Occurrence of Free and Conjugated Mycotoxins in Aromatic and Medicinal Plants and Dietary Exposure Assessment in the Moroccan Population
Abstract
:1. Introduction
2. Results and Discussion
2.1. Validation
2.2. Natural Occurrence of Mycotoxins
2.2.1. Aflatoxins (AFG1 and AFG2)
2.2.2. Fusarium Toxins (ZEN and HT-2)
2.2.3. Emerging Mycotoxins (ENA1 and ENB)
2.2.4. Alternaria Toxins (AOH and TENT)
2.3. Co-Occurrence of Mycotoxins in AMP
2.4. Conjugated Mycotoxins in AMP
2.5. Dietary Exposure
3. Conclusions
4. Material and Methods
4.1. Chemicals and Reagents
4.2. Plant Sampling
4.3. Mycotoxin Extraction Procedure
4.4. Analysis of Mycotoxins by LC-MS/MS
4.5. Method Validation
4.6. Analysis of Mycotoxin Metabolites by LC-QTOF-MS
4.7. Risk of Dietary Exposure
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Conflicts of Interest
List of Abbreviations
AFB1 | aflatoxin B1 |
AFB2 | aflatoxin B2 |
AFG1 | aflatoxin G1 |
AFG2 | aflatoxin G2 |
AFs | aflatoxins |
AMP | aromatic and medicinal plants |
AOH | alternariol |
BEA | beauvericin |
DON-3-G | DON-3-glucoside |
DON | deoxynivalenol |
PDI | probable daily intake |
ENA | enniatin A |
ENA1 | enniatin A1 |
ENB | enniatin B |
ENB1 | enniatin B1 |
ENs | enniatins |
OTA | ochratoxin A |
TENT | tentoxin |
TDI | tolerable daily intake |
ZEN | zearalenone |
α-ZEL | α-zearalenol |
β-ZEL | β-zearalenol |
ZEN-4-Glc | zearalenone-4-glucoside |
ZEN-4-Sulf | zearalenone-4-sulfate |
α-ZEL-14-Glc | α–ZEL-14-glucoside |
β-ZEL-14-Glc | β–ZEL-14-glucoside |
ZEN-16-Glc | zearalenone-16-glucoside |
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Mycotoxin | Repeatability | Sensitivity | ||
---|---|---|---|---|
R ± SD (%) | ME ± SD a (%) | LOD (ng/g) | LOQ (ng/g) | |
AFB1 | 102 ± 1.4 | 9 ± 1.9 | 4.05 | 13.52 |
AFB2 | 112 ± 5.1 | 9.9 ± 3.8 | 3.30 | 11.00 |
AFG1 | 86 ± 0.5 | 13.5 ± 4.8 | 1.22 | 4.06 |
AFG2 | 97 ± 0.02 | 8.1 ± 2.4 | 2.07 | 6.90 |
OTA | 92 ± 13 | 4.5 ± 0.96 | 0.93 | 3.11 |
BEA | 85 ± 16 | 7.2 ± 0.3 | 0.37 | 1.22 |
ENA | 100 ± 0.1 | 4.5 ± 0.7 | 0.29 | 0.95 |
ENA1 | 125 ± 14 | 3.6 ± 0.5 | 0.29 | 0.95 |
ENB | 103 ± 4.5 | 4.5 ± 0.6 | 0.09 | 1.02 |
ENB1 | 112 ± 13 | 5.4 ± 1.2 | 0.52 | 1.74 |
ZEN | 100 ± 2.5 | 3.6 ± 0.9 | 12.90 | 43.00 |
AOH | 109 ± 0.8 | 4.5 ± 0.7 | 0.20 | 6.67 |
TENT | 87 ± 6 | 5.4 ± 0.8 | 0.28 | 0.92 |
T-2 | 99 ± 0.01 | 7.2 ± 1.2 | 4.75 | 15.84 |
HT-2 | 81 ± 4.3 | 10.3 ± 1.3 | 1.59 | 5.30 |
Analyte | Rt a (min) | Parent Ion Q1 (m/z) | Product Ions Q3 | DP c | CEP c | |||||
---|---|---|---|---|---|---|---|---|---|---|
Q (m/z) b | CE c | CXP c | q (m/z) b | CEc | CXP c | |||||
AFB1 | 7.8 | 313.1 [M+H]+ | 241 | 41 | 4 | 285 | 39 | 4 | 46 | 18 |
AFB2 | 7.7 | 315.1 [M+H]+ | 259 | 39 | 6 | 287 | 33 | 6 | 81 | 18 |
AFG1 | 7.6 | 329.1 [M+H]+ | 311 | 29 | 6 | 243 | 39 | 6 | 76 | 18 |
AFG2 | 7.5 | 331.1 [M+H]+ | 245 | 39 | 6 | 313 | 27 | 6 | 61 | 18 |
