Author(s):
Deep Shikha Sharma, Sheetu Wadhwa, Sachin Kumar Singh, Arya Ramanunny, Rajan Kumar
Email(s):
singhsachin23@gmail.com
DOI:
10.5958/0974-360X.2020.00750.7 
Address:
Deep Shikha Sharma, Sheetu Wadhwa, Sachin Kumar Singh*, Arya Ramanunny, Rajan Kumar
School of Pharmaceutical Sciences, Lovely Professional University, Phagwara, Punjab (India)–144411.
*Corresponding Author
Published In:
Volume - 13,
Issue - 9,
Year - 2020
ABSTRACT:
The main intention of the current work is the development of a suitable, simple, precise, reproducible, and accurate high-performance liquid chromatography reversed-phase validated method which can be used for the estimation of 5-Fluorouracil.The estimation was done by high-performance liquid chromatography (HPLC) method. In this suitable stationary phase and optimum composition of the mobile phase was selected which provides good resolution and short run time for the estimation of 5-Fluorouracil (5-FU). Stationary phase used was Nucleodur C18 column (Reverse phase, 250mm × 4.6mm i.d., particle having 5 micron size) and mobile phase consists of combination of Ortho-Phosphoric Acid (OPA) (0.5%) and methanol having ratio 95:5, v/v were used in an isocratic mode of elution. The mobile phase used with a flow rate of 0.8 mL/min and volume of the injection for 5-FU was 20µL. The eluent observed at 266nm for measurement of 5-FU. The validation of this method was carried out with the help of various parameters such as sensitivity, selectivity, system suitability, precision (inter-day and intra-day), accuracy, and linearity according to International Conference on Harmonization guidelines i.e. ICH Q2 (R1). The 5-FU showing retention time at 7.2 min. The responses showing linearity in the concentration range between 2-10µg/mL with correlation coefficient 0.99. The % mean recovery of 5-FU was calculated at three different levels i.e Lower Quality control Concentration (LQC), Middle Quality control Concentration (MQC), High Quality control Concentration (LQC) whose results falls within the range i.e. 95% to 105%, which indicates the accuracy of this method. The % relative standard deviation (RSD) precision (intraday and intermediate) at 3 different levels was < 2% which indicated the precision of this method. LOD i.e. Limit of Detection and LOQ i.e. Limit of Quantification was found to be 0.870277 and 2.637202 respectively for 5-FU.
Cite this article:
Deep Shikha Sharma, Sheetu Wadhwa, Sachin Kumar Singh, Arya Ramanunny, Rajan Kumar. Estimation of 5-fluorouracil by high-performance liquid Chromatography Reversed-phase Validated method. Research J. Pharm. and Tech 2020; 13(9):4249-4254. doi: 10.5958/0974-360X.2020.00750.7
Cite(Electronic):
Deep Shikha Sharma, Sheetu Wadhwa, Sachin Kumar Singh, Arya Ramanunny, Rajan Kumar. Estimation of 5-fluorouracil by high-performance liquid Chromatography Reversed-phase Validated method. Research J. Pharm. and Tech 2020; 13(9):4249-4254. doi: 10.5958/0974-360X.2020.00750.7 Available on: https://rjptonline.org/AbstractView.aspx?PID=2020-13-9-38
REFERENCES:
1. Parrozzani R, Frizziero L, Trainiti S, and Testi L, et al. Topical 1% 5-fluoruracil as a sole treatment of corneo conjunctival ocular surface squamous neoplasia: long-term study. British Journal of Ophthalmology. 2017; 101:1094–1099.
2. Youssof AM, Salem-Bekhit MM, Shakeel F, Alanazi FK, Haq N. Analysis of anti-neoplastic drug in bacterial ghost matrix, w/o/w double nanoemulsion and w/o nanoemulsion by a validated ‘green’ liquid chromatographic method. Talanta. 2016; 154:292-2981. DOI: 10.1016/j.talanta.2016.03.086
3. Mishra V, Thakur S, Patil A, Shukla A. Quality by design (QbD) approaches in current pharmaceutical set-up. Expert Opinion on Drug Delivery. 2018; 15 (8):737-758. doi: 10.1080/17425247.2018.1504768.
4. Cohn NS. Similar cytological effects of hydroxylamine and 5-FUDR, agents with different modes of action. Experientia. 1964; 20 (3): 158-61,March 1964.
