CN1070411A

CN1070411A - From disabled person's urine, extract the method for urinary trypsin inhibitor and G CFS

Info

Publication number: CN1070411A
Application number: CN 91108119
Authority: CN
Inventors: 张博达
Original assignee: Nanjing University
Current assignee: Nanjing University
Priority date: 1991-07-24
Filing date: 1991-07-24
Publication date: 1993-03-31
Anticipated expiration: 2006-07-24
Also published as: CN1031577C

Abstract

Extract the method for urinary trypsin inhibitor (UTI) and G CFS (CSF) a kind of disabled person's urine after having extracted urokinase (UK) simultaneously, this method is to make sorbent material with kaolin, under certain pH value, adsorb UTI and CSF simultaneously, the characteristics of this method are that technology is simple, cost is low, can utilize the production unit of former extraction UK to extract UTI and CSF.

Description

From disabled person's urine, extract the method for urinary trypsin inhibitor and G CFS

The present invention is a production method of extracting urinary trypsin inhibitor (being called for short UTI) and G CFS (being called for short CSF) a kind of disabled person's urine after having extracted urokinase (being called for short UK) simultaneously.

UTI is a kind of glycoprotein, suppresses trypsinase specifically, can be used for treating diseases such as acute pancreatitis, chronic recidivity pancreatitis, acute circulatory disorder.CSF has the effect that stimulates scavenger cell, can be used for treating diseases such as granulocytopenia, cancer.About from people's urine, extracting the method for UTI, the method (seeing Chinese patent CN86104618A) that Nippon Chemical Research K.K.'s good fortune justice is arranged at present and wait the people to invent, this method is the people to be urinated under condition of neutral pH foam, gather foam concentrate liquid, from weakly acidic concentrated solution, adsorb UTI and UKN(urine Pancreatic Kininogenase simultaneously with chitosan then), reach the purpose of extracting UTI and two products of UKN simultaneously.The advantage of this method is once to obtain UTI and two products of UKN from people urine, shortcoming be to equipment requirements than higher, one-time investment is bigger, the chitosan price is also somewhat expensive in addition, therefore, the production cost of UTI and UKN is than higher.Extracting CSF and UTI from people's urine has the patented method (the clear 61-59610 of Japanese Patent) of Japanese MORINAGA MILK INDUSTRY Co., LTD..This method is earlier the people to be urinated ultrafiltration and concentration, then with weak base anion exchanger absorption, uses the damping fluid branch wash-out of different concns again, obtains two products.The advantage of this method is the ion-exchanger repeated use of can regenerating, and can obtain two products of CSF and UTI respectively, but will urinate ultrafiltration and concentration, and handling a large amount of urine needs very big ultrafiltration apparatus, invests quite big.About the disabled person's urine after extracting UK, extract the method for UTI and two products of CSF more simultaneously, be not seen in report as yet.

The objective of the invention is to make full use of disabled person's urine of extracting behind the UK, therefrom once extract two products of UTI and CSF simultaneously.

Method of the present invention is to have extracted pH regulator to 5.5～6.5 of the disabled person's urine behind the urokinase with the diatomite adsorption method, allow disabled person's urine fully contact with the kaolin sorbent material, after treating that urinary trypsin inhibitor and G CFS are adsorbed onto on the kaolin, the useless urine of venting, wash sorbent material with water, the urinary trypsin inhibitor and the G CFS that will be adsorbed onto on the kaolin with ammonia soln elutes then, in elutriant, add ammonium sulfate precipitation, after leaving standstill, precipitation is used diatomite filtration, obtains containing diatomaceous urinary trypsin inhibitor and G CFS crude product.

In the invention process of present method, the inventor has done a large amount of cut-and-try works.In the screening of kaolin sorbent material, the inventor finds that K-6, K-7, K-8, K-9 and K-90 kaolin sorbent material that China Kaolin Co., Ltd produces can be used for extracting simultaneously UTI and CSF from disabled person's urine, be best with K-8 and two kinds of kaolin sorbent materials of K-9 especially wherein, these two kinds of kaolin sorbent materials are that China Kaolin Co., Ltd improves production according to inventor's requirement and suggestion.

The kaolin sorbent material has very strong adsorptive power, and at pH hour, the impurity of absorption is more, and at pH less than 5 o'clock, UTI and CSF easily are degraded to small molecules.The present invention has selected the adsorption conditions of pH5.5～6.5.Greater than 5.5 o'clock, the adsorptive capacity of UTI reduced at pH, and Xi Fu impurity is also less simultaneously, was main if only need to extract the CSF product, and pH can regulate more greatly, but can not be greater than 6.5, and pH was greater than 6.5 o'clock, and the adsorptive capacity of CSF also will reduce.In the time of need extracting two products simultaneously, be 5.5 the bests then with pH.

Kaolin adsorbs UTI and CSF from disabled person's urine speed is than faster.The speed of absorption is relevant with temperature, the temperature height, rate of adsorption is fast, generally whip attachment is 1～1.5 hour, summer the room temperature height, whip attachment 1 hour, winter temperature is low, whip attachment 1.5 hours.

The molecular weight of UTI is about 68000, and the molecular weight of CSF is about 42000, and under unsuitable working condition, the both easily is degraded to small molecules.The mode of production of the present invention causes under the situation of UTI and CSF macromolecules degradation in that people urine is rotten, and the macromole content that can guarantee UTI and CSF in the product is more than 80%.

