• GIRI PRASAD GORUMUTCHU Department of Chemistry, Acharya Nagarjuna University, Nagarjuna Naga, Andhra Pradesh, India. http://orcid.org/0000-0003-0925-1132
  • VENKATA NADH RATNAKARAM Department of Chemistry, GITAM University, Bengaluru, Karnataka, India.
  • KIRAN KUMAR KATARI Department of Chemistry, Kakarparthi Bhavanarayana College, Vijayawada, Andhra Pradesh, India.




Validation, Piperacillin, Cobalt thiocyanate, Ion pair extraction, Assay


Objective: The objective of the study was to develop a simple, validated, and affordable visible spectrophotometric method for determination of piperacillin (PIP) present in bulk and powder for injection formulation.

Methods: In the present method, cobalt thiocyanate (CTC) was used as a chromogenic reagent where it forms 2:1 ion pair complex at pH 2 with PIP which is having secondary and tertiary amine groups.

Results: The formed bluish-green colored ion pair between PIP and CTC is quantitatively extractable into nitrobenzene with an absorption maximum of 665 nm. Regression analysis (r=0.9996) shows that the plotted calibration curve exhibits good linearity in the studied range of concentration (3–18 μg/mL). Low values of relative standard deviation (<2%) were observed indicating that the proposed method is reproducible, accurate, and precise.

Conclusions: As per the existing guidelines of ICH (international council for harmonization of technical requirements for pharmaceuticals for human use), various parameters of the proposed method were tested for validation and can be used method of choice for routine analysis in industrial quality control laboratories, especially in developing countries.


Download data is not yet available.

Author Biography

GIRI PRASAD GORUMUTCHU, Department of Chemistry, Acharya Nagarjuna University, Nagarjuna Naga, Andhra Pradesh, India.

Head - Chemistry, Research Coordinator and Dy.Director - Admissions


Tan JS, File TM Jr. Antipseudomonal penicillins. Med Clin North Am 1995;79:679-93.

Hauser AR. Antibiotic Basics for Clinicians. 2nd ed. Philadelphia, PA: Wolters Kluwer; 2013. p. 26-7.

Scheer F, Krämer I. A liquid chromatography assay for the simultaneous quantification of piperacillin and ciprofloxacin in human plasma and dialysate in critically III patients undergoing continuous renal replacement therapy. Int J Anal Mass Spectrom Chromatogr 2014; 2:43 51.

Barco S, Risso FM, Bruschettini M, Bandettini R, Ramenghi LA, Tripodi G, et al. A validated LC–MS/MS method for the quantification of piperacillin/tazobactamon dried blood spot. Bioanalysis 2014; 6:2795 802.

Li Z, Li Q, Yi W, Di C, Chen C. Determination of free and total piperacillin–tazobactam in plasma by HPLC–MS–MS an adapted method for neonates. Chromatographia 2012;75:533-39.

Veni PR, Sharmila N, Narayana KJ, Babu BH, Satyanarayana PV. Simultaneous determination of piperacillin and tazobactamin pharmaceutical formulations by RP-HPLC method. J Pharm Res 2013; 7:127-31.

Pai PN, Rao GK, Murthy MS, Prathibha H. Simultaneous estimation of piperacillin and tazobactam in injection formulations. Indian J Pharm Sci 2006;68:799-801.

Ramalingam P, Rajendran K. Detection of stability and degradation of piperacillin and tazobactam in injectables from in-patient wards and pharmacy by RP-HPLC method. Gulf Med J 2014;3:S132-41.

Rao AL, Krishna KS, Ch KK, Raja T. Simultaneous determination of piperacillin and tazobactum in bulk and pharmaceutical dosage forms by RP-HPLC. Int J Pharm J Pharm Sci 2011;3 Suppl 2:1033-40.

Augey V, Grosse PY, Albert G, Audran M, Bressolle F. High-performance liquid chromatographic determination of tazobactam and piperacillin in human plasma and urine. J Chromatogr B Biomed Appl 1996; 682:125-36.

Arzuaga A, Isla A, Gascón AR, Maynar J, Martín A, Solinís MA, et al. Quantitation and stability of piperacillin and tazobactam in plasma and ultrafiltrate from patients undergoing continuous venovenous hemo filtration by HPLC, Biomed Chromatogr 2005;19:570-8.

Veillette JJ, Winans SA, Forland SC, Maskiewicz VK. A simple and rapid RP-HPLC method for the simultaneous determination of piperacillin and tazobactam in human plasma. J Pharm Biomed Anal 2016; 131:80-6.

Sangeetha S, Kumar M, Kumudhavalli MV, Alexandar S, Jaykar B. Development and validation of UV spectrophotometric area under curve method for quantitative estimation of piperacillin and tazobactam. Int J Chem Tech Res 2017;10:988-94.

