Background
Methods
Chemicals and reagents
Marula stems, leaves and fruits collection and extraction
Marula stems collection and extraction
Marula oil production
Defatting of Marula stem extract
Concentration of actives of Marula stem extract
Defatting and concentration of actives of Marula stem extract
Determination of anti-elastase activity
Determination of anti-collagenase activity
UPLC-q-TOF-ms
MS conditions
Acquisition
Chemical profiling
Statistical procedures
Plant part | Elastase assay (% Inhibition ±SD) | Collagenase assay (% Inhibition ±SD) |
---|---|---|
Stems | 88.07 ± 2.26 | 76.92 ± 8.92 |
Leaves | 53.60 ± 1.06 | 21.23 ± 7.74 |
Fruits | 26.79 ± 6.61 | 25.89 ± 8.69 |
Controls | ||
10% MeOH | 1.45 ± 1.50 | 1.32 ± 0.73 |
EDTA | 79.88 ± 9.10 | |
Elafin | 93.09 ± 4.10 |
Extraction solvent used for the stems | Extraction yield (calculated from dry plant material) as w/w % | Elastase assay (% Inhibition ± SD) | Collagenase assay (% Inhibition ± SD) |
---|---|---|---|
Acetone | 3.36 | ** 91.25 ± 0.11 | 91.17 ± 3.59 |
Methanol: DCM (1:1) | 5.39 | ** 89.74 ± 0.32 | 93.75 ± 3.32 |
Ethanol | 3.09 | ** 93.35 ± 0.89 | 99.08 ± 3.57 |
Sequential extraction with hexane | 0.38 | ** 0 | ** 11.14 ± 2.51 |
Sequential extraction with DCM | 0.22 | ** 6.69 ± 1.36 | ** 19.62 ± 2.78 |
Sequential extraction with ethyl acetate | 0.87 | ** 65.02 ± 1.57 | 95.56 ± 1.68 |
Sequential extraction with methanol | 8.01 | 99.19 ± 0.18 | 94.46 ± 2.31 |
Controls | |||
10% MeOH | 3.28 ± 0.34 | 0 | |
10% DMSO | 4.70 ± 0.53 | 1.58 ± 0.60 | |
Elafin | 99.21 ± 0.05 | ||
EDTA | 95.66 ± 2.11 |
Results
Comparison of activity of different parts of S. birrea plant
Selection of the most suitable extraction solvent
Improving the quality of the ethanol extract as a cosmetic ingredient
Chemical characterization of the ethanol extract
Peak # | RT (min.) | Acquired [M-H]− m/z | Formula of possible structure | Theoretical [M-H]− m/z | Calculated accurate mass (Da) | Possible structure | Mass error (ppm) | MS/MS Data (fragments) | Confirmation with a standard | Ref | ||
---|---|---|---|---|---|---|---|---|---|---|---|---|
RT (min.) | [M-H]− m/z | |||||||||||
1 | 0.69 | 191.0564 | C7H12O6 | 191.0561 | 192.0634 | Quinic acid (Organic acid) | −1.57 | 173.0243 | [M-H]− -H2O | 0.69 | 191.0555 | |
127.0278 | [M-H]− - CO-2H2O | |||||||||||
85.0283 | [M-H]− - C3H6O3− - H2O | |||||||||||
71.0134 | [M-H]− - C4H8O3− - H2O | |||||||||||
59.0145 | [M-H]− - C5H8O4− | |||||||||||
2 | 2.45 | 289.0713 | C15H14O6 | 289.0717 | 290.0790 | Catechin (Flavonoid) | 1.38 | 109.0289 | [M-H]− - C10H12O3− | 2.45 | 289.0716 | [16] |
151.0336 | [M-H]− -C8H10O2− | |||||||||||
137.0209 | [M-H]− -C8H8O3− | |||||||||||
125.0248 | [M-H]-C9H10O−3 | |||||||||||
123.0416 | [M-H]− -C8H7O3− -H2O | |||||||||||
3 | 2.71 | 647.0876 | C28H24O18 | 647.0890 | 647.0963 | Unidentified | 2.16 | 495.0740 | ||||
343.0630 | ||||||||||||
325.0517 | ||||||||||||
169.0133 | ||||||||||||
125.0250 | ||||||||||||
4 | 2.93 | 457.0782 | C22H18O11 | 457.0776 | 458.0849 | Epigallocatechin-3-gallate (Flavonoid) | −1.31 | 305.0670 | [M-H]− - C7H4O4− | 2.93 | 457.0749 | |
289.0657 | [M-H]− - C7H4O5− | |||||||||||
169.0142 | [M-H]− - C15H12O6− | |||||||||||
125.0243 | [M-H]− - C16H13O8− | |||||||||||
5 | 3.17 | 729.1460 | C37H30O16 | 729.1461 | 730.1534 | Epicatechin-3-O-gallate-epicatechin (Proanthocyanidin | 0.14 | 577.1276 | [M-H]− - C7H4O4− | [16] | ||
451.0983 | [M-H]− - C7H4O4− -C8H8O3− - H2O | |||||||||||
407.0798 | [M-H]− -C15H12O6− -2H2O | |||||||||||
289.0717 | [M-H]− - C7H4O4− - C15H12O6− | |||||||||||
169.0147 | [M-H]− - C15H12O6− -2H2O-C15H13O−3 | |||||||||||
125.0240 | [M-H]− - C15H12O6− -2H2O-C15H13O3− -CO2 | |||||||||||
6 | 3.50 | 881.1577 | C44H34O20 | 881.1571 | 882.1644 | Procyanidin B2-3′3 di-O-gallate (Proanthocyanidin) | −0.68 | 729.1442 | [M-H]− - C7H4O−4 | |||
711.1376 | [M-H]− - C7H5O5− | |||||||||||
577.1229 | [M-H]− - C7H4O4− - C7H5O4− | |||||||||||
559.1210 | [M-H]− - C7H5O5− -C7H5O4− | |||||||||||
407.0760 | [M-H]− - C16H14O8− - C6H5O3− - H2O | |||||||||||
289.0716 | [M-H]− - C22H17O9− - C7H4O5− | |||||||||||
169.0142 | [M-H]− - C22H17O9− - C6H4O5− | |||||||||||
125.0232 | [M-H]− - C22H17O9− -C6H4O5− -H2O | |||||||||||
7 | 3.71 | 441.0817 | C22H18O10 | 441.0827 | 442.0900 | Epicatechin gallate (Flavonoid) | 2.27 | 289.0713 | [M-H]− - C7H4O4− | 3.74 | 441.0842 | [15] |
169.0138 | [M-H]− - C15H12O5− | |||||||||||
125.0234 | [M-H]− - C15H12O6− -CO2 |
Collagenase and elastase inhibition activity of pure standards
Comparison of chemical profiles of the Marula extract, defatted and concentrated fractions
Compound | RT (minutes) | [M-H]− m/z | Crude Ethanol extract | Defatted extract | Concentrated extract | Defatted and Concentrated fraction |
---|---|---|---|---|---|---|
Quinic acid | 0.69 | 191.0564 | present | present | absent | absent |
Catechin | 2.45 | 289.0713 | present | present | present | present |
Epigallocatechin gallate | 2.93 | 457.0782 | present | present | present | present |
Epicatechin gallate | 3.71 | 441.0817 | present | present | present | present |