- 239000000047 product Substances 0.000 abstract description 4
- 238000004821 distillation Methods 0.000 abstract description 3
- 238000004090 dissolution Methods 0.000 abstract description 2
- 239000002994 raw material Substances 0.000 abstract description 2
- 230000009286 beneficial effect Effects 0.000 abstract 1
- 238000000746 purification Methods 0.000 abstract 1
- 238000005406 washing Methods 0.000 abstract 1
- 238000002604 ultrasonography Methods 0.000 description 11
- 239000007924 injection Substances 0.000 description 8
- 238000002347 injection Methods 0.000 description 8
- 239000007788 liquid Substances 0.000 description 6
- 210000004204 blood vessel Anatomy 0.000 description 4
- 238000004255 ion exchange chromatography Methods 0.000 description 4
- 238000009938 salting Methods 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 2
- XSTXAVWGXDQKEL-UHFFFAOYSA-N Trichloroethylene Chemical compound ClC=C(Cl)Cl XSTXAVWGXDQKEL-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 230000003139 buffering effect Effects 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 229910052698 phosphorus Inorganic materials 0.000 description 2
- 239000011574 phosphorus Substances 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- JFDRXEGUWGGADQ-UHFFFAOYSA-N [Na+].[O-][N+]([O-])=O.OCC(O)CO Chemical compound [Na+].[O-][N+]([O-])=O.OCC(O)CO JFDRXEGUWGGADQ-UHFFFAOYSA-N 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000008081 blood perfusion Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000000084 colloidal system Substances 0.000 description 1
- 238000002086 displacement chromatography Methods 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 230000002526 effect on cardiovascular system Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 239000006210 lotion Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/22—Echographic preparations; Ultrasound imaging preparations ; Optoacoustic imaging preparations
- A61K49/222—Echographic preparations; Ultrasound imaging preparations ; Optoacoustic imaging preparations characterised by a special physical form, e.g. emulsions, liposomes
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Acoustics & Sound (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Radiology & Medical Imaging (AREA)
- Epidemiology (AREA)
- Physics & Mathematics (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The invention discloses a kind of preparation methods of acoustic contrast agent, belong to acoustic contrast agent preparing technical field.The present invention after chicken blood is freezed dissolution process repeatedly, is mixed with fatty with chicken blood for raw material with brine, and standing centrifuges to obtain supernatant, adds in phosphate buffered saline solution mixing and lower sediment thing is collected by centrifugation, obtain chicken blood extract after purification；Fat is chosen again adds in lipase enzymolysis, centrifuge to obtain supernatant, distillation is mixed after concentration with sodium nitrate, collect fraction glycerine, it is mixed with chicken blood extract, ammonium sulfate and phosphate buffered saline solution, sediment is centrifuged to obtain, is mixed again with phosphate buffered saline solution after washing, handled through ultrasonic disperse, centrifuge obtained acoustic contrast agent.The beneficial effects of the invention are as follows：Products obtained therefrom coating is moderate, and elasticity is good, and the residence time in blood circulation is long, and bubble size is uniform, is not easily broken；Pulmonary circulation can be passed through during use, effectively enhancing developing time is longer, imaging effect is good.
Description
Technical field
The present invention relates to a kind of preparation methods of acoustic contrast agent, belong to acoustic contrast agent preparing technical field.
Background technology
Acoustic contrast agent abbreviation UCA is a kind of chemicals that can significantly increase ultrasonic backscattering intensity.Its mainly into
It is microbubble to divide, and general a diameter of 2~10um can pass through pulmonary circulation.Earliest acoustic contrast agent is carbonated oxygen
Or the microbubble of air, mainly by hand shake physiological saline acquisition be simply possible to use in the right side feel concerned about system imaging.By improving purpose
The acoustics contrast of tissue and its surrounding tissue, and then improve the diagnostic reagent of medical ultrasonic echo-signal.Typically by outer
All veins, which inject contrast agent, enters the in vivo circulatory system, enters each tissue of whole body and organ, the acoustics of blood with blood
Gas component is completely different in resistance value and acoustic contrast agent, and ultrasonic wave cannot be caused linearly or nonlinearly ultrasonic by microvesicle
Scattering can improve the echo-signal of blood flow, so as to reflect the blood perfusion situation of cardiovascular organization organ and tumor tissues.
