• ILMA NUGRAHANI School of Pharmacy, Bandung Institute of Technology, Bandung, Indonesia
  • RIZKA FAUZIA Pyridam Farma Pharmaceutical Company, Cianjur, Indonesia




kanamycin sulfate, Quantitative FTIR, Vibrational spectrophotometry, Derivative, Visible spectrophotometry, Ninhydrin-colorimetry


Objective: The purpose of this study was to develop and validate Fourier Transform Infra-Red (FTIR) method for a non-chromophore antibiotic, kanamycin sulfate. Afterward, it was compared to other quantitative methods, namely colorimetry using ninhydrin.

Methods: FTIR was used to determine the level of kanamycin sulfate. Firstly, the absorbance spectra were measured. To increase the specificity, it used mathematical derivation, which would separate the active compound's spectrum clearer from the other component of the dosage form. The clearest absorbance spectra was found at 1540-1480 cm-1, as the N-H bending, which then was used as the base for the quantitative analysis. This method was then applied to determine the kanamycin sulfate content in the commercial reconstitution-injection dosage form. Afterward, the analysis performance compared to another method which had established, namely colorimetry. The counterpart method was using ninhydrin as the chemical reagent to produce a chromophore, which able to be measured by visible spectrophotometry. Validation test checked the parameters such as linearity, LOD, LOQ, range, accuracy, and precision, continuing to evaluate and to compare the time and material involved  with the colorimetry as the counterpart.

Results: The results showed that FTIR for kanamycin sulfate lavel determination gave linearity within the range 0.12–1.5% w/w, with the r = 0.99975. Meanwhile, the LOD and LOQ were 0.04% and 0.12% w/w. There were no significant differences between the results of content determination of the antibiotic from the two methods. However, FTIR shows more advantages on time reducing, more straightforward preparation, solvents reducing, less costly, and more convenient.

Conclusion: FTIR is suitable to be an alternative method for quantifying kanamycin sulfate in a dosage form accurately, sensitive, and economist. This method is superior in its accuracy, precision, and simplicity compared to the colorimetry method using UV spectrometer as the counterpart.


McEvoy GK, Snow E. AHFS drug information handbook. Maryland, United States: American Society of Health-System Pharmacists; 2011.

Alldredge BK, Corell RL, Ernst ME. Koda-Kimble and young's applied therapeutics: the clinical use of drugs. 10th ed. Philadelphia, USA: Lippincott Williams and Wilkins; 2013.

Sweetman SC. Martindale the complete drug reference. 36th ed. London, UK: Pharmaceutical Press; 2009.

Brunton LL, Lazo JS, Parker KL. Goodman and gilman are the pharmacological basis of therapeutics. 11th ed. New York, United States: McGraw-Hill; 2006.

Indonesian Pharmacist Institution. Informasi Spesialite Obat Indonesia. Vol. 50th. Jakarta: Indonesian Pharmacist Institution; 2016.

Ministry of Health of the Republic of Indonesia. Pharmacopoeia Indonesia. 5th ed. Jakarta: Ministry of Health of the Republic of Indonesia; 2014.

The British Pharmacopoeia Commission. British Pharmacopeia. London: British Pharmacopoeia Commission; 2013.

United States Pharmacopoeia Conv. USP 37/NF 32. Washington DC: US Pharmacopeia; 2014.

Ministry of Health of the Republic of Indonesia. Pharmacopoeia Indonesia, 4th ed. Jakarta: Ministry of Health of the Republic of Indonesia; 1995.

Ahuja S, Dong MW. Handbook of pharmaceutical analysis by HPLC. New York, United States: Elsevier Inc; 2005.

Coates J. Interpretation of infrared spectra, a practical approach. In: RA Meyers. (Ed.) Encyclopedia of Analytical Chemistry. Canada: John Wiley and Sons Ltd; 2014.

Griffiths PR. Chemical infrared fourier transform 43. Toronto: John Willey and SMS; 1975.

