DEVELOPMENT AND VERIFICATION OF UV SPECTROPHOTOMETRIC TECHNIQUE FOR DETERMINING N-ACETYLCYSTEINE IN TABLET FORMULATIONS

RAMA RAO NADENDLA; MADAMANCHI MOUNIKA; VENKATA SAI RANJITHADUBBA; MANDAVA JAHNAVI; SOWJANYASONTI; YAKSHITHAYARRA; VENKATA SURESH P.; SIVA PRASAD MORLA

doi:10.22159/ijpps.2024v16i10.51004

Authors

RAMA RAO NADENDLA Department of Pharmaceutics, Chalapathi Institute of Pharmaceutical Sciences, Chalapathi Nagar, Lam-522034, Guntur, Andhra Pradesh, India
MADAMANCHI MOUNIKA Department of Pharmaceutics, Chalapathi Institute of Pharmaceutical Sciences, Chalapathi Nagar, Lam-522034, Guntur, Andhra Pradesh, India https://orcid.org/0009-0006-8870-1072
VENKATA SAI RANJITHADUBBA Department of Pharmaceutics, Chalapathi Institute of Pharmaceutical Sciences, Chalapathi Nagar, Lam-522034, Guntur, Andhra Pradesh, India
MANDAVA JAHNAVI Department of Pharmaceutics, Chalapathi Institute of Pharmaceutical Sciences, Chalapathi Nagar, Lam-522034, Guntur, Andhra Pradesh, India
SOWJANYASONTI Department of Pharmaceutics, Chalapathi Institute of Pharmaceutical Sciences, Chalapathi Nagar, Lam-522034, Guntur, Andhra Pradesh, India
YAKSHITHAYARRA Department of Pharmaceutics, Chalapathi Institute of Pharmaceutical Sciences, Chalapathi Nagar, Lam-522034, Guntur, Andhra Pradesh, India
VENKATA SURESH P. Department of Pharmaceutical Analysis, Chalapathi Institute of Pharmaceutical Sciences, Chalapathi Nagar, Lam-522034, Guntur, Andhra Pradesh, India https://orcid.org/0000-0002-7232-3322
SIVA PRASAD MORLA Department of Pharmaceutical Analysis, Chalapathi Institute of Pharmaceutical Sciences, Chalapathi Nagar, Lam-522034, Guntur, Andhra Pradesh, India

DOI:

https://doi.org/10.22159/ijpps.2024v16i10.51004

Keywords:

N-acetylcysteine, UV-Visiblespectroscopy, ICH guideline, Dosage form, Validation

Abstract

Objective: This study aimed to determine the concentration of N-Acetyl Cysteine (NAC) in tablet formulations using UV-Visiblespectroscopy.

Methods: A precise and accurate UV spectrophotometric method was developed for determining N-acetylcysteine in tablets, using 0.1N NaOH as a diluent. The drug's purity was assessed using UV-visible spectrophotometry, which validated linearity, accuracy, precision, specificity, limit of detection, and quantification.

Results: The calibration curve had a high correlation coefficient (r² = 0.9992) and maximum absorbance at 235 nm. There was no interference in dosage. The results showed 100.17% mean % recovery and 100.27% tablet assay, with a precision of 0.60% and 0.57%, respectively. The method was robust and rugged.

Conclusion: The method was evaluated using statistical parameters, including precision, accuracy, linearity, recovery, and robustness. The results showed no significant differences compared to other methods. The method can be used to analyse pharmaceutical formulations quickly, and no significant differences in mean values and standard deviations were found.

Downloads

Download data is not yet available.

References

Williams DA, Lenke TL. Foye’s principles of medicinal chemistry. 6th ed, Lippincott Williams and Williams. Ltd. Vol. 41(II). India: Wolters Kluwer Health, Pvt; 2008. p. 1127-46.

https://pubchem.ncbi.nlm.nih.gov/compound/acetylcysteine [Last accessed on 15 Jan 2024].

Ooi SL, Green R, Pak SC. N-acetylcysteine for the treatment of psychiatric disorders: a review of current evidence. Biomed Res Int. 2018;2018:2469486. doi: 10.1155/2018/2469486, PMID 30426004.

Kelly GS. Clinical applications of N-acetylcysteine. Altern Med Rev. 1998;3(2):114-27. PMID 9577247.

Ayene SI, Kale RK, Srivastava PN. Radioprotective effect of 2-mercaptopropionyl glycine on radiation-induced lipid peroxidation and enzyme release in erythrocytes. Int J Radiat Biol Relat Stud Phys Chem Med. 1988;53(4):629-39. doi: 10.1080/09553008814550951, PMID 3258297.

