Characterization of an Unknown Impurity in Glucosamine Sulfate Sodium Chloride by HPLC-Q-TOF MS and NMR

Author(s): Miao Zhang, Peixi Zhu, Yue Chen, Weifang Ni, Yu Li*, Liya Hong*.

Journal Name: Current Pharmaceutical Analysis

Volume 15 , Issue 6 , 2019

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Graphical Abstract:


Background: Glucosamine sulfate sodium chloride (glucosamine-SP) is mainly used for the treatment of osteoarthritis. During quality control of glucosamine-SP capsules, an unknown impurity was detected. Another unknown degradation product was generated together with above-mentioned impurity in heat condition.

Objective: The study aimed to characterize an unknown impurity in glucosamine-SP capsules.

Methods: A new volatile HPLC method compatible with mass spectrometry detection was set up. An amino column at 35 °C with a mobile phase consisting of water and acetonitrile (20: 80, v/v) was used at a flow rate of 1.5 ml/min at 297 nm. High-performance liquid chromatography quadrupole time-offlight mass spectrometry (HPLC-Q-TOF MS) was used to identify the impurity with the electrospray ionization (ESI) source in the positive ionization mode.

Results: The results of HPLC-Q-TOF MS analysis indicated that the protonated molecule ions [M + H]+ of the unknown impurity and the novel degradation product were both at m/z 287. Preparative LC method was put into practice with a Prep-C18 column with a mobile phase consisting of water and acetonitrile (99: 1, v/v) at a flow rate of 20.0 ml/min at 297 nm. The assignment of the 1D and 2D NMR signals was performed for the unknown impurity. In addition, the formation of impurities was also studied.

Conclusion: An unknown impurity and a degradation product in glucosamine-SP capsules were characterized. They were assigned as (1R, 2S, 3R)-1-(5-((S, E)-3, 4-dihydroxybut-1-en-1-yl) pyrazin-2-yl) butane-1, 2, 3, 4-tetraol and (1R, 2S, 3R)-1-(5-((S, Z)-3, 4-dihydroxybut-1-en-1-yl) pyrazin-2-yl) butane- 1, 2, 3, 4-tetraol.

Keywords: Glucosamine sulfate sodium chloride, HPLC-Q-TOF mass spectrometry, unknown impurity, degradation product, preparative liquid chromatography, nuclear magnetic resonance (NMR).

Byron, D.H. Storage-stable glucosamine sulphate oral dosage forms and methods for their manufacture European Patent, EP 0 444 000 A2, February 12, 1991.
European Pharmacopeia, 9th ed; European Directorate for the Quality of Medicines & Healthcare: Strasbourg, 2016.
Yu, X.; Wang, F.; Li, J.; Shan, W.; Zhu, B.; Wang, J. Separation and characterization of unknown impurities and isomers in flomoxef sodium by LC-IT-TOF MS and study of their negative-ion fragmentation regularities. J. Pharm. Biomed. Anal., 2017, 140, 81-90.
Wang, J.; Xu, Y.; Wen, C.; Wang, Z. Application of a trap-free two-dimensional liquid chromatography combined with ion trap/time-of-flight mass spectrometry for separation and characterization of impurities and isomers in cefpiramide. Anal. Chim. Acta, 2017, 992, 42-54.
Ramachandra, B. Development of impurity profiling methods using modern analytical techniques. Crit. Rev. Anal. Chem., 2017, 47(1), 13.
Nagpal, S. Karan; Upadhyay, A.; Bhardwaj, T. R.; Thakkar, A. A review on need and importance of impurity profiling. Curr. Pharm. Anal., 2011, 7(1), 62-70.
Rahman, N.; Azmi, S.N.H.; Wu, H. The importance of impurity analysis in pharmaceutical products: An integrated approach. Accredit. Qual. Assur., 2006, 11, 69-74.
AlOthman, Z.A.; Rahman, N.; Siddiqui, M.R. Review on pharmaceutical impurities, stability studies and degradation products: an analytical approach. Rev. Adv. Sci. Eng., 2013, 2, 155-166.
Siddiqui, M.R.; AlOthman, Z.A.; Rahman, N. Analytical techniques in pharmaceutical analysis: A review. Arabian. J. Chem., 2017, 10, S1409-S1421.
Ukawala, R.D.; Ghate, M.; Vyas, V.K. Recent advances in characterization of impurities - use of hyphenated LC-MS technique. Curr. Pharm. Anal., 2010, 6(4), 299-306.
Patel, P.N.; Kalariya, P.D.; Gananadhamu, S.; Srinivas, R. Forced degradation of fingolimod: Effect of co-solvent andcharacterization of degradation products by UHPLC-Q-TOF–MS/MS and 1H NMR. J. Pharm. Biomed. Anal., 2015, 115, 388-394.
Pandey, A.K.; Rapolu, R.; Raju, C.K.; Sasalamari, G.; Goud, S.K.; Awasthi, A.; Navalgund, S.G.; Surendranath, K.V. The novel acid degradation products of losartan: Isolation and characterization using Q-TOF, 2D-NMR and FTIR. J. Pharm. Biomed. Anal., 2015, 120(12), 65-71.
Maggio, R.M.; Calvo, N.L.; Vignaduzzo, S.E.; Kaufman, T.S. Pharmaceutical impurities and degradation products: Uses and applications of NMR techniques. J. Pharm. Biomed. Anal., 2014, 101, 102-122.
United States Pharmacopeia, 41 ed.; The United States Pharmacopeial Convention: Rockville 2017.
Liu, H. HPLC determination of related substances in aciclovir glucose injection. Chin. J. Pharm. Anal, 2005, 25(6), 724-726.
Kompantsev, D. Stability of glucosamine dosage forms. Russ. J. Gen. Chem., 2012, 82(3), 579-585.
Chang, J. Principle and analysis of spectrum, 3rd ed; Science Press: Beijing, 2012.
Zhu, A.; Huang, J.; Clark, A.; Romero, R.; Petty, H.R. 2,5-Deoxyfructosazine, a D-glucosamine derivative, inhibits T-cell interleukin-2 production better than D-glucosamine. Carbohydr. Res., 2007, 342(18), 2745-2749.
Jia, L.; Zhang, Z.; Qiao, Y.; Pedersen, C.M.; Ge, H.; Wei, Z.; Deng, T.; Ren, J.; Liu, X.; Wang, Y.; Hou, X. Product distribution control for glucosamine condensation: nuclear magnetic resonance (NMR) investigation substantiated by density functional calculations. Ind. Eng. Chem. Res., 2017, 56, 2925-2934.
Wu, M.; Ma, H.; Ma, Z.; Jin, Y.; Chen, C.; Guo, X.; Qiao, Y.; Pedersen, C.M.; Hou, X.; Wang, Y. Deep eutectic solvents: green solvents and catalysts for the preparation of pyrazine derivatives by self-condensation of D-glucosamine. ACS Sustainable. Chem.& Eng., 2018, 6, 9434-9441.

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Article Details

Year: 2019
Page: [650 - 660]
Pages: 11
DOI: 10.2174/1573412915666190222160957
Price: $65

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