Antioxidant Activity and UHPLC-MS/MS Characterization of Polyphenol and Nicotine Content in Nicotiana Glauca Leaf Extracts: A Comparative Study of Conventional and Deep Eutectic Solvent Extraction Methods
Abstract
:1. Introduction
2. Materials and Methods
2.1. Plant Material
2.2. Methanol Conventional Extraction (MC)
2.3. Deep Eutectic Solvents (DES) Extraction
2.4. Determination of Total Phenolic
2.5. Determination of Total Flavonoids
2.6. In-Vitro Antioxidant Activity
2.7. UHPLC-MS/MS Methodology
Instrumentation and MS Parameters
2.8. Nicotine Content Determination
3. Results
3.1. Total Phenol Content
3.2. Total Flavonoid Content
3.3. Antioxidant Activity (DPPH Assay) for N. glauca Leaf Extracts
3.4. Identification of Phenols Using UHPLC-MS/MS Analysis
3.4.1. Methanol Conventional Extraction (MC)
3.4.2. Deep Eutectic Extraction (DES)
3.4.3. Identification and Quantification of Nicotine
4. Discussion
5. Conclusions
Author Contributions
Funding
Data Availability Statement
Conflicts of Interest
Appendix A
Appendix B
References
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Extraction Media | mg/mL ± SD (Equivalent to Gallic Acid) |
---|---|
DES 30% | 0.326 ± 0.11 |
DES 70% | 0.300 ± 0.03 |
DES 90% | 0.311 ± 0.02 |
MC | 0.119 ± 0.01 |
Extraction Media | mg/mL ± SD (Equivalent to Rutin) |
---|---|
DES 30% | 0.128 ± 0.03 |
DES 70% | 0.115 ± 0.14 |
DES 90% | 0.928 ± 0.09 |
MC | 0.011 ± 0.01 |
# | Rt [min] | m/z Meas. | M Meas. | Ions | Name | Molecular Formula |
---|---|---|---|---|---|---|
1 | 0.61 | 131.04612 | 132.05339 | [M–H]− | L-Asparagine | C4H8N2O3 |
2 | 0.62 | 114.05616 | 115.06344 | [M–H]− | Proline | C5H9NO2 |
3 | 0.97 | 180.06594 | 181.07322 | [M–H]− | L-Tyrosine | C9H11NO3 |
4 | 1 | 117.01933 | 118.02661 | [M–H]− | Succinic acid | C4H6O4 |
5 | 1.28 | 147.04508 | 148.05236 | [M–H]− | Cinnamic acid | C9H8O2 |
6 | 1.28 | 164.0718 | 165.07908 | [M–H]− | (±)-Phenylalanine | C9H11NO2 |
7 | 2.03 | 203.08226 | 204.08954 | [M–H]− | (±)-Tryptophan | C11H12N2O2 |
8 | 2.9 | 191.05607 | 192.06335 | [M–H]− | Quinic acid | C7H12O6 |
9 | 2.96 | 355.10248 | 354.09521 | [M+H]+ | Chlorogenic acid | C16H18O9 |
10 | 3.05 | 179.03487 | 180.04214 | [M–H]− | Caffeic Acid | C9H8O4 |
11 | 5.07 | 163.03979 | 162.03251 | [M+H]+ | Umbelliferone | C9H6O3 |
12 | 5.12 | 203.08228 | 204.08955 | [M–H]− | (±)-Tryptophan | C11H12N2O2 |
13 | 5.57 | 609.1455 | 610.15278 | [M–H]− | Quercetin 3-rutinoside | C27H30O16 |
14 | 5.61 | 303.05014 | 302.04287 | [M+H]+ | Robinetin | C15H10O7 |
15 | 5.61 | 465.10293 | 464.09566 | [M+H]+ | Hyperoside | C21H20O12 |
16 | 5.62 | 611.16099 | 610.15353 | [M+H]+, [M+Na]+ | Rutin | C27H30O16 |
17 | 6.31 | 179.05592 | 180.0632 | [M–H]− | Starch | C6H12O6 |
18 | 6.37 | 287.0557 | 286.04842 | [M+H]+ | 3,6,2′,4′-Tetrahydroxyflavone | C15H10O6 |
19 | 9.1 | 315.05061 | 316.05788 | [M–H]− | 3-O-Methyl Quercetin | C16H12O7 |
20 | 21.09 | 478.28898 | 477.2817 | [M+H]+ | 1-Hydroxy-2-(9Z,12Z-octadecadienoyl)-sn-glycero-3-phosphoethanolamine (NMR) | C23H44NO7P |
21 | 22.31 | 478.28916 | 477.28189 | [M+H]+ | 1-(9Z,12Z-Octadecadienoyl)-2-hydroxy-sn-glycero-3-phosphoethanolamine (NMR) | C23H44NO7P |
22 | 22.49 | 471.35137 | 470.34409 | [M+H]+ | 18-Beta-glycyrrhetinic acid | C30H46O4 |
23 | 28.62 | 221.15517 | 222.16244 | [M–H]− | Histamine | C10H18N6 |
