Identifying Key Markers for Monofloral (Eucalyptus, Rosemary, and Orange Blossom) and Multifloral Honey Differentiation in the Spanish Market by UHPLC-Q-Orbitrap-High-Resolution Mass Spectrometry Fingerprinting and Chemometrics
Abstract
:1. Introduction
2. Materials and Methods
2.1. Honey Samples
2.2. Sample Preparation and Extraction
2.3. Honey Fingerprinting by UHPLC-Q-Orbitrap-HRMS Untargeted Analysis
2.4. Processing Honey Omics Data: Designed Untargeted Metabolomics Workflow Using Compound DiscovererTM Software
2.5. Multivariate Data Analysis
2.6. Workflow to Identify Honey Markers
- VIP score cut-off > 1.00;
- Benjamini–Hochberg-adjusted p-values < 0.05;
- FC cut-off > 1.10;
- Two characteristic ions (precursor and at least one fragment ion) with mass accuracy ≤ 5 ppm;
- Fully overlapping chromatographic peaks of the ions at the metabolite retention time (RT ± 0.10 min);
- Study of the potential logical occurrence of the proposed marker in the study matrix based on the relevant published literature and food databases (if available).
3. Results and Discussion
3.1. Unsupervised Multivariate Data Analysis
3.1.1. Overview of Monofloral (Eucalyptus, Rosemary, and Orange Blossom) Honey
3.1.2. Overview of Multifloral vs. Monofloral Honey
3.2. Supervised Multivariate Data Analysis
3.2.1. Experiment 1: OPLS-DA to Discriminate Monofloral Samples (Eucalyptus, Rosemary, and Orange Blossom Honey)
3.2.2. Experiment 2: OPLS-DA to Discriminate Multifloral and Monofloral Honey
3.3. Identification of Honey Markers
3.3.1. Markers to Discriminate Monofloral Samples (Eucalyptus, Rosemary, and Orange Blossom Honey)
3.3.2. Markers to Discriminate Multifloral vs. Monofloral Honey
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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No. | RT (min) | Marker Name | Molecular Formula | VIP Score | p-Value 1 | Log2(FC) 2 |
---|---|---|---|---|---|---|
Rosemary vs. eucalyptus honey | ||||||
1 | 1.039 | L-proline | C5H9NO2 | 2.39 ± 1.06 | 1.2 × 10−8 | −0.37 |
2 | 1.083 | L-pyroglutamic acid | C5H7NO3 | 1.17 ± 0.74 | 8.9 × 10−9 | −0.85 |
3 | 1.133 | N-(1-deoxy-1-fructosyl)phenylalanine | C15H21NO7 | 2.45 ± 0.71 | 2.9 × 10−3 | −1.12 |
4 | 1.207 | L-phenylalanine | C9H12NO2 | 3.50 ± 1.88 | 2.9 × 10−3 | 0.27 |
Rosemary vs. orange blossom honey | ||||||
5 | 1.039 | L-proline | C5H9NO2 | 2.25 ± 1.36 | 2.8 × 10−2 | −0.15 |
6 | 1.062 | Raffinose | C18H32016 | 1.05 ± 0.55 | 9.4 × 10−7 | 0.34 |
7 | 1.207 | L-phenylalanine | C9H12NO2 | 5.55 ± 2.08 | 5.0 × 10−3 | 0.27 |
Orange blossom vs. eucalyptus honey | ||||||
8 | 1.039 | L-proline | C5H9NO2 | 1.36 ± 0.66 | 3.7 × 10−3 | −0.22 |
9 | 1.040 | N-(1-deoxy-1-fructosyl)proline | C11H19NO7 | 1.97 ± 0.30 | 8.9 × 10−9 | −2.02 |
10 | 1.083 | L-pyroglutamic acid | C5H7NO3 | 1.19 ± 0.80 | 1.8 × 10−7 | −0.59 |
11 | 1.089 | N-(1-deoxy-1-fructosyl)isoleucine | C12H23NO7 | 1.95 ± 0.52 | 8.7 × 10−9 | −2.10 |
Multifloral vs. monofloral honey | ||||||
12 | 1.035 | Trigonelline | C7H7NO2 | 1.45 ± 0.34 | 1.4 × 10−3 | 0.97 |
13 | 1.089 | N-(1-deoxy-1-fructosyl)isoleucine | C12H23NO7 | 2.49 ± 1.40 | 6.3 × 10−5 | 1.70 |
14 | 1.102 | L-isoleucine | C6H13NO2 | 1.42 ± 0.57 | 3.9 × 10−4 | 0.83 |
15 | 1.133 | N-(1-deoxy-1-fructosyl)phenylalanine | C15H21NO7 | 4.88 ± 1.12 | 7.3 × 10−3 | 1.62 |
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Rivera-Pérez, A.; Navarro-Herrera, A.M.; Garrido Frenich, A. Identifying Key Markers for Monofloral (Eucalyptus, Rosemary, and Orange Blossom) and Multifloral Honey Differentiation in the Spanish Market by UHPLC-Q-Orbitrap-High-Resolution Mass Spectrometry Fingerprinting and Chemometrics. Foods 2024, 13, 2755. https://doi.org/10.3390/foods13172755
Rivera-Pérez A, Navarro-Herrera AM, Garrido Frenich A. Identifying Key Markers for Monofloral (Eucalyptus, Rosemary, and Orange Blossom) and Multifloral Honey Differentiation in the Spanish Market by UHPLC-Q-Orbitrap-High-Resolution Mass Spectrometry Fingerprinting and Chemometrics. Foods. 2024; 13(17):2755. https://doi.org/10.3390/foods13172755
Chicago/Turabian StyleRivera-Pérez, Araceli, Alba María Navarro-Herrera, and Antonia Garrido Frenich. 2024. "Identifying Key Markers for Monofloral (Eucalyptus, Rosemary, and Orange Blossom) and Multifloral Honey Differentiation in the Spanish Market by UHPLC-Q-Orbitrap-High-Resolution Mass Spectrometry Fingerprinting and Chemometrics" Foods 13, no. 17: 2755. https://doi.org/10.3390/foods13172755