A Novel Stability-Indicating Method for Simultaneous Estimation of Glycopyrronium Bromide and Neostigmine Methyl Sulfate Injectable Formulations by RP-HPLC

Chetankumar Mashru; Jayesh Dhalani

doi:10.22146/ijc.106427

A Novel Stability-Indicating Method for Simultaneous Estimation of Glycopyrronium Bromide and Neostigmine Methyl Sulfate Injectable Formulations by RP-HPLC

https://doi.org/10.22146/ijc.106427

Chetankumar Mashru⁽¹⁾, Jayesh Dhalani^(2*)

(1) Department of Chemistry, School of Science, RK University, Rajkot-Bhavnagar Highway, Rajkot, Gujarat 360020, India
(2) Department of Chemistry, School of Science, RK University, Rajkot-Bhavnagar Highway, Rajkot, Gujarat 360020, India
(*) Corresponding Author

Abstract

Glycopyrronium bromide and neostigmine methyl sulfate drugs combination is used in treatment as an injectable formulation. The present study proposes the development and validation of an accurate, selective, and sensitive RP-HPLC method, which will enable the detection of degradation products for the combined estimation of glycopyrronium bromide and neostigmine methyl sulfate in both the parental dosage form and bulk samples. The optimum chromatographic condition was achieved using an octadecylsilane column, such as the Inertsil ODS-3 column with dimensions of 150 × 4.6 µm, 3 µm, at a flow rate of 1.00 mL/min with gradient elution. Phosphate buffer (pH 2.50) and acetonitrile were used as solutions A and B, respectively. Chromatography was achieved with an injection volume of 10 µL and a detection wavelength of 220 nm. The proposed study found a linear relationship between the ranges of 25–75 and 125–375 ppm for glycopyrronium and neostigmine methyl sulfate, respectively. The limit of detection and limit of quantification were achieved at 0.15 and 0.5 ppm for glycopyrronium bromide and neostigmine methyl sulfate, respectively. The validation of the method was carried out with a short run time of 20 min and a recovery rate of more than 99%.

Keywords

glycopyrronium bromide; neostigmine methylsulfate; RP-HPLC; validation

Full Text:

Full Text PDF

References

[1] Chawla, R.K., Rao, G.S.N.K., Kulandaivelu, U., Panda, S.P., and Alavala, R.R., 2021, A selective and sensitive method development and validation of 1,1-dimethyl-3-hydroxy-pyrrolidinium bromide impurity in glycopyrrolate oral solution by liquid chromatography-tandem mass spectroscopy, J. Chromatogr. Sci., 59 (6), 566–575.

[2] Kerwin, E., Feldman, G., Pearle, J., De La Cruz, L., Edwards, M., Beaudot, C., and Georges, G., 2021, Efficacy and safety of inhaled glycopyrronium bromide in COPD: A randomized, parallel group, dose-ranging study (GLIMMER), COPD, 18 (2), 181–190.

[3] Santus, P., Radovanovic, D., Cristiano, A., Valenti, V., and Rizzi, M. 2017, Role of nebulized glycopyrrolate in the treatment of chronic obstructive pulmonary disease, Drug Des., Dev. Ther., 11, 3257–3271.

[4] Ulrik, C.S., 2015, Once-daily glycopyrronium bromide (Seebri Breezhaler®) for the treatment of chronic obstructive pulmonary disease (COPD), Expert Opin. Pharmacother., 16 (17), 2653–2659.

[5] Prakash, A., Babu, K.S., and Morjaria, J.B., 2015, Profile of inhaled glycopyrronium bromide as monotherapy and in fixed-dose combination with indacaterol maleate for the treatment of COPD, Int. J. Chronic Obstruct. Pulm. Dis., 10 (1), 111–123.

[6] Parab, M., Shirsat, V.A., Kodgule, Y.M., and Kodgule, M., 2021, A RP-HPLC method for the analysis of neostigmine methylsulfate and process-related impurities, forced degradation studies in the injection formulation, Int. J. Anal. Chem., 2021 (1), 5570173.