OTA | 8.7 | 404.1 [M+H]+ | 102 | 97 | 6 | 239 | 27 | 6 | 55 | 21 |
ENA | 10.3 | 699.4 [M+NH4]+ | 228 | 59 | 16 | 210 | 35 | 14 | 66 | 30 |
ENA1 | 10.1 | 685.4 [M+NH4]+ | 214 | 59 | 10 | 210 | 37 | 8 | 66 | 30 |
ENB | 9.7 | 657.3 [M+NH4]+ | 214 | 59 | 10 | 196 | 39 | 8 | 51 | 29 |
ENB1 | 9.9 | 671.2 [M+NH4]+ | 228 | 57 | 12 | 214 | 61 | 10 | 66 | 29 |
AOH | 8.5 | 259.0 [M+H]+ | 184 | 42 | 3 | 128 | 65 | 3 | 39 | 16 |
TENT | 7.8 | 415.0 [M+H]+ | 256 | 39 | 2 | 312 | 29 | 2 | 55 | 21 |
BEA | 10 | 801.2 [M+NH4]+ | 244 | 39 | 6 | 784 | 27 | 10 | 116 | 33 |
HT-2 | 8.2 | 442.1 [M+NH4]+ | 215 | 19 | 8 | 263 | 19 | 4 | 21 | 22 |
T-2 | 8.4 | 484.1 [M+NH4]+ | 215 | 29 | 4 | 185 | 22 | 4 | 21 | 23 |
ZEN | 8.9 | 319.1 [M+H]+ | 282 | 19 | 4 | 301 | 15 | 10 | 26 | 18 |
AMP | Detected Mycotoxins | Incidence (%) | Mp ± SD (ng/g) | Range (ng/g) |
---|---|---|---|---|
Origanum vulgare (n = 12) | AOH | 9 (75) | 174 ± 96 | 8.6–309 |
ZEN | 3 (25) | 72 ± 29 | 86.6–91 | |
Rosmarinus officinalis (n = 7) | AFG2 | 3(43) | 27.7 ± 2.1 | 26.2–41 |
ZEN | 6 (86) | 45 ± 21 | 33.7–88 | |
AOH | 6(86) | 38 ± 16 | 10.9–53 | |
Myrtus communis (n = 5) | AFG1 | 5 (100) | 6.4 ± 1.3 | 4.9–9 |
AOH | 4 (80) | 40.7 ± 16 | 34.5–72 | |
TENT | 5 (100) | 1.7 ± 2 | 0.7–4.5 | |
Verveine officinale (n = 4) | ENA1 | 1 (25) | 0.3 | LOD–0.3 |
ENB | 1 (25) | 0.1 | LOD–0.1 | |
HT-2 | 1 (25) | 2.9 | LOD-2.0 | |
AOH | 4 (100) | 199.3 ± 70 | 124.6–293 | |
Mentha spicata (n = 2) | ZEN | 2 (100) | 95.7 ± 27 | 76.7–115 |
AOH | 2 (100) | 138.9 ± 1.2 | 138.1–140 | |
Lavandula intermedia (n = 3) | AFG1 | 2 (67) | 7.1 ± 1.5 | 6–8 |
ENB | 3 (100) | 0.2 ± 0.1 | LOD–0.4 | |
AOH | 3 (100) | 53.3 ± 21 | 29.8–69 | |
TENT | 2 (67) | 1.6 ± 0.1 | 1.5–1.6 | |
Artemisia absinthium (n = 2) | AOH | 1 (50) | 2.3 | LOD–2.3 |
Matricaria chamomilla (n = 5) | AOH | 5 (100) | 204.6 ± 102 | 30.9–279 |
Mycotoxin | Incidence (%) | Range Levels (ng/g) | Mp ± SD a (ng/g) | Mt ± SD (ng/g) | PDI (ng/kg b.w./day) [%TDI] | TDI (ng/kg b.w./day) | ||
---|---|---|---|---|---|---|---|---|
LB b | UB c | LB | UB | |||||
AFG1 | 7 (17.5) | 4.9–8.6 | 4.6 ± 1.4 | 1.16 ± 4.7 | 3.97 ± 1.35 | - | - | - |
AFG2 | 3 (7.5) | 26.2–41.1 | 27.7 ± 2.1 | 2.42 ± 8.8 | 7.27 ± 8.8 | - | - | - |
AFs | - | - | - | - | - | - | - | - |
ENA1 | 1 (2.5) | 0.35 | 0.16 ± 0.3 | 0.019 ± 0.05 | 1.3 ± 0.32 | - | - | - |
ENB | 1 (10) | LOD (0.1) d | 0.05 ± 0.1 | 0.02 ± 0.063 | 1.01 ± 0.06 | - | - | - |
ENs | - | - | - | - | - | - | - | - |
ZEN | 11 (27.5) | 33.7–114.7 | 55.5 ± 26 | 18.2 ± 32.7 | 37.8 ± 32.7 | 0.67 [0.56] | 1.39 [0.82] | 250 |
AOH | 34 (85) | 2.3–309.5 | 126.2 ± 40.4 | 99.7 ± 97.8 | 116.2 ± 97 | - | - | - |
TENT | 7 (17.5) | 0.7–4.5 | 1.47 ± 0.8 | 0.29 ± 1.2 | 0.88 ± 0.3 | - | - | - |
HT-2 | 1 (2.5) | LOD–LOQ (2.9) e | 1.47 ± 2.6 | 0.072 ± 2.05 | 5.49 ± 0.5 | 0.003 | 0.203 | - |
