CN100348201C - Methy cobalamine dispersion tablet and preparing method - Google Patents

Methy cobalamine dispersion tablet and preparing method Download PDF

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CN100348201C
CN100348201C CNB2005101057972A CN200510105797A CN100348201C CN 100348201 C CN100348201 C CN 100348201C CN B2005101057972 A CNB2005101057972 A CN B2005101057972A CN 200510105797 A CN200510105797 A CN 200510105797A CN 100348201 C CN100348201 C CN 100348201C
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mecobalamin
dissolution
magnesium stearate
dispersible
test
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CN1742748A (en
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周卓和
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Jiangsu Sihuan Biological Pharmaceutical Co., Ltd.
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周卓和
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Abstract

The present invention provides a mecobalamin dispersion tablet and a preparation method thereof. 1000 dispersion tablets contain the following components: 0.5 g of mecobalamin, 5 to 10 g of sodium carboxymethyl starch, 15 to 30 g of low-replace hydroxypropyl cellulose, 50 to 65 g of microcrystalline cellulose, 1 to 3 g of polyvidone, 5 g of gum arabic, 5 g of steviosin, 1 g of magnesium stearate and 1 g of talcum powder. The dispersion tablet is prepared under a light avoiding condition. The raw material of mecobalamin is sieved by a sieve with 100 meshes, auxiliary materials are sieved by a sieve with 80 meshes, and the mecobalamin and the auxiliary materials are weighed and are uniformly mixed according to an isodose progressive increase principle. A proper amount of alcohol whose concentration is 30% is added to prepare a soft material which is sieved by a nylon sieve with 20 meshes so as to manufacture granules. The granules are dried at 50 to 60 DEG C and are sieved by a sieve with 20 meshes, and the granules are finished. 1g of magnesium stearate and 1g of talcum powder are weighed and are sieved and uniformly mixed with the granules, and tabletting operation is carried out. After being checked, if qualified, the products are packed. The mecobalamin dispersion tablet has the advantages of good dispersivity uniformity and dissolution, more rapid absorption, higher biological availability and more convenient administration, and is particularly suitable for old people, children and patients having difficulty in swallow solids.

Description

Mecobalamin Dispersible tablet and its preparation method
Technical field
The present invention relates to a kind of Mecobalamin Dispersible tablet and its preparation method, belong to field of pharmaceutical preparations.
Background technology
Along with improving constantly of China people's living standard, the sickness rate of diabetes is progressively rising.According to World Health Organization's statistics, the diabetics of China has reached 2,000 ten thousand people from 1998 to 2000, and ascendant trend is very obvious, if effectively do not control, will reach 6,000 ten thousand people by 2010.Peripheral neuropathy is one of modal severe complications of diabetes, and sickness rate can reach about 80%.The change of diabete peripheral herve sexually transmitted disease (STD) can make the patient produce numb limbs and tense tendons, limbs ulcer, amyotrophy and violent pain can occur when serious, even cause that the necrosis of limbs causes the maimed person.
Mecobalamin is the medicine of peripheral neuropathy, belongs to vitamin B 12Class is called and is the methyl vitamin B 12It is synthetic by nucleic acid and protein and neural myelin in the promotion neurocyte, thus the peripheral nerve of repairing damage.Its chemical name is α-(5,6-dimethylbenzimidazole base)-Co-first cobalt amide, English chemistry α by name-(5,6-dinethylbenzimidazolyl)-Co-methyl-cobamide, molecular formula is C 63H 91CoN 13O 14P, molecular weight 1344.38, the chemical constitution of main component is
Figure C20051010579700041
From people such as Spies in 1948 vitamin B 12Since drug research, vitamin B 12Research obtain development at full speed, up to the present, as the vitamin B of medicine 12Mainly contain: cobalamin, hydroxocobalamin, cobamamide and mecobalamin.Preceding two kinds of vitamin Bs 12Do not have biological activity in human body: cobalamin (vitamin B12) is taken in cell through combine formation Tcs-VB12 complex with cobalt amine transporter in vivo, discharge hydroxocobalamin by the lysosomal protein enzyme then, the hydroxocobalamin generation mecobalamin that in Cell sap, methylates.The two kinds of coenzyme type vitamin Bs in back 12Realized synthetic, this treatment for the peripheral neuropathy patient provides powerful mean.Repairing aspect the impaired nervous tissue, cobamamide must be converted into mecobalamin earlier could participate in the one carbon unit circulation as coenzyme, thereby promotes the synthetic of nucleic acid, protein and lecithin.
Mecobalamin mainly is present in biological blood, the marrow liquid, compares with cobalamin, and it has good transitivity to nervous tissue, promotes nucleic acid, protein, lipid metabolism by the methyl conversion reaction, repairs the nervous tissue that is damaged.
Mecobalamin is the medicine of peripheral neuropathy, belongs to vitamin B 12Class is called and is the methyl vitamin B 12It is synthetic by nucleic acid and protein and neural myelin in the promotion neurocyte, thereby the peripheral nerve of repairing damage has been widely used in clinical at present.Clinical test results shows that mecobalamin is a kind of effective and safe drug of treatment diabetic neuropathy; Mecobalamin and peripheral facial paralysis all had good therapeutical effect, and curative effect is better than vitamin B12; Mecobalamin has facilitation to cubital tunnel syndrome operation back functional rehabilitation, and its curative effect is better than vitamin B12; Mecobalamin stops the glaucoma visual field to worsen and promote the effect that improve in its visual field in addition.
Tablet formulation makes the medicine stripping fast, rapid-action, and taking convenience can be directly oral, also can be scattered in the warm water to take, and is particularly suitable for the old man and swallows inconvenient patient, based on the such advantage of dispersible tablet, is necessary to provide mecobalamin dispersible tables.
Summary of the invention
The object of the present invention is to provide a kind of mecobalamin dispersible tables.
Another object of the present invention is to provide the preparation method of above-mentioned mecobalamin dispersible tables.
Per 1000 of dispersible tablet of the present invention contains following component:
Mecobalamin 0.5g
Carboxymethyl starch sodium 5-10g
L-hydroxypropyl cellulose 15-30g
Microcrystalline Cellulose 50-65g
Polyvidone 1-3g
Micropowder silica gel 5g
Steviosin 5g
Magnesium stearate 1g
Pulvis Talci 1g
Wherein:
1. carboxymethyl starch sodium is good intumescent disintegrating agent, and the disintegrate effect is obvious, and is noiseless to the mensuration of mecobalamin.
2. low-substituted hydroxypropyl cellulose is a kind of novel adjuvant, has disintegration, can increase dispersive fineness after the tablet disintegrate, and does not influence the mensuration of mecobalamin.
3. microcrystalline Cellulose has good disintegrate effect not disturb the mensuration of principal agent also as the disintegrating agent of tablet.
4.PVP as binding agent.
5. stevioside is a correctives.
Preparation method of the present invention is: under the lucifuge condition, the mecobalamin raw material is crossed 100 mesh sieves, carboxymethyl starch sodium, L-hydroxypropyl cellulose, microcrystalline Cellulose, polyvidone, micropowder silica gel, steviosin, magnesium stearate, Pulvis Talci are crossed 80 mesh sieves, take by weighing following amount:
Mecobalamin 0.5g
Carboxymethyl starch sodium 5-10g
L-hydroxypropyl cellulose 15-30g
Microcrystalline Cellulose 50-65g
Polyvidone 1-3g
Micropowder silica gel 5g
Steviosin 5g
Progressively increase principle with adjuvant and mecobalamin mix homogeneously according to equivalent; add an amount of 30% ethanol and make soft material, cross 20 order nylon mesh, make granule; and in 50～60 ℃ of dryings; cross 20 mesh sieves, granulate takes by weighing about 1g magnesium stearate and 1g Pulvis Talci; sieve and with the granule mix homogeneously; tabletting, is packed totally after the assay was approved by 1000.
The specification of mecobalamin dispersible tables of the present invention is preferably 500 μ g/ sheets, and content limit is for containing mecobalamin (C 63H 91CoN 13O 14P) should be 90.0～110.0% of labelled amount.
