CN102103133A - Method for measuring related substances in glucosamine by using high performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) - Google Patents
Method for measuring related substances in glucosamine by using high performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) Download PDFInfo
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- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
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- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
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Abstract
The invention discloses a method for detecting related substances in glucosamine. In the method, an evaporative light scattering detector (ELSD) is adopted, water added with 0.1 percent formic acid and 0.1 percent ammonia water, and acetonitrile in a ratio of (1-40):(20-70) are taken as a mobile phase, and a liquid chromatographic column for zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC, 150mm*4.6mm, and 5mu m) is taken as a chromatographic column. By the method for measuring the related substances in the glucosamine and a glucosamine-containing related preparation, the defect that the glucosamine is not absorbed in an ultraviolet region and is difficult to detect is overcome, and the impurities and degradation product of the glucosamine can be quickly and accurately detected. The method has the advantages of good separation effect, high sensitivity, high analysis speed, high accuracy and the like, the detection level of the related substances in glucosamine raw material and preparation is improved, and the quality control of the glucosamine raw material and preparation is enhanced.
Description
Technical field
The present invention relates to a kind of Glucosamine and contain the related substance detection method of the related preparations of Glucosamine, belong to pharmaceutical preparation analyzing and testing field.
Background technology
Glucosamine, a kind of food additives are to remove the osteoarthropathy pain, help to improve people's quality of life.The incidence of disease of osteoarthritis in the elderly is very high, in Beijing up to 38.7%, Shanghai 13%, and this disease does not still have desirable methods of treatment at present.For this reason, the ailing key that becomes osteoarthritis treatment of prevention and alleviation.At present, the prevention of using always in the world and the functional food ingredient of relief from osteoarthritis mainly contain Glucosamine, hyaluronic acid, collagen peptide etc.Glucosamine is at first found in 1969 by German doctor the curative effect of osteoarthritis.In recent years studies show that it is connective tissue and cartilage cell's a principal ingredient, is the elementary cell of glycoprotein and glycosaminoglycan chain in the articular cartilage matrix.Additional Glucosamine can promote the glycoprotein and the glycosaminoglycan of cartilage cell's synthesising physiological, to repair impaired cartilage.In addition, Glucosamine also can prevent the generation of the superoxide radical that is harmful to, studies show that it has the effect of anti-inflammatory and pain relieving, therefore is used as to prevent and improve degenerate effective ancillary drug of the various diseases that caused of cartilaginous tissue.Glucosamine also has the intestine immunity of promotion, strengthens brain memory function equivalent fruit except that helping alleviating the slight illness of joint.Animal experiment shows that with the forage feed mouse that contains 5% Glucosamine, its memory surpasses control group with learning ability.This means that the prevention to memory of elderly person decline and dementia also has certain help.Its effect of improving brain function is not second to DHA ginkgo leaf extract.Because Glucosamine is nontoxic, rare bad reaction, Japan, the U.S. extensively with it as food ingredient, with prevention or improve the symptom of osteoarthropathy.The long-term edible joint that can make keeps active, walks easily.Glucosamine is the most stable under acidity, therefore is suitable for most food, especially citrus food such as fruit juice, sour milk.Be not difficult to find out the welcome degree of Glucosamine from market demand.Latest data showed in 2006, and the American market demand reaches the 5000-6000 ton, and the Japanese market total sales volume reaches 2,000,000,000 yen, and its end product is 20 times of raw material up to 40,000,000,000 yen.Now, existing more than 100 tame larger food enterprises of Japan and pharmaceutical manufacturer are being engaged in the sale Glucosamine, and the market demand further increases.Glucosamine is to do raw material with crusts such as shrimp crabs, through decalcification, deacetylated after as shitosan, right hand hydrolysis forms.At present, China can produce Glucosamine in a large number, and especially coastland such as Zhejiang, Shandong except that medicine is made in a small amount of supply, much is with every kilogram of 3000-3500 yen outlet U.S. and Japan.
