Measurement of Saccharin and trans-Resveratrol Metabolites in Urine as Adherence Markers for Small Quantity Lipid-Based Nutrient Supplement Consumption.

Valerie Weinborn,Alyson E Mitchell,Marjorie J Haskell,Fanny B Morel,Arlie L Lehmkuhler,Mamane Zeilani,Sarah J Zyba

doi:10.1021/acs.jafc.0c06144

Abstract

Saccharin and trans-resveratrol were incorporated into small quantity lipid-based nutritional supplements (SQ-LNS) to be evaluated as the markers of consumption for nutritional intervention studies. Forty-seven healthy women consumed a single supplement with either 8.6 mg of saccharin or 5 mg of trans-resveratrol, and urine was collected for 4 h. A rapid 11 min method employing multiple reaction monitoring and ultrahigh performance liquid chromatography coupled to a triple quadrupole mass spectrometer was developed to measure saccharin and resveratrol metabolites in urine simultaneously. The linear dynamic range of the method was from 3 to 1000 ng mL–1, with the correlation coefficient of 0.999 and limits of quantification from 15.28 to 53.03 ng mL–1. Sample preparation was simple dilution with an average recovery of 97.8%. Ion suppression was observed with urine concentrations >10%. Mean levels of saccharin and resveratrol-3-O-sulfate in urine were 5.481 ± 4.359 and 3.440 ± 4.160 nmol L–1, respectively. We developed and validated a method to measure saccharin and trans-resveratrol metabolites in urine to objectively corroborate the consumption of SQ-LNS for the first time in nutrition intervention studies.

Highlights

Small quantity lipid-based nutrient supplements (SQ-LNS) are food-based supplements typically based on vegetable fat, skimmed milk powder, peanut paste, sugar, and a complex of minerals and vitamins, formulated to meet the needs of specific target populations.[1]
Blank urine samples were diluted 10:90 v/v with methanol/water 1:1 v/v and spiked at 10, 100, and 800 ng mL−1 with trans-resveratrol-3-O-β-D-glucuronide, transresveratrol-4′-O-D-glucuronide, resveratrol-3-O-sulfate, and saccharin to study the recovery of each analyte
The average content of resveratrol metabolites in urine at baseline was 0 ± 0, 15.50 ± 53.44, and 4.89 ± 22.39 nmol L−1 for transresveratrol-4′-O-D-glucuronide, trans-resveratrol-3-O-β-D-glucuronide, and resveratrol-3-O-sulfate, respectively, which increased to 507.82 ± 768.80, 913.63 ± 927.51, and 3439.96 ± 4160.05 nmol L−1 in the 4 h pooled sample after LNS2 consumption

Summary

■ INTRODUCTION

Small quantity lipid-based nutrient supplements (SQ-LNS) are food-based supplements typically based on vegetable fat, skimmed milk powder, peanut paste, sugar, and a complex of minerals and vitamins, formulated to meet the needs of specific target populations.[1]. Blank urine samples were diluted 10:90 v/v with methanol/water 1:1 v/v and spiked at 10, 100, and 800 ng mL−1 with trans-resveratrol-3-O-β-D-glucuronide, transresveratrol-4′-O-D-glucuronide, resveratrol-3-O-sulfate, and saccharin to study the recovery of each analyte. The urine concentrations of saccharin and resveratrol metabolites were measured in response to a single dose of SQ-LNS containing the marker compounds. The average content of resveratrol metabolites in urine at baseline was 0 ± 0, 15.50 ± 53.44, and 4.89 ± 22.39 nmol L−1 for transresveratrol-4′-O-D-glucuronide, trans-resveratrol-3-O-β-D-glucuronide, and resveratrol-3-O-sulfate, respectively, which increased to 507.82 ± 768.80, 913.63 ± 927.51, and 3439.96 ± 4160.05 nmol L−1 in the 4 h pooled sample after LNS2 consumption. A UHPLC(ESI)-MS/MS method was developed for the quantification of saccharin and trans-resveratrol metabolites in urine simultaneously, or independently, to monitor the consumption of SQ-LNS.

■ ACKNOWLEDGMENTS

Findings

■ REFERENCES