
Figure 1: HPLC-FT-ICR MS base peak intensity chromatograms (BPCs) of the test samples in negative-ion mode. A: GR extract; B: pGR extract; C: the peels extract; the compound numbers (1- 47) are the same as those in Table 1
Compound |
Regression equationa |
Correlation coefficient (r) |
Linear range (μg/mL) |
LOD (μg/mL) |
LOQ |
Xanthotoxol |
y= 35221x+ 53381 |
0.9996 |
0.25 ~ 4.00 |
0.10 |
0.35 |
Marmesin |
y= 197101x+ 23307 |
0.9995 |
0.25 ~ 8.00 |
0.10 |
0.40 |
Bergapten |
y= 272875x+ 1614.8 |
0.9996 |
0.20 ~ 1.80 |
0.05 |
0.18 |
Imperatorin |
y= 158572x- 1516.2 |
0.9996 |
0.20 ~1.10 |
0.05 |
0.17 |
Table 1: Linear regression, LOD, and LOQ of the standard compounds
ax is the concentraction of each compound in μg/mL; y is the peak area of the respective compound detected at 320 nm.
No. |
tR (min) |
[M - H]- |
|
|
[M + H]+ |
|
|
MS/MS |
Formula |
Identification |
GR |
pGR |
peels |
Observed Mass (Da) |
Calculated Mass (Da) |
Error (ppm) |
Observed Mass (Da) |
Calculated Mass (Da) |
Error (ppm) |
||||||||
1 |
6.86 |
329.08752 |
329.08781 |
0.88 |
- |
- |
- |
- |
C14H18O9 |
vanillic acid 4-O-β-D- glucopyranoside |
+ |
+ |
+ |
2 |
6.88 |
167.03474 |
167.03498 |
1.45 |
- |
- |
- |
- |
C8H8O8 |
vanillic acid |
+ |
+ |
+ |
3 |
8.33 |
369.08211 |
369.08272 |
1.65a |
325.09079 |
325.09179 |
3.08 |
- |
C15 H16O4 |
umbelliferone 7-O-D- glucopyranoside |
+ |
+ |
+ |
4 |
8.82 |
353.08753 |
353.08781 |
0.79 |
355.10117 |
355.10236 |
3.34 |
- |
C16H18O 9 |
scopolin |
+ |
+ |
+ |
5 |
9.38 |
551.17674 |
551.17701 |
0.49 |
- |
- |
- |
- |
C26H32o13 |
glehlinoside C |
+ |
+ |
+ |
6 |
9.71 |
463.08844 |
463.08820 |
-0.52 |
- |
- |
- |
- |
C21H20O12 |
isoquercetin |
- |
- |
+ |
7 |
9.92 |
505.17194 |
505.17154 |
-0.80 |
- |
- |
- |
- |
C25 H30 O11 |
2,3-E-2,3-dihydro-2- (3'-methoxy-4'-hydroxy |
- |
- |
+ |
8 |
10.13 |
485.16555 |
485.16645 |
1.85a |
- |
- |
- |
- |
C21 H28 O10 |
7-O-methylpeucedanol 3' |
+ |
- |
+ |
9 |
10.17 |
505.16951 |
505.17154 |
4.01 |
- |
- |
- |
- |
C25 H30 O11 |
glehlinoside I |
- |
- |
+ |
10 |
10.18 |
179.03476 |
179.03498 |
1.24 |
- |
- |
- |
135.04494 |
C9H8 O4 |
caffeic acid |
+ |
- |
+ |
11 |
10.25 |
571.13210 |
571.13046 |
-2.87a |
- |
- |
- |
- |
C23 H26 O14 |
bergaptol 5-O-β |
- |
- |
+ |
12 |
10.47 |
523.21772 |
523.21849 |
1.47 |
- |
- |
- |
- |
C26H36O11 |
(–)-secoisolariciresinol 4- |
+ |
+ |
+ |
13 |
10.48 |
361.16533 |
361.16566 |
0.92 |
363.17900 |
363.18022 |
3.34 |
|
C20H26 O6 |
(–)-secoisolariciresinol |
- |
- |
+ |
14 |
10.56 |
553.22858 |
553.22905 |
0.85 |
- |
- |
- |
- |
C27 H38 O12 |
glehlinoside G |
+ |
- |
+ |
15 |
10.58 |
469.13496 |
469.13515 |
0.40a |
- |
- |
- |
- |
C20O10 |
hydroxymarmesin 4'-O-β- D-glucopyranoside |
+ |
- |
+ |
16 |
11.13 |
609.14574 |
609.14611 |
0.60 |
- |
- |
- |
301.0362 |
C27H30O16 |
rutin |
+ |
- |
- |
17 |
11.19 |
585.18177 |
585.18249 |
1.24 |
- |
- |
- |
- |
C26 H34 O15 |
oxymarmesin 5'-O-β- D-glucopyranoside |
- |
- |
+ |
18 |
11.28 |
585.18020 |
585.18249 |
3.92a |
- |
- |
- |
- |
C25 H32 O13 |
marmesin 4'-O-β-D- apiofuranosyl-(1→6) |