Glycosaminoglycan is a kind of natural carbohydrate that derives from chitin; be the principal ingredient in the cartilage matrix; by changing its side-chain structure; easilier in articular cartilage combine with water; the maintenance articular cavity lubricates the effect with compensator or trimmer pressure; impaired cartilage cell is had the certain protection effect, can synthetic use, existing commonly used have Glucosamine, aminoglucose hydrochloride, Glucosamine Sulphate sodium chloride double salt or potassium chloride double salt etc.Glucosamine is one of least unit of forming poly glucosamine, and it and uronic acid have been formed poly glucosamine, comprise chondroitin sulfate, keratan sulfate, dermatan sulfate and hyaluronic acid.
Anti-inflammatory drug only can improve the symptom of Osteoarthritis, and also may command symptom development of glycosaminoglycan helps the reparation of cartilage.Preparation at U.S.'s glycosaminoglycan and chondroitin sulfate has been added in the food, has very high safe reliability.Oral such medicine can reach therapeutic purposes, the patient of, pulmonary disease poor with circulation system disease, hepatic and renal function, diabetes for some, has security equally, for the preparation of taking metallic elements such as adding magnesium, zinc, selenium, should note producing behind these electrolytic etching of metal to cardiovascular inhibiting effect.Oral dose reached 1500mg and can reach result of treatment every day, also wanted boost to 100～2500mg for the patient of fat or long-term oral diuretics, just can reach result of treatment after confirming in the experiment to take for 4～6 weeks, so should adhere to taking at least 1 month.Subsidiary reaction after taking for a long time is less, and is also gentle.
The cartilage protection effect of glycosaminoglycan and chondroitin sulfate also is embodied in and can improves synovial membrane and produce hyaluronic ability, and normal hyaluronic acid level can reduce the regression of cartilage surface in the articular cavity, alleviates the symptom of Osteoarthritis.When this class Chondroprotective agents applies to the cartilage cell of in vitro culture; synthetic and the gathering of proteoglycan all raises; the activity of clostridiopetidase A reduces; glycosaminoglycan and chondroitin sulfate are the synthetic base substances of proteoglycan, and sulfate group is to keep the stable critical network structure of cartilage cell epimatrix simultaneously.In animal model, glycosaminoglycan also has the potential of anti-inflammatory, although the activity influence to Cycloxygenase in the inflammatory reaction or proteolytic enzyme is little, owing to increased the synthetic of proteoglycan, but brought into play the effect and the tytosis epimatrix of stabilizing cell membrane, play the effect of anti-inflammatory indirectly, this is with NSAID s class mechanism of drug action difference, but this two classes drug combination can play synergy, reduce NSAID s class amount of drug, alleviate subsidiary reaction; Chondroitin sulfate also may have certain anti-inflammatory property, play a role at the aspects such as taxis, phagocyte and activity of lysozyme that suppress inflammatory mediator, but the anti-inflammatory ability is low than medicines such as indocin, brufens.Security and validity that a lot of experiment confirm glycosaminoglycan and chondroitin sulfate are also arranged clinically, Rovati is in a randomized double-blind experiment, the patient of 252 routine knee joint osseous arthritis, take glycosaminoglycan (each 500mg respectively, every day 3 times) and contrast take placebo treatment 4 time-of-weeks, patient's gonalgia of 55%, swelling, limitation of activity symptom obviously alleviate, and control group only is 38%.Eichelt etc. are in one group of patient, intramuscular injection CSSO3 400mg, 2 times weekly, administration group (55%) has the conspicuousness meaning with placebo (33%) difference on relief of symptoms when treating for 6 weeks, Buesi finds in the research of observing chondroitin sulfate extract for treating Osteoarthritis: each oral 800mg, every day 2 times, after taking 6 months continuously, patient's symptom of 43% is obviously improved, only have 3% to improve and take placebo, and can significant prolongation suffer from the limb travel time, symptom alleviate also be continue (taking back 1 respectively, 3, reach 15% in 6 months, 24% and 37%).Unite and take these two kinds of medicines (glycosaminoglycan 1500mg/d, chondroitin sulfate 110mg/d) and can reach the obvious treatment effect equally.In with the comparative study of NSAID s class medicine, find to reach and take pain relieving and the antiphlogistic effects of brufen during 1 week taking cartilage protection class medicine 2 Zhou Houke; as if brufen 2 all backs symptoms are stable taking; and continue to take the trend that symptom behind the chondroitin sulfate still has further improvement; final result of treatment is: the brufen group reaches 52%; chondroitin sulfate group 48%; but the incidence brufen group of subsidiary reaction is 35%, and the chondroitin sulfate group only is 6%.Therefore NSAID s class agents alleviate symptom is rapider, though and cartilage protection class medicine is low slightly on result of treatment, the onset gentleness, it is less to take the back subsidiary reaction for a long time.