Barbey F, Joly D, Rieu P, Méjean A, Daudon M, Jungers PP. Medical treatment of cystinuria: critical reappraisal of long-term results. J Urol. 2000;163(5):1419-23. doi: 10.1016/s0022-5347(05)67633-1, PMID 10751848.

Holdiness MR. Clinical pharmacokinetics of N-acetylcysteine. Clin Pharmacokinet. 1991;20(2):123-34. doi: 10.2165/00003088-199120020-00004, PMID 2029805.

Haggag R. Derivatization with 4-Chloro-7-Nitro-2,1,3-benzoxadiazole for the spectrophotometric and differential pulse polarographic determination of acetylcysteine and captopril. Sci Pharm. 2008;76(1):33-48. doi: 10.3797/scipharm.0711-02.

Toussaint B, Pitti C, Streel B, Ceccato A, Hubert P, Crommen J. Quantitative analysis of N-acetylcysteine and its pharmacopeial impurities in a pharmaceutical formulation by liquid chromatography-UV detection-mass spectrometry. J Chromatogr A. 2000;896(1-2):191-9. doi: 10.1016/S0021-9673(00)00741-X, PMID 11093654.

Lu C, Liu G, Jia J, Gui Y, Liu Y, Zhang M. Liquid chromatography-tandem mass spectrometry method for determination of N-acetylcysteine in human plasma using an isotope-labeled internal standard. Biomed Chromatogr. 2011;25(4):427-31. doi: 10.1002/bmc.1465, PMID 21374646.

Celma C, Allue JA, Prunonosa J, Peraire C, Obach R. Determination of N-acetylcysteine in human plasma by liquid chromatography coupled to tandem mass spectrometry. J Chromatogr A. 2000 Feb;870(1-2):13-22. doi: 10.1016/s0021-9673(99)01078-x, PMID 10722057.

Barricklow J, Ryder TF, Furlong MT. Quantitative interference by cysteine and N-acetylcysteine metabolites during the LC-MS/MS bioanalysis of a small molecule. Drug Metab Lett. 2009 Aug;3(3):181-90. doi: 10.2174/187231209789352148, PMID 19702543.

Longo A, Di Toro M, Galimberti C, Carenzi A. Determination of N-acetylcysteine in human plasma by gas chromatography-mass spectrometry. J Chromatogr. 1991;562(1-2):639-45. doi: 10.1016/0378-4347(91)80614-i, PMID 2026726.

Magesh AR, Dhanaraju MD. Simultaneous determination of n-acetyl cysteine and taurine by HPTLC method in active pharmaceutical ingredient and pharmaceutical dosage form. Am J Anal Chem. 2017;8(11):742-51. doi: 10.4236/ajac.2017.811054.

Abdelaleem EA, Naguib IA, Zaazaa HE, Hussein EA. Development and validation of HPLC and HPTLC methods for determination of cefoperazone and its related impurities. J Chromatogr Sci. 2016;54(2):179-86. doi: 10.1093/chromsci/bmv125, PMID 26306573.

Ourique AF, Coradini K, Chaves PS, Garcia SC, Pohlmann AR, Guterres SS. A LC-UV method to assay N-acetylcysteine without derivatization: analyses of pharmaceutical products. Anal Methods. 2013;5(13). doi: 10.1039/c3ay40426a.

Gowda AP, Schaefer AD, Schuck TK. A simple RP-HPLC method for the stability-indicating determination of N-acetyl-L-cysteine and N, N’-diacetyl-L-cystine in cell culture media. Cell Gene Therapy Insights. 2020;6(2):303-23. doi: 10.18609/cgti.2020.041.

Mathew EM, Ravi A, Rameshwar N, Sudheer M, Krishnamurthy B. Development and validation of an analytical method for related substances in N-acetyl–L-cysteine effervescent tablets by RP-HPLC. Indian J Pharm Educ Res. 2017;51(4):626-35. doi: 10.5530/ijper.51.4.93.

Vamseekrishna G, Sanjeev G, Ramanjaneya Varma KJ, Umesh K, Mahesh U, Jayadev B. Stability indicating RP-HPLC method for simultaneous estimation of N-acetylcysteine and ascorbic acid. Int J Pharm Pharm Res. 2022;24(2):153-70.

Petrlova J, Mikelova R, Stejskal K, Kleckerova A, Zitka O, Petrek J. Simultaneous determination of eight biologically active thiol compounds using gradient elution-liquid chromatography with coul-array detection. J Sep. 2006;29(8):1166-73. doi: 10.1002/jssc.200500425, PMID 16830732.

Ercal N, Oztezcan S, Hammond TC, Matthews RH, Spitz DR. High-performance liquid chromatography assay for N-acetylcysteine in biological samples following derivatization with N-(1-pyrenyl)maleimide. J Chromatogr B Biomed Appl. 1996;685(2):329-34. doi: 10.1016/s0378-4347(96)00196-x, PMID 8953175.

