IMPURITY PROFILING IMPURITY PROFILING OF THIAMINE HYDROCHLORIDE INJECTION BY RP-HPLC AND CHARACTERIZATION OF DEGRADATION PRODUCT BY LC-MS/MS/QTOF

CHARACTERIZATION OF DEGRADATION PRODUCT BY LC-MS/MS/QTOF

  • Srinivasu Kondra College of Pharmaceutical Sciences, Andhra University, Visakhapatnam, Andhra Pradesh
  • Dr.Bapuji APL Research Center, Aurobindo Pharma Limited, Hyderabad, Telangana, India
  • Dr. Gowri Sanker College of Pharmaceutical Sciences, Andhra University, Visakhapatnam, AP, India
  • Mr Murali Krishnam Raju APL Research Center, Aurobindo Pharma Limited, Hyderabad, Telangana, India

Abstract

Objective: To propose comprehensive, simple and affordable RP-HPLC method for impurity profiling and characterization of unknown degradation product of Thiamine hydrochloride injectable formulation.


Methods:


The chromatographic separation employs gradient mode using octadecyl silane column using mobile phase consist of Phosphate buffer with ion pair reagent, acetonitrile and methanol delivered flow rate at 1.2 ml/min. The detection was carried out at 248 nm using empower software. LC-MS/MS/QTOF hyphenated technique used for isolation and characterization of unknown degradation impurity. The performance of the method was systematically validated as per ICH Q2 (R2) guidelines.


Results:


Degradation product observed in accelerated stability was characterized by LC-MS/MS/QTOF hyphenated technique and found m/z value 351.1604 and postulated as oxidative degradation product of Thiamine due to excipient interaction.  The validated method was sensitive, selective and specific data proves the method is precise and accurate from LOQ to 150% level and results are within 95-108% and less than 4.5% RSD. Method is linear from 0.03-58.83 µg/ml with correlation coefficient more than 0.990 and LOD and LOQ value ranged from 0.03 to 1.51 μg/ml.


Conclusion:


A simple, selective and efficient RP-HPLC method for impurity profiling of Thiamine injectable formulation was successfully developed and validated. Moreover, degradant unknown product observed above identification threshold was identified and characterized by LC-MS/MS/QTOF hyphenated techniques. The method is systematically validated and found to be specific, precise, Linear, accurate, stable, rugged and robust and promises to be comprehensive simple affordable technique for impurity profiling in Thiamine hydrochloride injectable formulation in quality control department.

Keywords: Thiamine hydrochloride Injection, RP-HPLC, Identification, characterization, degradation products, validation, LC-MS-MS.

