hindawi publishing corporationdownloads.hindawi.com/journals/omcl/2019/5465463.f… · web viewthe...
TRANSCRIPT
Supplementary Material
Phytochemical and antioxidant studies on a rare Rheum cordatum
Losinsk. species from Kazakhstan
Gulsim Zhumashova, Wirginia Kukula-Koch,* Wojciech Koch, Tomasz Baj, Galiya Sayakova, Alma Shukirbekova, Kazimierz Głowniak and Zuriyadda Sakipova1
Figure S1. Compositional variation of the root extracts in relation to the collection date
Figure S2. The differences in the composition of ethanolic extracts in various organs of Rheum cordatum (TIC chromatograms recorded in the negative ionisation mode)
Figure S3. The influence of extractant on the composition of the Rheum cordatum root extract (1- 98% ethanol, 2- 50% ethanol, 3-dichloromethane, 4-water, 5-chloroform extracts)
Figure S4. The MS/MS spectra of the selected major constituents of the extractsAcetyl aloe-emodin
2x10
00.5
11.5
22.5
33.5
44.5
55.5
66.5
77.5
88.5
9-ESI Product Ion (rt: 28.032 min) Frag=170.0V [email protected] (311.0608[z=1] -> **) 2R0518_20_neg.d
311.
0579
269.
0477
224.
0539
248.
0819
169.
0687
298.
0645
197.
3009
255.
0881
114.
6740
129.
0348
238.
6512
158.
0525
230.
5508
361.
7164
149.
0927
324.
2470
216.
2753
338.
3396
Counts vs. Mass-to-Charge (m/z)120 140 160 180 200 220 240 260 280 300 320 340 360
Aloeemodin3x10
0
0.5
1
1.5
2
2.5
3-ESI Product Ion (rt: 29.850 min) Frag=170.0V [email protected] (269.0493[z=1] -> **) 2R0518_20_neg.d
269.
0456
225.
0517
341.
0795
180.
0617
125.
0250
485.
2378
829.
1705
927.
7012
392.
1866
1052
.135
3
433.
1833
762.
6471
Counts vs. Mass-to-Charge (m/z)100 200 300 400 500 600 700 800 900 1000 1100
Apigenin galloyl glucose3x10
00.20.40.60.8
11.21.41.61.8
22.22.42.62.8
33.2
-ESI Product Ion (rt: 24.641 min) Frag=170.0V [email protected] (583.1185[z=1] -> **) 2R0518_20_neg.d
583.
1060
169.
0111
269.
0388
435.
0940
341.
0273
311.
9866
125.
0168
417.
0839
225.
0238
651.
2875
151.
0375
187.
0416
Counts vs. Mass-to-Charge (m/z)100 150 200 250 300 350 400 450 500 550 600 650
Apigenin glucoside3x10
0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
5-ESI Product Ion (rt: 18.507 min) Frag=170.0V [email protected] (431.0978[z=1] -> **) 2RCE_20_neg.d
311.
0582
431.
1011
341.
0669
283.
0588
323.
0557
413.
1027
191.
0417
353.
0742
458.
3453
161.
0170
440.
1874
269.
0418
383.
0870
Counts vs. Mass-to-Charge (m/z)80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480
Catechin Chrysophanol
3x10
00.25
0.5
0.751
1.251.5
1.75
22.25
2.52.75
3
3.253.5
-ESI Product Ion (rt: 13.485 min) Frag=160.0V [email protected] (289.0698[z=1] -> **) 2RCE_20_neg.d
289.
0687
245.
0792
179.
0300
125.
0250
151.
0319
109.
0339
203.
0698
230.
0588
165.
0197
425.
9288
Counts vs. Mass-to-Charge (m/z)100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500
3x10
0
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
1.8
2
-ESI Product Ion (rt: 25.933 min) Frag=170.0V [email protected] (253.0548[z=1] -> **) 2R0518_20_neg.d
253.
0508
225.
0555
139.
1122
145.
0375
182.
0392
245.
3283
207.
0473
196.
0511
276.
7628
129.
5038
263.
2006
221.
6105
310.
8081
Counts vs. Mass-to-Charge (m/z)120 130 140 150 160 170 180 190 200 210 220 230 240 250 260 270 280 290 300 310
Emodin4x10
0
0.20.40.60.8
11.2
1.41.61.8
2
-ESI Product Ion (rt: 33.622 min) Frag=170.0V [email protected] (269.0493[z=1] -> **) 2R0518_20_neg.d
269.
0518
225.
0598
Counts vs. Mass-to-Charge (m/z)100 200 300 400 500 600 700 800 900 1000 1100
Emodin acetyl hexose3x10
00.10.20.30.40.50.60.70.80.9
11.11.21.31.41.51.61.7
-ESI Product Ion (rt: 25.443 min) Frag=170.0V [email protected] (473.1166[z=1] -> **) 2R0518_20_neg.d
473.
