The Identification of Metabolites and Effects of Albendazole in Alfalfa (Medicago sativa)
Abstract
:1. Introduction
2. Results and Discussion
2.1. Effect of ABZ on Germination of Alfalfa Seeds
2.2. Effect of ABZ on Proline Accumulation in Alfalfa Regenerants
2.3. ABZ Biotransformation in Alfalfa Regenerants
3. Materials and Methods
3.1. Chemicals
3.2. Plant Material and Their Cultivation
3.3. ABZ Phytotoxicity
3.4. ABZ Uptake and Biotransformation
3.5. UHPLC-MS/MS Conditions
3.6. Quantification of ABZ, ABZSO and ABZSO2
3.7. Statistical Analysis
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
ABZ | albendazole |
ABZSO | albendazole sulphoxide |
ABZSO2 | albendazole sulphone |
DME | drug-metabolizing enzyme |
DMSO | dimethyl sulfoxide |
DW | weight of dry tissue |
ESI | electrospray ionization |
M | metabolite |
MBZ | mebendazole |
MS | mass spectrometry |
N/A | not available |
UHPLC | ultrahigh-performance liquid chromatography |
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tR [min] | Theoretical m/z Values of [M+H]+ Ions | Molecular Formula | Description of Metabolite Formation | Product Ions of [M+H]+, m/z | Metabolite Designation | |
---|---|---|---|---|---|---|
Phase I | Phase II | |||||
1.96 | 444.1435 | C18H25N3O8S | S-oxidation | N-glucosidation | 282, 240, 208, 191, 159 | M1 |
2.04 | 460.1384 | C18H25N3O9S | 2*S-oxidation, hydrolysis, hydroxylation | Glucosidation, O-acetylation | 240, 198 | M31 |
2.09 | 460.1384 | C18H25N3O9S | 2*S-oxidation, hydrolysis, hydroxylation | Glucosidation, O-acetylation | 240 | M2 |
2.18 | 444.1435 | C18H25N3O8S | S-oxidation | N-glucosidation | 282, 240, 208, 191, 159 | M30 |
2.24 | 444.1435 | C18H25N3O8S | S-oxidation | N-glucosidation | 282, 240, 208, 191, 159 | M29 |
2.25 | 460.1384 | C18H25N3O9S | 2*S-oxidation, hydrolysis, hydroxylation | Glucosidation, O-acetylation | 240, 198 | M4 |
2.30 | 460.1384 | C18H25N3O9S | 2*S-oxidation, hydrolysis, hydroxylation | Glucosidation, O-acetylation | 240 | M33 |
2.44 | 314.0805 | C12H15N3O5S | 2*S-oxidation, hydroxylation | − | 238, 159 | M6 |
2.46 | 444.1435 | C18H25N3O8S | S-oxidation | N-glucosidation | 282, 240, 208 | M5 |
2.61 | 460.1384 | C18H25N3O9S | 2*S-oxidation, hydrolysis, hydroxylation | Glucosidation, O-acetylation | 240, 198 | M32 |
2.86 | 460.1384 | C18H25N3O9S | 2*S-oxidation | N-glucosidation | 298, 266, 224, 191, 159 | M7 |
2.99 | 460.1384 | C18H25N3O9S | 2*S-oxidation, hydrolysis, hydroxylation | Glucosidation, O-acetylation | 240, 133 | M8 |
3.22 | 282.0907 | C12H15N3O3S | S-oxidation | − | 240, 208, 191,159 | M10 |
3.36 | 460.1384 | C18H25N3O9S | 2*S-oxidation | N-glucosidation | 298, 266, 224, 191, 159 | M9 |
3.56 | 370.1431 | C16H23N3O5S | Hydrolysis | N-glucosidation | 208, 166 | M11 |
4.50 | 370.1431 | C16H23N3O5S | Hydrolysis | N-glucosidation | 208, 166 | M25 |
4.74 | 428.1486 | C18H25N3O7S | +O, hydrolysis | Glucosidation, O-acetylation | 208 | M16 |
5.00 | 298.0856 | C12H15N3O4S | 2*S-oxidation | − | 266, 224, 159 | M14 |
