Imperial College London

Emeritus ProfessorJeremyNicholson

Faculty of MedicineDepartment of Metabolism, Digestion and Reproduction

Emeritus Professor of Biological Chemistry



+44 (0)20 7594 3195j.nicholson Website




Ms Wendy Torto +44 (0)20 7594 3225




Office no. 665Sir Alexander Fleming BuildingSouth Kensington Campus






BibTex format

author = {Whiley, L and Chekmeneva, E and Berry, DJ and Jimenez, B and Yuen, AHY and Salam, A and Hussain, H and Witt, M and Takats, Z and Nicholson, JK and Lewis, MR},
doi = {10.1021/acs.analchem.9b00241},
journal = {Analytical Chemistry},
pages = {8873--8882},
title = {Systematic isolation and structure elucidation of urinary metabolites optimized for the analytical-scale molecular profiling laboratory},
url = {},
volume = {91},
year = {2019}

RIS format (EndNote, RefMan)

AB - Annotation and identification of metabolite biomarkers is critical for their biological interpretation in metabolic phenotyping studies, presenting a significant bottleneck in the successful implementation of untargeted metabolomics. Here, a systematic multi-step protocol was developed for the purification and de novo structural elucidation of urinary metabolites. The protocol is most suited for instances where structure elucidation and metabolite annotation are critical for the downstream biological interpretation of metabolic phenotyping studies. First, a bulk urine pool was desalted using ion-exchange resins enabling large-scale fractionation using precise iterations of analytical scale chromatography. Primary urine fractions were collected and assembled into a “fraction bank” suitable for long-term laboratory storage. Secondary and tertiary fractionations exploited differences in selectivity across a range of reversed-phase chemistries, achieving the purification of metabolites of interest yielding an amount of material suitable for chemical characterisation. To exemplify the application of the systematic workflow in a diverse set of cases, four metabolites with a range of physico-chemical properties were selected and purified from urine and subjected to chemical formula and structure elucidation by respective magnetic resonance mass spectrometry (MRMS) and NMR analyses. Their structures were fully assigned as teterahydropentoxyline, indole-3-acetic-acid-O-glucuronide, p-cresol glucuronide, and pregnanediol-3-glucuronide. Unused effluent was collected, dried and returned to the fraction bank, demonstrating the viability of the system for repeat use in metabolite annotation with a high degree of efficiency.
AU - Whiley,L
AU - Chekmeneva,E
AU - Berry,DJ
AU - Jimenez,B
AU - Yuen,AHY
AU - Salam,A
AU - Hussain,H
AU - Witt,M
AU - Takats,Z
AU - Nicholson,JK
AU - Lewis,MR
DO - 10.1021/acs.analchem.9b00241
EP - 8882
PY - 2019///
SN - 0003-2700
SP - 8873
TI - Systematic isolation and structure elucidation of urinary metabolites optimized for the analytical-scale molecular profiling laboratory
T2 - Analytical Chemistry
UR -
UR -
UR -
VL - 91
ER -