Current acoustic contrast agent mainly with experiment dyestuff, physiological saline, lotion, colloid and contains free bubble
Liquid is material preparation, but since its coating is thicker, poor flexibility, and also the air that wraps up is soluble easily in water etc., in blood circulation
In residence time is of short duration, bubble size not enough uniformly, be easily broken, it is impossible to pass through pulmonary circulation, effectively enhancing development
The shortcomings of time is shorter, imaging effect is poor so as to limit the time observed and diagnosed in clinical practice, could not be also reached in clinic
Upper realization is widely applied.
The content of the invention
The technical problems to be solved by the invention：For current acoustic contrast agent since its coating is thicker, poor flexibility, and
And the air of package is soluble easily in water etc., the residence time in blood circulation is of short duration, bubble size not enough uniformly, be easily broken,
Cause it that cannot pass through pulmonary circulation, effectively enhance that developing time is shorter, the drawbacks of imaging effect is poor, provide it is a kind of with
Chicken blood and fat is raw materials, after chicken blood is freezed dissolution process repeatedly, mixs with brine, phosphate buffered saline solution through centrifuging, is pure
Chicken blood extract is obtained after change；Fat is chosen again and adds in lipase enzymolysis, is centrifuged to obtain supernatant, is mixed distillation after concentration with sodium nitrate
Glycerine is obtained, is mixed with chicken blood extract, ammonium sulfate and phosphate buffered saline solution, then is mixed with phosphate buffered saline solution, through super
The method that acoustic contrast agent is made in sound decentralized processing, centrifugation.Preparation method of the present invention is simple, products obtained therefrom size uniform, uses
Effectively enhancing developing time is longer afterwards, imaging effect is good.
In order to solve the above technical problems, the present invention is using technical solution as described below：
（1）Fresh chicken blood is collected, is put at -20~-15 DEG C 2~3h of freezing, after the completion of to be frozen, put it into 25~
Melt naturally at 32 DEG C, after it melts completely, place into 2~3h of freezing at -20~-15 DEG C, so cycle 3~4 times, obtain pre-
The chicken blood of processing；
（2）By brine and container volume than 1:3, the brine that mass fraction is 0.75~0.9% is fitted into container, simultaneously
The chicken blood of above-mentioned pretreatment is added in, addition is 1 with brine quality ratio:2,1~2h is stood, then places it in refrigerated centrifuge
In, it is 3~9 DEG C in temperature, rotating speed is that 10~15min is centrifuged under 4000r/min, obtains supernatant；
（3）By phosphate buffered saline solution and supernatant volume than 1:20, concentration is molten for the phosphate-buffered salt of 0.01mol/L
Liquid is added dropwise to the speed of 2~3 drops/s in the supernatant of above-mentioned gained, and after stirring is sufficiently mixed it, 1 is stood at 3~5 DEG C
Then~2h is 3~9 DEG C in temperature, rotating speed is that 15~25min is centrifuged under 4000r/min, lower sediment thing is collected, through ion
Displacement chromatography is purified, and it is spare to obtain chicken blood extract；
（4）The fat newly extracted is chosen, by fat and hermetically sealed can volume ratio 2:3, fat is put into hermetically sealed can, while to
The lipase of fat mass 3~5% is wherein added, in the case where temperature is 37~40 DEG C, 2~3h is digested, waits after the completion of digesting, by enzyme
Solution object is placed in 8~10min of centrifugation in the centrifuge of 2000~3000r/min, collects supernatant；
（5）The supernatant of above-mentioned collection is rotated to the half for being evaporated to original volume, Ran Houxiang at 40~50 DEG C
Sodium nitrate is wherein added in, addition is 2~3mg/L, after stirring makes it be sufficiently mixed uniformly, is put into the distillation with condenser pipe and burns
In bottle, it is carried out to be heated to 80~90 DEG C, reacts 2~3h, collects the fraction in condenser pipe, i.e. glycerine；
（6）While stirring to step（3）The glycerine of above-mentioned gained is added in spare chicken blood extract, then is divided thereto
Not Jia Ru glycerine quality 3% ammonium sulfate and glycerine quality 3% concentration be 0.01mol/L phosphate buffered saline solution, stir
It mixes after it is made fully to dissolve, stands 2~3h, be subsequently placed in 20~30min of centrifugation in the centrifuge of 3000r/min, collect lower floor
Sediment；
（7）The lower sediment thing of collection is washed with deionized 3~4 times, then adding in concentration thereto is
The phosphate buffered saline solution of 0.01mol/L, addition are 1 with lower sediment thing mass ratio:25, after mixing, it is placed in ultrasound
1~2h of decentralized processing in ripple places into centrifuge collected after centrifugation lower sediment thing to get a kind of acoustic contrast agent.
