• SK MANIRUL HAQUE Department of Chemical and Process Engineering Technology, Jubail Industrial College, P. O. Box 10099, Jubail Industrial City 31961, Saudi Arabia http://orcid.org/0000-0003-2646-4164




Ampicillin, Potassium iodate, Potassium iodide, Spectrophotometer, Method development, Validation, Tablets


Objective: A simple, precise, and accurate spectrophotometric method has been developed to determine Ampicillin in pharmaceutical formulations.

Methods: The proposed method, based on the carboxylic acid group reaction, present in Ampicillin with a mixture of KIO3 and KI, form a yellow-colored product in an aqueous medium. The response was allowed to proceed at 25±1 °C, and absorbance measured after 5 min against a reagent blank prepared simultaneously using a UV-Vis spectrophotometer. The parameters verified were specificity, linearity, linearity range, accuracy, precision, detection limit, quantitation limit, robustness, and ruggedness.

Results: The yellow-colored product was measured at 352 nm against the reagent blank using UV–Vis spectrophotometer. The linear dynamic range of concentration was 0.25–2.5 µg/ml with a correlation coefficient of 0.9999. The LOD, LOQ values to be 0.086 and 0.261 µg/ml, respectively, for the proposed method. The percentage of recoveries was 98.27–100.89% with an acceptable relative standard deviation (±2%). The robustness and ruggedness values were excellent.

Conclusion: The ICH guidelines for pharmaceuticals and human use were followed and applied to validate the proposed method. The method was compared with available literature and found similar results that confirmed the reliability and effective way for Ampicillin's determination.


Download data is not yet available.


Tadesse DA, Zhao S, Tong E, Ayers S, Singh A, Bartholomew MJ, et al. Antimicrobial drug resistance in escherichia coli f+ rom humans and food animals, United States, 1950–2002. Emerg Infect Dis 2012;18:741–9.

Ahmed MS, Mohammed RH, Fadhil MA. RP-HPLC method validation for the determination of Ampicillin in Iraqi healthy volunteers serum. Int J Res Pharm Chem 2017;7:360–6.

Barot TG, Patidar K, Kshartri N, Vyas N. Development and validation of LC method for the determination of Ampicillin and dicloxacillin in pharmaceutical formulation using an experimental design. E J Chem 2009;6:955–64.

Simon B, Hancu G, Gyeresi A. Application of capillary electrophoresis to the simultaneous determination and stability study of four extensively used penicillin derivatives. Brazilian J Pharma Sci 2014;50:521–7.

Long C, Deng B, Sun S, Meng S. Simultaneous determination of chlortetracycline, Ampicillin and sarafloxacin in milk using capillary electrophoresis with electrochemiluminescence detection. Food Additives Contaminants Part A Chem Anal Control Exp Risk Assess 2017;34:24–31.

Totoli EG, Salgado HRN. Development and validation of the quantitative analysis of ampicillin sodium in powder for injection by fourier-transform infrared spectroscopy (FT-IR). Phys Chem 2012;2:103–8.

Chen L, Wang B, Diao Z, Zhao M, Xie K, Zhang P, Wang X, Zhang T, Wang J. Development and validation of an HPLC-ESI/MS/MS method for the determination of amoxicillin, its major metabolites, and ampicillin residues in chicken tissues. Molecules 2019;24:2652.

Parker SL, Adnan S, Meija JLO, Paterson DL, Lipman J, Roberts JA, et al. A UHPLC–MS/MS method for the simultaneous determination of Ampicillin and sulbactam in human plasma and urine. Bioanal 2015;7:2311–9.

Pallavi K, Babu PS, Babu GK. Development and validation of UV spectrophotometric method and RP-HPLC method for estimation of capecitabine in bulk and tablet dosage forms. Int J Appl Pharm 2016;8:24–9.

Madur S, Matole V, Kalshetti M. UV visible spectrophotometric method development and validation of dasatinib in bulk and solid dosage form. Int J Curr Pharm Res 2020;12:90–3.

Rai G, Kumar A. Screening and analysis of FTO gene polymorphism in north UP (India). J Critical Rev 2020;7:2274–80.

Supriya P, Madhavi LN, Rohith KBV, Ramana GV, Harini U, Pawar AKM. Development and validation of UV spectrophotometric and reversed-phase high-performance liquid chromatography‑PDA methods for the estimation of alogliptin benzoate. Asian J Pharm Clin Res 2016;9:282-7.

Khanchandani SS, Galgatte UC, Chaudhari PD. Development and validation of UV-visible spectroscopic method for estimation of rizatriptan benzoate in bulk and tablet dosage form. Asian J Pharm Clin Res 2013;6:113-6.

Ezeanokete CC, Ngwoke KG, Okoye FBC, Osadebe PO. Spectrophotometric determination of Ampicillin and cloxacillin in pure and fixed dosage forms through charge transfer complexation. Eur Chem Bull 2013;2:1009–12.

Misic IR, Miletic G, Mitic S, Mitic M, Marinkovic EP. A simple method for the ampicillin determination in pharmaceuticals and human urine. Chem Pharma Bull 2013;61:913–9.

Ahmed AJ, Abdulla S, Yunusa U, Omar A, Haque SM. Development, computational studies and validation of spectrophotometric method of metformin hydrochloride in pharmaceutical formulations. J Chilean Chem Soc 2020;65:4895–9.

United States Food and Drug Administration. Guideline for industry: text on validation of analytical procedures: ICH Q2A; 1995.

United States Food and Drug Administration. Guidance for industry: Validation of analytical procedures: methodology, Q2B; 1996.

International conference on the harmonization of the technical requirements for registration of pharmaceuticals for human use, ICH harmonized tripartite guideline, validation of analytical procedures: text and methodology, Q2 (R1); 1996.

Feigl F. Spot tests in the organic analysis. Chapter 3: Preliminary (Exploratory tests). 6th edition. Elsevier publishing company, Amsterdam; 1960. p. 117–8.



How to Cite

HAQUE, S. M. “OPTIMIZED AND VALIDATED SPECTROPHOTOMETRIC METHOD FOR THE DETERMINATION OF AMPICILLIN IN PHARMACEUTICAL FORMULATIONS”. International Journal of Pharmacy and Pharmaceutical Sciences, vol. 13, no. 2, Feb. 2021, pp. 34-38, doi:10.22159/ijpps.2021v13i2.40025.



Original Article(s)