CN101019889A

CN101019889A - Prepn process of brain protein hydrolysate injection

Info

Publication number: CN101019889A
Application number: CNA2007100554214A
Authority: CN
Inventors: 石海
Original assignee: Individual
Current assignee: Individual
Priority date: 2007-03-16
Filing date: 2007-03-16
Publication date: 2007-08-22

Abstract

The brain protein hydrolysate injection is prepared with health swine brain and contains 16 kinds of free amino acids and small amount of peptides. It is used in treating sequelae of craniocerebral trauma and cerebrovascular disease, improving cerebral function, etc. It is prepared through washing, homogenizing, thermal changing, enzyme hydrolysis, freezing, ultrafiltering, sterilizing and other steps.

Description

A kind of preparation process of brain protein hydrolysate injection

Technical field:

The invention belongs to medical technical field

Background technology

Brain protein hydrolysate injection is the sterile water solution that healthy Medulla sus domestica makes through enzymolysis, containing free amino acid among every 1ml is 28.08-42.14mg, contain total nitrogen (N) and should be 5.49-6.71mg, be used for craniocerebral trauma, apoplexy sequela is concentrated the doing well,improving of obstacle with hypomnesis and attention.Brain insufficiency is had the auxiliary improvement effect, also be used for potein deficiency, patient neurasthenia is to use the case to general protein digestibility malabsorption.

This kind is the earliest by Austrian import, domesticly afterwards produces successively.Its technical process bibliographical information is a lot, and its process is roughly defat, hydrolysis, ultrafiltration, chromatography, the amino acid whose process of allotment.But we can not produce qualified products by the technological operation of report in practice, mainly are that other materials are greater than 15% in standard " inspection " item, and color does not especially reach the requirement of light yellow clear liquid yet in storage process.Defatting step needs between hoolivan in addition, has increased cost on the one hand, does not need really in the technology on the one hand.The chromatography process is also unrealistic in practice.

Summary of the invention:

Process of the present invention is:

A, the cleaning of thawing: get the Medulla sus domestica that meets quality standard and took out from freezer the day before yesterday that feeds intake, room temperature is thawed, is cleaned blood and slime and dirt with drinking water, removes epithelium, with injection washing secondary.

B, homogenate: Medulla sus domestica is added 1.5 times of fresh water for injection, rub with colloid mill, make homogenate with 3 grades of fineness.

C, pepsin hydrolysis: steam-jacked kettle internal heating to 85 ℃ maintenance 15 minutes, cooling are then inserted in homogenate.When temperature is 42 ± 2 ℃, in homogenate, add hydrochloric acid and regulate about pH3.5, press Medulla sus domestica weight 3% then and add pepsin, hydrolysis is 6 hours under 42 ± 2 ℃ of conditions, be sub-packed in the stainless steel cask freezing, taking-up room temperature slowing down after deep colling next day.

The supernatant in d, pancreatin hydrolysis: c step is used about NaOH solution accent pH to 8.5, and pancreatin is added the dissolving of injection water, and average 1g pancreatin adds water 2ml, and adding calcium chloride is to 0.025N, and room temperature or 0-4 ℃ activates about 1 hour.Above-mentioned pancreatin is pressed 2% of Medulla sus domestica weight add in the supernatant, in 42 ± 2 ℃ of hydrolysis about 4 hours, hydrolyzed solution boiled 20 minutes in 100 ℃, and the cold water cooling promptly gets supernatant.

E, ultrafiltration and nanofiltration: the supernatant of d step is crossed the ultrafiltration post of 10,000 molecular weight, gets ultrafiltrate and then above-mentioned ultrafiltrate is used the nanofiltration of the daltonian nanofiltration device of molecular weight 300--1000, leaches about half of original volume, gets trapped fluid or claims the nanofiltration trapped fluid.The ultrafilter of again the nanofiltration trapped fluid being crossed 10,000 molecular weight once.

F, e is gone on foot ultrafiltrate carry out peptide figure and determined amino acid, determine extension rate, replenish each seed amino acid and to standard, limit by the sample preparation, add the sodium sulfite of final volume below 2/1000ths simultaneously, add water and supply volume, regulate pH, detect the index of refraction and the nitrogen content of solution

G, above-mentioned solution is after 10,000 molecular weight ultrafiltration posts, aseptic filtration, and embedding, sterilization, lamp inspection, detection promptly get brain protein hydrolysate injection.