Johannson M, Lenngren S. Determination of cysteine, glutathione and N-acetylcysteine in plasma by ion-pair reversed-phase liquid chromatography and post-column derivatization. J Chromatogr. 1988;432:65-74. doi: 10.1016/s0378-4347(00)80634-9, PMID 3220916.

Kumar CH, Uma G, Arcot S. Method development and validation for simultaneous assessment of clomiphene citrate and n-acetyl cysteine in mixed tablet dosage form using RP-UPLC. World J Pharm Pharm Sci. 2014;3:1773-80.

Jyothi NN, Pasha SI. Development and validation of a new RP-HPLC method for simultaneous estimation of n-acetylcysteine and l–arginine in combined dosage form. Orient J Chem. 2014;30(3):1371-8. doi: 10.13005/ojc/300357.

Dabir J, Mathew EM, Moorkoth S. Analytical method development and validation of RP-HPLC method for simultaneous estimation of n-acetyl cysteine and cefexime from its fixed-dose combination. Res J Pharm Technol X. 2016;9(7):835-42. doi: 10.5958/0974-360X.2016.00158.X.

Sana S, Rajania A, Sumedhab N, Pravin P, Shripad N. Development and validation of RP-HPLC method for the estimation of N-acetylcysteine in wet cough syrup. Int J Drug Dev Res. 2012;4:284-93. doi: 10.5937/arhfarm1403271i.

Farquhar J, Finlay G, Ford PA, Martin Smith M. A reversed-phase high-performance liquid chromatographic assay for the determination of N-acetylcysteine in aqueous formulations. J Pharm Biomed Anal. 1985;3(3):279-85. doi: 10.1016/0731-7085(85)80033-9, PMID 16867688.

Ourique AF, Coradini K, Chaves PS, Garcia SC, Pohlmann AR, Guterres SS. A LC-UV method to assay N-acetylcysteine without derivatization: analyses of pharmaceutical products. Anal Methods. 2013;5(13):3321-7. doi: 10.1039/C3AY40426A.

Acheampong A, Gyasi WO, Darko G, Apau J, Addai Arhin S. Validated RP-HPLC method for simultaneous determination and quantification of chlorpheniramine maleate, paracetamol and caffeine in tablet formulation. Springerplus. 2016;5:625. doi: 10.1186/S40064-016-2241-2, PMID 27330891.

Mamolo MG, Vio L, Maurich V. Simultaneous quantitation of paracetamol, caffeine and propyphenazone by high-pressure liquid chromatography. J Pharm Biomed Anal. 1985;3(2):157-64. doi: 10.1016/0731-7085(85)80019-4, PMID 16867698.

Houze P, Gamra S, Madelaine I, Bousquet B, Gourmel B. Simultaneous determination of total plasma glutathione, homocysteine, cysteinylglycine, and methionine by high-performance liquid chromatography with electrochemical detection. J Clin Lab Anal. 2001;15(3):144-53. doi: 10.1002/jcla.1018, PMID 11344530.

Raggi MA, Cavrini V, Di Pietra AM. Colorimetric determination of acetylcysteine, penicillamine, and mercaptopropionylglycine in pharmaceutical dosage forms. J Pharm Sci. 1982;71(12):1384-6. doi: 10.1002/jps.2600711218, PMID 7153888.

Ogwu V, Cohen G. A simple colorimetric method for the simultaneous determination of N-acetylcysteine and cysteine. Free Radic Biol Med. 1998;25(3):362-4. doi: 10.1016/s0891-5849(98)00024-0, PMID 9680182.

Gomes GC, Salgado HR. Validation of UV spectrophotometric method for determination of lomefloxacin in pharmaceutical dosage form. Acta Farm Bonaerense. 2005;243:406.

Kukoc-Modun L, Radić N. Spectrophotometric determination of N-Acetyl-L-cysteine and N-(2-Mercaptopropionyl)-glycine in pharmaceutical preparations. Int J Anal Chem. 2011. doi: 10.1155/2011/140756.

Agawane SS, Ashpak M, Tamboli MS, Patil SS, Swati T Mane. Simultaneous equation method for the estimation of clomiphene citrate and acetylcysteine by UV-visible spectrophotometry. Int J Pharm Res Appl. 2022;7(2 Mar-Apr):963-71.

Biocic M, Karabatić D, Tomic D, Kukoc Modun L. Spectrophotometric determination of N-acetyl-L-cysteine in pharmaceutical formulations by flow injection and sequential injection analysis: comparison of the methods. Croat Chem Acta. 2023;96(2):91-7. doi: 10.5562/cca4016.