References

REFERENCES
1. Singleton CK, Martin PR. Molecular mechanisms of thiamine utilization. Curr Mol Med 2001; 1:197–207.
2. Abdou E, Hazell AS. Thiamine deficiency: an update of pathophysiologic mechanisms and future therapeutic considerations. Neurochem Res 2015; 40:353–361.
3. Teigen LM, Twernbold DD, Miller WL. Prevalence of thiamine deficiency in a stable heart failure outpatient cohort on standard loop diuretic therapy. Clin Nutr 2016; 35:1323–1327.
4. Kerns JC, Arundel C, Chawla LS. Thiamin deficiency in people with obesity. Adv Nutr 2015; 6:147–153.
5. Jain A, Mehta R, Al-Ani M, Hill JA, Winchester DE. Determining the role of thiamine deficiency in systolic heart failure: a meta-analysis and systematic review. J Card Fail 2015; 21:1000–1007.
6. Shah S, Wald E. Type B lactic acidosis secondary to thiamine deficiency in a child with malignancy. Pediatrics 2015; 135:e221–4.
7. Al-Daghri NM, Alharbi M, Wani K, Abd-Alrahman SH, Sheshah E, Alokail MS. Biochemical changes correlated with blood thiamine and its phosphate esters levels in patients with diabetes type 1 (DMT1). Int J Clin Exp Pathol 2015; 8:13483–13488.
8. Page GL, Laight D, Cummings MH. Thiamine deficiency in diabetes mellitus and the impact of thiamine replacement on glucose metabolism and vascular disease. Int J Clin Pract 2011; 65:684–690.
9. Abdel Rehman S T, Elbashir A A, El-Mukhtar M, and Ibrahim M M. Application of spectrophotometric methods for the determination of thiamine (vb1) in pharmaceutical formulations using 7-chloro-4-nitrobenzoxadiazole (NBD– Cl). Journal of Analytical & Pharmaceutical Research 2016; 2: 18-
10. Lopez-de-Alba P L, Lopez-Martinez L, Cerda V, Amador-Hernandez. Simultaneous determination and classification of riboflavin, thiamine, nicotinamide and pyridoxine in pharmaceutical formulations, by UV-visible spectrophotometry and multivariate analysis. Journal of Brazilian Chemical Society. 2006;17 (4): 715–722
11. Abdel-Maaboud M, Horria M, Niveen M A, Marwa E.-Z. Chemometric methods for the simultaneous determination of some water-soluble vitamins. Journal of AOAC International. 2011; 94: 467–481.
12. Chen QY, Li DH, Yang HH, Zhu Q Z, Zheng H, Xu JG. Novel spectrofluorimetric method for the determination of thiamine with iron (iii) tetrasulfonatophthalocyanine as a catalyst. Analyst. 1999; 124: 771–775
13. Zhu H, Chen H, Zhou Y. Determination of thiamine in pharmaceutical preparations by sequential injection renewable surface solid-phase spectrofluorometry. Analytical Sciences. 2003; 19: 289–294
14. Alonso A, Almendral M J, Porras M J, Curto Y. Flowinjection solvent extraction without phase separation. fluorimetric determination of thiamine by the thiochrome method. Journal of Pharmaceutical and Biomedical Analysis . 2006; 42: 171–177.
15. Tabrizi A B. A cloud point extraction-spectrofluorimetric method for determination of thiamine in urine. Bulletin of Korean Chemical Society 2006; 27: 1604–1608.
16. Amjadi M, Manzoori J L, M. Orooji. use of crude extract of kohlrabi (brassica oleracea gongylodes) as a source of peroxidase in the spectrofluorimetric determination of thiamine. Bulletin of Korean Chemical Society 2007; 28: 246–250.
17. Sun J, Liu L, Ren C, Chen X, Hu Z. A feasible method for the sensitive and selective determination of vitamin B1 with CdSe quantum dots. Microchimica Acta 2008; 163: 271–276
18. Khan M A, Jin S O, Lee SH, Chung HY. Spectrofluorimetric determination of vitamin B1 using horseradish peroxidase as catalyst in the presence of hydrogen peroxide. Luminescence 2009; 24: 73–78.
19. Li Y, Wang P, Cao M et al., An immediate luminescence enhancement method for determination of vitamin B1 using long-wavelength emitting water-soluble CdTe nanorods. Microchimica Acta 2010; 169: 65–71.
20. Zeeb M, Ganjali MR, Norouzi P. Dispersive liquid-liquid microextraction followed by spectrofluorimetry as a simple and accurate technique for determination of thiamine (vitamin B1). Microchimica Acta 2010; 168: 317–324.
21. Mohamed A M, Mohamed HA, Abdel-Latif NM, Mohamed MR. Spectrofluorimetric determination of some water-soluble vitamins. Journal of AOAC International 2011; 94: 1758–1769.