1163
269.
0467
577.
4017
254.
0474
311.
0512
282.
0512
186.
0601
413.
0831
431.
1480
456.
1063
349.
9640
495.
4271
201.
0663
397.
2875
508.
5674
128.
6230
542.
1300
146.
8610
160.
0873
328.
5281
559.
4084
375.
2971
Counts vs. Mass-to-Charge (m/z)125 150 175 200 225 250 275 300 325 350 375 400 425 450 475 500 525 550 575 600
Emodin galloylglucose3x10
00.20.40.60.8
11.21.41.61.8
22.22.42.62.8
33.2
-ESI Product Ion (rt: 24.641 min) Frag=170.0V [email protected] (583.1185[z=1] -> **) 2R0518_20_neg.d
583.
1060
169.
0111
269.
0388
435.
0940
341.
0273
311.
9866
125.
0168
417.
0839
225.
0238
651.
2875
151.
0375
187.
0416
Counts vs. Mass-to-Charge (m/z)150 200 250 300 350 400 450 500 550 600 650 700
Emodin glucoside3x10
0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
5
5.5
-ESI Product Ion (rt: 23.986 min) Frag=170.0V [email protected] (431.1022[z=1] -> **) 2RST_20_neg.d
431.
1008
269.
0461
311.
0530
413.
1082
293.
0537
224.
0439
Counts vs. Mass-to-Charge (m/z)80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480
Emodin malonyl hexose3x10
0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
5
5.5
6
-ESI Product Ion (rt: 25.402 min) Frag=160.0V [email protected] (517.1041[z=1] -> **) 2RST_20_neg.d
473.
0726
517.
0615
269.
0252
431.
0502
Counts vs. Mass-to-Charge (m/z)125 150 175 200 225 250 275 300 325 350 375 400 425 450 475 500 525 550 575 600
Epicatechin3x10
0
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
1.8
2
2.2
-ESI Product Ion (rt: 15.743 min) Frag=160.0V [email protected] (289.0749[z=1] -> **) 2R0518_20_neg.d
289.
0740
245.
0795
205.
0528
181.
0293
123.
6484
139.
0424
165.
0176
221.
1489
109.
0301
150.
7214
355.
2125
229.
0402
301.
9061
101.
2901
Counts vs. Mass-to-Charge (m/z)80 100 120 140 160 180 200 220 240 260 280 300 320 340
Epicatechin gallate Epigallocatechin gallate
3x10
00.5
11.5
22.5
33.5
44.5
5-ESI Product Ion (rt: 29.630 min) Frag=170.0V [email protected] (451.1285[z=1] -> **) 2R0518_20_neg.d
451.
1127
407.
1273
305.
0673
1089
.745
7
167.
0493
125.
0251
242.
0424
Counts vs. Mass-to-Charge (m/z)100 200 300 400 500 600 700 800 900 1000 1100
3x10
0
1
2
3
4
5
6
-ESI Product Ion (rt: 16.227 min) Frag=160.0V [email protected] (457.0837[z=1] -> **) 2R0518_20_neg.d
169.
0140
457.
0782
305.
0666
125.
0250
261.
0771
331.
0559
Counts vs. Mass-to-Charge (m/z)150 200 250 300 350 400 450 500 550 600 650 700 750
Gallic acid3x10
00.250.5
0.751
1.251.5
1.752
2.252.5
-ESI Product Ion (rt: 3.696 min) Frag=160.0V [email protected] (169.0170[z=1] -> **) 2R0518_20_neg.d
125.
0238
169.
0113
227.
9971
Counts vs. Mass-to-Charge (m/z)80 100 120 140 160 180 200 220 240 260 280
Gallic acid hexoside3x10
00.25
0.50.75
11.25
1.51.75
22.25
2.52.75
33.25
3.53.75
44.25
-ESI Product Ion (rt: 4.585 min) Frag=160.0V [email protected] (331.0701[z=1] -> **) 2RST_20_neg.d
169.
0125
331.
0639
125.
0267
Counts vs. Mass-to-Charge (m/z)80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480
Kaempferol3x10
00.250.5
0.751
1.251.5
1.752
2.252.5
-ESI Product Ion (rt: 26.902 min) Frag=170.0V [email protected] (285.0385[z=1] -> **) 2RCE_20_neg.d
285.
0353
151.
0021
213.
0501
257.
0364
107.
0137
240.
0416
135.
0108
195.
0415
173.
1125
516.
6141
Counts vs. Mass-to-Charge (m/z)100 150 200 250 300 350 400 450 500
Quercetin3x10
0
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
1.8
2
2.2
-ESI Product Ion (rt: 24.288 min) Frag=170.0V [email protected] (301.0331[z=1] -> **) 2RCE_20_neg.d
151.
