Parakat Tanmai Meenakshi¹, Gunturu Jaya Syamala¹*, Y Ankamma Chowdary², Gorijavolu Vamseekrishna³ and Thanneeru Venkata Sowjanya⁴

¹Master of Pharmacy, Department of Pharmaceutical Quality Assurance, School of Pharmaceutical Sciences, Dr. RVR NRI Institute of Technology, Deemed to be University, Pothavarappadu (V), Agiripalli (M), Vijayawada Rural, Eluru District, Andhra Pradesh, India
²Assistant Professor, Department of Pharmaceutical Analysis, School of Pharmaceutical Sciences, Dr. RVR NRI Institute of Technology, Deemed to be University, Pothavarappadu (V), Agiripalli (M), Vijayawada Rural, Eluru District, Andhra Pradesh, India
³Dean, School of Pharmaceutical Sciences, Dr. RVR NRI Institute of Technology, Deemed to be University, Pothavarappadu (V), Agiripalli (M), Vijayawada Rural, Eluru District, Andhra Pradesh, India
⁴Professor, Department of Pharmaceutics, School of Pharmaceutical Sciences, Dr. RVR NRI Institute of Technology, Deemed to be University, Pothavarappadu (V), Agiripalli (M), Vijayawada Rural, Eluru District, Andhra Pradesh, India
⁵Research Scholar, Department of Pharmaceutical Analysis, School of Pharmaceutical Sciences, Dr. RVR NRI Institute of Technology, Deemed to be University, Pothavarappadu (V), Agiripalli (M), Vijayawada Rural, Eluru District, Andhra Pradesh, India

*Corresponding Author: Gunturu Jaya Syamala, Assistant Professor, Department of Pharmaceutical Analysis, School of Pharmaceutical Sciences, Dr. RVR NRI Institute of Technology, Deemed to be University, Pothavarappadu (V), Agiripalli (M), Vijayawa- da Rural, Eluru District-521212, Andhra Pradesh, India.

Received: September 08, 2026; Published: October 13, 2026

Abstract

Introduction: Simultaneous estimation of citicoline and piracetam, a fixed-dose nootropic combination widely used for cerebrovascular and cognitive disorders, is essential to ensure the quality of combined pharmaceutical formulations. This study aimed to develop and validate a simple, rapid, accurate, precise, robust and economical reversed-phase high-performance liquid chromatographic (RP-HPLC) method for the simultaneous estimation of citicoline and piracetam in bulk drug and tablet dosage form in accordance with International Council for Harmonisation (ICH) guidelines.

Materials and Methods: Chromatographic separation was carried out on a Hypersil BDS C18 column (250 × 4.6 mm, 5 μm) using ammonium dihydrogen orthophosphate buffer (pH 3.5, adjusted with ortho-phosphoric acid) as the mobile phase in isocratic mode, at a flow rate of 1.0 mL/min with ultraviolet detection at 210 nm. The method was validated for system suitability, specificity, linearity, accuracy, precision, robustness, limit of detection (LOD), limit of quantitation (LOQ) and assay as per ICH Q2 guidelines.

Results: The optimized method produced well-resolved, symmetrical peaks for citicoline (retention time 3.197 min) and piracetam (retention time 10.723 min) (resolution, Rs = 9.42; tailing factor ≤ 1.4; theoretical plates > 6700) within a 20 min isocratic run. Linearity was established over 25-75 μg/mL for citicoline and 40-120 μg/mL for piracetam, with correlation coefficients (R²) of 0.9997 and 0.9996, respectively (n = 5 concentrations, six replicate injections each). Mean percentage recoveries were 99.52 ± 0.13% for citicoline and 98.98 ± 0.23% for piracetam (n = 9 per drug), with %RSD values below 2.0% for all precision parameters (repeatability, intermediate precision and reproducibility). The LOD and LOQ, calculated from the residual standard deviation of the regression line and its slope, were 0.06 and 0.18 μg/mL for citicoline, and 0.09 and 0.26 μg/mL for piracetam, respectively. Peak area %RSD remained ≤ 0.36% under small, deliberate variation of flow rate, detection wavelength and mobile-phase pH, confirming robustness.

Discussion: All validation parameters satisfied the acceptance criteria of the ICH Q2(R2) guideline, and no co-eluting peak from tablet excipients or from acid-, base-, peroxide-, thermal- or photolytically generated degradation products was observed at the retention times of either analyte, confirming both the specificity and the stability-indicating capacity of the method.

Conclusion: The developed RP-HPLC method is simple, rapid, accurate, precise, robust and cost-effective, and is suitable for the routine simultaneous quality-control estimation of citicoline and piracetam in bulk drug and pharmaceutical dosage forms.

Keywords: Citicoline; Piracetam; RP-HPLC; Method Development; Method Validation; Simultaneous Estimation; ICH Guidelines

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Citation

Citation: Gunturu Jaya Syamala., et al. “Rp-Hplc Method Development and Validation for the Simultaneous Estimation of Citicoline and Piracetam in Bulk Drug and Tablet Dosage Form". Acta Scientific Pharmaceutical Sciences 10.6 (2026): 48-56.

Copyright

Copyright: © 2026 Gunturu Jaya Syamala., et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.