# | Rt [min] | m/z Meas. | M Meas. | Ions | Name | Molecular Formula |
---|---|---|---|---|---|---|
1 | 1.65 | 133.00998 | 134.0174 | [M–H]−, [M–H H2O]− | Malic acid | C4H6O5 |
2 | 1.91 | 59.01128 | 60.01856 | [M–H]− | Acetic acid | C2H4O2 |
3 | 2.39 | 350.14397 | 351.15125 | [M–H]− | (E)-Ribosylzeatin | C15H21N5O5 |
4 | 2.96 | 117.01473 | 118.022 | [M–H]− | Succinic acid | C4H6O4 |
5 | 3.32 | 147.04052 | 148.0478 | [M–H]− | Cinnamic acid | C9H8O2 |
6 | 3.35 | 87.04212 | 88.04939 | [M–H]− | 2-Methylpropanoic acid | C4H8O2 |
7 | 3.36 | 164.06681 | 165.07409 | [M–H]− | (±)-Phenylalanine | C9H11NO2 |
8 | 3.69 | 163.038920 | 162.031640 | [M+H]+ | Umbelliferone | C9H6O3 |
9 | 3.7 | 355.102500 | 354.095190 | [M+H]+, [M+K]+, [M+Na]+ | Chlorogenic acid | C16H18O9 |
10 | 3.79 | 173.04114 | 174.04841 | [M–H]− | Shikimic acid | C7H10O5 |
11 | 3.8 | 199.057710 | 198.050440 | [M+H]+ | Syringic acid | C9H10O5 |
12 | 4.6 | 193.049450 | 192.042170 | [M+H]+ | Scopoletin | C10H8O4 |
13 | 4.76 | 465.102820 | 464.095540 | [M+H]+ | Hyperoside | C21H20O12 |
14 | 4.77 | 303.050010 | 302.042740 | [M+H]+ | Robietin | C15H10O7 |
15 | 4.77 | 611.160240 | 610.152970 | [M+H]+ | Luteolin-7,3′-di-O-glucoside | C27H30O16 |
16 | 4.91 | 609.1281 | 610.13537 | [M–H]− | Prodelphinidin B3 | C30H26O14 |
17 | 5 | 221.05958 | 222.06686 | [M–H]− | Flavone | C15H10O2 |
18 | 5.15 | 595.165470 | 594.158150 | [M+H]+, [M+Na]+ | Saponarin | C27H30O15 |
19 | 5.74 | 195.065060 | 194.057780 | [M+H]+ | 3-Hydroxy-4-methoxycinnamic acid (isoferulic acid) | C10H10O4 |
20 | 7.39 | 163.039120 | 162.031850 | [M+H]+ | (4 or 7) Hydroxy-Coumarin Plus Hydrate | C9H6O3 |
21 | 7.41 | 177.054460 | 176.047190 | [M+H]+ | 4-Methylumbelliferone | C10H8O3 |
22 | 8.54 | 495.125630 | 494.118360 | [M+H]+ | Salvianolic acid A | C26H22O10 |
23 | 11.6 | 249.148000 | 248.140730 | [M+H]+ | 3-Oxocostusic acid | C15H20O3 |
Rt [min] | m/z Meas. | M Meas. | Ions | Name | Molecular Formula |
---|---|---|---|---|---|
2.89 | 163.12293 | 324.2313 | [M+H+H]2+ | Nicotine | C10H14N2 |
Sample | MC Extract | DES Extract |
---|---|---|
Area of Nicotine in Sample | 2,526,713 | 4,192,477 |
Concentration of Nicotine | 635.07 ppm | 1194.91 ppm |
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Issa, R.; Al-Akayleh, F.; Alnsour, L.; Al-Sammarraie, T.R.; Omari, K.W.; Awwad, S.H. Antioxidant Activity and UHPLC-MS/MS Characterization of Polyphenol and Nicotine Content in Nicotiana Glauca Leaf Extracts: A Comparative Study of Conventional and Deep Eutectic Solvent Extraction Methods. Plants 2024, 13, 2240. https://doi.org/10.3390/plants13162240
Issa R, Al-Akayleh F, Alnsour L, Al-Sammarraie TR, Omari KW, Awwad SH. Antioxidant Activity and UHPLC-MS/MS Characterization of Polyphenol and Nicotine Content in Nicotiana Glauca Leaf Extracts: A Comparative Study of Conventional and Deep Eutectic Solvent Extraction Methods. Plants. 2024; 13(16):2240. https://doi.org/10.3390/plants13162240
Chicago/Turabian StyleIssa, Reem, Faisal Al-Akayleh, Lilian Alnsour, Tabarak R. Al-Sammarraie, Khaled W. Omari, and Shady H. Awwad. 2024. "Antioxidant Activity and UHPLC-MS/MS Characterization of Polyphenol and Nicotine Content in Nicotiana Glauca Leaf Extracts: A Comparative Study of Conventional and Deep Eutectic Solvent Extraction Methods" Plants 13, no. 16: 2240. https://doi.org/10.3390/plants13162240