Validated Ultra-High-Performance Liquid Chromatography Hybrid High-Resolution Mass Spectrometry and Laser-Assisted Rapid Evaporative Ionization Mass Spectrometry for Salivary Metabolomics.
2020
Whereas urine and
blood are typically targeted in clinical research,
saliva represents an interesting alternative because its intrinsic
metabolome is chemically diverse and reflective for various biological
processes. Moreover, saliva collection is easy and noninvasive, which
is especially valuable for cohorts in which sample collection is challenging,
for example, infants and children. With this rationale, we established
a validated ultra-high-performance liquid chromatography high-resolution
mass spectrometry (UHPLC-HRMS) method for salivary metabolic profiling
and fingerprinting. Hereby, 450 μL of saliva was centrifuged
and passed over a 0.45-μm polyamide membrane filter, after which
the extract was subjected to chromatographic analysis (HSS T3 column)
and Q-Exactive Orbitrap-MS. For the majority of the profiled metabolites,
good linearity (R2 ≥ 0.99) and
precision (coefficient of variance ≤ 15%) was achieved. The
fingerprinting performance was evaluated based on the complete metabolome
(11 385 components), whereby 76.8% was found compliant with
the criteria for precision (coefficient of variance ≤ 30%)
and 82.7% with linearity (R2 ≥
0.99). In addition, the method was proven fit-for-purpose for a cohort
of 140 adolescents (6–16 years, stratified according to weight),
yielding relevant profiles (45 obesity-related metabolites) and discriminative
fingerprints (Q2 of 0.784 for supervised
discriminant analysis). Alternatively, laser-assisted rapid evaporative
ionization mass spectrometry (LA-REIMS) was established for rapid
fingerprinting of saliva, thereby using a Nd:YAG laser and Xevo G2-XS
QToF-MS. With an acquisition time of 0.5 min per sample, LA-REIMS
offers unique opportunities for point-of-care applications. In conclusion,
this work presents a platform of UHPLC-HRMS and LA-REIMS, complementing
each other to perform salivary metabolomics.
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