Check other routine examinations of having worked out related substance, dissolution and tablet in the item.According to the regulation of in " chemical drugs pharmacy part guideline " related substance being checked, the tablet of making by formulation and technology is carried out the related substance testing result be respectively 1.70%, 1.62%, 1.48% by Self-control method calculating, stability study (accelerated test, long term test 9 months) is the result show, the related substance limit of product of the present invention is decided to be and is not more than 3%.
Product dissolution determination of the present invention adopts little agar diffusion method, 75 rev/mins of oar methods, high effective liquid chromatography for measuring, the stripping homogeneity is better, three batch sample dissolution determinations meansigma methods as a result are respectively 100.59%, 100.56%, 100.16%, and the stripping limit is decided to be and is not less than 80% of labelled amount.
Assay adopts high effective liquid chromatography for measuring.By the methodology test, precision is better, and RSD is all less than 2%; Average recovery rate is 99.61 ± 0.90% (n=9), and RSD is that the assay result of 0.90%, three batch sample is respectively: 100.24%, 101.36%, 99.14%, and it is 90.0%～110.0% that content limit is ordered.
In the preliminarily stabilised investigation, factorial experiments is the result show, mecobalamin dispersible tables is responsive to light, related substance increases, high temperature (60 ℃) condition is influential slightly to the mecobalamin dispersible tables related substance, and sheet heavily increases comparatively obviously under the super-humid conditions, but does not influence other quality index; Accelerated test and long-term the placement were tested after 9 months, and the quality of product does not have obvious variation, and every index is still in the claimed range of quality standard.Therefore, product of the present invention should be avoided high temperature also strict lucifuge, airtight preservation, and effect duration is 2 years.
Mecobalamin is the medicine of peripheral neuropathy, belongs to vitamin B 12Class is called and is the methyl vitamin B 12It is synthetic by nucleic acid and protein and neural myelin in the promotion neurocyte, thereby the peripheral nerve of repairing damage has been widely used in clinical at present.Mecobalamin can be treated the diabetes nerve systemic disease; Alleviate numbness and pain, alleviate neuralgia rapidly; Open clinical trial shows at random, and mecobalamin can quicken the recovery of facial nerve function; The multi-center clinical trial of Hanai tissue shows that nervous symptoms such as pain that mecobalamin improves cervical spondylosis and cause, paraesthesia have better curative effect; The recovery double-blind trial of doigte shows that mecobalamin plays an important role to the neurological functional recovery of patients with finger replantation after the mecobalamin promotion replantation of amputated finger; A double-blind study shows, and is more effective than other vitamin to the treatment mecobalamin of peripheral neuropathy.Domestic clinical research shows that also the mecobalamin group is better than other vitamin B complex in curative effect aspect the treatment diabetic neuropathy.
Clinical safety is estimated: in people's such as Chen Jialun the clinical research, mecobalamin side effect incidence rate is 5.6%, and none example withdraws from research because of side effect.
Preparation method technology of the present invention is simple, reasonable, and the every index of the dispersible tablet of making all meets the requirements.
The specification of mecobalamin dispersible tables is the 0.5mg/ sheet.
The mensuration of disperseing the time limit is to place 20 ± 1 ℃ 100ml water jolting extremely to disperse required time fully in tablet.(two appendix IA of Chinese Pharmacopoeia version in 2000)
(4500 ± 5001x) mecobalamins are extremely unstable, shine 4 hours, and the catabolite of mecobalamin is increased to 4.26% at strong illumination.Do following test (producing 100 for every batch) by No. 2 prescription preparation methylcobalamin tablets.
Carry out 4.1 all operate under the natural light condition, be pressed into agreement that contracts a film or TV play to an actor or actress from preparing burden to 3 hours.
Carry out under the lucifuge condition 4.2 all operation is strict, use weak red illumination in case of necessity.
According to aforementioned prescription and prepared mecobalamin dispersible tables, extract a certain amount ofly as sample, placed under the hot conditions (60 ℃) 10 days, investigate its mass change.The results are shown in Table 1.
Table 1 high temperature is to the influence of mecobalamin dispersible tables quality
Time 0 day 5 days 10 days
Outward appearance Bright and clean Bright and clean Bright and clean
Sheet is heavy 0.1056 0.1039 0.1046
Disperse the time limit 53″ 56″ 55″
Related substance 1.34％ 1.81％ 1.92％
Content 100.01％ 99.00％ 98.74％
Conclusion: investigate 10 days under high temperature (60 ℃) condition, the related substance of mecobalamin dispersible tables changes little, and mecobalamin dispersible tables is stable with this understanding.
Get mecobalamin dispersible tables and place (RH92.5%) under the super-humid conditions, placed 10 days, investigate its mass change.The results are shown in Table 2.
Table 2 high humidity is to the influence of mecobalamin dispersible tables quality
Time 0 day 5 days 10 days
Outward appearance Bright and clean Rough surface Rough surface
Sheet is heavy 0.1056 0.1209 0.1368
Disperse the time limit 53″ 49″ 46″
Related substance 1.34％ 1.53％ 1.55％
Content 100.01％ 99.38％ 99.10％
Conclusion: methylcobalamin tablet (RH92.5%) under super-humid conditions was placed 10 days, and related substance does not have to change substantially, and sheet is heavy obviously to be increased, and illustrates that the mecobalamin dispersible tables hygroscopicity is stronger, and packing should be noted airtight.
Get mecobalamin dispersible tables, placed under the high light condition (45001x) 5 hours, investigate the variation of mecobalamin dispersible tables quality.The results are shown in Table 3.
Table 3 strong illumination is to the influence of mecobalamin dispersible tables quality
Time Before the investigation After the investigation
Outward appearance Bright and clean Bright and clean
Sheet is heavy 0.1056 0.1052
Disperse the time limit 53″ 55″
Related substance 1.34％ 4.06％
Content 100.01％ 95.14％
Conclusion: mecobalamin dispersible tables (45001x) under the high light condition was placed 5 hours, and related substance changes greatly, and mecobalamin dispersible tables quality instability under illumination condition is described, packing and storage should be noted lucifuge.
Because dispersible tablet of the present invention sees that light easily decomposes, and has certain moisture, therefore, packaging material are must the lucifuge performance good, and airtight performance is good.Get mecobalamin dispersible tables, adopt two aluminum packings, under the environment of illumination 45001x, humidity 92.5%, placed 30 days, investigate the mass change of mecobalamin dispersible tables.The results are shown in Table 4.
Table 4 packing is to the influence of mecobalamin dispersible tables quality
Time 0 day 30 days
Outward appearance Bright and clean Bright and clean
Sheet is heavy 0.1056 0.1058
Disperse the time limit 53″ 56″
Related substance 1.34％ 1.54％
Content 100.01％ 100.00％
Therefore, dispersible tablet of the present invention adopts two aluminum packings.
The present invention is making things convenient under the prerequisite of sufferer, provide mecobalamin dispersible tables this dosage form, with respect to conventional tablet, good dispersion, homogeneity is good, dissolution is good, (refer to foregoing dissolution determination: adopt little agar diffusion method, 75 rev/mins of oar methods, high effective liquid chromatography for measuring, the stripping homogeneity is better, three batch sample dissolution determinations meansigma methods as a result are respectively 100.59%, 100.56%, 100.16%) make absorption more rapid, bioavailability is higher, takes more conveniently, especially is fit to old, the children and the patient of solid difficulty that swallows.
