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  • Development and Validation of an HPLC-UV Method for Determination of Eight Phenolic Compounds in Date Palms.

Development and Validation of an HPLC-UV Method for Determination of Eight Phenolic Compounds in Date Palms.

Journal of AOAC International (2015-11-04)
Fuad Al-Rimawi, Imad Odeh
ABSTRACT

A simple, precise, accurate, and selective method was developed and validated for determination of eight phenolic compounds (gallic acid, p-hydroxybenzoic acid, vanilic acid, caffeic acid, syringic acid, p-coumaric acid, ferulic acid, and sinapic acid) in date palms. Separation was achieved on an RP C18 column using the mobile phase methanol-water with 2% acetic acid (18+82, v/v). This method was validated according to the requirements for new methods, which include accuracy, precision, selectivity, robustness, LOD, LOQ, linearity, and range. The method demonstrated good linearity over the range 1-1000 ppm of gallic acid, p-hydroxybenzoic acid, vanilic acid, caffeic acid, and syringic acid with r2 greater than 0.99, and in the range of 3-1000 ppm for p-coumaric acid, ferulic acid, and sinapic acid with r2 greater than 0.99. The recovery of the eight phenolic compounds ranged from 97.1 to 102.2%. The method is selective because adjacent peaks of phenolic compounds were well separated with good resolution. The degree of reproducibility of the results obtained as a result of small deliberate variations in the method parameters and by changing analytical operators proved that the method is robust and rugged.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Acetic acid-12C2, 99.9 atom % 12C
Sigma-Aldrich
Methanol solution, contains 0.50 % (v/v) triethylamine
Sigma-Aldrich
Methanol solution, NMR reference standard, 4% in methanol-d4 (99.8 atom % D), NMR tube size 3 mm × 8 in.
Sigma-Aldrich
Sinapic acid, ≥98%, powder
Sigma-Aldrich
p-Coumaric acid, ≥98.0% (HPLC)
Sigma-Aldrich
Acetic acid, natural, ≥99.5%, FG
Sigma-Aldrich
Acetic acid, ≥99.5%, FCC, FG
Sigma-Aldrich
Methanol, NMR reference standard
Sigma-Aldrich
Methanol-12C, 99.95 atom % 12C
Sigma-Aldrich
4-Hydroxybenzoic acid, ReagentPlus®, ≥99%
Sigma-Aldrich
Methanol, anhydrous, 99.8%
Sigma-Aldrich
trans-Ferulic acid, 99%
Sigma-Aldrich
trans-Ferulic acid, ≥99%
Sigma-Aldrich
4-Hydroxybenzoic acid, ReagentPlus®, 99%
Sigma-Aldrich
Methanol, purification grade, 99.8%
Sigma-Aldrich
Methanol, ACS reagent, ≥99.8%
Sigma-Aldrich
Acetic acid, glacial, ACS reagent, ≥99.7%
Sigma-Aldrich
Acetic acid solution, suitable for HPLC
Sigma-Aldrich
Methanol, BioReagent, ≥99.93%
Sigma-Aldrich
Acetic acid, glacial, puriss., 99-100%
Sigma-Aldrich
Acetic acid, glacial, puriss., meets analytical specification of Ph. Eur., BP, USP, FCC, 99.8-100.5%
Sigma-Aldrich
Methanol, ACS reagent, ≥99.8%
Sigma-Aldrich
Acetic acid, glacial, ≥99.99% trace metals basis
Sigma-Aldrich
Methanol, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.8% (GC)
Sigma-Aldrich
Acetic acid, glacial, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.8%
Sigma-Aldrich
Methanol, puriss., meets analytical specification of Ph Eur, ≥99.7% (GC)
Sigma-Aldrich
Acetic acid, glacial, ReagentPlus®, ≥99%
Sigma-Aldrich
Methanol, ACS reagent, ≥99.8%
Sigma-Aldrich
Methanol, ACS spectrophotometric grade, ≥99.9%
Sigma-Aldrich
Gallic acid, 97.5-102.5% (titration)