MilliporeSigma
  • Home
  • Search Results
  • Development of a targeted method for twenty-three metabolites related to polyphenol gut microbial metabolism in biological samples, using SPE and UHPLC-ESI-MS/MS.

Development of a targeted method for twenty-three metabolites related to polyphenol gut microbial metabolism in biological samples, using SPE and UHPLC-ESI-MS/MS.

Talanta (2014-07-26)
Mattia Gasperotti, Domenico Masuero, Graziano Guella, Fulvio Mattivi, Urska Vrhovsek
ABSTRACT

An increasing number of studies have concerned the profiling of polyphenol microbial metabolites, especially in urine or plasma, but only a few have regarded their accurate quantification. This study reports on a new ultra-performance liquid chromatography tandem mass spectrometry method with electrospray ionisation (UHPLC-ESI-MS/MS) using a simple clean-up step with solid phase extraction (SPE) and validation on different biological matrices. The method was tested with spiked samples of liver, heart, kidneys, brain, blood and urine. The purification procedure, after the evaluation of three different cartridges, makes it possible to obtain cleaner samples and better quantification of putative trace metabolites, especially related to dietary studies, with concentrations below ng/g in tissue and for urine and blood, starting from ng/ml. Limits of detection and linear range were also assessed using mixed polyphenol metabolite standards. Short chromatographic separation was carried out for 23 target compounds related to the polyphenol microbial metabolism, coupled with a triple quadrupole mass spectrometer for their accurate quantification. By analysing different spiked biological samples we were able to test metabolite detection in the matrix and validate the overall recovery of the method, from purification to quantification. The method developed can be successfully applied and is suitable for high-throughput targeted metabolomics analysis related to nutritional intervention, or the study of the metabolic mechanism in response to a polyphenol-rich diet.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
3-Hydroxy-4-methoxycinnamic acid, predominantly trans, 97%
Sigma-Aldrich
4-Hydroxybenzoic acid, ReagentPlus®, 99%
Sigma-Aldrich
1,2-Dihydroxybenzene, ReagentPlus®, ≥99%
Supelco
4-Hydroxybenzoic acid, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
4-Hydroxybenzoic acid, ReagentPlus®, ≥99%
Sigma-Aldrich
trans-Ferulic acid, 99%
Sigma-Aldrich
4-Hydroxybenzoic acid, ≥99%, FG
Sigma-Aldrich
trans-Ferulic acid, ≥99%
Supelco
trans-Ferulic acid, matrix substance for MALDI-MS, ≥99.0% (HPLC)
Sigma-Aldrich
3-(4-Hydroxy-3-methoxyphenyl)propionic acid, ≥96.0% (T)
Supelco
Caffeic acid, matrix substance for MALDI-MS, ≥99.0% (HPLC)
Supelco
trans-Sinapic acid, analytical standard
Supelco
trans-Isoferulic acid, analytical standard
Supelco
trans-Ferulic acid, certified reference material, TraceCERT®
Supelco
trans-p-Coumaric acid, analytical standard
Supelco
Methanol solution, contains 0.10 % (v/v) formic acid, UHPLC, suitable for mass spectrometry (MS), ≥99.5%
Sigma-Aldrich
Caffeic acid, ≥98.0% (HPLC)
Sigma-Aldrich
Acetonitrile, anhydrous, 99.8%
Sigma-Aldrich
Acetonitrile, suitable for HPLC, gradient grade, ≥99.9%
Sigma-Aldrich
Acetonitrile, HPLC Plus, ≥99.9%
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, NMR reference standard, 4% in methanol-d4 (99.8 atom % D), NMR tube size 5 mm × 8 in.
Sigma-Aldrich
Formic acid, ACS reagent, ≥96%
Sigma-Aldrich
Acetonitrile, suitable for HPLC, gradient grade, ≥99.9%
Supelco
Acetonitrile, HPLC grade, ≥99.93%
Supelco
Acetonitrile, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Formic acid solution, BioUltra, 1.0 M in H2O
Sigma-Aldrich
Formic acid, ≥95%, FCC, FG
USP
Residual Solvent Class 2 - Acetonitrile, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
Formic acid, reagent grade, ≥95%