Description of drawings
Fig. 1 is a flow chart of the present invention
Fig. 2 is a standard curve
Fig. 3 is with the stripping curve figure of batch sample 6 times
The stripping homogeneity curve chart of Fig. 4 three batch samples
The specific embodiment
Embodiment 1
Under the lucifuge condition, the mecobalamin raw material is crossed 100 mesh sieves, carboxymethyl starch sodium, L-hydroxypropyl cellulose, microcrystalline Cellulose, polyvidone, micropowder silica gel, steviosin, magnesium stearate, Pulvis Talci are crossed 80 mesh sieves, take by weighing following amount:
Mecobalamin 0.5g
Carboxymethyl starch sodium 5g
L-hydroxypropyl cellulose 15g
Microcrystalline Cellulose 65g
Polyvidone 3g
Micropowder silica gel 5g
Steviosin 5g
Progressively increase principle with adjuvant and mecobalamin mix homogeneously according to equivalent; add an amount of 30% ethanol and make soft material, cross 20 order nylon mesh, make granule; and in 50 ℃ of dryings; cross 20 mesh sieves, granulate takes by weighing about 1g magnesium stearate and 1g Pulvis Talci; sieve and with the granule mix homogeneously; tabletting, is packed totally after the assay was approved by 1000.
Embodiment 2
Under the lucifuge condition, the mecobalamin raw material is crossed 100 mesh sieves, carboxymethyl starch sodium, L-hydroxypropyl cellulose, microcrystalline Cellulose, polyvidone, micropowder silica gel, steviosin, magnesium stearate, Pulvis Talci are crossed 80 mesh sieves, take by weighing following amount:
Mecobalamin 0.5g
Carboxymethyl starch sodium 7g
L-hydroxypropyl cellulose 20g
Microcrystalline Cellulose 60g
Polyvidone 2g
Micropowder silica gel 5g
Steviosin 5g
Progressively increase principle with adjuvant and mecobalamin mix homogeneously according to equivalent; add an amount of 30% ethanol and make soft material, cross 20 order nylon mesh, make granule; and in 55 ℃ of dryings; cross 20 mesh sieves, granulate takes by weighing about 1g magnesium stearate and 1g Pulvis Talci; sieve and with the granule mix homogeneously; tabletting, is packed totally after the assay was approved by 1000.
Embodiment 3
Under the lucifuge condition, the mecobalamin raw material is crossed 100 mesh sieves, carboxymethyl starch sodium, L-hydroxypropyl cellulose, microcrystalline Cellulose, polyvidone, micropowder silica gel, steviosin, magnesium stearate, Pulvis Talci are crossed 80 mesh sieves, take by weighing following amount:
Mecobalamin 0.5g
Carboxymethyl starch sodium 9g
L-hydroxypropyl cellulose 25g
Microcrystalline Cellulose 55g
Polyvidone 2g
Micropowder silica gel 5g
Steviosin 5g
Progressively increase principle with adjuvant and mecobalamin mix homogeneously according to equivalent; add an amount of 30% ethanol and make soft material, cross 20 order nylon mesh, make granule; and in 55 ℃ of dryings; cross 20 mesh sieves, granulate takes by weighing about 1g magnesium stearate and 1g Pulvis Talci; sieve and with the granule mix homogeneously; tabletting, is packed totally after the assay was approved by 1000.
Embodiment 4
Under the lucifuge condition, the mecobalamin raw material is crossed 100 mesh sieves, carboxymethyl starch sodium, L-hydroxypropyl cellulose, microcrystalline Cellulose, polyvidone, micropowder silica gel, steviosin, magnesium stearate, Pulvis Talci are crossed 80 mesh sieves, take by weighing following amount:
Mecobalamin 0-5g
Carboxymethyl starch sodium 10g
L-hydroxypropyl cellulose 30g
Microcrystalline Cellulose 50g
Polyvidone 1g
Micropowder silica gel 5g
Steviosin 5g
Progressively increase principle with adjuvant and mecobalamin mix homogeneously according to equivalent; add an amount of 30% ethanol and make soft material, cross 20 order nylon mesh, make granule; and in 60 ℃ of dryings; cross 20 mesh sieves, granulate takes by weighing about 1g magnesium stearate and 1g Pulvis Talci; sieve and with the granule mix homogeneously; tabletting, is packed totally after the assay was approved by 1000.
Comparative example 1
This experimental example carries out the mensuration of disintegration and dispersing uniformity to the product of embodiment 1-4, illustrates that dispersible tablet of the present invention has good disintegrate dispersive property.
Disintegration: pressing 2000 editions two appendix disintegration time mensuration methods of Chinese Pharmacopoeia and measure, is solvent with 20 ℃ ± 1 ℃ water, the complete disintegrate of planted agent in 3 minutes.
Dispersing uniformity: get two, place 100 milliliters of 20 ℃ ± 1 ℃ water joltings, the required time that writes down whole disintegrates and can pass through to sieve for No. 2.
Table 5:
The embodiment product Disintegration Dispersing uniformity
1 Disintegrate is complete in 2 minutes 15 seconds Disintegrate is complete in 3 minutes, and evenly suspendible is crossed sieve No. 2
2 Disintegrate is complete in 2 minutes 28 seconds Disintegrate is complete in 3 minutes, and evenly suspendible is crossed sieve No. 2
3 Disintegrate is complete in 2 minutes 15 seconds Disintegrate is complete in 3 minutes, and evenly suspendible is crossed sieve No. 2
4 Disintegrate is complete in 2 minutes 18 seconds Disintegrate is complete in 3 minutes, and evenly suspendible is crossed sieve No. 2
Experimental example 1
This experimental example is the test of embodiment 1-3 product mecobalamin dispersible tables quality standard research.
Content limit: this product is a mecobalamin dispersible tables.Specification is every and contains mecobalamin (C 63H 91C 0O 14P) 500 μ g, embodiment 1-3 product content measurement result is 100.24%, 101.36%, 99.14%.The result shows, the content of three batch samples is all between 90.0%～110.0%, so the content limit of this product is decided to be 90.0%～110.0% of labelled amount.
Character: odorless, tasteless, color depth is red, and adjuvant is white, after the compacting in flakes, shows pale red.So this product character is light red color chips.Three batch sample character see Table 6.
Table 6 three batch sample character check results
Embodiment Character
1 Light red color chips
2 Light red color chips
3 Light red color chips
Differentiate:
Spectrographic method is differentiated: the lucifuge operation.Get product, porphyrize adds water and makes the solution that every 1ml contains mecobalamin 50 μ g approximately, filter, get filtrate and measure according to spectrophotography (two appendix VIA of Chinese Pharmacopoeia version in 2000), visible this product has been located absorption maximum about wavelength 266,342,522nm, the results are shown in following table 7
Table 7 three batch sample maximum absorption wavelength measurement results
Embodiment Lot number Wavelength 1 Wavelength 2 Wavelength 3
1 030805 267 342 523
2 030806 266 342 522
3 030807 267 342 521
According to The above results, 266 ± 1nm, 342 ± 1nm, 522 ± 1nm can be used for product as characteristic absorption wavelength and differentiate.
High performance liquid chromatography is differentiated: in the chromatogram that writes down under the assay item, the retention time of need testing solution main peak is consistent with the retention time of reference substance main peak, and the retention time of the three batch sample need testing solution main peaks all retention time with the reference substance main peak is consistent.
The chemical colour reaction reaction is differentiated: get this product fine powder an amount of (being equivalent to mecobalamin 1mg approximately), add potassium acid sulfate 50mg, put in the crucible, calcination is to fusion, puts coldly, adds water 3ml, boil and make dissolving, add 1 of instructions phenolphthalein solution, dropping sodium solution is after show pale red, add sodium acetate 0.5g, spirit of vinegar 0.5ml and 1-Nitroso-2-naphthol-3.6-sodium disulfonate 0.5ml promptly shows red or orange red, adds hydrochloric acid 0.5ml, boiled 1 minute, color does not disappear.Three batch samples all show above-mentioned reaction.
The mixture of getting adjuvant is an amount of, with the method operation, does not show above-mentioned reaction, shows that adjuvant does not disturb this discriminating.
Check:, formulate the inspection item of this product according to requirement under two tablet general rules of Chinese Pharmacopoeia version in 2000 item.
Related substance: the related substance of this product detects the HPLC method that adopts.Self-control method is calculated, and chromatographic condition and system suitability are with " assay " item.
Lowest detectable limit: with reference to method among two appendix X of Chinese Pharmacopoeia version in 2000 VIII A, the amount that is about 3: 1 corresponding injection mecobalamin with signal to noise ratio is determined detectability.