Vedangkinjawadekara. Snehalathaboddua, deepalijadhava, sudharathod. Absorption correction method for the simultaneous estimation of n-acetyl-L-cysteine and ambroxol hydrochloride in bulk and in combined tablet dosage form. Int J Pharm Pharm Sci. 2016;8(5):191-5.

Hasan S, Sultan S. Utility of 2,2׳-bipyridyl reagent in spectrophotometric assay of acetylcysteine in pure form and in its pharmaceutical preparations. JES. 2022;31(3):42-53. doi: 10.33899/edusj.2022.134059.1246.

Li H, Du J. Sensitive chemiluminescence determination of three thiol compounds based on Cu(II)-catalyzing luminol reaction in the absence of an oxidant. Anal Lett. 2009 Aug 21;42(13):2131-40. doi: 10.1080/00032710903082754.

Samadi Maybodi A, Akhoondi R. Trace analysis of N-acetyl-L-cysteine using luminol-H2O2 chemiluminescence system catalyzed by silver nanoparticles. Luminescence. 2015 Sep 1;30(6):775-9. doi: 10.1002/bio.2819, PMID 25428294.

Bondarenko NY, Blazheyevskiy MY. Determination of N-acetylcysteine in tablets by means of chemiluminescence inhibition method. MOCA. 2018;13(3):110-4. doi: 10.17721/moca.2018.110-114.

Foroughi MM, Beitollahi H, Tajik S, Akbari A, Hosseinzadeh R. Electrochemical determination of N-acetylcysteine and folic acid in pharmaceutical and biological samples using a modified carbon nanotube paste electrode. International Journal of Electrochemical Science. 2014 Dec 1;9(12):8407-21. doi: 10.1016/S1452-3981(23)11056-X.

Da Silva IS, Araújo MF, Ferreira HA, Varela Jde J, Tanaka SM, Tanaka AA. Quantification of N-acetylcysteine in pharmaceuticals using cobalt phthalocyanine modified graphite electrodes. TalantaTalanta. 2011;83(5):1701-6. doi: 10.1016/j.talanta.2010.11.070, PMID 21238771.

Toito Suarez W, Marcolino LH, Fatibello Filho O. Voltammetric determination of N-acetylcysteine using a carbon paste electrode modified with copper(II) hexacyanoferrate(III). Microchem J. 2006;82(2):163-7. doi: 10.1016/j.microc.2006.01.007.

Ensafi AA, Karimi Maleh H, Mallakpour S, Hatami M. Simultaneous determination of N-acetylcysteine and acetaminophen by voltammetric method using N-(3,4-dihydroxyphenethyl)-3,5-dinitrobenzamide modified multiwall carbon nanotubes paste electrode. Sens Actuators B. 2011;155(2):464-72. doi: 10.1016/j.snb.2010.12.048.

Shahrokhian S, Kamalzadeh Z, Bezaatpour A, Boghaei DM. Differential pulse voltammetric determination of N-acetylcysteine by the electrocatalytic oxidation at the surface of carbon nanotube-paste electrode modified with cobalt salophen complexes. Sens Actuators B. 2008;133(2):599-606. doi: 10.1016/j.snb.2008.03.034.

Shaidarova LG, Gedmina AV, Zhaldak ER, Chelnokova IA, Budnikov GK. Voltammetric determination of acetylcysteine in drugs using an electrode modified by an osmium hexacyanocobaltate film. Pharm Chem J. 2014;47(12):670-4. doi: 10.1007/s11094-014-1029-3.

Santos VB, 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. J Braz Chem Soc. 2011;22(10):1968-78, doi: 10.1590/S0103-50532011001000019.

Dette C, Watzig H. Separation of enantiomers of N-acetylcysteine by capillary electrophoresis after derivatization by o-phthaldialdehyde. Electrophoresis. 1994;15(6):763-8. doi: 10.1002/elps.11501501106, PMID 7982397.

Jaworska M, Szulinska G, Wilk M, Tautt J. Capillary electrophoretic separation of N-acetylcysteine and its impurities as a method for quality control of pharmaceuticals. J Chromatogr A. 1999;853(1-2):479-85. doi: 10.1016/s0021-9673(99)00727-x, PMID 10486756.

Zinellu A, Sotgia S, Scanu B, Usai MF, Fois AG, Spada V. Simultaneous detection of N-acetyl-L-cysteine and physiological low molecular mass thiols in plasma by capillary electrophoresis. Amino Acids. 2009;37(2):395-400. doi: 10.1007/s00726-008-0167-x, PMID 18695935.

ICH Q2. Validation of analytical procedures: text and methodology. In: Proceedings of the international conference on harmonization. Vol. R1. Geneva, Switzerland; 2005. p. 1-13.