The application process of the present invention：In B ultrasound, acoustic contrast agent produced by the present invention is injected in blood vessel, injection rate is
1~3mg every time can obtain very strong B ultrasound echo after injection, on the image clearer display vessel position and size.This is super
Residence time of the sound contrast agent in blood circulation and effective enhancing developing time are long, imaging effect is good, and enhancing efficiency is higher than it
His acoustic contrast agent more than 21.6%, is worthy to be popularized and uses.
Compared with other methods, advantageous effects are the present invention：
（1）Products obtained therefrom coating of the present invention is moderate, and elasticity is good, and the residence time in blood circulation is long, and bubble size is equal
It is even, it is not easily broken；
（2）Pulmonary circulation can be passed through during use, effectively enhancing developing time is longer, imaging effect is good.
Specific embodiment
Fresh chicken blood is collected first, is put into 2~3h of freezing at -20~-15 DEG C, after the completion of to be frozen, is put it into 25
Melt naturally at~32 DEG C, after it melts completely, place into 2~3h of freezing at -20~-15 DEG C, so cycle 3~4 times, obtain
The chicken blood of pretreatment；Then by brine and container volume than 1:3, the brine that mass fraction is 0.75~0.9% is packed into container
In, while the chicken blood of above-mentioned pretreatment is added in, addition is 1 with brine quality ratio:2,1~2h is stood, is then placed it in cold
Freeze in centrifuge, be 3~9 DEG C in temperature, rotating speed is that 10~15min is centrifuged under 4000r/min, obtains supernatant；Delay again by phosphoric acid
Salting liquid is rushed with supernatant volume than 1:20, by concentration be 0.01mol/L phosphate buffered saline solution with the speed of 2~3 drops/s
It is added dropwise in the supernatant of above-mentioned gained, after stirring is sufficiently mixed it, 1~2h is stood at 3~5 DEG C, be then 3 in temperature
~9 DEG C, rotating speed is that 15~25min is centrifuged under 4000r/min, collects lower sediment thing, is purified, obtained through ion-exchange chromatography
Chicken blood extract is spare；The fat newly extracted is chosen, by fat and hermetically sealed can volume ratio 2:3, fat is put into hermetically sealed can, together
When add the lipase of fat mass 3~5% thereto, in the case where temperature is 37~40 DEG C, digest 2~3h, wait after the completion of digesting,
Zymolyte is placed in 8~10min of centrifugation in the centrifuge of 2000~3000r/min, is collected supernatant；By the supernatant of above-mentioned collection
Liquid rotates the half for being evaporated to original volume at 40~50 DEG C, then adds in sodium nitrate thereto, addition for 2~
3mg/L after stirring makes it be sufficiently mixed uniformly, is put into the distilling flask with condenser pipe, it is carried out to be heated to 80~90
DEG C, 2~3h is reacted, collects the fraction in condenser pipe, i.e. glycerine；It is added in while stirring into spare chicken blood extract above-mentioned
The glycerine of gained, then the ammonium sulfate of glycerine quality 3% is separately added into thereto and the concentration of glycerine quality 3% is
The phosphate buffered saline solution of 0.01mol/L, stirring it is made fully to dissolve after, stand 2~3h, be subsequently placed in 3000r/min from
20~30min is centrifuged in scheming, collects lower sediment thing；The lower sediment thing of collection is finally washed with deionized 3~4
It is secondary, the phosphate buffered saline solution that concentration is 0.01mol/L is then added in thereto, and addition is 1 with lower sediment thing mass ratio:
25, after mixing, 1~2h of decentralized processing in ultrasonic wave is placed in, places into collected after centrifugation lower sediment thing in centrifuge, i.e.,
Obtain a kind of acoustic contrast agent.
