Study On The Quality Of Glucosamine Sulfate Preparation

Glucosamine is a natural amino monosaccharide,which is generally obtained from chitin hydrolyzation and then purified.However,glucosamine is very unstable.It needs to be preserved in the form of salt.Glucosamine sulfate is a kind of the salt,but it is hygroscopic and is easy to be oxidated.It is possible to stabilize by the formation of the double salt form such as glucosamine sulfate sodium chloride and glucosamine sulfate potassium chloride.Glucosamine sulfate preparation is prepared from glucosamine sulfate,glucosamine sulfate sodium chloride or glucosamine sulfate potassium chloride.The dosage forms include capsules,tablets,granules,and powders.As a health food and a common medicine for treating arthritis,glucosamine sulfate preparetion is widely used in the middle-aged and elderly population,with large demand and high market share.China has become the largest producer and exporter of this products.However,the impurity profiles are different because of the different production process of glucosamine sulfate and various domestic enterprises.Furthermore,the ultraviolet end absorption makes the detection of glucosamine sulfate more difficult.The quality control methods for key items such as related substances,dissolution and content in the current drug registration standards are different and unsatisfactory.There is no unified national standard,leading to poor product quality control.So,there might be security risks of drug quality,and the standards are urgently to be further studied.In this project,the glucosamine sulfate capsules with most approval numbers were used as research objects,and the quality control techniques of key items such as impurity profiles,dissolution and content were studied.The structure of an unknown impurity in the capsules was confirmed.Current quality standards have been improved and unified,the drafted national standard was proposed.It could provide reference for the revision of quality standards for other dosage forms of glucosamine sulfate,and provide technical support for supervision and quality control.In the first part of the thesis,the impurity profile of glucosamine sulfate capsules was studied.First,the process impurities and degradation impurities of glucosamine sulfate capsules were studied with the current liquid chromatography method.Then a novel impuritiy was found.Meanwhile,the impurity was produced together with another unknown impurity after heat degradation.Afterwards,a new liquid chromatography method adapted to MS was studied.With high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(HPLC-Q-TOF MS),the masses and fragments of two unknown impurities were obtained and their structures were conducted.A new preparative liquid chromatography method was established and the detected unknown impurity was prepared.Furthermore,the structure of the unknown impurity was confirmed by nuclear magnetic resonance(NMR).It was named(1R,2S,3R)-1-(5-((S,E)-3,4-dihydroxybut-1-en-1-yl)pyrazin-2-yl)butane-1,2,3,4-tetraol.Another unknown impurity was conducted with the data of MS and UV spectra.It was the isomers named(1R,2S,3R)-1-(5-((S,Z)-3,4-dihydroxybut-1-en-1-yl)pyrazin-2-yl)butane-1,2,3,4-tetraol.At last,the mechanism of the generation of unknown impurities was proposed.The two unknown impurities might be produced from impurity C in European Pharmacopoeia in heat condition.Some suggestions were also given for the storage of glucosamine sulfate capsules.In the second part of the thesis,the methods for the determination of the content and dissolution of glucosamine sulfate capsules by post-column derivatization high performance liquid chromatography with fluorescence spectroscopy were studied and methodological verification was carried out separately.One part is content determination method.Its injection volume is 10 μL.The linear concentration of glucosamine sulfate is in the range of 0.25 mg/mL ~ 2.45 mg/mL.The linear equation is y = 887656 x – 12079(r = 0.9999).The limit of quantitation is 0.049 mg/mL,which was equivalent to 3.9% of the concentration of the test solution for content assay.The average recovery is 99.3%(RSD = 1.1%,n = 9).Another part is dissolution determination method.Its injection volume is 50 μL.The linear concentration of glucosamine sulfate is in the range of 0.028 mg/mL ~ 0.56 mg/mL.The linear equation is y = 4326341x-5904.5(r = 0.9999).The limit of quantitation is 0.009 mg/mL,which was equivalent to 3.3% of the concentration of the test solution for dissolution determination.The average recovery is 100.0%(RSD = 0.4%,n = 9).Compared with the current standard method,the new method has lower detection limit,stronger speifcity,more stable baseline,weaker baseline noise,better repeatability,better instrument precision and shorter running time for per injection.Thus,the new methods are suitable for the test of large-scale samples.Meanwhile,the two liquid chromatography methods were unifine.Dissolution curves of two companies’ products were determined.The dissolution curves of two companies’ products is similar.