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Deciphering the human urine matrix: a new approach to simultaneously quantify the main ions and organic compounds by ion chromatography/mass spectrometry (IC-MS)

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Abstract

Analyzing the composition of (human) urine plays a major role in the fields of biology and medicine. Organic molecules (such as urea, creatine) and ions (such as chloride, sulfate) are the major compounds present in urine, the quantification of which allows for the diagnosis of a subject’s health condition. Various analytical methods have been reported for studying urine components and validated on the basis of known and referenced compounds. The present work introduces a new method able to simultaneously determine both major organic molecules and ions contained in urine, by combining ion chromatography using a conductimetric detector with mass spectroscopy. The analysis of organic and ionized compounds (anionic and cationic) was achieved in double injections. For quantification, the standard addition method was used. Human urine samples were pre-treated (diluted and filtered) for IC-CD/MS analysis. The analytes were separated in 35 min. Calibration ranges (0–20 mg.L−1) and correlation coefficients (> 99.3%) as well as detection (LODs < 0.75 mg.L−1) and quantification (LOQs < 2.59 mg.L−1) limits were obtained for the main organic molecules (lactic, hippuric, citric, uric, oxalic acids, urea, creatine, and creatinine) and ions (chloride, sulfate, phosphate, sodium, ammonium, potassium, calcium, and magnesium) contained in urine. The intra- and inter-day accuracies of the analytes consistently ranged from 0.1 to 5.0%, and the precision was within 4.0%. For all analytes, no significant matrix effects were observed, and recoveries ranged from 94.9 to 102.6%. Finally, quantitative results of analytes were obtained from 10 different human urine samples.

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Abbreviations

DP:

Dual pump

ESI:

Electrospray ionization

HPLC:

High-performance liquid chromatography

IC-CD/MS:

Ion chromatography–conductivity detector/mass spectroscopy

ICH:

International Council for Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use

LC:

Liquid chromatography

LOD:

Limit of detection

LOQ:

Limit of quantification

ME:

Matrix effect

RE:

Relative error

RF:

Radio frequency

RSD:

Relative standard deviation

SD:

Standard deviation

UHPLC:

Ultra-high-performance liquid chromatography

a.u. :

Arbitrary unit

B:

Magnetic field (T)

K1 and K2 :

Calculated ratios to validate the human urine presence in a sample (dimensionless)

\({~}^{\mathrm{m}}\!\left/ \!{~}_{\mathrm{z}}\right.\) :

Mass-to-charge ratio

n :

Number of repetitions (dimensionless)

N :

Size of the population

Tr :

Retention time (min)

R 2 :

Correlation coefficient (dimensionless)

x i :

Each value from the population

μ :

Population mean

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Acknowledgements

The authors would like to thank the volunteers for their urine donations.

Funding

This work was supported by the French National Research Agency (proposal HYUREA ANR-19-CE04-0009).

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Authors and Affiliations

  1. Laboratoire de Génie Chimique, Université de Toulouse, CNRS, INPT, UPS, Toulouse, France

    Guillaume Hopsort, Laure Latapie, Karine Groenen Serrano, Karine Loubière & Theodore Tzedakis

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  1. Guillaume Hopsort
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  2. Laure Latapie
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  5. Theodore Tzedakis
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Contributions

Guillaume Hopsort: conceptualization; methodology; validation; formal analysis; investigation; data curation; writing - original draft; and visualization. Laure Latapie: conceptualization; methodology; validation; formal analysis; investigation; resources; writing - review and editing; and funding acquisition. Karine Groenen Serrano: writing - review and editing, and supervision. Karine Loubière: writing - review and editing, and supervision. Théodore Tzedakis: writing - review and editing; supervision; project administration; and funding acquisition.

Corresponding authors

Correspondence to Guillaume Hopsort or Theodore Tzedakis.

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Ethical approval

This is an observational study; no ethical approval was needed for the study. Before gathering any information, we ensured to obtain written approvals from each participant with notice of information and consent form. All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments. This study was conducted with healthy individuals aged 18 and above, who had no previous record of kidney-related ailments. The study excluded those who were currently suffering from a urinary tract infection. Furthermore, female participants were instructed to undertake the test on non-menstrual days.

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Hopsort, G., Latapie, L., Groenen Serrano, K. et al. Deciphering the human urine matrix: a new approach to simultaneously quantify the main ions and organic compounds by ion chromatography/mass spectrometry (IC-MS). Anal Bioanal Chem 415, 5337–5352 (2023). https://doi.org/10.1007/s00216-023-04808-2

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