Example 1
Fresh chicken blood is collected first, is put at -15 DEG C and freezes 2h, after the completion of to be frozen, is put it into natural at 25 DEG C
Melt, after it melts completely, place at -15 DEG C and freeze 2h, so Xun Huan 3 times, obtain the chicken blood of pretreatment；Then brine is pressed
With container volume than 1:3, the brine that mass fraction is 0.75% is fitted into container, while the chicken blood of above-mentioned pretreatment is added in, add
It is 1 to enter amount with brine quality ratio:2,1h is stood, is then placed it in refrigerated centrifuge, is 3 DEG C in temperature, rotating speed is
10min is centrifuged under 4000r/min, obtains supernatant；Again by phosphate buffered saline solution and supernatant volume than 1:20, be by concentration
The phosphate buffered saline solution of 0.01mol/L is added dropwise to the speed of 2 drops/s in the supernatant of above-mentioned gained, and stirring makes it fully mixed
After conjunction, 1h is stood at 3 DEG C, is then 3 DEG C in temperature, rotating speed is that 15min is centrifuged under 4000r/min, collects lower sediment thing,
It is purified through ion-exchange chromatography, it is spare to obtain chicken blood extract；The fat newly extracted is chosen, by fat and hermetically sealed can volume ratio
2:3, fat is put into hermetically sealed can, while the lipase of fat mass 3% is added thereto, and in the case where temperature is 37 DEG C, enzymolysis
2h is waited after the completion of digesting, and zymolyte is placed in the centrifuge of 2000r/min and is centrifuged 8min, collected supernatant；By above-mentioned collection
Supernatant the half for being evaporated to original volume is rotated at 40 DEG C, then add in sodium nitrate thereto, addition is
2mg/L after stirring makes it be sufficiently mixed uniformly, is put into the distilling flask with condenser pipe, it is carried out to be heated to 80 DEG C, instead
2h is answered, collects the fraction in condenser pipe, i.e. glycerine；While stirring the third of above-mentioned gained is added in into spare chicken blood extract
Triol, then the phosphorus of the ammonium sulfate of glycerine quality 3% and the concentration of glycerine quality 3% for 0.01mol/L is separately added into thereto
Acid buffering salting liquid after stirring makes it fully dissolve, stands 2h, is subsequently placed in the centrifuge of 3000r/min and centrifuges 20min,
Collect lower sediment thing；Finally the lower sediment thing of collection is washed with deionized 3 times, then adding in concentration thereto is
The phosphate buffered saline solution of 0.01mol/L, addition are 1 with lower sediment thing mass ratio:25, after mixing, it is placed in ultrasound
Decentralized processing 1h in ripple places into centrifuge collected after centrifugation lower sediment thing to get a kind of acoustic contrast agent.
In B ultrasound, acoustic contrast agent produced by the present invention is injected in blood vessel, injection rate is each 1mg, can be obtained after injection
It is clearer on the image to show vessel position and size to very strong B ultrasound echo.The acoustic contrast agent is in blood circulation
Residence time and effective enhancing developing time are long, imaging effect is good, and enhancing efficiency is worth higher than other acoustic contrast agents 21.69%
It promotes and uses.