Hydrophilic interaction liquid chromatography (HILIC) is to separate polytype polar compound and the most effective chromatographic technique of hydrophilic compounds.Simply say, HILIC be normal-phase chromatography separate a kind of, but its use anti-phase in the normal moving phase of using.Generally speaking, the eluting order of HILIC is opposite with RPLC, and the charging property and the water wettability that keep along with analyte increase and strengthen.This has guaranteed the direct separation of compound without reserve on the RPLC chromatographic column.
HILIC is a chromatographic column with hydrophilic stationary phase.Can make water, damping fluid and at high proportion with the miscible organic phase of water as moving phase.Typical HILIC uses the buffer solution system that can with an organic solvent account for 50%-95%, and buffer salt should have good dissolubility in organic solvent.
The HILIC chromatographic column is to analyze solute by sample introduction to keep at the water layer subregion of moving phase and HILIC stationary phase water-wetted surface at present.The analyte water wettability is strong more, the easy more water layer that turns to stationary phase of partition balancing, so the reservation of analyte is also strong more.The object of the present invention is to provide a kind of HPLC-ELSD of employing that Glucosamine and the related preparations that contains Glucosamine are carried out the assay method that related substance detects.
The realization of related substance detection method of the present invention may further comprise the steps:
(1) chromatographic condition: detecting device adopts evaporative light-scattering detector (ELSD), and the drift tube temperature scope is 60-140 ℃, and the optimum detection temperature is 100 ℃; Nitrogen flow rate is 2.0-2.5L/min, and the moving phase of chromatographic condition is made up of water-acetonitrile, and its optimal proportion is (30: 70), and adds 0.1% formic acid and 0.1% ammoniacal liquor in moving phase, regulates pH scope 1～5, and its optimal pH is about 1.8; Chromatogram column temperature is a room temperature, and flow velocity is 1.0ml/min.Theoretical cam curve is calculated by Glucosamine and is not less than 2000.
(2) preparation of need testing solution: get Glucosamine or contain the related preparations of Glucosamine an amount of, add the solution that moving phase is mixed with 0.3mg/ml, as need testing solution
(3) preparation of contrast solution: it is an amount of to measure above-mentioned (2) need testing solution, adds moving phase and is diluted to the solution that contains 3 μ g among every 1ml approximately, solution in contrast.
(4) assay method:
Get contrast solution 20 μ l and inject liquid chromatograph, regulate detection sensitivity, make major component chromatogram peak height be about 20% of full scale, get need testing solution 20 μ l again and inject liquid chromatograph, the record chromatogram is to 3 times of the Glucosamine retention time, in the need testing solution chromatogram if any impurity peaks, measure each impurity peak area and, must not be greater than the peak area (1.0%) of contrast solution main peak.