Toral MI, Nova-Ramirez F, Nacaratte F. Simultaneous determination of piperacillin and tazobactam in the pharmaceutical formulation tazonam by derivative spectrophotometry. J Chil Chem Soc 2012;57:1189-93.

Xiaoping B, Yongli BX, Xiaolei W. Content determination of sodium piperacillin and sodium tazobactam for injection by double wavelength UV spectrophotometry. J Chin Pharm Affairs 2003;8:27.

Zhang AP, Bai XH, Tong LG. Determination of piperacillin in compound injection by double wavelength ultraviolet spectrophotometry. J Shanxi Med Univ 2004;6:15.

Her Majesty’s Stationery Office. British Pharmacopoeia. London, UK: Her Majesty’s Stationery Office; 1993. p. 77.

Amin AS, Aly HM. Utilization of colorimetric and atomic absorption spectrometric determination of aztreonam through ion pair complex formation. Int J Pharm 2000;205:35-41.

El-Kousy N, Bebawy LI. Determination of some antihistaminic drugs by atomic absorption spectrometry and colorimetric methods. J Pharm Biomed Anal 1999;20:671-9.

Job P. Formation and stability of inorganic complexes in solution. Ann Chim 1928;2:113

Yoe J, Jones A. Colorimetric determination of iron with disodium-1, 2-dihydroxybenzene-3, 5-disulfonate. Ind Eng Chem Anal Ed 1944;16:111-5.

Brashy EL, El-Sayed M, Abdallah MF, Sepai EL. Spectrophotometric determination of some fluoroquinolone antibacterials by ion-pair complex formation with cobalt (II) tetrathiocyanate. J Chin Chem Soc 2005; 52:77-84.

Abdel-Gawad FM, El-Guindi NM. Spectrophotometric determination of metoclopramide and oxybuprocaine through ion pair formation with thiocyanate and molybdenum (V) or cobalt (II). Anal Lett 1995; 28:1437-47.

Fekria M. Determination of amineptine and amprolium hydrochlorides through ion associates with cobalt (II) thiocyanate. Sci Pharm 2017; 70:355-66.

Sethi PD. HPLC Quantitative Analysis of Pharmaceutical Formulations. India: CBS Publications; 2001.

ICH Guidelines. Validation of analytical procedures. Text Methodol 2015; 2:8-13.

Kiran KK, Nadh RV, Nagoji KEV. Extractive spectrophotometric determination of nicergoline through ion-pair complexation reaction. Orient J Chem 2013;29:263-9.

Sudhir MS, Nadh RV. Simple and validated ultraviolet spectrophotometric method for the estimation of febuxostat in bulk and pharmaceutical dosage forms. Orient J Chem 2013;29:1507-14.

Kiran KK, Nadh RV, Nagoji KE. Determination of bendamustine hydrochloride in pure and dosage forms by ion-associative complex formation reaction. Orient J Chem 2014;30:905-10.

Gorumutchu GP, Ratnakaram GV. Oxidative coupling: A tranquil approach for determination of selexipag by visible spectrophotometry. Orient J Chem 2018;34.

Gorumutchu GP, Ratnakaram VN. Determination of mianserine using tropaeoline-OOO by ion pair formation. Int J Appl Pharm 2019;1:11.

Gorumutchu GP, Ratnakaram VN. Determination of mianserine using Fe3+-phenanthroline by visible spectrophotometry. Res J Pharm Techn 2019;1:12.

Manasi BK, Anagha MJ. An experimental design approach for optimization of modified colorimetric first-order derivative method for estimation of serralysin in bulk and pharmaceutical formulation. Asian J Pharm Clin Res 2018;11:293-300.

Akram M, Monuir ZE, Nora O, Saleem S. Extractive-spectrophotometric determination of some antimuscarinic antagonist in tablet formulations using eriochrome cyanine R. Int J Pharm Pharm Sci 2018;4:22-8.

Sharma DK, Singh J, Raj P. Spectrophotometric determination of propranolol hydrochloride and metoprolol tartrate in pharmaceutical dosage forms, spiked water and biological fluids. Int J Pharm Pharm Sci 2018;10:107-15.

Godambe RD, Disouza JI, Jamkhandi CM, Kumbhar PS. Development of spectrophotometric and fluorometric methods for estimation of darunavir using Qbd approach. Int J Curr Pharm Res 2018;10:13-9.



How to Cite

GORUMUTCHU, G. P., V. N. RATNAKARAM, and K. KUMAR KATARI. “PIPERACILLIN ESTIMATION BY ION-ASSOCIATIVE COMPLEX FORMATION”. Asian Journal of Pharmaceutical and Clinical Research, vol. 12, no. 3, Mar. 2019, pp. 159-63, doi:10.22159/ajpcr.2019.v12i3.29289.



Original Article(s)