0022
178.
9969
301.
0299
107.
0116
273.
0358
121.
0313
215.
0286
227.
0297
240.
0295
190.
0167
257.
0383
143.
0721
Counts vs. Mass-to-Charge (m/z)120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480
Tetrahydroxystilbene-O-(acetyl)-hexose3x10
00.20.40.60.8
11.21.41.61.8
22.22.42.62.8
33.23.4
-ESI Product Ion (rt: 24.617 min) Frag=160.0V [email protected] (447.1324[z=1] -> **) 2RST_20_neg.d
285.
0738
447.
1281
242.
0530
227.
0326
270.
0489
144.
9902
402.
6246
349.
0662
Counts vs. Mass-to-Charge (m/z)125 150 175 200 225 250 275 300 325 350 375 400 425 450 475 500 525 550 575
Table S1: Content of the main constituents determined in the particular plant parts, depending on the solvents used.
Leaves Seeds stems Radix 2017 Radix04.2018
Radix 05.2018
Etha
nol e
xtra
ct
Emodin 0.0614±0.0009 0.0694±0.0017 0.6581±0.0056 0.5174±0.0043 0.3626±0.003 0.8023±0.0152
Aloeemodin
0.0037±0.00006
0.0029±0.00002 0.0135±0.00008 0.0889±0.0021 0.0049±0.0007 0.0089±0.0001
ECG 0.8552±0.0264 0.7824±0.0147 0.0509±0.0011 2.93±0.0921 2.5052±0.0276 2.38±0.0316
EGCG 0.0151±0.0002 0.24±0.0064 0.0001±0.000003 0.4394±0.0034 0.2467±0.0035 0.1265±0.005EC 0.0304±0.0009 0.0736±
0.00270.0116±0.00008 0.0274±0.0012 0.0114±0.0002 0.1100±0.0042
Rutoside 2.108±0.0244 1.614±0.0361 0.1504±0.0026 0.0327±0.0002 0.0155±0.0003 0.0160±0.0003
Kaempferol glucoside
0.0410±0.0004 0.0032±0.0001 0.0043±0.00006 0.0433±0.001 0.0428±0.0006 0.0225±0.0001
DPPH <LOQ 89.80±1.97 <LOQ 92.05±0.39 92.3±0.69 68.13±4.18TPC <LOQ 460±24.70 <LOQ 530.2±37.65 430.2±28.67 720.3±34.22
Etha
nol:w
ater
Emodin 0.5420±0.0114 0.1416±0.0012 0.4482±0.0194 1.29±0.0068 0.9650±0.027 0.7956±0.022Aloeemodin
0.0025±0.00002
<LOQ 0.0293±0.0003 0.1273±0.0007 0.0539±0.0036 0.017±0.0003
ECG 2.1589±0.0131 2.0829±0.0031 1.6876±0.0132 3.084±0.0319 3.3669±0.0262 5.004±0.1265
EGCG 0.2350±0.0017 0.6432±0.0195 0.0363±0.0003 0.2212±0.0028 0.3596±0.0026 0.24±0.0015EC 0.0464±0.0005 0.0416±0.0004 0.0873±0.005 0.0213±0.0000
40.0222±0.0005 0.081±0.0033
Rutoside 0.0757±0.0009 2.0235±0.0106 0.8525±0.009 0.0232±0.00003
0.4144±0.0064 0.022±0.0005
Kaempferol glucoside
0.1564±0.0051 0.0466±0.0001 0.1408±0.0021 0.0549±0.0017 0.0159±0.0007 0.021±0.0003
DPPH 89.96±0.4559 89.06±1.54 92.74±0.9577 88.48±0.68 89.69±1.63 83.7±2.08TPC 719.8±26.61 669.8±19.86 540.2±30.74 480.3±30.07 462±23.56 620.7±39.93
Dich
loro
met
hene
Emodin 0.1241±0.0005 0.0004±0.00001 1.1747±0.0354 0.2855±0.0009 0.7823±0.0180 1.0646±0.0287Aloeemodin
0.0002±0.00002
<LOQ <LOQ <LOQ <LOQ <LOQ
ECG 0.0032±0.0002 0.0022±0.00002 0.0403±0.0021 0.0019±0.00004
0.0045±0.00009 0.0014±0.00015
EGCG <LOQ 0.0003±0.00002 0.0008±0.00005 0.0002±0.000006
0.0003±0.00002 0.00009±0.00001
EC 0.0007±0.00002
0.0002±0.000002
0.0006±0.00004 0.0004±0.00003
0.0003±0.00002 0.0003±0.00002
Rutoside 0.0106±0.0002 0.0014±0.00005 0.0059±0.0001 0.0003±0.00003
0.0018±0.00001 <LOQ
Kaempferol glucoside
<LOQ 0.0005±0.00001 0.0019±0.00006 <LOQ <LOQ <LOQ
DPPH <LOQ <LOQ <LOQ <LOQ <LOQ <LOQTPC <LOQ <LOQ <LOQ <LOQ <LOQ <LOQEmodin 0.0197±0.0001 0.1898±0.001 0.06±0.0008 0.8431±0.0308 0.5222±0.0337 0.3369±0.011