22. Tarigh G D, Shemirani F. Simultaneous in situ derivatization and ultrasound-assisted dispersive magnetic solid phase extraction for thiamine determination by spectrofluorimetry. Talanta 2014; 123: 71–77.
23. Zhu J, Liu S, Liu Z, Liu Y, Qiao M, Hu X. Enhanced spectrofluorimetric determination of hypochlorite based on the catalytic oxidation of thiamine to thiochrome in the presence of trace ferrocyanide. RSC Advances 2014; 4: 5990–5994.
24. Tan H, Li Q, Zhou Z et al. A sensitive fluorescent assay for thiamine based on metal-organic frameworks with intrinsic peroxidase-like activity. Analytica Chimica Acta 2015; 856: 90–95.
25. Restani M, Neubert RH. Thiamine analysis in biological media by capillary zone electrophoresis with a high-sensitivity cell. Journal of Chromatography A 2000; 871:351-356
26. Herve C, Beyne P, Delacoux E. Determination of thiamine and its phosphate esters in human erythrocytes by high-performance liquid chromatography with isocratic elution. J Chromatogr B Biomed Appl. 1994; 653: 217–20.
27. Baines M: Improved high performance liquid chromatographic determination of thiamin diphosphate in erythrocytes. ClinChimActa. 1985; 153:43–8.
28. Lynch PL, Trimble ER and Young IS: High-performance liquid chromatographic determination of thiamine diphosphate in erythrocytes using internal standard methodology. J Chromatogr B Biomed Sci Appl. 1997; 701:120 –3
29. Losa R, Sierra MI, Fernandez A, Blanco D and Buesa JM: Determination of thiamine and its phosphorylated forms in human plasma, erythrocytes and urine by HPLC and fluorescence detection: a preliminary study on cancer patients. J Pharm Biomed Anal. 2005; 37:1025–9.
30. Bettendorff L, Peeters M, Jouan C, Wins P and Schoffeniels E: Determination of thiamin and its phosphate esters in cultured neurons and astrocytes using an ion-pair reversed-phase high-performance liquid chromatographic method, Anal Biochem. 1991; 198:52–9.
31. Tang X, Cronin D A, Brunton N P. A simplified approach to the determination of thiamine and riboflavin in meats using reverse phase HPLC. Journal of Food Composition and Analysis. 2006; 19: 831–837
32. Ake M, Soko YN, Malan KA. Liquid chromatographic determination of free thiamine and its esters in whole blood. Dakar Medical 2006; 51: 172–177
33. Zafra-Gomez A, Garballo A, Morales J C, Garcia-Ayuso LE. Simultaneous determination of eight water-soluble vitamins in supplemented foods by liquid chromatography. Journal of Agriculture and Food Chemistry 2006; 54 : 4531–4536
34. Lebiedzinska A, Marszall ML, Kuta J, Szefer P. Reversed-phase high-performance liquid chromatography method with coulometric electrochemical and ultraviolet detection for the quantification of vitamins B(1) (thiamine), B(6) (pyridoxamine, pyridoxal and pyridoxine) and B(12) in animal and plant foods. Journal of Chromatography A 2007; 1173: 71–80.
35. N. Yanith, D. Widowati, Wartini, and T. Aryani, “Validation of HPLC method for determination of thiamine hydrochloride, riboflavin, nicotinamide, and pyridoxine hydrochloride in syrup preparation,” Canadian Journal on Scientific and Industrial Research, vol. 2, no. 7, pp. 269–278, 2011.
36. Wang X X, Hu, Sun YZ, Bie M J, Sun CJ. Simultaneous determination of five water-soluble vitamins in human serum by high performance liquid chromatography. Journal of Sichuan University, Medical Science Edition 2010; 41: 158–161
37. Tan J, Li R, Jiang ZT. Determination of thiamine (vitamin B1) in pharmaceutical tablets and human urine by titania-based ligand-exchange hydrophilic interaction chromatography. Analytical Methods, vol. 3, pp. 1568–1573, 2011.
38. Dinc E, Kokdil G, Onur F. A comparison of matrix resolution method, ratio spectra derivative spectrophotometry and HPLC method for the determination of thiamine HCl and pyridoxine HCl in pharmaceutical preparation. Journal of Pharmaceutical and Biomedical Analysis 2000 ; 22: 915–923
39. Hampel D, York E R, Allen LH. Ultra-performance liquid chromatography tandem mass-spectrometry (UPLC– MS/MS) for the rapid, simultaneous analysis of thiamin, riboflavin, flavin adenine dinucleotide, nicotinamide and pyridoxal in human milk,” Journal of Chromatography B Analytical Technologies in the Biomedical and Life Sciences 2012; 903: 7–13
