J Food Bioact

Journal of Food Bioactives, ISSN 2637-8752 print, 2637-8779 online

Journal website www.isnff-jfb.com

Original Research

Volume 10, June 2020, pages 77-85

The chemical composition, antioxidant activity, and antiproliferative activity of selected seed flours

Figure 1. Total phenolic content (TPC) of seed flour samples (mg GAE/g seed flour). Values marked by different letters are significantly different (P < 0.05).

Figure 2. ABTS⁺ scavenging capacities of seed flour samples (µmol TE/g seed flour). Values marked by different letters are significantly different (P < 0.05).

Figure 3. Relative DPPH^• scavenging capacities (RDSC) of seed flour samples (µmol TE/g seed flour). Values marked by different letters are significantly different (P < 0.05).

Figure 4. Cell toxicity in 3T3-L1 preadipocytes for selected seed flour extracts. Different letters within the same day represent significant difference (P < 0.05).

Figure 5. Antiproliferation of SW480 colon cancer cells treated with seed flour extracts. Different letters within the same day represent significant difference (P < 0.05).

**Table 1.** Characterization of chemical compounds present in the broccoli seed flour extract using UHPLC-MS
Peak ID	t_{R (min)}	Chemical Formula	Tentatively Identification	Theoretical [M-H]⁻	Experimental [M-H]⁻	MS² Ions	References
^1,2,3Represented the three greatest peaks based on the typical UHPLC chromatogram peak area.
1	3.62	C₁₂H₂₃NO₁₀S₃	Glucoraphanin isomer	436.0406	436.0407	372.0422, 259.0999	Rochfort et al., 2008
2	3.99	C₁₂H₂₃NO₁₀S₃	Glucoraphanin isomer	436.0406	436.0408	372.0489, 259.1865
3	4.22	C₁₂H₂₃NO₁₀S₃	Glucoraphanin isomer	436.0406	436.0400	372.0485, 291.1234, 259.1592
4	9.87	C₁₂H₂₃NO₉S₃	Glucoerucin	420.0457	420.0457	340.2986, 275.0135, 259.0399, 242.0890	Rochfort et al., 2008
5	11.61	C₁₇H₂₂O₁₀	Sinapoylhexose	385.1135	385.1136	223.0611, 205.0510	Shao et al., 2014
6	21.38	C₃₄H₄₂O₁₉	Disinapoylgentiobiose¹	753.2242	753.2244	529.1563, 223.0612	Shao et al., 2014
7	25.89	C₂₈H₃₂O₁₄	1,2-Disinapoylglucoside³	591.1714	591.1708	367.1031, 223.0611, 205.0509	Shao et al., 2014
8	26.86	C₄₅ H₅₂ O₂₃	1,2,2′-Trisinapoylgentiobioside²	959.2821	959.2809	735.1401, 529.1462	Velasco et al., 2011

Table 1. Characterization of chemical compounds present in the broccoli seed flour extract using UHPLC-MS

Peak ID

t_{R (min)}

Chemical Formula

Tentatively Identification

Theoretical [M-H]⁻

Experimental [M-H]⁻

MS² Ions

References

^1,2,3Represented the three greatest peaks based on the typical UHPLC chromatogram peak area.