It is an amount of to get reference substance, and accurate the title decides, and the solution that every 1ml contains mecobalamin 0.181 μ g/ml is made in serial dilution, sample introduction 20 μ l, and observation main constituent peak height is about 3 times of baseline noise, draws the mecobalamin lowest detection with this and is limited to 3.62ng.
Specificity:
The interference test of adjuvant: because this product content is extremely low, only be 500 μ g/ sheets, the sample major part is an adjuvant, should investigate the mensuration that whether influences related substance.We press tablet formulation, take by weighing right amount of auxiliary materials (the adjuvant amount that is equivalent to 4 of mecobalamin dispersible tables approximately), put in the 10ml measuring bottle, add the mobile phase dissolving and are diluted to scale, filter, and get subsequent filtrate 10 μ l sample introductions.Measure down and the record chromatogram according to the related substance item, do not have assorted peak substantially and occur.
Test accelerates the failure: be the impurity of verifying that can this method detect the related substance in the sample and degrade and produce when placing, we have carried out destructive testing under conditions such as strong illumination, high temperature, high humidity and acid, alkali, oxidation to sample, by detecting wherein impurity to determine the specificity of method.
1) 105 ℃ of samples of placing 40 hours are got in hot test, according to the operation down of determination of related substances item.
2) exposure experiments to light 2 days the sample of 45001x illumination of learning from else's experience is according to the operation down of determination of related substances item.
3) high wet test is taken at the sample of placing under the RH92.5% condition 10 days, according to the operation down of determination of related substances item.
4) sour failure test is got the fine powder an amount of (being equivalent to mecobalamin 4mg approximately) of this product porphyrize, put in the 10ml measuring bottle, add 0.1mol/L hydrochloric acid solution 5ml, shake up, placed 6 hours, the reuse sodium hydroxide solution is regulated about pH value to 6.0, adds mobile phase and is diluted to scale, shake up, according to the operation down of determination of related substances item.
5) the alkali failure test is got the fine powder an amount of (being equivalent to mecobalamin 4mg approximately) of this product porphyrize, put in the 10ml measuring bottle, add 0.1mol/L sodium hydroxide solution 5ml, shake up, placed 1 hour, the reuse hydrochloric acid solution is regulated about pH value to 6.0, adds mobile phase and is diluted to scale, shake up, according to the operation down of determination of related substances item.
6) oxidation test is got the fine powder an amount of (being equivalent to mecobalamin 4mg approximately) of this product porphyrize, puts in the 10ml measuring bottle, adds concentration and be 3% hydrogen peroxide 5ml, shakes up, and places 6 hours, adds mobile phase and is diluted to scale, shakes up, and operates down according to the determination of related substances item.
Algoscopy: lucifuge operation.Get this product, porphyrize, it is an amount of that precision takes by weighing fine powder, adds mobile phase jolting dissolving and dilution and make the solution that every 1ml contains mecobalamin 400 μ g approximately, filters, and gets subsequent filtrate as need testing solution; Precision is measured need testing solution 3.0ml and is put in the 100ml volumetric flask, adds mobile phase and is diluted to scale, shakes up, in contrast solution.Precision is measured contrast solution 10 μ l and is injected chromatograph of liquid, regulates detection sensitivity, and making the main constituent peak height is 10～20% of full scale.In addition precision is measured need testing solution and each 10 μ l of contrast solution respectively, sample introduction, and the record chromatogram is to 3 times of the retention time at main constituent peak.Each related substance peak area summation in the need testing solution all must not be greater than contrast solution main peak area, and three batch sample measurement results see Table 8.
Table 8 three batch sample determination of related substances results
Embodiment The impurity peaks gross area Contrast solution main peak area Its related substances
1 22658 39977 1.70％
2 20832 38523 1.62％
3 19981 40487 1.48％
In the related substance %=need testing solution related substance peak area and/contrast solution main peak area * 3%
According to above testing result, the limit of this product related substance must not be decided to be 3.0%.Uniformity of dosage units: belong to low dose of tablet, even for guaranteeing content, meet the regulation of Chinese Pharmacopoeia version in 2000, should do the uniformity of dosage units inspection.Press two appendix V of Chinese Pharmacopoeia version in 2000 D high effective liquid chromatography for measuring, chromatographic condition is seen " assay " item.
The preparation lucifuge operation of algoscopy reference substance solution.It is an amount of to get 105 ℃ of mecobalamin reference substances that are dried to constant weight, and accurate the title decides, and adds mobile phase jolting dissolving and dilution and makes the solution that every 1ml contains mecobalamin 50 μ g approximately, promptly.
The preparation lucifuge operation of need testing solution.Get 1 of this product, put in the 10ml measuring bottle, it is an amount of to add mobile phase, and jolting makes the mecobalamin dissolving and is diluted to scale, filters, and gets subsequent filtrate as need testing solution.Essence is got reference substance solution and each 10 μ l of need testing solution, injects the high performance liquid chromatogram chromatograph of liquid respectively, with calculated by peak area content, should meet the regulation of two appendix X of Chinese Pharmacopoeia version in 2000 E by external standard method, the results are shown in Table 9.
Table 9 mecobalamin dispersible tables Determination of Content Uniformity result
Embodiment 1 2 3
Every indicates content (%) 1 100.38 106.59 102.39
2 99.05 99.89 99.63
3 99.62 100.75 97.25
4 101.66 97.05 99.58
5 103.05 105.40 100.21
6 104.99 98.86 97.78
7 104.31 103.61 102.49
8 102.48 102.67 104.00
9 101.34 102.02 100.36
10 104.13 98.90 106.05
Meansigma methods 102.10 101.57 100.97
S 2.042 3.054 2.749
A+1.80S 5.776 7.067 5.918
Conclusion Qualified Qualified Qualified
Dissolution
Assay method intends adopting high-efficient liquid phase technique to measure, and carries out with reference to content assaying method.
Recovery test
Precision takes by weighing mecobalamin an amount of (high, medium and low three groups) respectively, press recipe quantity and add corresponding each adjuvant, put in the 100ml stripping rotor, oar method 75 is changeed, stirred 10 minutes, get solution and filter in right amount, and be mixed with 9 parts of solution of high, medium and low three groups of concentration (be equivalent to respectively measure concentration 5 μ g/ml 120%, 100% and 80%); It is an amount of that precision takes by weighing the mecobalamin reference substance in addition, adds mobile phase and quantitatively dilute the reference substance solution of making 5 μ g/ml.Get each 20 μ l of need testing solution and reference substance solution respectively and inject high performance liquid chromatograph, measure in accordance with the law, press external standard method with calculated by peak area, the response rate.The results are shown in Table 10.
Table 10 mecobalamin dissolution determination method is learned the recovery test result
Addition (mg) Peak area The amount of recording (mg) The response rate (%) Average recovery rate (%) RSD (％)
Low concentration 10.3 11.8 11.1 18525 20923 19231 10.50 11.86 10.91 100.99 100.55 98.25 100.33±1.35 1.35
Middle concentration 16.6 17.9 17.6 29846 31513 31114 16.92 17.87 17.64 99.96 101.83 100.25
High concentration 20.2 20.8 20.6 35927 36081 36650 20.37 20.46 20.78 100.86 98.37 100.89
Reference substance 5.28 μ g/ml 37244
It is higher that this law is measured the dissolution accuracy.
The selection of leaching condition
Determining of little agar diffusion method
Because of the labelled amount of mecobalamin dispersible tables is 500 μ g, content is less, so we adopt the cuvette method to measure the dissolution of mecobalamin, and quantity of solvent is 100ml.
Determining of dissolution medium
Mecobalamin is molten in the water part omitted, is gastric soluble tablet, so we have selected the hydrochloric acid of water, 0.1mol/L to test as solvent, and rotating speed is 75 rev/mins, sampling respectively in 3,5,10 minutes, and measurement result sees Table 11, table 12.