Advantage of the present invention is:

A, freeze deeply behind pepsin hydrolysis, it is qualified to help the final clarity of preparation.

Other materials have been removed in the nanofiltration post nanofiltration of b, usefulness molecular weight 300-1000, and inorganic salt etc. make the index of refraction of preparation qualified.

C, add sodium sulfite below 2 ‰ according to practical situation during the course, guaranteed preparation and be light yellow, prevented the oxidation of active ingredient in effect duration.

D, as a complete technological, can produce the qualified products that meet quality standard (national drug standards WS-XG-025-2000).

Concrete embodiment

One, gets fresh Medulla sus domestica 80kg, remove fascia, after cleaning, the water for injection that adds 1.5 times is used colloid mill homogenate, places jacketed pan, be heated to 85 ℃ and keep 15 minutes postcooling to 42 ℃, regulate pH value about 3.5 with hydrochloric acid, add pepsin and stir evenly for 2.4 kilograms, be sub-packed in the stainless steel cask, hydrolysis is about 6 hours in 42 ± 2 ℃ water bath, taking-up is positioned over the deep colling of spending the night in-20 ℃ of freezers, and it is saturating to take out the room temperature naturalization next day, draws supernatant.Get the 1.6kg pancreatin and add water 3.2kg dissolving, add 7g calcium chloride, stir evenly in room temperature and placed 1 hour, in 42 ± 2 ℃ of hydrolysis 4 hours.Place folder room pot to be heated to boil said hydrolyzed liquid and continue 20 minutes, cooling promptly gets supernatant rapidly.Above-mentioned supernatant is crossed 10,000 molecular weight ultrafiltration post ultrafiltration once, get the about 160kg of solution, through the daltonian nanofiltration device nanofiltration of molecular weight 300---1000, leach volume 80kg again, keep trapped fluid 80kg, again through 10,000 molecular weight ultrafilter ultrafiltration.Various amino acid concentrations of sampling and measuring and peptide figure, test through sample, determine 2.4 times of dilutions, add the 0.85g sodium sulfite for every liter, supply each seed amino acid by limit in the concentration simultaneously by final volume, the adjusting final volume is 192 liters and gets final product, cross 10,000 molecular weight ultrafiltration posts once, transfer pH7.2, aseptic filtration, embedding, 105 ℃ of sterilizations in 40 minutes promptly get brain protein hydrolysate injection.

Claims

1, a kind of brain protein hydrolysate injection preparation technology, its process is

A, cleaning, homogenate: get the fresh and healthy Medulla sus domestica, clean,, add 1.5 times of waters for injection, use colloid mill homogenate with injection washing secondary.

B, pepsin hydrolysis: homogenate is heated to 85 ℃ and kept 15 minutes, is cooled to 42 ± 2 ℃ with about hydrochloric acid adjusting pH3.5, and press Medulla sus domestica weight 3% and add pepsin, freezing after hydrolysis finishes, room temperature slowing down after the deep colling is drawn supernatant.

C, above-mentioned supernatant transfer pH to alkalescence, press Medulla sus domestica weight 2% and add with 60 minutes pancreatin of 0.025N calcium chloride activation, and pH neutrality is transferred in hydrolysis, is heated to and boils, and cooling is drawn supernatant and filtered.

D, above-mentioned clear liquid are crossed 10,000 molecular weight ultrafiltration posts.

E, above-mentioned ultrafiltrate are crossed the nanofiltration device, keep a certain proportion of trapped fluid.

Extension rate is determined in f, detection, and supplies each seed amino acid by the concentration center line.By the following sodium sulfite that adds of 2/1000ths (W/V) of final volume, regulate index of refraction and nitrogen content simultaneously.

G, above-mentioned solution are crossed 10,000 molecular weight ultrafiltration posts, and embedding, sterilization, lamp inspection, detection promptly get brain protein hydrolysate injection.

It is characterized in that:

Freezing behind a, the pepsin hydrolysis, room temperature is thawed after the deep colling, sucts clearly, and it is qualified to help the final products clarity.

Adopted nanofiltration in b, the technology, promptly molecular weight is the daltonian Filter column of 300---1000, has removed other materials in the raw material, and inorganic salt etc., make the index of refraction of preparation qualified simultaneously.

Added sodium sulfite in c, the raw material, prevented the oxidation of composition in production and the storage process, made " character " item be light yellow clear liquid before the deadline.

D, be mainly said process, can produce the brain protein hydrolysate injection that conforms with quality standard fully as a complete technological.