Shehzadi N, Hussain K, Khan MT. Development of a validated colorimetric assay method for estimation of amikacin sulfate in bulk and injectable dosage form. J Chem Soc Pak 2016;38:63-9.

Das V, Kheeci S, Inda SS. Analytical method development and its validation for estimation of kanamycin sulfate by UV-Visible spectrophotometry as bulk and in the dosage form. Int J Pharm Res Bio Sci 2017;6:19-26.

Nugrahani I, Dillen N. rapid assay development of diclofenac sodium coated tablet assay using ftir compared to HPLC method. Int J Appl Pharm 2018;10:43-50.

Chan CC, Herman L, Lee YCX. Analytical method validation and instrument performance verification. Canada: John Wiley and Sons Inc; 2014.

Dean JA. Lange’s Handbook of Chemistry. 15th ed. USA: McGraw-Hill, Inc; 1999.

Furniss BS, Hannaford AJ, Smith PW. Vogel’s textbook of practical organic chemistry. 5th ed. New York, United States: John Wiley and Sons; 1989.

Silverstein RM, Webster FX, Kiemle DJ. Spectrometric identification of organic compounds. USA: John Wiley and Sons, Inc; 2005.

Harvey D. Modern analytical chemistry. New York: McGraw-Hill; 2000.

Settle F. Handbook of instrumental techniques for analytical chemistry. New Jersey: Prentice-Hall; 1997.

Skoog DA. Principles of instrumental analysis. 6th ed. USA: Thomson Higher Education; 2007.

Bhattacharjee, Mrinal K. Chemistry of antibiotics and related drugs. switzerland: Springer International Publishing; 2016.

Carlson J, Wysoczanski A, Voigtman E. Limits of quantitation-yet another suggestion. Spectrochimica Acta Part B: Atomic Spectroscopy 2014;96:69-73.

Alfa Aesar by Thermo Fisher Scientific, 39794 Potassium bromide, FTIR Grade. Available from: https://www.alfa.com/ en/catalog/039794/. [Last accessed on 02 Oct 2017]

Sigma-Aldrich. Acetone Laboratory Reagent. Available from: https://www.sigmaaldrich.com/chemistry/solvents/products.html?TablePage=17292082. [Last accessed on 02 Oct 2017].

Sigma-Aldrich. Ninhydrin ACS Reagent. Available from: https://www.sigmaaldrich.com/catalog/product/sial/151173?lang=enandregion=ID). [Last accessed on 02 Oct 2017]

Manyasree D, Kiran MP, Ravikumar R. CuO nanoparticles: synthesis, characterization and their bactericidal efficacy. Int J Appl Pharm 2017;9:71-4.

Arslan FN, Çaglar F. Attenuated total reflectance–fourier transform infrared (ATR–FTIR) spectroscopy combined with chemometrics for rapid determination of cold-pressed wheat germ oil adulteration. Food Anal Meth 2018;12:355-70.

Ethiraj R, Thiruvengadam E, Sampath VS, Vahid A, Raj J. Development and validation of stability indicating spectroscopic method for content analysis of ceftriaxone sodium in pharmaceuticals. Int School Res Notices 2014. http://dx.doi.org/10.1155/2014/278173

Weisstein EW. Student's t-Distribution. From MathWorld-a wolfram web resource. http://mathworld.wolfram.com/Studentst-Distribution.html. [Last accessed on 17 Apr 2019]

Papanna RK, Krishnegowda JB, Nagaraja P. Spectrophotometric method for the determination of cefepime, cefazolin sodium and cefalothin sodium in pure and pharmaceutical dosage forms by using ninhydrin. Int J Pharm Pharm Sci 2015;7:194-9.



How to Cite

NUGRAHANI, I., & FAUZIA, R. (2019). QUANTITATIVE VIBRATIONAL METHODS DEVELOPMENT AND ITS PERFORMANCE COMPARISON TO COLORIMETRY ON THE ASSAY OF KANAMYCIN SULFATE. International Journal of Applied Pharmaceutics, 11(4), 426-435. https://doi.org/10.22159/ijap.2019v11i4.32991



Original Article(s)