CN101259142A

CN101259142A - Bone peptide injection preparation

Info

Publication number: CN101259142A
Application number: CNA2007100553866A
Authority: CN
Inventors: 韩英
Original assignee: Individual
Current assignee: Individual
Priority date: 2007-03-08
Filing date: 2007-03-08
Publication date: 2008-09-10

Abstract

Bone peptide injection liquid is an injection liquid which is made by being extracted from porcine or fetal calf limb bone and is used for accelerating fracture healing. The present invention introduces a manufacturing method, the process of which is thermal extraction, enzymolysis and acid hydrolysis deposition, basic hydrolysis deposition, hyperfiltration, aseptic filtration and encapsulation sterilization. The process is simple and practical, reserves the nutrient component of the bone and has high yield.

Description

A kind of bone peptide injection preparation

Technical field:

The invention belongs to medical technical field.

Background technology:

Bone peptide injection is the aseptic aqueous solution that the bones of limbs of fresh or refrigerated pig or tire cattle is made through extraction.Its Main Ingredients and Appearance is organic calcium, phosphorus, inorganic calcium, inorganic salt, trace nothing element, aminoacid etc.This technical process does not still have open report, and is easy, practical, stable, is suitable for big commercial production.

Summary of the invention:

Process of the present invention is:

A, extraction, degrease: get healthy fresh pig or fetal bovine limb bone, clean, smash, weigh, add water, filtered through gauze is used in hot pressing, and the bone slag adds water again, the same method operation.Filter, merge filtrate twice, place jacketed pan lead to cooling water immediately, be chilled to room temperature, be positioned in the freezer, to be cooled changing in the cold house to 0--5 ℃ left standstill, and skims upper strata fat, heats to make to freeze the shape thing and melt into liquid, concentrates, must concentrated solution.

B, hydrolysis: concentrated solution is transferred about pH3 with hydrochloric acid, pressed 1-3% and add pepsin, hydrolysis 2-4 hour.

C, acid precipitation, alkali deposited: aforesaid liquid is heated to 100 ℃ in pH, continues 30 minutes, cooling sucts clearly, and reuse NaOH transfers about pH to 8.5, is heated to 100 ℃, continues 30 minutes, and cooling sucts clearly.

D, filtration and ultrafiltration: above-mentioned clear liquid is filtered to clearly,, promptly gets bone peptide solution after 10,000 molecular weight ultrafiltration posts three times.

E, preparation: bone peptide stock solution is pressed the quality standard method measure content of peptides, add water move to every milliliter greater than 5mg, transfer pH to 5.5-7.5, degerming, filtration, embedding, sterilization are promptly.

Advantage of the present invention:

1, no any antiseptic in the whole technical process, thus make preparation safer.

2, at utmost kept the bone collagen composition, the nutrition of bone and providing of active ingredient have been provided.

3, removed macromolecular substances, made whole preparation component molecules amount, guaranteed that preparation is not irritated all less than 10,000.

4, whole technology is simple, and easily row is reliable and stable, is particularly useful for suitability for industrialized production.

Detailed process process of the present invention:

1, extraction, degrease, concentrated: get healthy fresh pig bones of limbs, clean, smash and weigh.Every 75kg raw material adds water 75kg, in 1.2kg/cm ²Hot pressing 2 hours is filtered with double gauze, and the bone slag adds water 75kg again, and the same operation hot pressing 1 hour is filtered, and merges filtrate twice., be positioned in-20 to-30 ℃ of freezers to room temperature in the jacketed pan internal cooling, to be cooled changing over to 0-5 ℃ left standstill 12-24 hour in the cold house, skim upper strata fat, heats to make to freeze the shape thing and melt into liquid.

2, hydrolysis: the said extracted thing is transferred about pH3, pressed 1-3% and add pepsin, hydrolysis 4 hours.

3, acid precipitation: above-mentioned liquid in 100 ℃ of heating 30 minutes, is continued acid hydrolysis, cooling sucking-off supernatant.

4, alkali deposited: above-mentioned supernatant is transferred about pH8.5, is heated to boiling, continues 30 minutes, continues the basic hydrolysis cooling, sucts clearly.

5, ultrafiltration: above-mentioned supernatant is transferred pH to 7.4, is filtered to clearly, crosses 10,000 molecular weight ultrafiltration three times, promptly gets bone peptide stock solution, gets 180000 milliliters of stock solutions.

6, preparation: bone peptide stock solution is pressed biuret method measure content of peptides, add water move to 5.0-6.0mg/ml, PH5.5-7.5, aseptic filtration, embedding is sterilized 40 minutes promptly for 105 ℃.

Experiment:

1, yield experiment:

Precision is measured this product 2ml, put in the test tube, the accurate biuret test solution that adds (is got copper sulfate 1.50g and Tartaric acid potassium sodium 5.0g, add water 500ml and make dissolving, add 10% sodium hydroxide solution 300ml while stirring, be diluted with water to 1000ml, mixing) 4ml, mixing, room temperature was placed 30 minutes, according to spectrophotography, wavelength place at 540nm measures trap, it is an amount of that precision takes by weighing the casein reference substance in addition, adds the 0.05mol/L sodium hydroxide test solution and make the solution that contains 7mg among every 1ml approximately, and precision is measured 2ml, in the self-purchased test tube ... rise, with the method operation, measure trap, calculate.

Experimental result is as follows:

2, allergic experiment, the thermal source experiment, the undue toxicity presses quality standard WS1-XG-035---2000 and checks all up to specification.

3, all other indexs all meet injection general rule and WS1-XG-035---2000 bone peptide injection quality standard.

Claims

1. bone peptide injection preparation, its process is:

(1), extraction, degrease: get healthy fresh pig or fetal bovine limb bone, clean, smash, weigh, add water, filtered through gauze is used in hot pressing, and filtrate is chilled to room temperature, is put in the freezer, changes in the cold house after cooling and leaves standstill, and skims upper strata fat, promptly gets the grease removal extracting solution.

(2), the said extracted thing is transferred about PH3, by volume heavy 1-3% adds pepsin, in 37 ℃-42 ℃ hydrolysis 2-4 hour.

(3), acidic precipitation, alkaline sedimentation: said hydrolyzed liquid under agitation adds acid to be transferred about pH to 4.0, and in 100 ℃ of heating, the cooling precipitation sucts clearly, and reuse NaOH transfers about pH8.5, and in 100 ℃ of heating, the cooling precipitation sucts clearly.

(4), above-mentioned supernatant is filtered clarification, the ultrafiltration post after 10,000 molecular weight promptly gets the bone peptide intermediate.

(5), measure content of peptides, and transfer to desired concn, embedding, sterilization is promptly.

It is characterized in that:

(1), adopted pepsin hydrolysis, acid hydrolysis, basic hydrolysis and thermal denaturation, make preparation clarification, clarity is good, also makes the injection preparation become possibility.

(2), at utmost kept the osteotrophy composition, made greater than the polypeptide of 10,000 molecular weight and remove fully, do not lost again simultaneously less than the polypeptide of 10,000 molecular weight and other such as nutritions such as inorganic salts, guaranteed that simultaneously the pyrogen of injection is qualified, anaphylaxis is qualified.