Example 2
Fresh chicken blood is collected first, is put at -18 DEG C and freezes 2.5h, after the completion of to be frozen, is put it at 28 DEG C certainly
So melt, after it melts completely, place at -18 DEG C and freeze 2.5h, so Xun Huan 4 times, obtain the chicken blood of pretreatment；Then press
Brine is with container volume than 1:3, the brine that mass fraction is 0.83% is fitted into container, while adds in the chicken of above-mentioned pretreatment
Blood, addition are 1 with brine quality ratio:2,1.5h is stood, is then placed it in refrigerated centrifuge, is 6 DEG C in temperature, rotating speed
To centrifuge 13min under 4000r/min, supernatant is obtained；Again by phosphate buffered saline solution and supernatant volume than 1:20, be by concentration
The phosphate buffered saline solution of 0.01mol/L is added dropwise to the speed of 3 drops/s in the supernatant of above-mentioned gained, and stirring makes it fully mixed
After conjunction, 1.5h is stood at 4 DEG C, is then 6 DEG C in temperature, rotating speed is that 20min is centrifuged under 4000r/min, collects lower sediment
Object is purified through ion-exchange chromatography, and it is spare to obtain chicken blood extract；The fat newly extracted is chosen, by fat and sealed shell of tank
Product ratio 2:3, fat is put into hermetically sealed can, while the lipase of fat mass 4% is added thereto, and in the case where temperature is 39 DEG C, enzyme
2.5h is solved, is waited after the completion of digesting, zymolyte is placed in the centrifuge of 2500r/min and centrifuges 9min, collected supernatant；It will be above-mentioned
The supernatant of collection rotates the half for being evaporated to original volume at 45 DEG C, then adds in sodium nitrate, addition thereto
For 3mg/L, after stirring makes it be sufficiently mixed uniformly, it is put into the distilling flask with condenser pipe, it is carried out to be heated to 85 DEG C,
2.5h is reacted, collects the fraction in condenser pipe, i.e. glycerine；While stirring above-mentioned gained is added in into spare chicken blood extract
Glycerine, then be separately added into thereto glycerine quality 3% ammonium sulfate and glycerine quality 3% concentration be 0.01mol/L
Phosphate buffered saline solution, stirring it is made fully to dissolve after, stand 2.5h, be subsequently placed in the centrifuge of 3000r/min and centrifuge
25min collects lower sediment thing；Finally the lower sediment thing of collection is washed with deionized 4 times, is then added in thereto dense
The phosphate buffered saline solution for 0.01mol/L is spent, addition is 1 with lower sediment thing mass ratio:25, after mixing, it is placed in
Decentralized processing 1.5h in ultrasonic wave places into centrifuge collected after centrifugation lower sediment thing to get a kind of acoustic contrast agent.
In B ultrasound, acoustic contrast agent produced by the present invention is injected in blood vessel, injection rate is each 2mg, can be obtained after injection
It is clearer on the image to show vessel position and size to very strong B ultrasound echo.The acoustic contrast agent is in blood circulation
Residence time and effective enhancing developing time are long, imaging effect is good, and enhancing efficiency is worth higher than other acoustic contrast agents 22.78%
It promotes and uses.
