CN103852362B - The preparation method of the freeze-drying rabbit brain tissue factor - Google Patents
The preparation method of the freeze-drying rabbit brain tissue factor Download PDFInfo
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- CN103852362B CN103852362B CN201410093404.XA CN201410093404A CN103852362B CN 103852362 B CN103852362 B CN 103852362B CN 201410093404 A CN201410093404 A CN 201410093404A CN 103852362 B CN103852362 B CN 103852362B
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Abstract
The invention discloses the preparation method of the freeze-drying rabbit brain tissue factor, the steps include: to get fresh rabbit brain, peel off meninx and blood vessel, discard brain stem, clean with physiological saline; Be placed in clean mortar, add acetone to firm submergence rabbit brain; With grinding hammer grinding, remove acetone, three times repeatedly; With filter paper elimination acetone; Be placed in 37 DEG C of baking boxs and spread out, acetone evaporated is dried; Add physiological saline, put into the pre-hot activation of 37 DEG C of waters bath with thermostatic control after stirring evenly, hatch and stir; Centrifuging; After getting supernatant liquid filtering, obtain rabbit brain F III tissue factor suspension, add in bottle, freeze-drying, bleed, pressure bottle, namely make the freeze-drying rabbit brain tissue factor ,-20 DEG C of preservations.Adopt such scheme, technology is more ripe, quality is more stable, the shelf-life is longer, uses more convenient, has high medical teaching, scientific research, clinic diagnosis and economic worth.
Description
Technical field
The invention belongs to the technical field of biomedical preparation.More particularly, the present invention relates to a kind of preparation method of the freeze-drying rabbit brain tissue factor.
Background technology
Last century, the seventies, in order to study disseminated intravascular coagulation (DIC), and developed rabbit (dog) the brain powder (F III: the semifinished product of tissue factor) obtained.The detection that rabbit (dog) brain powder (F III: the semifinished product of tissue factor) starts the factor and DIC hematology diagnosis index prothrombin time (PT) and activated partial thromboplastin time (APTT) as disseminated intravascular coagulation (DIC) animal model has important value.
But in the prior art, rabbit brain thromboplastin (F III: tissue factor) only has pulvis, the prothrombin time (PT) made with rabbit brain thromboplastin is only had to measure kit.During use, need through pre-hot activation one hour, then the process such as centrifugal, filtration, can use.
In addition, the shelf-life of preparation of the prior art is shorter, only has 8 months.
But in the prior art, commercially available only have rabbit brain powder agent (in fact just F III: the semifinished product of tissue factor).During use, need through pre-hot activation, hatch 60min, then after the processes such as centrifugal, filtration, could use, use and comparatively bother.In addition, the technical quality of rabbit brain powder agent in prior art is unstable, the shelf-life is shorter, and-20C preserves the shelf-life and only has 6 months.
Summary of the invention
The invention provides the preparation method of the freeze-drying rabbit brain tissue factor, its objective is that quality is more stable, it is more convenient to use, the shelf-life is longer.
To achieve these goals, the technical scheme that the present invention takes is:
The preparation method of the freeze-drying rabbit brain tissue factor of the present invention, the steps include:
A, get fresh rabbit brain, peel off meninx and blood vessel, discard brain stem, clean with physiological saline; Be placed in clean mortar, add acetone to firm submergence rabbit brain;
B, with grind hammer grinding, then, remove acetone; Acetone is removed again after adding the grinding of new acetone again; So three times repeatedly;
C, use filter paper elimination acetone;
D, the rabbit brain after elimination acetone be placed in 37 DEG C of baking boxs and spread out, acetone evaporated being dried, makes rabbit brain powder, be i.e. rabbit brain tissue factor semifinished product, its vigor PT≤13S;
E, get by the obtained rabbit brain tissue factor of above-mentioned steps, i.e. rabbit brain powder, adds physiological saline, puts into the pre-hot activation of 37 DEG C of waters bath with thermostatic control, hatches 60min, every 15min stirring once after stirring evenly;
F, then centrifuging; After getting the supernatant liquid filtering that centrifuging obtains, namely obtain rabbit brain tissue factor suspension.
G, get rabbit brain tissue factor suspension and add in bottle, freeze-drying, bleed, pressure bottle, namely make the freeze-drying rabbit brain tissue factor ,-20 DEG C of preservations.
In described step f, the parameter of centrifuging is 1000 turns/min, and the time is 5min.
In described step e, get rabbit brain powder 400mg, add physiological saline to 10ml.
In described step g, described bottle specification is 5ml or 10ml, namely makes freeze-drying rabbit brain tissue factor 5ml/ bottle or 10ml/ bottle lyophilized formulations.
During use, by freeze-drying rabbit brain tissue factor 5ml/ bottle adding distil water 5ml, or 10ml/ bottle freeze-drying rabbit brain tissue factor adding distil water 10ml, namely make the rabbit brain tissue factor suspension of 5ml or 10ml respectively.