Glucosamine of the present invention and contain the related substances detection method of Glucosamine, the preparation of need testing solution in above-mentioned (2): for satisfying the needs of need testing solution detectable concentration, it is an amount of to get Glucosamine, being mixed with concentration with moving phase is the solution that every 1ml contains 0.3mg, as need testing solution.Be diluted to the solution of a series of variable concentrations more respectively with moving phase, sample introduction 1 μ l makes it to produce the signal that main peak is three times of baseline noises respectively.Through test, detectability is 0.3ng (S/N 〉=3), if the concentration 0.3mg/ml calculating when checking with related substance, its limit of detection is 0.1%.
Glucosamine of the present invention and contain the related substance detection method of the related preparations of Glucosamine, the preparation of contrast solution in above-mentioned (3): precision is measured above-mentioned need testing solution 1ml, put in the 50ml measuring bottle, add the moving phase dissolving and be diluted to scale, solution in contrast.When this contrast solution concentration range was in 1-5 μ g/ml, peak area became good linear relationship with concentration, and its linear dependence property coefficient is 0.9999.
Glucosamine of the present invention and contain the related substance detection method of the related preparations of Glucosamine, its related substance detects in the high-efficient liquid phase chromatogram, occur if any impurity peaks, measure each impurity peak area and, should be greater than the main peak area (1%) of contrast solution.
Glucosamine of the present invention and contain the related substance detection method of the related preparations of Glucosamine, be applicable to that various needs carry out Glucosamine that related substance detects and the related preparations that contains Glucosamine, as tablet, capsule, granule, injection class preparation, controlled release and sustained release preparation and the compound preparation that contains Glucosamine.
The present invention's great advantage compared with the prior art is:
1, use evaporative light-scattering detector that this product is carried out related substance and detect, having overcome this product raw material does not have chromophoric group, the shortcoming that is difficult to detect with UV-detector.Classic method is used derivatization method always, and the present invention adopts evaporative light-scattering to need not to carry out complicated derivative reaction, and has effectively improved its detection sensitivity.
2, the present invention is applicable to that various needs carry out Glucosamine that related substance detects and the related preparations that contains Glucosamine, as tablet, capsule, granule, injection class preparation, controlled release and sustained release preparation and the compound preparation that contains Glucosamine.
3, detection method of the present invention is accurate, simple to operation, and is convenient, favorable reproducibility, highly sensitive, can fully satisfy the requirement that related substance inspection and catabolite are measured, control the special impurities in the sample preferably, guarantee product quality, practical in real work.
Embodiment
Embodiment one:
1, chromatographic condition and system suitability test
1.1, the selection of detecting instrument: owing to do not have chromophoric group in this product structure, so this product does not absorb in the ultraviolet region.In order better to detect impurity, better control product quality, we have selected evaporative light-scattering detector for use.
1.2, the selection of chromatographic condition
Instrument: SHIMADZU LC-10AT VP, HPLC 1A/ELSD (SEDEX 75)
(5) ELSD drift tube temperature scope is 60-140 ℃, and its optimum detection temperature is 100 ℃; Nitrogen flow rate is 2.0-2.5L/min.Liquid-phase chromatographic column hydrophilic interaction liquid chromatography (ZIC-HILIC, 150mm * 4.6mm, 5 μ m), with reference to pertinent literature and in conjunction with the test concrete condition, successively select multiple conditions such as methanol-water, methanol-water-glacial acetic acid, acetonitrile-0.3% trifluoroacetic acid solution for use, finally definite moving phase is made up of water-acetonitrile, its optimal proportion is (30: 70), and in moving phase, add 0.1% formic acid and 0.1% ammoniacal liquor, and regulate pH scope 1～5, its optimal pH is about 1.8; Chromatogram column temperature is a room temperature, and flow velocity is 1.0ml/min.
Under this chromatographic condition, the Glucosamine retention time is moderate, and peak shape is better.