+ |
- |
- |
19 |
11.61 |
453.13985 |
453.14024 |
0.84a |
409.14853 |
409.14931 |
1.91 |
247.09641; |
C20 H24 O9 |
marmesinin (nodakenin) |
+ |
+ |
+ |
20 |
11.64 |
617.20815 |
617.20871 |
0.91a |
- |
- |
- |
- |
C26H36 O14 |
7-O-methylpeucedanol 3'- |
+ |
- |
+ |
21 |
11.94 |
599.19706 |
599.19814 |
1.81a |
- |
- |
- |
- |
C26 H34 O13 |
osthenol 7-O-β-D- gentiobioside |
+ |
- |
+ |
22 |
12.34 |
687.22781 |
687.22944 |
2.37 |
- |
- |
- |
- |
C34 H40 O15 |
glehlinoside D |
+ |
- |
+ |
23 |
12.44 |
191.03474 |
191.03498 |
1.27 |
193.04931 |
193.04954 |
1.16 |
- |
C10 H8 O4 |
scopoletin |
+ |
- |
+ |
24 |
12.58 |
161.02417 |
161.02442 |
1.53 |
- |
- |
- |
- |
C9 H6 O3 |
umbelliferone |
+ |
- |
- |
25 |
13.10 |
703.25956 |
703.26074 |
1.69 |
705.27363 |
705.27530 |
2.36 |
- |
C35 H44 O15 |
glehlinoside B |
+ |
- |
+ |
26 |
13.21 |
519.18677 |
519.18719 |
0.80 |
- |
- |
- |
- |
C26 H32 O11 |
glehlinoside J |
- |
- |
+ |
27 |
13.25 |
685.24752 |
685.25018 |
3.88 |
687.26254 |
687.26473 |
3.19 |
- |
C35 H42 O14 |
glehlinoside F |
- |
- |
+ |
28 |
13.49 |
673.24896 |
673.25018 |
1.81 |
675.26304 |
675.26473 |
2.51 |
- |
C34 H42 O14 |
glehlinoside A |
+ |
+ |
+ |
29 |
14.38 |
699.26414 |
699.26583 |
2.41 |
701.27773 |
701.28038 |
3.78 |
- |
C36H44O14 |
glehlinoside E |
+ |
- |
+ |
30 |
15.30 |
245.08171 |
245.08193 |
0.91 |
247.09583 |
247.09649 |
2.67 |
- |
C14 H14 O4 |
marmesin |
+ |
+ |
+ |
31 |
15.75 |
193.05036 |
193.05063 |
1.41 |
- |
- |
- |
- |
C10 H10 O4 |
ferulic acid |
+ |
- |
+ |
32 |
16.29 |
231.02964 |
231.02990 |
0.83a |
- |
- |
- |
- |
C11 H6 O3 |
psoralen |
+ |
- |
+ |
33 |
18.01 |
303.08717 |
303.08741 |
0.43a |
- |
- |
- |
- |
C15 H14 O4 |
7-O- (3,3-dimethylallyl) |
+ |
- |
+ |
34 |
18.88 |
- |
- |
- |
217.04921 |
217.04954 |
1.49 |
202.02555 |
C12 H8 O4 |
xanthotoxin |
+ |
+ |
+ |
35 |
20.22 |
- |
- |
- |
247.06003 |
247.06010 |
0.28 |
232.03650; |
C13 H10 O5 |
isoimpinellin |
+ |
+ |
+ |
36 |
20.28 |
- |
- |
- |
217.04944 |
217.04954 |
0.44 |
202.02604 |
C12 H8 O4 |
bergapten |
+ |
+ |
+ |
37 |
22.65 |
229.08678 |
229.08702 |
1.02 |
231.10141 |
231.10157 |
0.70 |
- |
C14 H14 O3 |
demethylsuberosin |
+ |
+ |
+ |
38 |
23.25 |
269.08168 |
269.08193 |
0.60 |
- |
- |
- |
- |
C16 H14 O4 |
8-(1,1-dime thylallyl)- 5-hydroxypsorolen |
+ |
+ |
+ |
39 |
24.57 |
299.09221 |
299.09250 |
0.96 |
301.10636 |
301.10705 |
2.29 |
- |
C17 H16 O5 |
cnidilin |
+ |
+ |
+ |
40 |
24.60 |
269.08174 |
269.08193 |
0.73 |
- |
- |
- |
254.05898; |
C16 H14 O4 |
alloisoimperatorin |
+ |
+ |
+ |
41 |
25.05 |
269.08169 |
269.08193 |
0.50 |
271.09571 |
271.09649 |
2.87 |
203.03440 |
C16 H14 O4 |
imperatorin |
+ |
+ |
+ |
42 |
25.06 |
- |
- |
- |
339.15904 |
339.15909 |
0.13 |
- |
C21 H22 O4 |
8-geranyloxypsoralen |
- |
- |
+ |
43 |
27.09 |
269.08170 |
269.08193 |
0.86 |
271.09619 |
271.09649 |
1.09 |
203.03364 |
C16 H14 O4 |
isoimperatorin |
+ |
+ |
+ |
44 |
27.10 |
201.01909 |
201.01933 |
1.19 |
203.03368 |
203.03389 |
0.99 |
147.04402 |
C11 H6 O4 |
xanthotoxol |
+ |
+ |
+ |
45 |
28.65 |
259.17014 |
