MilliporeSigma
  • Home
  • Search Results
  • Determination of polyphenolic compounds in Cirsium palustre (L.) extracts by high performance liquid chromatography with chemiluminescence detection.

Determination of polyphenolic compounds in Cirsium palustre (L.) extracts by high performance liquid chromatography with chemiluminescence detection.

Talanta (2014-12-02)
Edyta Nalewajko-Sieliwoniuk, Julita Malejko, Monika Mozolewska, Elżbieta Wołyniec, Jolanta Nazaruk
ABSTRACT

The first method for the simultaneous determination of polyphenolic antioxidants in extracts from leaves of Cirsium palustre based on high performance liquid chromatography combined with flow injection chemiluminescence detection (HPLC-FI-CL) has been developed. The extracts were prepared by using methanol as extraction medium and two types of extraction methods (reflux and ultrasound assisted extraction). The post-column CL determination of polyphenols was based on their enhancing effect on the chemiluminescence intensity generated in manganese(IV)-hexametaphosphate-formaldehyde system in a phosphoric acid medium. Main antioxidants determined in C. palustre leaves were eriodictyol-7-O-glucoside, luteolin-7-O-glucoside and 6-hydroxyluteolin-7-O-glucoside belonging to flavonoids, and chlorogenic acid belonging to phenolic acids. Chromatographic separation was carried out on a C18 column with gradient elution by using a mobile phase containing 0.25% (v/v) phosphoric acid in water (solvent A) and 100% methanol (solvent B). Under the optimized conditions of chromatographic separation and CL detection the validation of the method was performed. The calibration curves showed good linearity in the concentration range from 0.5 to 40 µg mL(-1). The HPLC-FI-CL method was successfully applied to the determination of four polyphenolic compounds in methanolic extracts from leaves of C. palustre. The accuracy of the developed method was confirmed by the comparison of the results with those obtained by an HPLC-PDA method. The relative error of determination does not exceed 6.1%. However, the HPLC-FI-CL method is characterized by 40-65 times higher sensitivity compared to the HPLC-PDA method.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Sodium formate, 99.998% trace metals basis
Sigma-Aldrich
5α-Androstan-17β-ol-3-one, ≥97.5%
Supelco
Acetonitrile, analytical standard
Sigma-Aldrich
Sodium formate, BioUltra, ≥99.0% (NT)
Sigma-Aldrich
Luteolin, ≥98% (TLC), powder
Sigma-Aldrich
Acetonitrile, ≥99.5%, ACS reagent
Sigma-Aldrich
Acetonitrile, anhydrous, 99.8%
Sigma-Aldrich
Acetonitrile, electronic grade, 99.999% trace metals basis
Supelco
5α-Androstan-17β-ol-3-one, VETRANAL®, analytical standard
Millipore
Bifido Selective Supplement B, suitable for microbiology
Sigma-Aldrich
Sodium hydroxide-16O solution, 20 wt. % in H216O, 99.9 atom % 16O
Sigma-Aldrich
Methanol-12C, 99.95 atom % 12C
Supelco
Acetonitrile, HPLC grade, ≥99.93%
Supelco
Acetonitrile, Pharmaceutical Secondary Standard; Certified Reference Material
USP
Residual Solvent Class 2 - Acetonitrile, United States Pharmacopeia (USP) Reference Standard
Luteolin, primary reference standard
USP
Methyl alcohol, United States Pharmacopeia (USP) Reference Standard
USP
Glacial acetic acid, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
Acetonitrile, HPLC Plus, ≥99.9%, poly-coated bottles
Supelco
Methanol solution, NMR reference standard, 4% in methanol-d4 (99.8 atom % D), NMR tube size 5 mm × 8 in.
Sigma-Aldrich
Methanol solution, NMR reference standard, 4% in methanol-d4 (99.8 atom % D), NMR tube size 3 mm × 8 in.
Supelco
Methanol solution, contains 0.10 % (v/v) formic acid, UHPLC, suitable for mass spectrometry (MS), ≥99.5%
Supelco
Luteolin, analytical standard
Supelco
Eriodictyol, analytical standard
Sigma-Aldrich
Sodium formate, ACS reagent, ≥99.0%
Sigma-Aldrich
Acetonitrile, puriss. p.a., ACS reagent, reag. Ph. Eur., ≥99.5% (GC)
Sigma-Aldrich
Acetonitrile, ReagentPlus®, 99%
Sigma-Aldrich
Sodium formate, reagent grade, 97%
Sigma-Aldrich
Acetonitrile, biotech. grade, ≥99.93%
Sigma-Aldrich
Acetonitrile, suitable for DNA synthesis, ≥99.9% (GC)