3.62

C₁₂H₂₃NO₁₀S₃

Glucoraphanin isomer

436.0406

436.0407

372.0422, 259.0999

Rochfort et al., 2008

3.99

C₁₂H₂₃NO₁₀S₃

Glucoraphanin isomer

436.0406

436.0408

372.0489, 259.1865

4.22

C₁₂H₂₃NO₁₀S₃

Glucoraphanin isomer

436.0406

436.0400

372.0485, 291.1234, 259.1592

9.87

C₁₂H₂₃NO₉S₃

Glucoerucin

420.0457

340.2986, 275.0135, 259.0399, 242.0890

Rochfort et al., 2008

11.61

C₁₇H₂₂O₁₀

Sinapoylhexose

385.1135

385.1136

223.0611, 205.0510

Shao et al., 2014

21.38

C₃₄H₄₂O₁₉

Disinapoylgentiobiose¹

753.2242

753.2244

529.1563, 223.0612

Shao et al., 2014

25.89

C₂₈H₃₂O₁₄

1,2-Disinapoylglucoside³

591.1714

591.1708

367.1031, 223.0611, 205.0509

Shao et al., 2014

26.86

C₄₅ H₅₂ O₂₃

1,2,2′-Trisinapoylgentiobioside²

959.2821

959.2809

735.1401, 529.1462

Velasco et al., 2011

**Table 2.** Characterization of chemical compounds present in the carrot seed flour extract using UHPLC-MS
Peak ID	tR (min)	Chemical Formula	Tentatively Identification	Theoretical [M-H]⁻	Experimental [M-H]⁻	MS² Ions	References
^1,2,3Represented the three greatest peaks based on the typical UHPLC chromatogram peak area.
1	7.60	C₂₁H₂₈O₁₄	Caffeoyldihexoside	503.1401	503.1400	341.1647, 179.0352,	Oszmiański et al., 2018
2	7.80	C₂₁H₂₈O₁₃	Cistanoside F	487.1452	487.1452	325.1151, 179.0351	Xiao-Long et al., 2019
3	8.00	C₂₂H₃₀O₁₄	Lycibarbarphenylpropanoid C	517.1557	517.1558	355.2340, 193.0508	Li et al., 2020; Xiao-Long et al., 2019
4	11.38	C₂₇H₃₀O₁₅	Kaempferol-3-O-rutinoside isomer	593.1506	593.1505	447.1625, 285.1403	Inbaraj et al., 2010
5	15.17	C₂₇H₃₀O₁₅	Kaempferol-3-O-rutinoside isomer¹	593.1506	593.1501
6	16.27	C₂₁H₂₀O₁₁	Kaempferol-3-O-glucoside isomer³	447.0927	447.0932	285.0400, 284.0326	Kelebek, 2016
7	18.08	C₂₇H₃₀O₁₄	Apigenin-7-O-β-D-rutinoside	577.1557	577.1558	269.0451, 227.1987	Wang et al., 2017
8	18.95	C₂₈H₃₂O₁₅	Diosmetin-7-rutinoside	607.1663	607.1661	299.1501	Gironés-Vilaplana et al., 2014
9	19.90	C₂₁H₂₀O₁₁	Kaempferol-3-O-glucoside isomer	447.0927	447.0940	285.0403	Kelebek, 2016
10	30.26	C₁₅H₁₀O₆	Luteolin²	285.0399	285.0402	217.1554, 199.0589, 175.0652, 133.0716	De la Torre-Carbot et al., 2005

Table 2. Characterization of chemical compounds present in the carrot seed flour extract using UHPLC-MS

Peak ID

tR (min)

Chemical Formula

Tentatively Identification

Theoretical [M-H]⁻

Experimental [M-H]⁻

MS² Ions

References

^1,2,3Represented the three greatest peaks based on the typical UHPLC chromatogram peak area.