The present invention adopts technique scheme, and the rabbit brain tissue factor (F III: tissue factor) is extracted from rabbit brain tissue, and brain tissue content is the highest; Make rabbit brain powder (semifinished product of tissue factor) and make rabbit brain tissue factor 5ml/ bottle or 10ml/ bottle lyophilized formulations again, therefore, technology is more ripe, quality is more stable, the shelf-life longer (>=Dec), use more convenient; Potency ratio is higher; As long as adding distil water dissolves and just can use during use, do not need through pre-hot activation, hatch, the process such as centrifugal, filtration, there is purposes widely in the fields such as hemopathic experimental study and experimental teaching such as clinical diagnose, treatment, disseminated intravascular coagulations (DIC), there is high medical teaching, scientific research, clinical diagnosis, treatment and economic worth.
Embodiment
Below by the description to embodiment, the specific embodiment of the present invention is described in further detail, has more complete, accurate and deep understanding to help those skilled in the art to inventive concept of the present invention, technical scheme.
The present invention is a kind of preparation method of the freeze-drying rabbit brain tissue factor, rabbit brain powder (F III: tissue factor semifinished product) is made 5ml/ bottle or 10ml/ bottle lyophilized formulations, technology is more ripe, quality is more stable, the shelf-life is longer, more convenient to use, can be widely used in medical teaching, scientific research and clinical conditions.
In order to solve prior art Problems existing and overcome its defect, realize using the goal of the invention more convenient, technology is more ripe, quality is more stable, the shelf-life is longer, the technical scheme that the present invention takes is:
The step of the preparation method of the above-mentioned freeze-drying rabbit brain tissue factor is:
A, get fresh rabbit brain, peel off meninx and blood vessel, discard brain stem, clean with physiological saline; Be placed in clean mortar, add acetone to firm submergence rabbit brain;
B, with grind hammer grinding, then, remove acetone; Acetone is removed again after adding the grinding of new acetone again; So three times repeatedly;
C, use filter paper elimination acetone;
D, the rabbit brain after elimination acetone be placed in 37 DEG C of baking boxs and spread out in case, acetone evaporated being dried, makes rabbit brain powder, be i.e. rabbit brain tissue factor semifinished product, its vigor PT≤13S;
E, to get by the obtained rabbit brain powder (tissue factor semifinished product) of above-mentioned steps, add physiological saline, put into the pre-hot activation of 37 DEG C of waters bath with thermostatic control after stirring evenly, hatch 60min, every 15min stirring once;
F, then centrifuging; After getting the supernatant liquid filtering that centrifuging obtains, obtain rabbit brain thromboplastin suspension, this brain thromboplastin suspension and F III tissue factor suspension;
G, get rabbit brain F III tissue factor suspension and add in bottle, freeze-drying, bleed, pressure bottle, namely make the freeze-drying rabbit brain tissue factor, guarantee the quality for subzero 20 DEG C, shelf-life >=Dec; The described freeze-drying rabbit brain tissue factor, i.e. rabbit brain thromboplastin is also F III tissue factor.
The rabbit brain F III tissue factor lyophilized formulations obtained by said method, technology of preparing is more ripe, quality is more stable, the shelf-life is longer, it is more convenient to use.
During use, do not need the pre-hot activation of pulvis sample, hatch, centrifugal, filter, as long as adding distil water dissolve can use.-20 DEG C of preservations, shelf-life >=Dec.And prior art-20 DEG C preservation, the shelf-life is only 6 months.
Therefore, no matter the development of rabbit brain F III tissue factor lyophilized formulations is at medical research teaching, clinic diagnosis, have important value economically.Practical value is widely had at the experimental study and experimental teaching etc. of clinic diagnosis, disseminated intravascular coagulation (DIC).
The technical parameter of embodiments of the invention is:
In described step f, the parameter of centrifuging is 1000 turns/min, and the time is 5min.
In described step e, get rabbit brain powder 400mg, add physiological saline to 10ml.
In described step g, described bottle specification is 5ml, namely makes rabbit brain tissue factor 5ml/ bottle lyophilized formulations.
Or in described step g, described bottle specification is 10ml, namely make rabbit brain tissue factor 10ml/ bottle lyophilized formulations.
During use, namely rabbit brain tissue factor 5ml/ bottle lyophilized formulations adding distil water 5ml be can be made into rabbit brain tissue factor suspension 5ml.Or, namely rabbit brain tissue factor 10ml/ bottle lyophilized formulations adding distil water 10ml be can be made into rabbit brain tissue factor suspension 10ml.
By said method, rabbit brain powder (F III: tissue factor semifinished product) can make rabbit brain F III tissue factor lyophilized formulations.