1.3, sensitivity determination
It is an amount of to get Glucosamine, and being mixed with concentration with moving phase is the solution that every 1ml contains 0.3mg, as need testing solution.Be diluted to the solution of a series of variable concentrations more respectively with moving phase, sample introduction 1 μ l makes it to produce the signal that main peak is three times of baseline noises respectively.Through test, detectability is 0.3ng (S/N 〉=3), if the concentration 0.3mg/ml calculating when checking with related substance, its limit of detection is 0.1%.The result proves that this method is highly sensitive, can fully satisfy the requirement that the related substance inspection is measured.
1.4, stability of solution
Get with a need testing solution, measure respectively at 0,2,4,8 hour difference sample introduction, its main peak peak area and determination of related substances result are basicly stable in 8 hours.
Above test findings shows, the easy sensitivity of the method, and favorable reproducibility can be carried out quite good detecting to the quality of Glucosamine in the sample preferably.
The preparation of 2 need testing solutions: get Glucosamine or contain the related preparations of Glucosamine an amount of, add the solution that moving phase is mixed with 0.3mg/ml, as need testing solution
The preparation of 3 contrast solutions: it is an amount of to measure above-mentioned (2) need testing solution, adds moving phase and is diluted to the solution that contains 3 μ g among every 1ml approximately, solution in contrast.
4, Glucosamine and contain the detection of the related preparations related substance of Glucosamine
Get contrast solution 20 μ l and inject liquid chromatograph, regulate detection sensitivity, make major component chromatogram peak height be about 20% of full scale, get need testing solution 20 μ l injecting chromatographs again, the record chromatogram is to 3 times of Glucosamine main peak retention time, the need testing solution chromatogram is if any impurity peaks, measure each impurity peak area and, must not be greater than the peak area of main peak in the contrast solution.
Embodiment two: the mensuration of related substance in the compound glucosamine sulfate parenteral solution
Get the little pin of this product, be diluted to moving phase and contain Glucosamine 0.3mg/ml, as need testing solution.Precision is measured in right amount, adds moving phase and is diluted to the solution that contains Glucosamine 3 μ g among every 1ml approximately, solution in contrast.Under following selected chromatographic condition: HPLC-ELSD detecting device (SHIMADZU LC-10AT VP, HPLC 1A/ELSD (SEDEX 75)), with hydrophilic interaction liquid chromatography (ZIC-HILIC, 150mm * 4.6mm, 5 μ m) be chromatographic column, moving phase is made up of water-acetonitrile, its optimal proportion is (30: 70), and in moving phase, add 0.1% formic acid and 0.1% ammoniacal liquor, and regulate pH scope 1～5, its optimal pH is about 1.8; Chromatogram column temperature is a room temperature, and flow velocity is 1.0ml/min.Theoretical cam curve is calculated by Glucosamine should be not less than 2000.Get contrast solution 1 μ l and inject liquid chromatograph, regulate detection sensitivity, make major component chromatogram peak height be about 20% of full scale, get need testing solution 1 μ l again and inject liquid chromatograph, the record chromatogram is to 3 times of the Glucosamine retention time, in the need testing solution chromatogram if any impurity peaks, measure each impurity peak area and, must not be greater than the peak area of contrast solution main peak.Three reply side's Glucosamine Sulphate Injection by HPLC-ELSD figure see accompanying drawing.
Table 1. compound glucosamine sulfate parenteral solution determination of related substances result
Embodiment three: the mensuration of compound glucosamine sulfate sheet related substance
It is an amount of to get this product, porphyrize, and precision takes by weighing fine powder (being equivalent to Glucosamine 400mg approximately), adds moving phase and is mixed with the solution that contains 0.3mg among every 1ml approximately, filters, and gets filtrate as need testing solution.Precision is measured in right amount, adds moving phase and is diluted to the solution that contains 3 μ g among every 1ml approximately, solution in contrast.Under following selected chromatographic condition: HPLC-ELSD detecting device (SHIMADZU LC-10AT VP, HPLC 1A/ELSD (SEDEX 75)), with hydrophilic interaction liquid chromatography (ZIC-HILIC, 150mm * 4.6mm, 5 μ m) be chromatographic column, moving phase is made up of water-acetonitrile, its optimal proportion is (30: 70), and in moving phase, add 0.1% formic acid and 0.1% ammoniacal liquor, and regulate pH scope 1～5, its optimal pH is about 1.8; Chromatogram column temperature is a room temperature, and flow velocity is 1.0ml/min.Theoretical cam curve is calculated by Glucosamine should be not less than 2000.Get contrast solution 20 μ l and inject liquid chromatograph, regulate detection sensitivity, make major component chromatogram peak height be about 20% of full scale, get need testing solution 1 μ l again and inject liquid chromatograph, the record chromatogram is to 3 times of the Glucosamine retention time, in the need testing solution chromatogram if any impurity peaks, measure each impurity peak area and, must not be greater than the peak area of contrast solution main peak.