Table 11 dissolution medium is the dissolution (%) of water
Dissolution time (minute) 1 2 3 4 5 6 On average
3 88.69 89.98 84.97 86.98 87.43 92.23 88.26
5 101.10 103.42 99.22 100.34 100.99 106.65 101.95
10 100.98 103.87 99.74 101.18 101.00 105.71 102.08
Table 12 dissolution medium is the dissolution (%) of 0.1mol/ml hydrochloric acid
Dissolution time (minute) 1 2 3 4 5 6 On average
3 93.17 85.25 91.86 91.15 88.45 88.29 89.69
5 104.56 96.10 99.42 99.56 95.87 98.58 99.02
10 104.85 96.64 100.58 100.92 98.12 102.04 100.52
Result of the test shows, mecobalamin dispersible tables is at the stripping of water and 0.1mol/l hydrochloric acid two media no significant difference as a result, and more convenient because of water, so we determine that water is dissolution medium.
The selection of stripping rotating speed
Our initial option 50 rev/mins, 75 rev/mins two kinds of rotating speeds, test, sampling was respectively measured at 3,5,10 minutes, measurement result sees Table 13, table 14.
Table 13 rotating speed is 50 rev/mins a dissolution (%)
Dissolution time (minute) 1 2 3 4 5 6 On average
3 81.27 85.98 76.94 80.07 77.85 75.31 79.57
5 86.71 89.71 89.71 88.53 90.96 89.12 89.12
10 105.30 97.42 104.62 102.14 99.18 98.13 101.13
Table 14 rotating speed is 75 rev/mins a dissolution (%)
Dissolution time (minute) 1 2 3 4 5 6 On average
3 94.50 86.63 89.43 90.17 84.27 86.98 88.66
5 102.79 104.82 103.61 102.41 96.91 98.48 101.50
10 100.02 103.42 106.05 105.71 98.20 103.27 102.78
The result shows, when rotating speed is 75 rev/mins, and basic stripping fully when average dissolution reached 88.66%, 5 minute in 3 minutes; Rotating speed is 50 rev/mins, and then dissolution is on the low side.For making test accurately, so determine that rotating speed is 75 rev/mins.
The stripping uniformity test
We are being dissolution medium with water, and rotating speed is under 75 rev/mins the condition, with three batch samples done with batch and batch between the stripping uniformity test.
5.3.3.1 uniformity test in batch:
Get embodiment 1 product, measure 6 times dissolution, the results are shown in following table 15.
Table 15 dissolution uniformity test result
First group
Sample time (branch) Dissolution (%)
1 2 3 4 5 6 On average
3 82.38 85.64 92.13 90.26 85.73 88.73 87.48±3.561
5 90.50 95.27 105.39 102.83 97.86 98.67 98.42±5.311
10 92.46 95.70 104.55 98.79 96.48 100.67 98.11±4.216
Second group
Sample time (branch) Dissolution (%)
1 2 3 4 5 6 On average
3 94.09 86.92 81.75 86.16 90.42 91.65 88.50±4.773
5 105.32 97.22 98.48 93.33 99.91 100.34 99.10±3.954
10 104.43 99.23 95.38 92.25 99.74 101.79 98.21±4.390
The 3rd group
Sample time (branch) Dissolution (%)
1 2 3 4 5 6 On average
3 88.77 86.16 90.68 91.47 90.58 83.65 88.55±3.066
5 99.08 93.61 103.36 102.73 97.20 95.47 98.57±3.916
10 98.24 94.62 102.91 101.54 96.48 94.85 98.11±3.476
The 4th group
Sample time (branch) Dissolution (%)
1 2 3 4 5 6 On average
3 88.78 91.00 88.53 92.42 85.45 90.24 89.40±2.413
5 99.41 106.40 101.82 104.11 94.16 100.86 101.13±4.214
10 100.94 100.46 105.82 101.99 94.50 99.83 100.59±3.664
The 5th group
Sample time (branch) Dissolution (%)
1 2 3 4 5 6 On average
3 94.69 90.15 89.48 91.48 85.01 88.28 89.85±3.230
5 107.56 101.44 102.39 101.72 94.73 96.43 100.71±4.590
10 106.31 102.86 103.58 101.30 95.61 97.99 101.27±3.900
The 6th group
Sample time (branch) Dissolution (%)
1 2 3 4 5 6 On average
3 82.53 87.34 92.51 95.59 91.48 91.92 90.23±4.602
5 92.44 99.89 105.95 103.31 105.84 104.24 101.94±5.154
10 92.82 100.24 105.29 101.59 104.85 103.74 101.42±4.638
Draw stripping curve by above-mentioned six groups of stripping results, see Figure 22.The result shows: this dissolution determination method repeatability is better.
5.3.3.2 stripping uniformity test between batch
Get three batch samples (embodiment 1,2,3) and carry out dissolution determination respectively 1 time, the results are shown in following table 16:
Table 16 mecobalamin dispersible tables is criticized a stripping uniformity test
Embodiment 1
Sample time (branch) Dissolution (%)
1 2 3 4 5 6 On average
3 88.78 91.00 88.53 92.42 85.45 90.24 89.40±2.413
5 99.41 106.40 101.82 104.11 94.16 100.86 101.13±4.214
10 100.94 100.46 105.82 101.99 94.50 99.83 100.59±3.664
Embodiment 2
Sample time (branch) Dissolution (%)
1 2 3 4 5 6 On average
3 86.66 91.59 91.17 84.99 90.25 85.74 88.40±2.934
5 98.90 101.07 102.33 99.07 106.42 98.96 102.33±2.945
10 99.61 99.35 105.00 98.07 104.55 96.77 100.56±3.423
Embodiment 3
Sample time (branch) Dissolution (%)
1 2 3 4 5 6 On average
3 87.46 88.47 84.94 92.42 91.68 90.68 89.27±2.841
5 97.99 98.10 97.38 105.43 99.63 101.99 100.09±3.098
10 98.21 100.45 98.08 103.72 99.57 100.93 100.16±2.088
Draw the stripping curve of three batch samples by The above results, see Figure 23.
Table 17 three batch samples batch between the stripping difference results
Sample time (branch) 3 5 10
Embodiment 1 89.40 101.13 100.59
Embodiment 2 88.40 102.33 100.56
Embodiment 3 89.27 100.09 100.16
Average stripping (%) 89.02±0.54 101.18±1.12 100.44±0.24
RSD(％) 0.61 1.10 0.24
The result shows: RSD is less than 5% as a result for the dissolution determination between different batches, and the production technology of this product is more stable.Dissolution method: lucifuge operation.Getting this product, according to dissolution determination method (two appendix XC of Chinese Pharmacopoeia version in 2000 three therapeutic methods of traditional Chinese medicine), is solvent with 100ml water, and rotating speed is 75 rev/mins, in accordance with the law operation.Through 10 minutes, it was an amount of to get solution, filtered, and got subsequent filtrate as need testing solution.It is an amount of that precision takes by weighing 105 ℃ of mecobalamin reference substances that are dried to constant weight in addition, add the mobile phase dissolving, and the solution that every 1ml contains mecobalamin 5 μ g, product solution in contrast made in dilution.Essence is got reference substance solution and each 20 μ l of need testing solution, injects high performance liquid chromatograph respectively, the record peak area, and by the stripping quantity of external standard method with every of calculated by peak area, the dissolution computing formula is dissolution (%)=(A Sample/ W Sample)/(A Mark/ W Mark).
Three batch sample dissolution determinations: according to said method, measure the dissolution of three batch samples, measurement result sees Table 18.
Table 18 three batch sample dissolution test results
Embodiment 1 2 3
Stripping quantity % 100.94 100.46 105.82 101.99 94.50 99.83 99.61 99.35 105.00 98.07 104.55 96.77 98.21 100.45 98.08 103.72 99.57 100.93
Meansigma methods 100.59％±3.664 100.56％±3.423 100.16％±2.088
According to the measurement result of above-mentioned sample, the stripping quantity of this product in the time of 10 minutes limited the quantity of, and to be decided to be 80% of labelled amount be comparatively suitable.
