Phytochemical Analysis of Pinus cembra Heartwood—UHPLC-DAD-ESI-MSn with Focus on Flavonoids, Stilbenes, Bibenzyls and Improved HPLC Separation
Abstract
:1. Introduction
2. Results and Discussion
2.1. UHPLC-DAD-ESI-MSn Analysis
2.2. Isolation of Major Compounds
2.3. Chromatographic Optimization for HPLC Separation of Isolated Compounds
2.4. Application to Extract Sample
3. Materials and Methods
3.1. Plant Material
3.2. Solvents and Reference Substances
3.3. Extraction and Fractionation
3.4. UHPLC-DAD-ESI-MSn
3.5. HPLC-DAD
3.6. Semipreparative HPLC
3.7. NMR
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Nr. | Rt [min] | Mol. Mass | [M-H]− | [M+H]+ | UVmax [nm] | Group | Substance |
---|---|---|---|---|---|---|---|
1 | 4.50 | 288 | 287 | 289 | 228, 291, 330 sh | Flavanonol | Aromadendrin * |
2 | 6.82 | 270 | 269 | 271 | 268, 335 | Flavone | Apigenin a* |
3 | 6.83 | 272 | 271 | 273 | 290, 330 sh | Flavanone | Naringenin * |
4 | 7.84 | 242 | 241 | 243 | 217, 237 sh, 307, 318 | Stilbene | Pinostilbene * |
5 | 8.50 | 214 | 213 | 215 | 201, 276 | Bibenzyl | Dihydropinosylvin [5,6,16] |
6 | 8.56 | 212 | 211 | 213 | 210, 228 sh, 300, 308 | Stilbene | Pinosylvin [5,6,16] |
7 | 9.60 | 254 | 253 | 255 | 269, 313 | Flavone | Chrysin a [4,5,6,16] |
8 | 9.91 | 256 | 255 | 257 | 290, 330 sh | Flavanone | Pinocembrin b [4,5,6,10,11,16] |
9 | 11.36 | 228 | 227 | 229 | 205, 276 | Bibenzyl | Dihydropinosylvin monomethyl ether b [5,9] |
10 | 11.36 | 226 | 225 | 227 | 211, 227 sh, 300, 308 | Stilbene | Pinosylvin monomethyl ether b [4,5,9,15] |
11 | 13.05 | 270 | 269 | 271 | 289, 330 sh | Flavanone | Pinostrobin a [5,6] |
12 | 13.05 | 268 | 267 | 269 | 268, 310 | Flavone | Tectochrysin b [4,5,6] |
13 | 14.77 | 240 | - 1 | 241 | 206, 229 sh, 300, 307 | Stilbene | Pinosylvin dimethyl ether * |
14 | 18.24 | 278 | 277 | 279 | 193 | Fatty acid | Linolenic acid [13] |
15 | 18.36 | 302 | 301 | 303 | 243 | Diterpenic acid | Abietic acid a [13,15] |
16 | 19.29 | 280 | 279 | - | 194 | Fatty acid | Linoleic acid a [13] |
17 | 20.78 | 282 | 281 | - | 195 | Fatty acid | Oleic acid a [13] |
CC Fraction | Isolated Substance (Quantity in mg) |
---|---|
F17–18 | 9 (1.5) |
F19 | 9 (49.3) |
F20–22 | 9 + 10 1 (22.9), 12 (2.9) |
F23–25 | 8 (7.6), 10 (12.2), 12 (1.1) |
F28 | 7 (9.5) |
HPLC Column | Mobile Phase B 1 | Substance | Rt [min] | Rs 2 |
---|---|---|---|---|
EclC18 | 20–100% | 9 + 10 | 11.460 | - 3 |
30% | 9 | 51.243 | ||
10 | 55.020 | 1.68 | ||
KtxC18 | 20–100% | 9 + 10 | 9.893 | - |
30% | 9 | 24.787 | ||
10 | 27.267 | 1.57 | ||
KtxPFP | 20–100% | 9 | 9.337 | |
10 | 9.607 | 1.90 | ||
40% | 9 | 5.620 | ||
10 | 6.237 | 2.55 |
HPLC Column | Substance | Rt [min] | Rs 1 |
---|---|---|---|
EclC18 | 7 | 9.673 | |
8 | 9.980 | 2.44 | |
9 + 10 | 11.433 | - 2 | |
11 + 12 | 13.127 | - | |
KtxC18 | 7 + 8 | 8.567 | - |
9 + 10 | 9.867 | - | |
11 | 11.303 | ||
12 | 11.487 | 0.88 | |
KtxPFP | 7 + 8 | 8.480 | - |
9 | 9.317 | ||
10 | 9.593 | 1.90 | |
11 | 10.810 | ||
12 | 10.967 | 1.25 |
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Alperth, F.; Schneebauer, A.; Kunert, O.; Bucar, F. Phytochemical Analysis of Pinus cembra Heartwood—UHPLC-DAD-ESI-MSn with Focus on Flavonoids, Stilbenes, Bibenzyls and Improved HPLC Separation. Plants 2023, 12, 3388. https://doi.org/10.3390/plants12193388
Alperth F, Schneebauer A, Kunert O, Bucar F. Phytochemical Analysis of Pinus cembra Heartwood—UHPLC-DAD-ESI-MSn with Focus on Flavonoids, Stilbenes, Bibenzyls and Improved HPLC Separation. Plants. 2023; 12(19):3388. https://doi.org/10.3390/plants12193388
Chicago/Turabian StyleAlperth, Fabian, Anna Schneebauer, Olaf Kunert, and Franz Bucar. 2023. "Phytochemical Analysis of Pinus cembra Heartwood—UHPLC-DAD-ESI-MSn with Focus on Flavonoids, Stilbenes, Bibenzyls and Improved HPLC Separation" Plants 12, no. 19: 3388. https://doi.org/10.3390/plants12193388