Table 2. compound glucosamine sulfate sheet has the substance-measuring result
Embodiment four: the mensuration of glucosamine sulfate tablet related substance
It is an amount of to get this product, porphyrize, and precision takes by weighing fine powder (being equivalent to Glucosamine 300mg approximately), adds moving phase and is mixed with the solution that contains 0.3mg among every 1ml approximately, filters, and gets filtrate as need testing solution.Precision is measured in right amount, adds moving phase and is diluted to the solution that contains 3 μ g among every 1ml approximately, solution in contrast.Under following selected chromatographic condition: HPLC-ELSD detecting device (SHIMADZU LC-10AT VP, HPLC 1A/ELSD (SEDEX 75)), with hydrophilic interaction liquid chromatography (ZIC-HILIC, 150mm * 4.6mm, 5 μ m) be chromatographic column, moving phase is made up of water-acetonitrile, its optimal proportion is (30: 70), and in moving phase, add 0.1% formic acid and 0.1% ammoniacal liquor, and regulate pH scope 1～5, its optimal pH is about 1.8; Chromatogram column temperature is a room temperature, and flow velocity is 1.0ml/min.Theoretical cam curve is calculated by Glucosamine should be not less than 2000.Get contrast solution 20 μ l and inject liquid chromatograph, regulate detection sensitivity, make major component chromatogram peak height be about 20% of full scale, get need testing solution 1 μ l again and inject liquid chromatograph, the record chromatogram is to 3 times of the Glucosamine retention time, in the need testing solution chromatogram if any impurity peaks, measure each impurity peak area and, must not be greater than the peak area of contrast solution main peak.
Table 3. glucosamine sulfate tablet has the substance-measuring result
Figure of description:
Fig. 1: compound glucosamine sulfate parenteral solution (20100101) HPLC-ELSD figure
Fig. 2: compound glucosamine sulfate parenteral solution (20100102) HPLC-ELSD figure
Fig. 3: compound glucosamine sulfate parenteral solution (20100103) HPLC-ELSD figure.
Claims (4)
1. a Glucosamine and the related substance detection method that contains the related preparations of Glucosamine is characterized in that detection method may further comprise the steps:
(1) chromatographic condition:
Do not have chromophoric group in this product bulk drug structure, therefore select evaporative light-scattering detector for use.Because Glucosamine polarity is big, adopt with water (0.1% formic acid+0.1% ammoniacal liquor): acetonitrile=30: 70 is a moving phase, and flow velocity is 1.0ml/min.Theoretical cam curve is calculated by Glucosamine and is not less than 2000.
(2) preparation of need testing solution:
Get Glucosamine or contain the related preparations of Glucosamine an amount of, add the solution that moving phase is mixed with 0.3mg/ml, as need testing solution.
(3) preparation of contrast solution
It is an amount of to measure above-mentioned (2) need testing solution, adds moving phase and is diluted to the solution that contains 3 μ g among every 1ml approximately, solution in contrast.
(4) assay method
Get contrast solution 1 μ l and inject liquid chromatograph, regulate detection sensitivity, make major component chromatogram peak height be about 20% of full scale, get need testing solution 1 μ l again and inject liquid chromatograph, the record chromatogram is to 3 times of the Glucosamine retention time, in the need testing solution chromatogram if any impurity peaks, measure each impurity peak area and, must not be greater than the peak area (1.0%) of contrast solution main peak.