Other animal except rabbit, as dog, large mouse, human brain pulvis and placenta make F III: tissue factor pulvis (F III: the thick product of tissue factor), then makes F III: the principle of tissue factor lyophilized formulations is identical substantially.
Above to invention has been exemplary description; obvious specific implementation of the present invention is not subject to the restrictions described above; as long as have employed the improvement of the various unsubstantialities that method of the present invention is conceived and technical scheme is carried out; or design of the present invention and technical scheme directly applied to other occasion, all within protection scope of the present invention without to improve.
Claims (4)
1. the preparation method of the freeze-drying rabbit brain tissue factor, is characterized in that described preparation method's step is:
A, get fresh rabbit brain, peel off meninx and blood vessel, discard brain stem, clean with physiological saline; Be placed in clean mortar, add acetone to firm submergence rabbit brain;
B, with grind hammer grinding, then, remove acetone; Acetone is removed again after adding the grinding of new acetone again; Above-mentionedly add acetone, grinding, the process repeatedly three times altogether of removing acetone;
C, use filter paper elimination acetone;
D, the rabbit brain after elimination acetone be placed in 37 DEG C of baking boxs and spread out, acetone evaporated being dried, makes rabbit brain powder, be i.e. rabbit brain tissue factor semifinished product, its prothrombin time PT≤13s;
E, get by the obtained rabbit brain tissue factor of above-mentioned steps, i.e. rabbit brain powder, adds physiological saline, puts into the pre-hot activation of 37 DEG C of waters bath with thermostatic control, hatches 60min, every 15min stirring once after stirring evenly;
F, then centrifuging; After getting the supernatant liquid filtering that centrifuging obtains, namely obtain rabbit brain tissue factor suspension;
G, get rabbit brain tissue factor suspension and add in bottle, freeze-drying, bleed, pressure bottle, namely make the freeze-drying rabbit brain tissue factor ,-20 DEG C of preservations.
2., according to the preparation method of the freeze-drying rabbit brain tissue factor according to claim 1, it is characterized in that:
In described step f, the parameter of centrifuging is 1000 turns/min, and the time is 5min.
3., according to the preparation method of the freeze-drying rabbit brain tissue factor according to claim 1, it is characterized in that:
In described step e, get rabbit brain powder 400mg, add physiological saline to 10ml;
In described step g, described bottle specification is 5ml or 10ml, namely makes freeze-drying rabbit brain tissue factor 5ml/ bottle or 10ml/ vial formulation.
4., according to the preparation method of the freeze-drying rabbit brain tissue factor according to claim 3, it is characterized in that:
During use, by freeze-drying rabbit brain tissue factor 5ml/ bottle adding distil water 5ml, or 10ml/ bottle freeze-drying rabbit brain tissue factor adding distil water 10ml, namely make the rabbit brain tissue factor suspension of 5ml or 10ml respectively.
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| CN105372424A (en) * | 2015-11-12 | 2016-03-02 | 武汉景川诊断技术股份有限公司 | Hepaplastin test determination kit and preparation method thereof |
| CN105368916B (en) * | 2015-11-12 | 2019-11-22 | 武汉景川诊断技术股份有限公司 | A kind of prothrombin time determination reagent box and preparation method thereof |
| CN116165039B (en) * | 2023-02-11 | 2023-09-22 | 保定天岳生物工程有限公司 | Preparation method of rabbit brain tissue replacing traditional rabbit brain powder |
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| JP2005345194A (en) * | 2004-06-01 | 2005-12-15 | Yamada Yoshihiko | Quick treatment method for hard tissue using ultrasonic wave |
| CN102565427A (en) * | 2011-12-28 | 2012-07-11 | 苏州良辰生物医药科技有限公司 | Preparation method for prothrombin time determination reagent |
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| JP2005345194A (en) * | 2004-06-01 | 2005-12-15 | Yamada Yoshihiko | Quick treatment method for hard tissue using ultrasonic wave |
| CN102608337A (en) * | 2011-04-22 | 2012-07-25 | 武汉塞力斯生物科技有限公司 | Prothrombin time test kit and preparation method thereof |
| JP2013088328A (en) * | 2011-10-19 | 2013-05-13 | Meiji Co Ltd | Preparation method for transmission electric microscope sample |
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Inventor after: Lu Xiaohua Inventor after: Lu Linjie Inventor after: Wang Guoguang Inventor after: Zhao Xue Inventor after: Wang Quanhai Inventor after: Xu Lei Inventor after: Li Wei Inventor after: Zhang Cui Inventor after: Li Hui Inventor after: Dong Juan Inventor after: Wu Juan Inventor after: Lu Ying Inventor before: Lu Xiaohua Inventor before: Wang Quanhai Inventor before: Xu Lei Inventor before: Wang Guoguang Inventor before: Li Wei Inventor before: Zhao Xue |
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