T2+HT2 | - | - | - | - | - | 0.003 | 0.203 | 100 |
Samples Code | Mycotoxins Conjugate and Metabolites | Molecular Formula | Precursor Ion Mass (m/z) | Exact Molecular Mass (Da) | Mass Error (ppm) | Purity Score | Area | Retention Time (min) |
---|---|---|---|---|---|---|---|---|
PAM 2.d | β-ZEL | C18 H24 O5 | 365.1616 | 320.1633 | 1.2 | 82.27 | 273957 | 9.1 |
PAM 3.d | C18 H24 O5 | 379.1771 | 320.1623 | 2.3 | 82.95 | 317981 | 9.1 | |
PAM 4.d | C18 H24 O5 | 379.1773 | 320.1623 | 1.3 | 98.15 | 312003 | 9.3 | |
PAM 5.d | C18 H24 O5 | 319.1553 | 320.1615 | 1.62 | 84.71 | 387715 | 9.3 | |
PAM 6.d | C18 H24 O5 | 379.1769 | 320.1630 | 0.9 | 98.23 | 326655 | 9.3 | |
PAM 9.d | C18 H24 O5 | 379.1769 | 320.1630 | 1.5 | 83.70 | 439643 | 9.3 | |
PAM 27.d | C18 H24 O5 | 379.1763 | 320.1627 | 2.4 | 98.06 | 599853 | 9.3 | |
PAM 28.d | C18 H24 O5 | 365.1614 | 320.1631 | −1.3 | 94.51 | 274979 | 9.2 | |
PAM 32.d | C18 H24 O5 | 365.1614 | 320.1633 | 1.04 | 80.27 | 14188 | 9.1 | |
PAM 8.d | α-ZEL | C18 H24 O5 | 319.1553 | 320.1629 | 1.3 | 96.50 | 450446 | 10.4 |
PAM 27.d | C18 H24 O5 | 319.1560 | 320.1636 | 1.4 | 97.38 | 185920 | 10.2 | |
PAM 2.d | ZEN-14-Glc * | C24 H32 O10 | 525.2144 | 480.1992 | 2.5 | 71.67 | 24470 | 6.8 |
PAM 3.d | C24 H32 O10 | 539.2156 | 480.2017 | 2.6 | 69.62 | 20691 | 6.5 | |
PAM 6.d | C24 H32 O10 | 525.1971 | 480.2028 | −2.3 | 73.19 | 9149 | 6.6 | |
PAM 8.d | C24 H34 O10 | 537.2128 | 481.2165 | 2.4 | 69.49 | 29254 | 6.3 | |
PAM 8.d | β-ZEL-14-Glc * | C24 H34 O10 | 481.2058 | 482.2139 | 2.05 | 67.21 | 9817 | 15.9 |
PAM 8.d | ZEN-4-Sulf * | C18 H22 O8 S | 399.2550 | 400.2624 | 1.1 | 71.02 | 32229 | 11.1 |
PAM 27.d | C18 H22 O8 S | 399.2555 | 400.2625 | −1.03 | 75.04 | 10390 | 11.1 | |
PAM 28.d | C18 H22 O8 S | 399.2539 | 400.2608 | −2.2 | 92.11 | 11117 | 11.1 | |
PAM 32.d | C18 H20 O8 S | 397.0968 | 398.1031 | −1.29 | 93.36 | 22950 | 13.5 |
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El Jai, A.; Zinedine, A.; Juan-García, A.; Mañes, J.; Etahiri, S.; Juan, C. Occurrence of Free and Conjugated Mycotoxins in Aromatic and Medicinal Plants and Dietary Exposure Assessment in the Moroccan Population. Toxins 2021, 13, 125. https://doi.org/10.3390/toxins13020125
El Jai A, Zinedine A, Juan-García A, Mañes J, Etahiri S, Juan C. Occurrence of Free and Conjugated Mycotoxins in Aromatic and Medicinal Plants and Dietary Exposure Assessment in the Moroccan Population. Toxins. 2021; 13(2):125. https://doi.org/10.3390/toxins13020125
Chicago/Turabian StyleEl Jai, Aicha, Abdellah Zinedine, Ana Juan-García, Jordi Mañes, Samira Etahiri, and Cristina Juan. 2021. "Occurrence of Free and Conjugated Mycotoxins in Aromatic and Medicinal Plants and Dietary Exposure Assessment in the Moroccan Population" Toxins 13, no. 2: 125. https://doi.org/10.3390/toxins13020125