Example 3
Fresh chicken blood is collected first, is put at -20 DEG C and freezes 3h, after the completion of to be frozen, is put it into natural at 32 DEG C
Melt, after it melts completely, place at -20 DEG C and freeze 3h, so Xun Huan 4 times, obtain the chicken blood of pretreatment；Then brine is pressed
With container volume than 1:3, the brine that mass fraction is 0.9% is fitted into container, while the chicken blood of above-mentioned pretreatment is added in, add
It is 1 to enter amount with brine quality ratio:2,2h is stood, is then placed it in refrigerated centrifuge, is 9 DEG C in temperature, rotating speed is
15min is centrifuged under 4000r/min, obtains supernatant；Again by phosphate buffered saline solution and supernatant volume than 1:20, be by concentration
The phosphate buffered saline solution of 0.01mol/L is added dropwise to the speed of 3 drops/s in the supernatant of above-mentioned gained, and stirring makes it fully mixed
After conjunction, 2h is stood at 5 DEG C, is then 9 DEG C in temperature, rotating speed is that 25min is centrifuged under 4000r/min, collects lower sediment thing,
It is purified through ion-exchange chromatography, it is spare to obtain chicken blood extract；The fat newly extracted is chosen, by fat and hermetically sealed can volume ratio
2:3, fat is put into hermetically sealed can, while the lipase of fat mass 5% is added thereto, and in the case where temperature is 40 DEG C, enzymolysis
3h is waited after the completion of digesting, and zymolyte is placed in the centrifuge of 3000r/min and is centrifuged 10min, collected supernatant；By above-mentioned receipts
The supernatant of collection rotates the half for being evaporated to original volume at 50 DEG C, then adds in sodium nitrate thereto, and addition is
3mg/L after stirring makes it be sufficiently mixed uniformly, is put into the distilling flask with condenser pipe, it is carried out to be heated to 90 DEG C, instead
3h is answered, collects the fraction in condenser pipe, i.e. glycerine；While stirring the third of above-mentioned gained is added in into spare chicken blood extract
Triol, then the phosphorus of the ammonium sulfate of glycerine quality 3% and the concentration of glycerine quality 3% for 0.01mol/L is separately added into thereto
Acid buffering salting liquid after stirring makes it fully dissolve, stands 3h, is subsequently placed in the centrifuge of 3000r/min and centrifuges 30min,
Collect lower sediment thing；Finally the lower sediment thing of collection is washed with deionized 4 times, then adding in concentration thereto is
The phosphate buffered saline solution of 0.01mol/L, addition are 1 with lower sediment thing mass ratio:25, after mixing, it is placed in ultrasound
Decentralized processing 2h in ripple places into centrifuge collected after centrifugation lower sediment thing to get a kind of acoustic contrast agent.
In B ultrasound, acoustic contrast agent produced by the present invention is injected in blood vessel, injection rate is each 3mg, can be obtained after injection
It is clearer on the image to show vessel position and size to very strong B ultrasound echo.The acoustic contrast agent is in blood circulation
Residence time and effective enhancing developing time are long, imaging effect is good, and enhancing efficiency is worth higher than other acoustic contrast agents 24.2%
It promotes and uses.
Claims (1)
1. a kind of preparation method of acoustic contrast agent, it is characterised in that specifically preparation process is：
（1）Fresh chicken blood is collected, 2~3h of freezing at -20~-15 DEG C is put into, after the completion of to be frozen, puts it into 25~32 DEG C
It is lower to melt naturally, after it melts completely, 2~3h of freezing at -20~-15 DEG C is placed into, so cycles 3~4 times, must pre-process
Chicken blood；
（2）By brine and container volume than 1:3, the brine that mass fraction is 0.75~0.9% is fitted into container, is added in simultaneously
The chicken blood of above-mentioned pretreatment, addition are 1 with brine quality ratio:2,1~2h is stood, is then placed it in refrigerated centrifuge,
It it is 3~9 DEG C in temperature, rotating speed is that 10~15min is centrifuged under 4000r/min, obtains supernatant；
（3）By phosphate buffered saline solution and supernatant volume than 1:20, by concentration be 0.01mol/L phosphate buffered saline solution with