5.54 | 428.1486 | C18H25N3O7S | − | N-glucosidation | 266, 234 | M20 |
6.06 | 428.1486 | C18H25N3O7S | +O, hydrolysis, | Glucosidation, O-acetylation | 208, 250 | M21 |
6.56 | 428.1486 | C18H25N3O7S | − | N-glucosidation | 266, 234 | M22 |
7.23 | 266.0958 | C12H15N3O2S | − | − | 234 | ABZ |
M. sativa Roots | M. sativa Leaves | ||||||||
---|---|---|---|---|---|---|---|---|---|
ABZ (µM) | ABZ (µM) | ||||||||
0.01 | 0.1 | 1.0 | 10 | 0.01 | 0.1 | 1.0 | 10 | ||
M1 | − | − | + | + | + | + | + | ||
M30 | − | − | − | + | − | − | − | − | |
M29 | − | − | − | + | − | − | − | + | |
M31 | − | − | − | + | − | − | − | + | |
M4 | − | − | − | + | − | − | − | + | |
M32 | − | − | − | + | − | − | − | − | |
M7 | − | − | − | + | − | − | − | + | |
M9 | − | − | − | + | − | − | − | + | |
M25 | − | − | − | + | − | − | − | − | |
M21 | − | − | − | + | − | − | − | − | |
M5 | − | − | − | + | − | − | − | − | |
M2 | − | − | − | + | − | − | − | + | |
M33 | − | − | − | + | − | − | − | − | |
M8 | − | + | + | + | − | − | − | + | |
M11 | − | − | − | + | − | + | + | + | |
M16 | − | − | − | + | − | − | − | − | |
M20 | − | − | − | + | − | − | − | − | |
M22 | − | − | − | + | − | − | − | − | |
M10 | + | + | + | + | + | + | + | + | |
M14 | + | + | + | + | + | + | + | + | |
M6 | − | − | − | + | − | − | − | + |
Exposition of Alfalfa to ABZ (µM) | |||||
---|---|---|---|---|---|
0.01 | 0.1 | 1.0 | 10 | ||
Roots | ABZ | 0.008 ± 0.002 | 0.019 ± 0.01 | 0.015 ± 0.005 | 0.162 ± 0.011 |
ABZSO | 1.9 ± 1.2 | 4.1 ± 1.1 | 113.6 ± 23.7 | 1073 ± 209 | |
ABZSO2 | 0.25 ± 0.12 | 0.54 ± 0.08 | 6.59 ± 1.37 | 42.7 ± 6.21 | |
Leaves | ABZ | 0.005 ± 0.002 | 0.005 ± 0.003 | 0.012 ± 0.005 | 0.006 ± 0.001 |
ABZSO | 0.444 ± 0.134 | 0.44 ± 0.19 | 3.25 ± 0.25 | 61.6 ± 2.19 | |
ABZSO2 | 0.094 ± 0.016 | 0.09 ± 0.02 | 0.64 ± 0.24 | 6.21 ± 0.10 |
Standard Curve (y = ax + b) | r2 | LOD (ng/mL) | LOQ (ng/mL) | |
---|---|---|---|---|
ABZ | Y = (16.9401)X + (0.892919) | 0.9959 | 0.0008 | 0.0028 |
ABZSO | Y = (0.731182)X + (0.0388037) | 0.9972 | 0.0104 | 0.0350 |
ABZSO2 | Y = (0.463911)X + (−0.00401502) | 0.9934 | 0.0110 | 0.0367 |
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Raisová Stuchlíková, L.; Navrátilová, M.; Langhansová, L.; Moťková, K.; Podlipná, R.; Szotáková, B.; Skálová, L. The Identification of Metabolites and Effects of Albendazole in Alfalfa (Medicago sativa). Int. J. Mol. Sci. 2020, 21, 5943. https://doi.org/10.3390/ijms21165943
Raisová Stuchlíková L, Navrátilová M, Langhansová L, Moťková K, Podlipná R, Szotáková B, Skálová L. The Identification of Metabolites and Effects of Albendazole in Alfalfa (Medicago sativa). International Journal of Molecular Sciences. 2020; 21(16):5943. https://doi.org/10.3390/ijms21165943
Chicago/Turabian StyleRaisová Stuchlíková, Lucie, Martina Navrátilová, Lenka Langhansová, Kateřina Moťková, Radka Podlipná, Barbora Szotáková, and Lenka Skálová. 2020. "The Identification of Metabolites and Effects of Albendazole in Alfalfa (Medicago sativa)" International Journal of Molecular Sciences 21, no. 16: 5943. https://doi.org/10.3390/ijms21165943