[7] Vasu Babu, P., and Akiladevi, D., 2025, A bioanalytical method development and validation for quantification of glycopyrrolate and neostigmine in rat plasma by LC-MS and its application to pharmacokinetic study, J. Appl. Pharm. Res., 13 (3), 119–128.

[8] Fadr, M., Amro, A.N., Ben Aoun, S., Ratrout, S., and Asfour, F., 2018, Voltammetric determination of glycopyrrolate in a pharmaceutical formulation, Turk. J. Chem., 42 (6), 1736–1743.

[9] Mishra, S., and Arora, V., 2016, Analytical method validation of HPLC method for assay of anticholinergic drug in parenteral formulation, Int. J. Pharm. Res. Scholars, 5 (4), 101–104.

[10] Panda, S.S., Kumar, B.V.V.R., and Swain, S., 2020, Analytical lifecycle management oriented development and validation of a new stability indicating ultra-fast liquid chromatographic method: Case study of glycopyrrolate, Braz. J. Pharm. Sci., 56, 18420.

[11] Erukulla, K.K., and Rengitham, S.S., 2018, Synchronous estimation of glycopyrrolate and formoterol in bulk and pharmaceutical dosage form by RP-HPLC method, Indian J. Pharm. Educ. Res., 52 (4s), S47–S55.

[12] Salem, Y.A., Hammouda, M.E.A., Abu El-Enin, M.A., and El-Ashry, S.M., 2019, Multiple analytical methods for determination of formoterol and glycopyrronium simultaneously in their novel combined metered dose inhaler, BMC Chem., 13 (1), 75.

[13] Jamadar, P., and Meshram, D., 2015, Isoabsorptive point method for the simultaneous estimation of neostigmine methyl sulphate and glycopyrrolate in injectable dosage form, FS J. Pharm. Res., 4 (1), 5–8.

[14] Agrawal, B., Jagdale, S., Kadam, P., Sakpal, P., Nalawade, S., Maske, S., and Dale, P., 2024, Development and validation of stability indicating RP-HPLC method for the estimation of glycopyrrolate and neostigmine in bulk and injection, J. Chromatogr. Sci., 62 (3), 213–221.

[15] Panchal, J., Dhaduk, B., and Dhalani, J., 2023, Stability indicating isocratic RP-HPLC and second derivative UV spectroscopic methods for simultaneous determination of remogliflozin etabonate and vildagliptin hydrochloride, Rasayan J. Chem., 16 (2), 579–587.

[16] Panchal, J., and Dhalani, J., 2024, An optimization through experimental design approach for simultaneous quantification of remogliflozin and teneligliptin by RP-UFLC, Res. J. Chem. Environ., 28 (11), 16–24.

[17] Shah, T., and Dhalani, J., 2023, Determination of four novel process-related genotoxic impurities in olmesartan medoxomil tablet by RP-HPLC, Rasayan J Chem. 16 (3), 1543–1552.

[18] Shah, T., Kapadiya, K., and Dhalani, J., 2024, Isolation, characterization and cumulative quantification approach for nine organic impurities in empagliflozin and linagliptin drugs, Indian J. Chem. Technol., 31, 470–482.

[19] Wardhana, S.B., Kolb, M., Bahadir, M., Purwono, B., and Wahyuningsih, T.D., 2014, Method development for the analysis of pharmaceuticals with acethylcholinesterase activity in water using HPLC-DAD and solid phase extraction, Indones. J. Chem., 14 (1), 22–31.

[20] Nurani, L.H., Edityaningrum, C.A., Irnawati, I., Putri, A.R., Windarsih, A., Guntarti, A., and Rohman, A., 2023, Chemometrics-Assisted UV-vis spectrophotometry for quality control of pharmaceuticals: A review, Indones. J. Chem., 23 (2), 542–567.

DOI: https://doi.org/10.22146/ijc.106427