A Non-Instrumental Green Analytical Method Based on Surfactant-Assisted Dispersive Liquid-Liquid Microextraction-Thin-Layer Chromatography-Smartphone-Based Digital Image Colorimetry(SA-DLLME-TLC-SDIC) for Determining Favipiravir in Biological Samples

Bharti Jain; Rajeev Jain; Prashant Kumar Jaiswal; Torki Zughaibi; Tanvi Sharma; Abuzar Kabir; Ritu Singh; Shweta Sharma

doi:10.3390/molecules28020529

A Non-Instrumental Green Analytical Method Based on Surfactant-Assisted Dispersive Liquid-Liquid Microextraction-Thin-Layer Chromatography-Smartphone-Based Digital Image Colorimetry(SA-DLLME-TLC-SDIC) for Determining Favipiravir in Biological Samples

Molecules. 2023 Jan 5;28(2):529. doi: 10.3390/molecules28020529.

Authors

Bharti Jain^{1

2}, Rajeev Jain¹, Prashant Kumar Jaiswal³, Torki Zughaibi^{4

5}, Tanvi Sharma², Abuzar Kabir^{6

7}, Ritu Singh³, Shweta Sharma²

Affiliations

¹ Central Forensic Science Laboratory, Dakshin Marg, Sector-36A, Chandigarh 160036, India.
² Institute of Forensic Science & Criminology, Panjab University, Chandigarh 160014, India.
³ School of Earth Sciences, Department of Environmental Sciences, Central University of Rajasthan, NH-8, Bandar Sindri, Ajmer 305817, India.
⁴ Department of Medical Laboratory Sciences, Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah 21589, Saudi Arabia.
⁵ King Fahd Medical Research Center, King Abdulaziz University, Jeddah 21589, Saudi Arabia.
⁶ International Forensic Research Institute, Department of Chemistry and Biochemistry, Florida International University, Miami, FL 33199, USA.
⁷ Department of Pharmacy, Faculty of Allied Health Science, Daffodil International University, Dhaka 1207, Bangladesh.

Abstract

Favipiravir (FAV) has become a promising antiviral agent for the treatment of COVID-19. Herein, a green, fast, high-sample-throughput, non-instrumental, and affordable analytical method is proposed based on surfactant-assisted dispersive liquid-liquid microextraction (SA-DLLME) combined with thin-layer chromatography-digital image colourimetry (TLC-DIC) for determining favipiravir in biological and pharmaceutical samples. Triton X-100 and dichloromethane (DCM) were used as the disperser and extraction solvents, respectively. The extract obtained after DLLME procedure was spotted on a TLC plate and allowed to develop with a mobile phase of chloroform:methanol (8:2, v/v). The developed plate was photographed using a smartphone under UV irradiation at 254 nm. The quantification of FAV was performed by analysing the digital images' spots with open-source ImageJ software. Multivariate optimisation using Plackett-Burman design (PBD) and central composite design (CCD) was performed for the screening and optimisation of significant factors. Under the optimised conditions, the method was found to be linear, ranging from 5 to 100 µg/spot, with a correlation coefficient (R²) ranging from 0.991 to 0.994. The limit of detection (LOD) and limit of quantification (LOQ) were in the ranges of 1.2-1.5 µg/spot and 3.96-4.29 µg/spot, respectively. The developed approach was successfully applied for the determination of FAV in biological (i.e., human urine and plasma) and pharmaceutical samples. The results obtained using the proposed methodology were compared to those obtained using HPLC-UV analysis and found to be in close agreement with one another. Additionally, the green character of the developed method with previously reported protocols was evaluated using the ComplexGAPI, AGREE, and Eco-Scale greenness assessment tools. The proposed method is green in nature and does not require any sophisticated high-end analytical instruments, and it can therefore be routinely applied for the analysis of FAV in various resource-limited laboratories during the COVID-19 pandemic.

Keywords: digital image colourimetry; favipiravir; surfactant-assisted dispersive liquid–liquid microextraction; thin-layer chromatography.

MeSH terms

COVID-19*
Chromatography, High Pressure Liquid
Chromatography, Thin Layer
Colorimetry
Humans
Limit of Detection
Lipoproteins
Liquid Phase Microextraction* / methods
Pandemics
Pharmaceutical Preparations
Pulmonary Surfactants*
Smartphone
Solvents
Surface-Active Agents

Substances

Surface-Active Agents
favipiravir
Solvents
Pulmonary Surfactants
Lipoproteins
Pharmaceutical Preparations

Grants and funding

Bharti Jain is thankful to the Department of Science & Technology (DST)-Inspire, New Delhi, grant no. DST/INSPIRE Fellowship/2019/IF190181. The author Torki Zughaibi gratefully acknowledges the technical and financial support from the Ministry of Education and the Deanship of Scientific Research (DSR), King Abdulaziz University (KAU), Jeddah, Saudi Arabia (IFPDP-68-22).