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Case Study

Study of urinary (poly)phenol biomarkers by LDTD-MS/MS

The Institute of Nutrition and Functional Foods (INAF) is a group of food and health researchers founded at Laval University in Quebec. In order to contribute to sustainable nutrition and health, Professor Yves Desjardins’ team mainly focuses on the research areas of food and well-being as well as the personalization of nutrition. The LDTD technology can be applied to the fields of clinical and food research by quantifying nutritional biomarkers present in biological matrices. Indeed, in a study by Lessard-Lord et al. (2023), the analysis of metabolites of poly(phenols), namely phenyl-y-valerolactones (PVL), biomarkers of flavan-3-ols in urine, can be carried out using the LDTD-MS/MS technology and the Azeo Liquid Handler.

Project Type

Analysis of urinary biomarkers (3-HPVL and 3,4-DHPVL) of flavan-3-ols, (poly)phenols, present, among others, in tea and berries such as cranberries. Additionally, tandem creatinine analysis allows normalization of biomarker concentrations in urine.

Challenges

Analysis of urinary biomarkers is generally performed by high-performance liquid chromatography (HPLC). However, with the evolution of technology, the use of ultra-high performance liquid chromatography (UPLC) makes it possible to reduce analysis times. Unfortunately, manual sample preparation and the use of UPLC do not enable high-speed analysis for large-scale epidemiological analysis.

Objectives

Objective 1

Enable large-scale epidemiological analysis using a large number of samples. By analyzing many different patient samples, it is possible to evaluate solutions to health problems through these epidemiological studies. Analyzing this high number of samples requires adopting a high-throughput approach, which means automating a time-consuming process to make it more efficient.

Objective 2

Reduce analysis times during clinical studies to obtain trial results more quickly, and therefore, iterate more rapidly.

Solutions

Increase high throughput analysis with LDTD technology to reduce analysis times.
Use a robot, the Azeo Liquid Handler, for sample preparation, thereby reducing sample preparation times.
Enable better sample traceability when analyzing large volumes in large-scale epidemiological studies.

Results

When analyzing urinary biomarkers by mass spectrometry, the LDTD-MS/MS is faster than UPLC-MS/MS by 50 times (10 seconds vs 8 minutes). This allows the analysis of 5000 samples in only 9 days with the Azeo Liquid Handler and the Luxon Ion Source, compared to the 100 days it takes with HPLC. The Azeo Liquid Handler allows a 90% recovery.

The precision on 3-HPVL, 3,4-DHPVL and creatinine ranged from 1.6% to 11.6% of the nominal concentration and the accuracy was between 90.3% and 112.8%, both using the LDTD-MS/MS technology.

50X

faster than UPLC

5000

samples in 9 days

90%

recovery with Azeo

References

Lessard-Lord, J., Auger, S., Demers, S., Plante, P.-L., Picard, P., Desjardins, Y. (2023). Automated High-Throughput Quantification of Phenyl-γ-valerolactones and Creatinine in Urine by Laser Diode Thermal Desorption.  J. Agric. Food Chem. 71, 16787−16796. https://doi.org/10.1021/acs.jafc.3c03888

https://www.inaf.ulaval.ca/a-propos/

https://www.inaf.ulaval.ca/membres/yves-desjardins/