Aloeemodin
<LOQ 0.0029±0.000009
0.0027±0.00005 0.0831±0.001 0.0257±0.0028 0.011±0.00008
ECG 1.424±0.0111 1.7212±0.0035
0.6364±0.0134 2.5042±0.0131 2.76±0.1116 2.29±0.0196
EGCG 0.1107±0.0005 0.7922±0.0097 0.0144±0.00004 0.1564±0.0004 0.2335±0.0014 0.1224±0.0028EC 0.0087±0.0000
40.0557±0.0021 0.0341±0.0002 0.0186±0.0000
90.0178±0.0003 0.0421±0.0009
Water
Rutoside 4.8664±0.0604 1.9050±0.0102 0.5992±0.0171 0.0211±0.0003 0.0157±0.0006 0.015±0.0004Kaempferol glucoside
0.1299±0.0016 0.0181±0.00005 0.0567±0.002 0.014±0.00006 0.011±0.0013 0.0049±0.00007
DPPH 45.9±1.38 88.23±1.26 21.83±8.42 82.79±2.07 91.59±1.11 90.6±1.00TPC 279.8±37.35 470.3±18.75 90.33±9.52 320±31.55 498±19.71 751.8±34.78
Chlo
rofo
rm
Emodin 0.0658±0.0026 0.0375±0.0006 0.37±0.0038 0.5656±0.0417 0.6566±0.0161 0.8611±0.0195Aloeemodin
<LOQ <LOQ <LOQ <LOQ <LOQ <LOQ
ECG 0.0024±0.00002
0.0019±0.00002 0.0035±0.00005 0.0034±0.00005
0.0011±0.00005 0.8647±0.0545
EGCG 0.0001±0.000005
0.0002±0.000001
0.0002±0.00001 0.0006±0.00004
0.0002±0.00002 0.0193±0.00005
EC 0.0001±0.00002
0.0008±0.00003 0.0006±0.0001 <LOQ 0.0003±0.00003 0.0459±0.0015
Rutoside 0.0007±0.0001 0.0005±0.00001 0.0005±0.00006 0.0004±0.00001
0.0039±0.0002 0.0023±0.00005
Kaempferol glucoside
0.0041±0.00003
<LOQ <LOQ 0.0042±0.00008
0.0102±0.0007 0.0076±0.0003
DPPH <LOQ <LOQ <LOQ <LOQ 21.71±7.36 <LOQTPC <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ
Table S2. The correlations between the single metabolites determined in the extracts
I% Emodin Aloeemodin ECG EGCG EC RutosideKaempferol glucoside
I% 1
Emodin-
0.0173 1
Aloeemodin 0.3448 0.259271 1
ECG 0.8852 -0.05801 0.256912 1
EGCG 0.7165 -0.12694 -0.11936 0.695647 1
EC 0.8673 -0.03761 0.589719 0.721823 0.48493 1
Rutoside 0.3956 -0.35367 -0.05216 0.538240.38586
4 0.256191 1
Kaempferol glucoside 0.5809 0.048616 0.424163 0.759295
0.094772 0.489354
0.415671 1
Table S3. The results of univariate analysis of variance
Groups Counter Sum Average Variance
Emodin 15 3.962698 0.26418 0.10624
Aloeemodin 15 0.0578 0.003853 6.15E-05
ECG 15 11.45336 0.763558 0.703273
EGCG 15 2.088774 0.139252 0.062664
EC 15 0.392295 0.026153 0.000861
Rutoside 15 14.21484 0.947656 1.864206
Kaempferol glucoside 15 0.603319 0.040221 0.003165
ANALYSIS OF VARIANCE
Source of variance SS df MS F p value Test F
between groups13.3592
7 6 2.226544 5.687277 4.18242E-05 2.192518
inside groups38.3665
7 98 0.391496
Sum51.7258
4 104
Figure S5. Emodin variation graph depending on the solvent used
Figure S6. Emodin variation plot depending on the part of the plant used.
Table S4. The list of secondary metabolites present in other species of Rheum or Polygonum gender, which presence was not confirmed in the herein studied extracts by HR-MS studies.
Name NamePhyscion GentisinRhein Aloeemodin acetyl-
hexosideChrysophanol hexoside HydroxyemodinDimer of rhein and emodin and acetyl-hexose
Rhapontigen hexoside
Rhein Resveratrol glucosideRhein hexoside Emodin malonyl-
glucosideChrysophanol acetyl-hexoside
Dimer of emodin and chrysophanol