7.60

C₂₁H₂₈O₁₄

Caffeoyldihexoside

503.1401

503.1400

341.1647, 179.0352,

Oszmiański et al., 2018

7.80

C₂₁H₂₈O₁₃

Cistanoside F

487.1452

325.1151, 179.0351

Xiao-Long et al., 2019

8.00

C₂₂H₃₀O₁₄

Lycibarbarphenylpropanoid C

517.1557

517.1558

355.2340, 193.0508

Li et al., 2020; Xiao-Long et al., 2019

11.38

C₂₇H₃₀O₁₅

Kaempferol-3-O-rutinoside isomer

593.1506

593.1505

447.1625, 285.1403

Inbaraj et al., 2010

15.17

C₂₇H₃₀O₁₅

Kaempferol-3-O-rutinoside isomer¹

593.1506

593.1501

16.27

C₂₁H₂₀O₁₁

Kaempferol-3-O-glucoside isomer³

447.0927

447.0932

285.0400, 284.0326

Kelebek, 2016

18.08

C₂₇H₃₀O₁₄

Apigenin-7-O-β-D-rutinoside

577.1557

577.1558

269.0451, 227.1987

Wang et al., 2017

18.95

C₂₈H₃₂O₁₅

Diosmetin-7-rutinoside

607.1663

607.1661

299.1501

Gironés-Vilaplana et al., 2014

19.90

C₂₁H₂₀O₁₁

Kaempferol-3-O-glucoside isomer

447.0927

447.0940

285.0403

Kelebek, 2016

30.26

C₁₅H₁₀O₆

Luteolin²

285.0399

285.0402

217.1554, 199.0589, 175.0652, 133.0716

De la Torre-Carbot et al., 2005

**Table 3.** Characterization of chemical compounds present in the milk thistle seed flour extract using UHPLC-MS
Peak ID	t_{R (min)}	Chemical Formula	Tentatively Identification	Theoretical [M-H]⁻	Experimental [M-H]⁻	MS² Ions	References
^1,2,3Represented the three greatest peaks based on the typical UHPLC chromatogram peak area. *CE stands for a chlorogenic acid equivalent. SE stands for a silibinin equivalent.
1	8.24	C₁₆H₁₈O₉	Chlorogenic acid isomer	353.0873	353.0875	191.0563, 179.0351, 135.0455, 111.0900	Willems et al., 2016
2	10.15	C₁₆H₁₈O₉	Chlorogenic acid isomer	353.0873	353.0876	191.0563, 173.0940, 127.0063, 111.1268
3	13.90	C₃₁H₃₄O₁₆	5-p-(6-caffeoyl-glucopyranosyl)-coumaroylquinic acid isomer	661.1769	661.1760	499.1453, 337.1499, 173.1541, 163.0531	Choe et al., 2019
4	15.32	C₃₁H₃₄O₁₆	5-p-(6-caffeoyl-glucopyranosyl)-coumaroylquinic acid isomer	661.1769	661.1764	499.1456, 337.1122, 173.0676, 163.1128
5	15.62	C₃₂H₃₆O₁₇	Methyl 5-(6-Caffeoyl-glucopyranosyl)-caffeoylquinic acid	691.1874	691.1886	661.1766, 499.1458, 173.0309	Choe et al., 2019
6	17.47	C₁₅H₁₂O₇	Taxifolin	303.0505	303.0511	285.0483, 177.0815, 124.9742	Álvarez-Ferná́ndez et al., 2016
7	22.83	C₂₅H₂₂O₁₀	Silychristin isomer	481.1135	481.1141	463.1035, 453.1453, 445.1099, 179.0201	Shibano et al., 2007
8	24.74	C₂₅H₂₂O₁₀	Silychristin isomer	481.1135	481.1142	463.1968, 453.1838, 419.2541, 355.1195, 179.0223
9	25.33	C₂₅H₂₂O₁₀	Silychristin isomer²	481.1135	481.1141	463.0804, 453.0892, 355.0830, 178.9522
10	26.04	C₂₅H₂₂O₁₀	Silychristin isomer¹	481.1135	481.1139	463.1879, 453.1561, 327.0796, 179.0533
11	36.87	C₂₅H₂₂O₁₀	Silybin A	481.1135	481.1142	463.0794, 453.1011, 355.1670, 300.9895	Shibano et al., 2007
12	37.35	C₂₅H₂₂O₁₀	Silybin B	481.1135	481.1141	463.1127, 453.1870, 355.1905, 300.9849
13	40.31	C₂₅H₂₂O₁₀	Isosilybin³	481.1135	481.1137	463.2560, 453.1837, 301.1072, 283.0059	Shibano et al., 2007

Table 3. Characterization of chemical compounds present in the milk thistle seed flour extract using UHPLC-MS

Peak ID

t_{R (min)}

Chemical Formula

Tentatively Identification

Theoretical [M-H]⁻

Experimental [M-H]⁻

MS² Ions

References

^1,2,3Represented the three greatest peaks based on the typical UHPLC chromatogram peak area. *CE stands for a chlorogenic acid equivalent. SE stands for a silibinin equivalent.

8.24

C₁₆H₁₈O₉

Chlorogenic acid isomer

353.0873

353.0875

191.0563, 179.0351, 135.0455, 111.0900

Willems et al., 2016

10.15

C₁₆H₁₈O₉

Chlorogenic acid isomer

353.0873

353.0876

191.0563, 173.0940, 127.0063, 111.1268

13.90

C₃₁H₃₄O₁₆

5-p-(6-caffeoyl-glucopyranosyl)-coumaroylquinic acid isomer

661.1769

661.1760

499.1453, 337.1499, 173.1541, 163.0531

Choe et al., 2019

15.32

C₃₁H₃₄O₁₆

5-p-(6-caffeoyl-glucopyranosyl)-coumaroylquinic acid isomer

661.1769

661.1764

499.1456, 337.1122, 173.0676, 163.1128

15.62

C₃₂H₃₆O₁₇

Methyl 5-(6-Caffeoyl-glucopyranosyl)-caffeoylquinic acid

691.1874

691.1886

661.1766, 499.1458, 173.0309

Choe et al., 2019

17.47

C₁₅H₁₂O₇

Taxifolin

303.0505

303.0511

285.0483, 177.0815, 124.9742

Álvarez-Ferná́ndez et al., 2016

22.83

C₂₅H₂₂O₁₀

Silychristin isomer

481.1135

481.1141

463.1035, 453.1453, 445.1099, 179.0201

Shibano et al., 2007

24.74

C₂₅H₂₂O₁₀

Silychristin isomer

481.1135

481.1142

463.1968, 453.1838, 419.2541, 355.1195, 179.0223

25.33

C₂₅H₂₂O₁₀

Silychristin isomer²

481.1135

481.1141

463.0804, 453.0892, 355.0830, 178.9522

26.04

C₂₅H₂₂O₁₀

Silychristin isomer¹

481.1135

481.1139

463.1879, 453.1561, 327.0796, 179.0533

36.87

C₂₅H₂₂O₁₀

Silybin A

481.1135

481.1142

463.0794, 453.1011, 355.1670, 300.9895

Shibano et al., 2007

37.35

C₂₅H₂₂O₁₀

Silybin B

481.1135

481.1141

463.1127, 453.1870, 355.1905, 300.9849

40.31

C₂₅H₂₂O₁₀

Isosilybin³

481.1135

481.1137

463.2560, 453.1837, 301.1072, 283.0059

Shibano et al., 2007