259.17035 |
0.81 |
261.18469 |
261.18491 |
0.83 |
- |
C17 H24 O2 |
panaxydiol |
+ |
+ |
- |
46 |
29.35 |
289.18066 |
289.18092 |
0.49a |
245.18978 |
245.18999 |
0.88 |
- |
C17 H24 O |
panaxynol |
+ |
+ |
+ |
47 |
36.81 |
259.17010 |
259.17035 |
0.53 |
261.18492 |
261.18491 |
-0.05 |
- |
C17 H24 O2 |
falcarindiol |
+ |
+ |
+ |
Table 2: The identified compounds in the test samples
a m/z [M+HCOO]-; + detectable; - not detectable
Sample |
Content (μg/g) |
||||
Xanthotoxol |
Marmesin |
Bergapten |
Imperatorin |
Total content |
|
GR |
13.723 ± 0.212 |
56.689 ± 0.505 |
8.099 ± 0.282 |
10.589 ± 0.236 |
89.100 ± 0.717 |
pGR |
8.782 ± 0.130 |
36.346 ± 0.435 |
4.314 ± 0.216 |
6.598 ± 0.286 |
56.040 ± 0.906 |
peels |
18.403 ± 0.100 |
62.902 ± 0.197 |
7.510 ± 0.180 |
12.057 ± 0.485 |
100.872 ± 0.502 |
Table 3: Contents of four furanocoumarins in the test samples (n = 3, mean ± SD)
Samples |
a |
IC50 (μg/mL) |
||||
10 (μg/mL) |
50 (μg/mL) |
100 (μg/mL) |
200 (μg/mL) |
500 (μg/mL) |
||
GR |
14.66 ± 0.94* |
16.58 ± 1.27* |
20.91 ± 0.34* |
39.58 ± 0.53* |
57.77 ± 0.86* |
461.02 |
pGR |
10.68 ± 0.73* |
14.04 ± 0.70* |
15.83 ± 0.84* |
17.59 ± 1.83* |
28.97 ± 2.62* |
> 500 |
peels |
14.92 ± 0.77* |
22.49 ± 2.81* |
31.25 ± 0.83* |
45.56 ± 1.00* |
65.67 ± 1.79* |
249.88 |
Table 4: DPPH scavenging ratios of the test samples
a Values represent the means ± SD in triplicates; *P< 0.001, compared with the untreated control group
Samples |
a |
IC50 (μg/mL) |
||||
10 (μg/mL) |
50 (μg/mL) |
100 (μg/mL) |
200 (μg/mL) |
500 (μg/mL) |
||
GR |
4.98 ± 1.61* |
10.47 ± 0.37* |
19.48 ± 0.65* |
40.07 ± 1.07* |
75.15 ± 0.16* |
253.25 |
pGR |
3.16 ± 1.73* |
7.52 ± 0.00* |
20.71 ± 2.27* |
35.09 ± 0.03* |
62.31 ± 0.61* |
335.47 |
peels |
7.33 ± 1.31* |
17.94 ± 0.38* |
33.93 ± 0.91* |
57.24 ± 0.13* |
74.54 ± 0.11* |
174.66 |
Table 5: ABTS+ scavenging ratios of the test samples
a Values represent the means ± SD in triplicates; *P< 0.001, compared with the untreated control group
Figure 1: HPLC-FT-ICR MS base peak intensity chromatograms (BPCs) of the test samples in negative-ion mode. A: GR extract; B: pGR extract; C: the peels extract; the compound numbers (1- 47) are the same as those in Table 1
Figure 2: The chemical structures of detected compounds. The compound numbers (1- 47) are the same as those in Table 1; Glc: β-D-glucopyranosyl, Api: β-D-apiofuranosyl, Rha: β-D-rhamnopyranosyl
Figure 3: The HPLC chromatograms of the test samples. a: a mixture of standard compounds; b: GR extract; c: pGR; d: the peels extract; 1:Xanthotoxol; 2: Marmesin; 3: Bergapten; 4: Imperatorin
Figure 4: Antiproliferative effects of the test samples. A: HT-29 cellular viability; B: HL-7702 cellular viability; Results are expressed as the means ± SD of three independent experiments; *p < 0.01, compared with untreated control group
Figure 5: Effects of the test samples on LPS-induced NO production in RAW264.7 cells. Results are expressed as the means ± SD of three independent experiments; *p < 0.001, compared with LPS group
Tables at a glance
Figures at a glance