Dispersing uniformity is according to the requirement inspection of " dispersible tablet " in " two appendix I of Chinese Pharmacopoeia version in 2000 A tablet ", and all whole disintegrates in 3 minutes of three batch samples are also sieved by No. 2.The results are shown in following table 19
Table 19 three batch sample dispersing uniformity check results
Embodiment 1 2 3
The result Disintegrate in 30 seconds in 1 fen Disintegrate in 45 seconds in 1 fen Disintegrate in 30 seconds in 1 fen
Conclusion Up to specification Up to specification Up to specification
The result shows that the dispersing uniformity of this product is up to specification.
Limit test of microbe: measure according to microbial limit test (two appendix XI of Chinese Pharmacopoeia version in 2000 J), three batch sample check results see the following form 20
Table 20 limit test of microbe result
Embodiment 1 2 3
Bacterial population (individual/g) fungi count is (individual/g) the escherichia coli demodicid mite conclusion of living 40＜10 do not detect up to specification 60＜10 do not detect up to specification 30＜10 do not detect 10 do not detect up to specification
Assay: adopt high performance liquid chromatography (two appendix VD of Chinese Pharmacopoeia version in 2000).
Chromatographic condition and system suitability octadecylsilane chemically bonded silica are filler; With acetonitrile-0.03mol/L potassium dihydrogen phosphate (18: 82), regulating pH to 4.5 with phosphoric acid or 0.2mol/L potassium hydroxide is mobile phase; The detection wavelength is 342nm.Strict lucifuge operation.Number of theoretical plate calculates with mecobalamin main constituent peak, should be not less than 2000, and the separating degree of main constituent peak and adjacent impurity peaks should meet the requirements.
By above-mentioned chromatographic condition, strict lucifuge operation detects three batch samples (embodiment 1,2,3), the record chromatogram, and theory of computation plate number is respectively n=3477,3480,3537, and separating degree R＞1.5 meet the requirements.
The system suitability replica test: get the mecobalamin solution 10 μ l of 200 μ g/ml, continuous sample introduction 5 times, the record peak area, RSD is 0.68% as a result, and is less than 2%, up to specification.
The checking of method
Linear relationship and scope: lucifuge operation.Precision takes by weighing mecobalamin reference substance 21.7mg, puts in the 50ml measuring bottle, adds the solution that the mobile phase dissolving is made into 434ug/ml, and accurate absorption 0.2,2,4,8,16,25ml place the 25ml measuring bottle, are diluted to scale with mobile phase.Be mixed with every 1ml and contain the solution of mecobalamin 3.472,34.72,69.44,138.88,277.76,434 μ g, essence is got 10 μ l sample introductions, each concentration sample introduction 2 times, average, with peak area (A) sample concentration (C) is made regression Calculation, regression equation the results are shown in Table 21.
The linear relationship of table 21 mecobalamin peak area and concentration
Concentration C (μ g/ml) 3.472 34.72 69.44 138.88 277.76 434
Peak area A 11672 116062 229544.5 457632.5 906422 1423171.5
Regression equation A＝3271.6C+1577.7 r＝0.9999
Standard curve is seen Fig. 2.
Recovery test: it is an amount of that precision takes by weighing mecobalamin respectively, press recipe quantity and add corresponding each adjuvant, with mobile phase dissolving, filter, and be mixed with 9 parts of solution of high, medium and low three groups of concentration (be equivalent to respectively measure concentration 200 μ g/ml 120%, 100% and 80%); It is an amount of that precision takes by weighing the mecobalamin reference substance in addition, adds mobile phase and quantitatively dilute the reference substance solution of making 200 μ g/ml.Get each 10 μ l of need testing solution and reference substance solution respectively and inject high performance liquid chromatograph, measure in accordance with the law, press external standard method with the calculated by peak area response rate.The results are shown in Table 22
Table 22 mecobalamin content assaying method is learned the recovery test result
Addition (mg) Peak area The amount of recording (mg) The response rate (%) Average recovery rate (%) RSD (％)
Low concentration 34.9 35.2 31 567859.5 589113.5 509594 34.35 35.63 30.82 98.42 101.23 99.43 99.61±0.90 0.90
Middle concentration 42.6 44.4 40.3 694672.5 732887.5 661477.5 59.27 58.73 58.82 98.78 101.25 99.19
High concentration 48.2 51.6 48.6 802864 847985 799837.5 48.56 51.29 48.38 100.75 99.40 99.54
Reference substance 202 μ g/ml 715821.5
The result shows that the content accuracy of this law mensuration mecobalamin is higher.Stability of solution: lucifuge operation.Compound concentration is the solution of 200 μ g/ml, at 0,4,8,24,48 hour difference sample introduction 10 μ l, and the record chromatogram, and calculate its relative standard deviation RSD, the results are shown in Table 23.
Table 23 stability of solution measurement result
Time (min) 0 30 60 90 120
Peak area A 621057.5 629182 623860.5 614323.5 601047
±SD 617894.1±10835.1
RSD％ 1.75％
Specificity compares the HPLC chromatogram of adjuvant and the HPLC collection of illustrative plates of sample, and the adjuvant of visible tablet does not disturb the assay of mecobalamin.
Algoscopy
The preparation lucifuge operation of reference substance solution.It is an amount of to get 105 ℃ of mecobalamin reference substances that are dried to constant weight, accurate claims surely, adds the mobile phase dissolving, and dilution makes every 1ml and contain the solution that mecobalamin is 200 μ g, promptly.
The preparation lucifuge operation of need testing solution.Get 20 of this product, accurate claim fixed, porphyrize, precision takes by weighing in right amount (being equivalent to mecobalamin 2mg approximately) and puts in the 10ml measuring bottle, adds the mobile phase dissolving and is diluted to scale, shakes up, filtration promptly gets every 1ml and contains the solution that mecobalamin is 200 μ g.
Algoscopy gets need testing solution respectively and each 10 μ l of reference substance solution inject high performance liquid chromatogram, measures in accordance with the law, presses external standard method with calculated by peak area, promptly.
Measure three batch samples and the results are shown in Table 24.
Table 24 mecobalamin dispersible tables three batch sample assay results
Embodiment Average sheet heavy (g) Content (labelled amount %)
1 0.1045 100.24
2 0.1073 101.36
3 0.1059 99.14
Conclusion: three batches of mecobalamin dispersible tables contain C 63H 91CoN 13O 14P is all in 90.0～110.0% regulations of labelled amount.Stripping curve figure with batch sample 6 times sees Fig. 3.
Three batch sample stripping homogeneity curve charts are seen Fig. 4.
Experimental example 2
This experimental example is the stability experiment data of product of the present invention.
1. sample: embodiment 1,2,3 products, specification: 500 μ g/ sheets.
2. investigation project and condition:
2.1 factor affecting test:
2.1.1 strong illumination test: get for a collection of (lot number: 030820), place airtight clean container, under 45001x ± 5001x illumination, place,, detect of test agent according to investigating content respectively at sampling in 0,5,10 day.
2.1.2 hot test: get for test agent, under 60 ℃ of conditions, place,, detect according to investigating content respectively at sampling in 0,5,10 day.
2.1.3 high humility test: get for test agent, opening places the clean container of airtight constant humidity, and (saturated KNO3 solution RH92.5%) and under RH75% ± 5% (the saturated NaCl solution) condition is placed at RH90.0% ± 5%, respectively at sampling in 0,5,10 day, detect according to investigating content.
2.2 accelerated test: 3 batches of test samples, simulation listing packing 40 ℃ ± 2 ℃ of temperature, was placed 6 months under the condition of relative humidity 75% ± 5%; At 0,1,2,3,6 the end of month of investigating, sampling is once carried out project according to the content of investigating and is detected respectively.
2.3 long term test: 3 batches of test samples, simulate the listing packing, 25 ℃ ± 2 ℃ of temperature, placed 12 months under the condition of relative humidity 60% ± 10%, take a sample once with 0,3,6 the end of month of investigating respectively, carry out project according to the content of investigating and detect.