The speed of 2~3 drops/s is added dropwise in the supernatant of above-mentioned gained, stirring be sufficiently mixed it after, at 3~5 DEG C stand 1~
Then 2h is 3~9 DEG C in temperature, rotating speed is that 15~25min is centrifuged under 4000r/min, collects lower sediment thing, is handed over through ion
It changes chromatography to be purified, it is spare to obtain chicken blood extract；
（4）The fat newly extracted is chosen, by fat and hermetically sealed can volume ratio 2:3, fat is put into hermetically sealed can, while thereto
The lipase of fat mass 3~5% is added, in the case where temperature is 37~40 DEG C, 2~3h is digested, waits after the completion of digesting, by zymolyte
8~10min of centrifugation in the centrifuge of 2000~3000r/min is placed in, is collected supernatant；
（5）The supernatant of above-mentioned collection is rotated to the half for being evaporated to original volume at 40~50 DEG C, then thereto
Sodium nitrate is added in, addition is 2~3mg/L, after stirring makes it be sufficiently mixed uniformly, is put into the distilling flask with condenser pipe
In, it is carried out to be heated to 80~90 DEG C, reacts 2~3h, collects the fraction in condenser pipe, i.e. glycerine；
（6）While stirring to step（3）The glycerine of above-mentioned gained is added in spare chicken blood extract, then is added respectively thereto
Enter the phosphate buffered saline solution of the ammonium sulfate of glycerine quality 3% and the concentration of glycerine quality 3% for 0.01mol/L, stirring makes
After it is fully dissolved, 2~3h is stood, 20~30min of centrifugation in the centrifuge of 3000r/min is subsequently placed in, collects lower sediment
Object；
（7）The lower sediment thing of collection is washed with deionized 3~4 times, it is 0.01mol/L's then to add in concentration thereto
Phosphate buffered saline solution, addition are 1 with lower sediment thing mass ratio:25, after mixing, it is placed in decentralized processing in ultrasonic wave
1~2h places into centrifuge collected after centrifugation lower sediment thing to get a kind of acoustic contrast agent.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004105802A1 (en) * | 2003-05-27 | 2004-12-09 | Philips Intellectual Property & Standards Gmbh | Tracer for medical imaging |
| CN1698652A (en) * | 2005-04-20 | 2005-11-23 | 沈阳斯佳科技发展有限公司 | Method for extracting substance with cell respiration stimulating activity from pig blood |
| CN101366727A (en) * | 2008-09-27 | 2009-02-18 | 高翔 | Method for preparing calf blood protein-removed extract and products thereby |
| CN101957366A (en) * | 2009-07-15 | 2011-01-26 | 英科新创(厦门)科技有限公司 | Human erythrocyte membrane antigen coated microsphere and application thereof |
| CN102466730A (en) * | 2010-11-19 | 2012-05-23 | 南京神州英诺华医疗科技有限公司 | Novel method for matching blood reverse typing reagent |
| US20140066761A1 (en) * | 2012-02-22 | 2014-03-06 | Alexander A. Oraevsky | Optoacoustic-Ultrasonic Contrast Agents with Enhanced Efficiency |
-
2015
- 2015-12-31 CN CN201511015312.0A patent/CN105561346B/en not_active Expired - Fee Related
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004105802A1 (en) * | 2003-05-27 | 2004-12-09 | Philips Intellectual Property & Standards Gmbh | Tracer for medical imaging |
| CN1698652A (en) * | 2005-04-20 | 2005-11-23 | 沈阳斯佳科技发展有限公司 | Method for extracting substance with cell respiration stimulating activity from pig blood |
| CN101366727A (en) * | 2008-09-27 | 2009-02-18 | 高翔 | Method for preparing calf blood protein-removed extract and products thereby |
| CN101957366A (en) * | 2009-07-15 | 2011-01-26 | 英科新创(厦门)科技有限公司 | Human erythrocyte membrane antigen coated microsphere and application thereof |
| CN102466730A (en) * | 2010-11-19 | 2012-05-23 | 南京神州英诺华医疗科技有限公司 | Novel method for matching blood reverse typing reagent |
| US20140066761A1 (en) * | 2012-02-22 | 2014-03-06 | Alexander A. Oraevsky | Optoacoustic-Ultrasonic Contrast Agents with Enhanced Efficiency |
Non-Patent Citations (4)
| Title |
|---|
| Microbubble contrast agents: a new era in ultrasound;Martin J K Blomley,et al,;《BMJ》;20010519;第322卷;第1222-1225页 * |
| Use of Ultrasound Contrast Agents for Gene or Drug Delivery in Cardiovascular Medicine;Raffi Bekeredjian，et al;《Journal of the American College of Cardiology》;20050201;第45卷(第3期);第329-335页 * |
| 超声造影剂靶向性作用机制的研究进展;景香香 等，;《临床超声医学杂志》;20131230;第5卷(第6期);第359-360页 * |
| 靶向微泡造影技术的研究进展;刘晓婷 等，;《中华医学超声杂志》;20140901;第11卷(第9期);第706-709页 * |
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