2. Glucosamine according to claim 1 or contain the related substance detection method of the related preparations of Glucosamine, it is characterized in that, wherein (1) detecting device adopts evaporative light-scattering detector, and the drift tube temperature scope is 60-140 ℃, and the optimum detection temperature is 100 ℃; Nitrogen flow rate is 2.0-2.5L/min, and the moving phase of chromatographic condition is made up of water-acetonitrile, and its optimal proportion is (30: 70), and adds 0.1% formic acid and 0.1% ammoniacal liquor in moving phase, regulates pH scope 1～5, and its optimal pH is about 1.8; Chromatogram column temperature is a room temperature, and flow velocity is 1.0ml/min.
3. Glucosamine according to claim 1 or contain the related substance detection method of the related preparations of Glucosamine, it is characterized in that: the wherein preparation of (3) contrast solution: because the detection limit of Glucosamine related substance is 1%, simultaneously for satisfying the needs of need testing solution detectable concentration, it is an amount of to get the Glucosamine related preparations, add the solution that moving phase is mixed with 0.3mg/ml, as need testing solution.Precision is measured test sample 1ml again, puts in the 50ml volumetric flask, adds the moving phase dissolving and is diluted to scale, solution in contrast.
4. according to claim 1,2,3 described Glucosamines or contain the detection method of the related substance of Glucosamine related preparations, it is characterized in that: this method is applicable to that various needs carry out Glucosamine that related substance detects and the related preparations that contains Glucosamine, as the compound preparation of tablet, capsule, granule, injection class preparation, controlled release and sustained release preparation and various Glucosamines.
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| CN102415596A (en) * | 2011-10-14 | 2012-04-18 | 宁波大学 | Carbonated beverage |
| CN102590410A (en) * | 2012-01-13 | 2012-07-18 | 亚宝药业太原制药有限公司 | Content detection method for glucosamine hydrochloride capsules |
| CN103760275A (en) * | 2013-11-13 | 2014-04-30 | 江苏正大清江制药有限公司 | Content determination method of glucosamine hydrochloride raw material |
| CN111796039A (en) * | 2014-11-13 | 2020-10-20 | 沃特世科技公司 | Liquid chromatography calibration method for rapidly-labeled N-glycan |
| CN114324659A (en) * | 2021-12-29 | 2022-04-12 | 江苏海悦康医药科技有限公司 | Method for detecting organic impurities in gamma cyclodextrin |
| CN114544788A (en) * | 2020-11-25 | 2022-05-27 | 中国科学院大连化学物理研究所 | Chromatographic separation method for chitosan oligosaccharide isomers at different acetylation sites |
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2010
- 2010-10-27 CN CN2010105212094A patent/CN102103133A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102415596A (en) * | 2011-10-14 | 2012-04-18 | 宁波大学 | Carbonated beverage |
| CN102590410A (en) * | 2012-01-13 | 2012-07-18 | 亚宝药业太原制药有限公司 | Content detection method for glucosamine hydrochloride capsules |
| CN103760275A (en) * | 2013-11-13 | 2014-04-30 | 江苏正大清江制药有限公司 | Content determination method of glucosamine hydrochloride raw material |
| CN111796039A (en) * | 2014-11-13 | 2020-10-20 | 沃特世科技公司 | Liquid chromatography calibration method for rapidly-labeled N-glycan |
| CN114544788A (en) * | 2020-11-25 | 2022-05-27 | 中国科学院大连化学物理研究所 | Chromatographic separation method for chitosan oligosaccharide isomers at different acetylation sites |
| CN114324659A (en) * | 2021-12-29 | 2022-04-12 | 江苏海悦康医药科技有限公司 | Method for detecting organic impurities in gamma cyclodextrin |
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