3. investigation content:
Appearance luster, dispersing uniformity, content, related substance, dissolution
4. result:
4.1 factor affecting test:
4.1.1 exposure experiments to light: illumination is after 10 days, and related substance rises to 10.61% from 1.68% in the mecobalamin dispersible tables, illumination after 10 days content drop to 78.13% from 100.37%, all great changes have taken place, dissolution has minimizing slightly, and other index does not have significant change, sees Table 25, Fig. 1～10.
4.1.2 hot test: 60 ℃ of hot conditionss were placed after 10 days, and the related substance of mecobalamin dispersible tables rises to 2.74% from 1.68%, and other indexs do not have significant change, saw Table 26, Figure 11～14.
4.1.3 high humility test: the RH92.5% super-humid conditions was placed after 5 days, and weight increases by 9.32%, changes into as requested under the RH75% high humidity and investigates, places after 10 days, and weight increase by 2.14%, the basic no change of other indexs sees Table 27, Figure 15～18.
4.2 accelerated test condition: the accelerated test condition was placed after 6 months, the mecobalamin related substance rises to 2.24%, 2.18%, 2.23% from 1.68%, 1.56%, 1.76% respectively in three batch samples, increase is arranged slightly, and other indexs such as content do not have significant change, see Table 28, Figure 19～42.
4.3 long term test: long term test has been carried out 9 months up till now, and is still continuing.Under the condition of this test, to place after 9 months, the basic no changes of index such as the related substance of mecobalamin dispersible tables, content see Table 29 in three batch samples; Figure 43～57.
5. conclusion: factorial experiments is the result show, mecobalamin dispersible tables is responsive to light, and related substance increases, and hot conditions is influential slightly to mecobalamin dispersible tables related substance, content, and the super-humid conditions sheet heavily increases comparatively obvious, but does not influence other quality index; Accelerated test and long-term the placement were tested after 9 months, and the quality of this product does not have obvious variation, and every index is still in the claimed range of quality standard.Therefore, this product should be kept away high temperature also strict lucifuge, airtight preservation, and effect duration is decided to be 2 years.
Table 25 illumination (45001x) result of the test
Embodiment 1
Time (my god) Appearance luster Related substance % Dissolution (%) Dispersing uniformity Content (labelled amount %)
0 5 10 Light red color chips, the complete light red color chips of smooth surface, the complete light red color chips of smooth surface, smooth surface is complete 1.68 5.16 10.61 Up to specification up to specification Up to specification up to specification 100.37 86.01 78.13
Table 26 high temperature (60 ℃) result of the test
Embodiment 2
Time (my god) Appearance luster Related substance % Dissolution (%) Dispersing uniformity Content (labelled amount %)
0 5 Light red color chips, the complete light red color chips of smooth surface, smooth surface is complete 1.68 2.73 Up to specification Up to specification 100.37 99.60
10 Light red color chips, smooth surface is complete 2.74 Up to specification Up to specification 101.15
Table 27 high humidity (RH75%) influences result of the test
Embodiment 3
Time (my god) Appearance luster Related substance % Dissolution (%) Dispersing uniformity Moisture absorption (%) Content (labelled amount %)
0 5 10 The smooth complete moisture absorption of smooth surface full surface fluffs 1.68 1.56 1.82 Up to specification up to specification Up to specification up to specification - 0.93 2.14 100.37 99.19 100.30
Table 28 accelerated test result
Embodiment Time (moon) Appearance luster Related substance (%) Dissolution (%) Dispersing uniformity Content (the cup amount of showing %)
1 0 1 2 3 6 Light red color chips, the complete light red color chips of smooth surface, the complete light red color chips of smooth surface, the complete light red color chips of smooth surface, the complete light red color chips of smooth surface, smooth surface is complete 1.68 1.66 1.71 2.04 2.24 Up to specification up to specification Up to specification up to specification 100.37 99.54 99.94 100.22 101.09
2 0 1 2 3 6 Light red color chips, the complete light red color chips of smooth surface, the complete light red color chips of smooth surface, the complete light red color chips of smooth surface, the complete light red color chips of smooth surface, smooth surface is complete 1.56 1.89 1.88 2.46 2.18 Up to specification up to specification Up to specification up to specification 98.46 98.87 98.25 99.18 99.39
3 0 1 2 3 6 Light red color chips, the complete light red color chips of smooth surface, the complete light red color chips of smooth surface, the complete light red color chips of smooth surface, the complete light red color chips of smooth surface, smooth surface is complete 1.76 1.90 1.81 1.80 2.23 Up to specification up to specification Up to specification up to specification 102.14 102.00 102.03 101.44 101.88
Table 29 long-term test results
Embodiment Time/moon Appearance luster Related substance % Dissolution (%) Dispersing uniformity Content (labelled amount %)
1 0 3 6 9 Light red color chips, the complete light red color chips of smooth surface, the complete light red color chips of smooth surface, the complete light red color chips of smooth surface, smooth surface is complete 1.68 1.74 1.66 1.60 Up to specification Up to specification 100.37 100.54 100.72 100.21
2 0 3 6 9 Light red color chips, the complete light red color chips of smooth surface, the complete light red color chips of smooth surface, the complete light red color chips of smooth surface, smooth surface is complete 1.56 1.86 1.56 1.58 Up to specification Up to specification 98.46 98.88 98.85 99.95
3 0 3 6 9 Light red color chips, the complete light red color chips of smooth surface, the complete light red color chips of smooth surface, the complete light red color chips of smooth surface, smooth surface is complete 1.78 1.77 1.81 1.92 Up to specification Up to specification 102.14 100.99 102.05 100.53
Experimental example 3
This experimental example is a product pharmacological research data of the present invention.
1 pharmacological action
1.1 mecobalamin is easily absorbed to Wistar Mus intramuscular injection mecobalamin and cobalamin by submicroscopic structure such as nucleus, the mitochondrion etc. of neurocyte, dispensing 2,5, behind the 18h Mus is put to death, observation demonstration mecobalamin can shift well in 5h and enter in the organelle, still keep high concentration during 18h, thereby bring into play pharmacological action effectively, cobalamin is then very poor.
1.2 mecobalamin promotes interior nucleic acid of neurocyte and proteinic synthesizing to raise with the B that is deficient in vitamin to rat 12With the diet of folic acid, by taking in 3The kinetics that the uridnine of H labelling is observed the spinal nerves cell RNA changes, and finds to compare with cobamamide group and normal diet group with the mecobalamin group, and the former RNA increases.
1.3 mecobalamin promotes myelin phospholipid synthetic with mecobalamin and other vitamin Bs 12Add respectively and observe neural myelin formation in the Mus cerebellum cell culture fluid, the neural myelin of the visible Mus of mecobalamin group forms and increases, and other vitamin B 12Group does not then have this phenomenon.
1.4 mecobalamin promotes axoplasmic transport and axon regeneration to cause the rat diabetes model with streptozotocin, rat axoplasmic transport speed reduces by 50%.Mecobalamin is proteinic synthetic by promoting, makes the axon skelemin carry normalization, thereby makes axoplasmic transport recover normal.The axoplasmic transport speed of mecobalamin group and matched group has significant difference.Cause Mus peripheral neuropathy (axonal degeneration) model with acrylamide, intramuscular injection mecobalamin group is compared with intramuscular injection normal saline group, and the number of the former aixs cylinder and myelin is more.Show that mecobalamin can stimulate axon regeneration.
1.5 after the normalization rat sciatic nerve extruding degeneration of acetyl choline content, wherein use mecobalamin for one group in recovery that mecobalamin acceleration synapse is transmitted and the brain, another group then need not.Measure end plate potential with microelectrode method in the born of the same parents, found that the mecobalamin group end plate potential recovery early, show that mecobalamin can quicken the synapse transmission.Experimentize with the feedstuff raising rat that lacks choline, find that mecobalamin can make acetyl choline content normalization in the low brain.