The characteristics of method of the present invention are that technology is simple, and can utilizing at present fully, therefore domestic many places need not to increase what and just invest and can increase new product on the basis of original production UK with the simple production equipment of diatomite extraction UK.The sorbent material that uses is that china natural resources is abundant, cheap kaolin, and production cost is low.And,,, also can directly from protoplast's urine, extract UTI and CSF with the identical processing condition of the present invention if do not produce UK.

Below be embodiments of the invention:

Embodiment 1: the absorption property of kaolin sorbent material is measured, and will transfer to 5.5 with the pH that diatomite extracts the disabled person's urine behind the UK.Get 1 liter of disabled person's urine (pH=5.5) and increase mountain range earth adsorbing 10 gram, stirred 1.5 hours, draining useless urine after, with 50 ml distilled waters washing 1 time, use 50 milliliter 3% ammoniacal liquor (pH10.5～12.0) wash-out then, the content of UTI and CSF the results are shown in Table 1 in the mensuration elutriant

The absorption property of table 1 kaolin sorbent material

The trade mark	UTI content (U) *	CSF content (Colonyu) * *
			K-6	4043	4.3×10 ⁵
K-7	4107	4.3×10 ⁵
			K-8	4385	4.6×10 ⁵
K-9	4413	4.6×10 ⁵
			K-90	3467	3.9×10 ⁵

^*The mensuration of UTI among the present invention is the U of 1UTI unit with the trypsinase of 4 units of every inhibition, measures and makes substrate with BAPNA.

^*The Colonyu that tires of CSF cultivates (individual layer method) with the external agar of bone marrow cells in mice and measures calculating.Protein content converts with the O.D.280 value.

Embodiment 2: under the different pH, and the absorption property of kaolin sorbent material

Take diatomite and extract 5 liters of disabled person's urines behind the UK, be divided into 5 parts, 1 liter every part, transfer pH to 5.0 respectively, 5.5,6.0,6.5,7.0, each adds 10 gram K-9 kaolin sorbent materials, whip attachment 1.5 hours, draining useless urine after, wash once with 50 ml distilled waters respectively, use 50 milliliter 3% ammoniacal liquor wash-out then, measure UTI in each elutriant, CSF and protein content the results are shown in Table 2.

Under the different PH of table 2, the absorption property of kaolin sorbent material

PH	5.0	5.5	6.0	6.5	7.0
						UTI(U)	4435	4423	4012	2715	1210
CSF(ColonyU)	4.6×10 ⁵	4.6×10 ⁵	4.6×10 ⁵	4.6×10 ⁵	4.1×10 ⁵
						Albumen egg (mg)	215	163	124	97	93

Embodiment 3: get 1 ton of disabled person's urine of extracting behind the UK respectively, 1 ton of Freshman urine stoste is transferred pH to 5.5 respectively, respectively add 10 kilograms in K-9 kaolin sorbent material, whip attachment 1.5 hours is behind the useless urine of venting, wash with water, add ammonium sulfate to saturation ratio and be 80% and saltout, after saltouing, precipitation is used diatomite filtration, obtains containing diatomaceous UTI and CSF crude product and is respectively 283 grams and 301 grams, behind crude product difference mixing, respectively take by weighing 2 grams, measure UTI respectively, CSF and Protein content the results are shown in Table 3.

UTI, CSF and protein that table 3 disabled person urine and former human urine extract

	UTI（U）	CSF（ColonyU)	Protein (mg)
				Disabled person's urine	23109	2.848×10 ⁶	216
Former human urine	20797	2.751×10 ⁶	228

Claims

1, a kind of method of from extracted the disabled person's urine behind the urokinase, extracting urinary trypsin inhibitor and G CFS simultaneously, it is characterized in that and to have extracted between the pH regulator to 5.5 and 6.5 of the disabled person's urine behind the urokinase, allow disabled person's urine fully contact with the kaolin sorbent material, after treating that urinary trypsin inhibitor and G CFS are adsorbed onto on the kaolin, the useless urine of venting, wash sorbent material with water, the urinary trypsin inhibitor and the G CFS that will be adsorbed onto on the kaolin with ammonia soln elutes then, in elutriant, add ammonium sulfate precipitation, after leaving standstill, precipitation is used diatomite filtration, obtains containing diatomaceous urinary trypsin inhibitor and G CFS crude product.

2, method according to claim 1 is characterized in that this method also can be used for extracting urinary trypsin inhibitor and G CFS from crude urine.

3, according to claim 1 and 2 described methods, it is characterized in that the kaolin sorbent material is K-6, K-7, K-8, K-9 and K-90 kaolin sorbent material, wherein best with the effect of K-8 and two kinds of kaolin sorbent materials of K-9.

4, according to claim 1 and 2 described methods, the pH value that it is characterized in that regulating useless urine is more higher, and the impurity absorption amount is less, and the while adsorptive capacity of urinary trypsin inhibitor also reduces, and need extract two products simultaneously, is best with pH5.5.

5,, it is characterized in that the kaolin sorbent material consumption and the ratio of urine amount are that 1 gram is optimum than 100 milliliters according to claim 1 and 2 described methods.

6,, it is characterized in that the ammonia soln of wash-out urinary trypsin inhibitor and G CFS is an optimum with pH10.5～12.0 from the kaolin sorbent material according to claim 1 and 2 described methods.

7, according to claim 1 and 2 described methods, the saturation ratio of ammonium sulfate is 80% when it is characterized in that saltouing.