5. Sameer A, Abdulrahman M. Sensitive and selective spectrophotometric assay of gabapentin in capsules using sodium 1, 2-naphthoquinone-4-sulfonate. Drug Testing and Analysis. 2011;10.
6. Saka, Chromatographic Methods for Determination of Drugs Used in Prostate Cancer in Biological and Pharmacological Samples. Critical Reviews in Analytical Chemistry. 2019; 49 (1) 78-99.https://doi.org/10.1080/10408347.2018.1487776
7. Ravisankar P, Navyal N, Pravallika V, NavyaSri D, A Review on Step-by-Step Analytical Method Validation. IOSR Journal Of Pharmacy. 2015; 5 (10):7-19.
8. Sharma P, Singh SK, Pandey N, Kumar NR, et al. Impact of solid carriers and spray drying on pre/post-compression properties, dissolution rate and bioavailability of solid self nano emulsifying drug delivery system loaded with simvastatin. Powder Technology. 2018: 836-846.
9. Mujeeb U, Rahman U., Rathore A, Siddqui AA, Parveen G, Yar MS, Determination of the Stabilities of New Quinazoline Derivatives by HPLC. Journal of Liquid Chromatography and Related Technologies. 2014; 29 (5): 733-43. doi: 10.3109/14756366
10. Jyoti J, Anandhakrishnan NK, Singh SK, Kumar B, et al. A three-pronged formulation approach to improve oral bioavailability and therapeutic efficacy of two lipophilic drugs with gastric lability. Drug Delivery and Translational. 2019:1-18. doi: 10.1007/s13346-019-00635-0
11. Hassain Y, Gupta VK. Analysis of Melatonin in Dosage Formulation by Capillary Electrophoresis, Journal of Liquid Chromatography and Related Technologies. 2007;30 (4):545-556.
12. Garg VK, Singh S, Pandey NK, Bhatia A, et al. Impact of spray drying over conventional surface adsorption technique for improvement in micromeritic and biopharmaceutical characteristics of self-nanoemulsifying powder loaded with two lipophilic as well as gastrointestinal labile drugs. Powder Technology. 2018; 326:425-442.
13. Jin Y, Rong LF, Lu XW, Huang Y, et al. Pharmacokinetics and tissue distribution of 5-fluorouracil encapsulated by galactosy lceramide liposomes in mice. Acta Pharmacologica Sinica, 2005; 26 (2) 250-6.
14. Mohanta S, Singh SK, Kumar B, Gulati M, et al. Solidification of liquid Modified Apple Polysaccharide by its adsorption on solid porous carriers through spray drying and evaluation of its potential as binding agent for tablets. International Journal of Biological Macromolecules, 2018; 120:1975-1998..
15. Mittal S, Alexander KS, Dollimore D. A High-Performance Liquid Chromatography Assay for Yohimbine HCl Analysis. Drug Development and Industrial Pharmacy. 2000;26 (10);1059-65.
16. Sharma AK, Ponery AS, Lawrence PA, Ahmed I, et al. Effect of α‐tocopherol supplementation on the ultrastructural abnormalities of peripheral nerves in experimental diabetes. Journal of the Peripheral Nervous System. 2001;6 (1):33-9.
17. Awad M, Hammad MA, Omar MA. Utility of Von Pechman synthesis of coumarin reaction for development of spectrofluorimetric method for quantitation of salmeterol xinafoate in pharmaceutical preparations and human plasma. Luminescence. 2018;33 (5): 913-918. doi: 10.1002/bio.3490.
18. Kim W, Yousaf AM, Li DX, et al. Development of RP-HPLC method for simultaneous determination of docetaxel and curcumin in rat plasma: Validation and stability. Asian Journal of Pharmaceutical Sciences. 2017;12(1):105-113.
19. Omar MA, Hammad MA, Awad M. Utility of Europium ion characteristic peak for quantitation of Fenoterol hydrobromide and Salmeterol xinafoate in different matrices; application to stability studies. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy. 2019;217:182-189.
20. Kamalakkannan V, Puratchikody A, Ramanathan L, Jayapraba S. Development and validation of a dissolution test with reversed-phase high performance liquid chromatographic analysis for Candesartan cilexetil in tablet dosage forms. Arabian Journal of Chemistry. 2016;9 (1): S867-S873.