1.6 mecobalamin has the function cut-out rabbit peroneal nerve that promotes neuranagenesis and is sewed up, to mecobalamin group (500ugd -1, 30d altogether) and the neurohistology inspection of matched group, comprise motor nerve conduction velocity and axoplasmic transport.Inject in the dorsal part nerve root 3The H-leucine is to observe axoplasmic transport speed.The myelinated nerve fiber quantity that experiment shows mecobalamin group and matched group all has growth when sewing up, but mecobalamin group showed increased, what leucine was transported in aixs cylinder in the mecobalamin group is more, the faster (P＜0.01=of speed.The motor nerve conduction velocity of mecobalamin group also have clear improvement (P＜0.01).
2 pharmacokinetics: rat is oral 57Co-CH 3B 12(25ugkg -1), behind the 72h, the concentration of kidney, adrenal gland, pancreas, liver, stomach is higher, and it is lower that muscle, testis, cranial nerve etc. are located concentration.Give the rat intravenous injection 57Co-CH 3B 12(10ugkg -1), behind the 24h, kidney, adrenal gland, intestinal, pancreas, concentration pituitary are higher, and it is low that eye, spinal cord, brain, muscle etc. are located concentration.Mainly from fecaluria, discharge.The mecobalamin tablet, the once oral 120ug of healthy people, 1500ug, all 3h reaches blood medicine peak value after administration, and its absorption is dose dependent.t 1/2Deng pharmacokinetic parameter shown in table 30.
The pharmacokinetic parameter of table 30 mecobalamin
Dosage (ug) T max(h) C max(pg·ml -1) ΔC max(pg·ml -1) ΔC max％ (％) ΔAUC 12 0(pg·h·ml -1) t 1/2(h)
120 2.8±0.2 743±47 37±15 5.1±2.1 168±58 Can not calculate
1500 3.6±0.5 972±55 255±51 36.0±7.9 2033±51 12.5
8h after taking medicine, total B in the urine 12Excretion be after the medication 24 h excretions 40%～80%.Healthy people uses 12 weeks, 1500ugd continuously -1The total B of serum in 4 weeks to the drug withdrawal 12Changing value as follows: after 4 weeks of administration, the total B of serum 12Changing value be about before the administration 2 times, increase gradually later on, to 12 week about 2.8 times of back, end 1.8 times of 4 weeks of administration.A healthy intramuscular injection of people or quiet notes mecobalamin 500ug, intramuscular injection T MaxBe 0.9 ± 0.1h, quiet notes T Max3min.The total B of highest serum 12Concentration increases part (C MaxBe respectively 22.4 ± 1.1,85.0 ± 8.9ngml -1144h after the administration, the AUC of intramuscular injection, quiet notes administration is respectively 204 ± 12.9,358.6 ± 34.4nghml -1The quiet notes mecobalamin of healthy people logotype 10d, the total B of serum 12Concentration (C Min) rise along with the increase of administration natural law.C after the administration in first day, second day, the 3rd day MinBe respectively 3.9 ± 1.2ngml -1, 5.3 ± 1.8ngml -1, 6.8 ± 1.5ngml -1, back one concentration is maintained to post-drug period.
Experimental example 4
This experimental example is a product toxicological study data of the present invention.
I. acute toxicity: its mouse oral is taken the LD50 of mecobalamin greater than 1000mg/kg.Rat oral is taken the LD50 of mecobalamin greater than 500mg/kg.
II. subacute toxicity: rat oral administration 1 month, dosage are respectively 0.2,2.0,20mg/kg/ day, and general symptom, body weight, blood, urine organ weights and histopathologic examination etc. all do not have special variation.
III. long term toxicity: rat oral administration 6 months, dosage are respectively 0.2,2.0,20mg/kg/ day, and general symptom, body weight, blood, urine, organ weights and histopathologic examination etc. all do not have special variation.
IV. genotoxicity: at the period of organogenesis of rat and mice oral administration, dosage is respectively 0.2,2.0,20mg/kg/ day, and the young and new cub of tire all occurs unusual, and also not seeing has the teratogenesis phenomenon to take place.
V. methylating of inorganic mercury: in vitro, this product and mercuric chloride reaction generate methyl mercury, but do not observe this reaction under the condition that protein such as human blood exist.
VI. in addition, during the feedstuff raising rat that adds with inorganic mercury, do not increase the methyl mercury of organism because of oral administration and this product.Further the feedstuff that adds with inorganic mercury is raised male rat, and oral administration and 6 months, give 1.5 respectively, 15mg/kg/ day mecobalamin, in the observation of general symptom, blood, urine examination and histopathology, its result there is no the influence that brings because of throwing simultaneously and inorganic mercury and mecobalamin.

Claims (4)

1, a kind of mecobalamin dispersible tables is characterized in that, per 1000 of described dispersible tablet contains following component:
Mecobalamin 0.5g
Carboxymethyl starch sodium 5-10g
L-hydroxypropyl cellulose 15-30g
Microcrystalline Cellulose 50-65g
Polyvidone 1-3g
Micropowder silica gel 5g
Steviosin 5g
Magnesium stearate 1g
Pulvis Talci 1g.
2, mecobalamin dispersible tables according to claim 1 is characterized in that, per 1000 of described dispersible tablet contains following component:
Mecobalamin 0.5g
Carboxymethyl starch sodium 5g
L-hydroxypropyl cellulose 15g
Microcrystalline Cellulose 65g
Polyvidone 3g
Micropowder silica gel 5g
Steviosin 5g
Magnesium stearate 1g
Pulvis Talci 1g.
3, mecobalamin dispersible tables according to claim 1 and 2 is characterized in that, described dispersible tablet specification is 500 μ g/ sheets.
4, the preparation method of mecobalamin dispersible tables, it is characterized in that, described method is: under the lucifuge condition, the mecobalamin raw material is crossed 100 mesh sieves, carboxymethyl starch sodium, L-hydroxypropyl cellulose, microcrystalline Cellulose, polyvidone, micropowder silica gel, steviosin, magnesium stearate, Pulvis Talci are crossed 80 mesh sieves, take by weighing following amount:
Mecobalamin 0.5g
Carboxymethyl starch sodium 5-10g
The low hydroxypropyl cellulose 15-30g that replaces
Microcrystalline Cellulose 50-65g
Polyvidone 1-3g
Micropowder silica gel 5g
Steviosin 5g
Progressively increase principle with adjuvant and mecobalamin mix homogeneously according to equivalent; add an amount of 30% ethanol and make soft material, cross 20 order nylon mesh, make granule; and in 50～60 ℃ of dryings; cross 20 mesh sieves, granulate takes by weighing 1g magnesium stearate and 1g Pulvis Talci; sieve and with the granule mix homogeneously; tabletting, is packed totally after the assay was approved by 1000.
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CN102716102B (en) * 2012-05-30 2014-02-05 杭州康恩贝制药有限公司 Mecobalamin tablet and preparation method thereof
CN104784143A (en) * 2014-01-17 2015-07-22 南京瑞尔医药有限公司 Mecobalamin tablet composition
CN104784049A (en) * 2014-01-17 2015-07-22 南京瑞尔医药有限公司 Preparation method for mecobalamin tablets
CN105997917A (en) * 2016-05-12 2016-10-12 扬子江药业集团南京海陵药业有限公司 Improved mecobalamine tablets and preparing method thereof
CN106236719B (en) * 2016-08-30 2021-01-19 扬子江药业集团南京海陵药业有限公司 Pharmaceutical composition containing mecobalamin and preparation method thereof
CN109820831A (en) * 2019-03-26 2019-05-31 江苏四环生物制药有限公司 A kind of method of granulating of mecobalamin dispersible tables
CN111044625A (en) * 2019-11-26 2020-04-21 卓和药业集团有限公司 Novel method for determining related substances in mecobalamin dispersible tablets
CN112121024A (en) * 2020-11-03 2020-12-25 卓和药业集团有限公司 Mecobalamin orally disintegrating tablet and preparation process thereof
CN112630366A (en) * 2020-12-18 2021-04-09 卓和药业集团有限公司 High performance liquid chromatography detection method for content of mecobalamin dispersible tablets
CN112730658A (en) * 2020-12-18 2021-04-30 卓和药业集团有限公司 Method for analyzing dissolution curve of mecobalamin tablets

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