40. Santos V B, Guerreiro TB, Suarez WT, Faria RC, Fatibello-Filho O. Evaluation of turbidimetric and nephelometric techniques for analytical determination of n-acetylcysteine and thiamine in pharmaceutical formulations employing a lab-made portable microcontrolled turbidimeter and nephelometer. Journal of the Brazilian Chemical Society 2011; 22: 1968–1978.
41. Akyilmaz E, Yasa I, Dinckaya E. Whole cell immobilized amperometric biosensor based on Saccharomyces cerevisiae for selective determination of vitamin B1 (thiamine). Analytical Biochemistry 2006; 354: 78–84.
42. Jiang X, Sun T. Indication ion square wave voltammetric determination of thiamine and ascorbic acid. Electrochemistry 2007; 40: 2589–2596.
43. Norouzi P, Garakani T M, Rashedi H, Zamani H A, Ganjali MR. Ultrasensitive flow-injection electrochemical method using fast fourier transform square-wave voltammetry for detection of vitamin B1. International Journal of Electrochemical Science 2010; 5: 639–652.
44. Bas B, Jakubowska M, Gorski L. Application of renewable silver amalgam annular band electrode to voltammetric determination of vitamins C, B1 and B2. Talanta, 2011; 84: 1032–1037.
45. Tyszczuk-Rotko K. New voltammetric procedure for determination of thiamine in commercially available juices and pharmaceutical formulation using a lead film electrode. Food Chemistry. 2012; 134: 1239–1243.
46. Brahman P K, Dar RA, Pitre KS. DNA functionalized electrochemical biosensor for detection of vitamin B1 using electrochemically treated multiwalled carbon nanotube paste electrode by voltammetric methods. Sensors and Actuators B: Chemical 2013; 177: 807–812
47. Antal IP, Bazel YR, Kormosh Z A. Electrochemical methods for determining group B vitamins. Journal of Analytical Chemistry 2013; 68: 565–576.
48. David IG, Florea M A, Cracea OG et al. Voltammetric determination of B1 and B6 vitamins using a pencil graphite electrode. Chemical Papers 2015; 69: 901–910
49. Dwivedi BK, Arnold RG, Libbey LM. Chemistry of Thiamine degradation. Mechanisms of thiamine degradation in a model system. Journal of Food Science 1972; 37: 689-92.
50. Mulley E A, Stumbo C R, Hunting WM. Kinetics f thiamine degradation by heat. Effect of pH and form of the vitamin on its rate of destruction. Journal of Food Science 1975; 40: 989-92.
51. Adrienne LV, Jenna M, Cordelia AR, Lynne ST, Lisa JM. Chemical stability and reaction kinetics of two thiamine salts (thiamine mononitrate and thiamine chloride hydrochloride) in solution. Food Research International 2018; 112:443-456.
52. British Pharmacopoeia, Monograph on 9iamine, Her Majesty’s Stationary Office, London, UK, 2016
53. United States Pharmacopeia 29, Monograph on Thiamine, United States Pharmacopeial Convention, Rockville, MD, USA, 2016
54 Guidance for Industry, ICH. Q3B (R2): Impurities in New drug product, Conference on Harmonization, Geneva (2006).
55 Srikanth G, Uttamkumar R, Murali N, Badrinadh Gupta P, Jagadeesh Kumar V, Satheesh D and Islam A: Identification, Isolation and characterization of process related impurities in ezetimibe. J. Pharm. Biomed. Anal. 2014; 88: 385-390.
56 ICH (2005) ICH Harmonized Tripartite Guidelines Q2 (R1): Validation of analytical procedures. ICH, Geneva.
57 FDA (2000), Guidance for Industry, Analytical Procedures and Method Validation (Chemistry, Manufacturing and Controls Documentation), Centre for Drug Evaluation and Research (CDER), Centre for Biologics Evaluation and Research (CBER), Rockville, USA.
58 ICH (2005) ICH Harmonized guideline Q1A (R2): Stability testing of new drug substances and products: text and methodology. ICH, Geneva.
59 Young PM and Gorenstein MV: Tryptic mapping by reversed-phase HPLC with photodiode-array detection incorporating the spectral-contrast technique. LC GC 1994;12 : 832–838.
60 Warren WJ, Stanick WA, Gorenstein MV and Young PM: HPLC analysis of synthetic
Oligonucleotides using spectral contrast techniques. Biotechniques 1995; 18: 282–287.
Statistics
8 Views | Downloads
How to Cite
Kondra, S., Akula , T. B., D, G. S., & Potturi, M. K. R. (2020). IMPURITY PROFILING IMPURITY PROFILING OF THIAMINE HYDROCHLORIDE INJECTION BY RP-HPLC AND CHARACTERIZATION OF DEGRADATION PRODUCT BY LC-MS/MS/QTOF . International Journal of Applied Pharmaceutics, 12(6). Retrieved from https://innovareacademics.in/journals/index.php/ijap/article/view/38283
Section
Original Article(s)