Antiproliferative Activity of Neem Leaf Extracts Obtained by a Sequential Pressurized Liquid Extraction
Abstract
:1. Introduction
2. Results and Discussion
2.1. Pressurized Liquid Extraction Process
2.2. Liquid Chromatography Analysis
2.3. Cytotoxicity Evaluation of Neem Leaves Extracts
3. Materials and Methods
3.1. Neem Samples
3.2. Pressurized Liquid Extraction Process
3.3. HPLC-PDA-ESI-MS Analysis
3.4. Cytotoxicity Assays
3.5. Statistical Analysis
4. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Neem Leaves (20 g) | One-Step Extraction (g) | Three-Step Extraction (g) |
---|---|---|
Hexane (SH) | ____ | 0.07 ± 0.01 b |
Ethyl acetate (SEA) | ____ | 0.06 ± 0.01 b |
Etanol 80% (SE) | ____ | 1.50 ± 0.12 a |
Etanol 80% (EE) | 1.58 ± 0.26 a | _____ |
Extract | Peak | tR (min) | Area | Compound | Observed Ions (m/z) |
---|---|---|---|---|---|
SH | 1 | 8.42 | 45724118 | Nimbandiol | 371, 401, 421, 425, 441, 444, 457 [M + H]+, 474 [M + H2O]+ |
SEA | 8.46 | 38701542 | |||
SE | 8.56 | 20010994 | |||
EE | 8.40 | 25006988 | |||
SH | 2 | 11.29 | 29873977 | 6-Deacetylnimbin | 389, 453, 467, 499 [MH]+, 516 [M + H2O]+ |
SEA | 11.42 | 22241890 | |||
SE | 11.32 | 2818278 | |||
EE | 11.46 | 14511268 | |||
SH | 3 | 12.77 | 81159631 | 2,3-Dihydronimbolide | 178, 315, 426, 433, 441, 450, 469 [MH]+, 486 [M + H2O]+ |
SEA | 12.75 | 37973099 | |||
SE | 12.76 | 28340365 | |||
EE | 12.84 | 36750310 | |||
SH | 4 | 13.91 | 13066767 | Rutin | 266, 480, 546, 558, 611 [M + H]+, 628 [M + H2O]+ |
SEA | 14.02 | 17659833 | |||
SE | 13.95 | 14990955 | |||
EE | 14.02 | 15674396 | |||
SH | 5 | 15.56 | 36960991 | Nimonol | 274, 293, 353, 421, 439, 453 [M + H]+, 470 [M + H2O]+ |
SEA | 15.59 | 31070403 | |||
SE | 15.67 | 15495619 | |||
EE | 15.52 | 20995715 | |||
SH | 6 | 16.39 | 70699349 | Nimbolide | 277, 435, 435, 467 [M + H]+,484 [M + H2O]+ |
SEA | 16.54 | 86571238 | |||
SE | 16.42 | 50917437 | |||
EE | 16.45 | 57586856 | |||
SH | 6 | 16.39 | 70699349 | 3-Deacetylsalannin | 555 [M + H]+, 572 [M + H2O]+ |
SEA | 16.54 | 86571238 | |||
SE | 16.42 | 50917437 | |||
EE | 16.45 | 57586856 | |||
SH | 7 | 18.22 | 32928497 | 6-Deacetylnimbinene | 363, 393, 409, 441 [M + H]+, 458 [M + H2O]+ |
SEA | 18.18 | 37398457 | |||
SE | 18.12 | 15714996 | |||
EE | 18.32 | 21712675 | |||
SH | 8 | 19.88 | 15628192 | Nimbanal | 221, 265, 339, 345, 405, 428, 451,453, 455, 471, 482,493, 511 [M + H]+, 528 [M + H2O]+ |
SEA | 19.93 | 23156736 | |||
SE | 19.87 | 6245251 | |||
EE | 19.74 | 11010022 | |||
SH | 9 | 24.96 | 14175318 | Salannin | 199, 230, 278, 319, 378, 481, 515, 571, 597 [M + H]+, 614 [M + H2O]+ |
SEA | 24.86 | 12952957 | |||
SE | 24.87 | 5526287 | |||
EE | 24.93 | 2517812 | |||
SH | 10 | 25.49 | 17132995 | Gedunin | 184, 259, 287, 344, 372, 405, 425, 451, 483 [M + H]+, 500 [M + H2O]+ |
SEA | 25.67 | 13462271 | |||
SE | 25.75 | 6429235 | |||
EE | 25.70 | 7359673 |
Lines | Extract (µg/mL) | Control (µg/mL) | |||
---|---|---|---|---|---|
SH | SEA | SE | EE | Ellipticine | |
MCF-7 | 188.8 ± 6.4 a | 82.3 ± 4.3 b | >250 c | >250 c | 0.9 ± 0.1 d |
NCI-H460 | 224.4 ± 14.4 a | 60.6 ± 4.3 b | >250 c | >250 c | 1.0 ± 0.1 d |
HeLa | 203.9 ± 13.6 a | 48.8 ± 4.3 b | >250 c | >250 c | 1.9 ± 0.1 d |
HepG2 | 115.5 ± 14.4 a | 52.3 ± 4.8 b | >250 c | >250 c | 1.1 ± 0.2 d |
PLP2 | >250 a | 201.3 ± 17.0 b | >250 a | >250 a | 3.2 ± 0.7 c |
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Santos, K.S.; Barbosa, A.M.; Freitas, V.; Muniz, A.V.C.S.; Mendonça, M.C.; Calhelha, R.C.; Ferreira, I.C.F.R.; Franceschi, E.; Padilha, F.F.; Oliveira, M.B.P.P.; et al. Antiproliferative Activity of Neem Leaf Extracts Obtained by a Sequential Pressurized Liquid Extraction. Pharmaceuticals 2018, 11, 76. https://doi.org/10.3390/ph11030076
Santos KS, Barbosa AM, Freitas V, Muniz AVCS, Mendonça MC, Calhelha RC, Ferreira ICFR, Franceschi E, Padilha FF, Oliveira MBPP, et al. Antiproliferative Activity of Neem Leaf Extracts Obtained by a Sequential Pressurized Liquid Extraction. Pharmaceuticals. 2018; 11(3):76. https://doi.org/10.3390/ph11030076
Chicago/Turabian StyleSantos, Klebson S., Andriele M. Barbosa, Victor Freitas, Ana Veruska C. S. Muniz, Marcelo C. Mendonça, Ricardo C. Calhelha, Isabel C. F. R. Ferreira, Elton Franceschi, Francine F. Padilha, Maria Beatriz P. P. Oliveira, and et al. 2018. "Antiproliferative Activity of Neem Leaf Extracts Obtained by a Sequential Pressurized Liquid Extraction" Pharmaceuticals 11, no. 3: 76. https://doi.org/10.3390/ph11030076