UPLC Analysis of Phenolic Compounds

The method describe earlier by Aqil et al. (2014) (link) was adapted with little modification. Briefly, methanolic extracts of MLwas analyzed for the phenolics on a Shimadzu ultra performance liquid chromatography (UPLC) system comprised of two LC-20AD-XR pumps, SIL-20A-XR autosampler, and SPD-M20A photodiode array detector (PDA) controlled by Class VP software (ver 7.4, SP3) attached to a Shim-pack XR-ODS-II column (3.0 mm × 150 mm; 2.2 μ). A linear gradient of 3.5% phosphoric acid (solvent A) and acetonitrile (solvent B) with flow rate of 0.7 ml/min was used. In the gradient, solvent B was initially 15% for 2 min and increased to 20% by 3 min. The solvent B concentration was further increased to 60% from 3 to 10 min and held for 1 min and returned to 15% by 14 min. The chromatogram was collected from 200 to 600 nm and general phenolics were analyzed at 280 nm.

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Husain F.M., Ahmad I., Al-thubiani A.S., Abulreesh H.H., AlHazza I.M, & Aqil F. (2017). Leaf Extracts of Mangifera indica L. Inhibit Quorum Sensing – Regulated Production of Virulence Factors and Biofilm in Test Bacteria. Frontiers in Microbiology, 8, 727.

Publication 2017

SPD-M20A XR-ODS-II column

Acetonitrile Held Liquid chromatography Methanolic Phosphoric acid Shim Solvent

Corresponding Organization : Aligarh Muslim University

Other organizations : King Saud University, Umm al-Qura University, University of Louisville

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Variable analysis

independent variables

Methanolic extracts of ML

dependent variables

Phenolics analysis

control variables

Shimadzu ultra performance liquid chromatography (UPLC) system comprised of two LC-20AD-XR pumps, SIL-20A-XR autosampler, and SPD-M20A photodiode array detector (PDA) controlled by Class VP software (ver 7.4, SP3)
Shim-pack XR-ODS-II column (3.0 mm × 150 mm; 2.2 μ)
Linear gradient of 3.5% phosphoric acid (solvent A) and acetonitrile (solvent B) with flow rate of 0.7 ml/min
Solvent B concentration: Initial 15% for 2 min, increased to 20% by 3 min, further increased to 60% from 3 to 10 min and held for 1 min, and returned to 15% by 14 min
Chromatogram collected from 200 to 600 nm, general phenolics analyzed at 280 nm

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