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Identification of chemical constituents in Scrophulariae Radix by HPLC-IT-TOF-MS

玄参化学成分的HPLC-ESI-IT-TOF-MS分析



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IdentificationofchemicalconstituentsinScrophulariae
RadixbyHPLCITTOFMS
WANGJingzhe,XUFeng,LIUZhen,MALiman,SHANGMingying,LIUGuangxue,CAIShaoqing
(SchoolofPharmaceuticalSciences,PekingUniversity,Beijing100191,China)
[Abstract] ThisexperimentwasperformedtoestablishaqualitativeanalysisonchemicalconstituentsofScrophulariaeRadixby
HPLCESIITTOFMS.TheanalysiswasconductedonaC18column(Kromasil1005,46mm×250mm,5μm)with01% formic
acidacetonitrileasthemobilephaseforgradientelution;ESIionsourcewasusedformassspectra,anddatawerecolectedinnegative
andpositivemodesTheresultsshowedthat64compoundsfromScrophulariaeRadixhadbeenidentifiedbyanalyzingnegativeionmass
dataincludingelementcompositionandbycomparingwithdatafromliteratureTwonewcompounds(4hydroxy6Omethylcatalpol
andacetylangorosideC)andseventeenknowncompoundsweredetectedfromScrophulariaeRadixforthefirsttimeSeventeenknown
compoundsincludedtwelveiridoidglycosides,threephenylpropanoidglycosidesandtwootherkindcompoundsThisstudywilprovide
chemicalbasisforelucidationoftheefectivesubstanceintheScrophulariaeRadix.
[Keywords] ScrophulariaeRadix;HPLCESIITTOFMS;iridoidglycosides;phenylpropanoidglycosides
doi:10.4268/cjcmm20160717
  
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Table1 MSdataofidentifiedconstituentsfromScrophulariaeRadix
No tR/min —ïó [M+HCOO]-/m/zÄÔ/ppm –+æ-žï/m/z _V˜®¼
P1 14990C15H22O10 4071177 -442  4071177[M+HCOO]-,3611138[M-H]-,1990617
[M-H-glucosyl]-,1810504[M-H-glucosyl-H2O]-,
1690520[M-H-glucosyl-HCHO]-,1630426[M-H-glu
cosyl-2H2O]-,1510504[M-H-glucosyl-H2OHCHO]-
catalpol[14]
P2 20498C16H26O11 4391438 -433  4171382[M+Na]+,4391438[M+HCOO]-,3931369
[M-H]-,2310888[M-H-glucosyl]-,2130782[M-H-
glucosyl-H2O]-,1950626[M-H-glucosyl-2H2O]-,
1830616[M-H-glucosyl-HCHO]-,1630452[M-H-glu
cosyl-2H2OCH3OH]-
4hydroxy6Omethylcatalpol
P3 20757C15H22O9 3911230 -409  3691146[M+Na]+,3911230[M+HCOO]-,3451208
[M-H]-,1830671[M-H-glucosyl]-,1650671[M-H-
glucosyl-H2O]-,1350581[M-H-glucosyl-H2OHCHO]-
aucubin[15]
P4 21298C21H32O14 5531747 -488  5531747[M+HCOO]-,5071705[M-H]-,3451203
[M-H-glucosyl]-[aucubin-H]-,3271006[M-H-glu
cosyl-H2O]-
6Oβglucosylaucubin[16]
P5 21782C21H32O14 5531761 -235  5311640[M+Na]+,5531761[M+HCOO]-,5071681
[M-H]-,3230981[2glucosyl-H]-,1830330[M-H-
2glucosyl]-[aucubinHglucosyl]-,1790563[glucose-H]-,
1610463[glucosyl-H]-
6′Oβglucosylaucubin[17]
P6 22240C15H26O10 4111490 -438  3891418[M+Na]+,4111490[M+HCOO]-,3651440
[M-H]-[hargagideH+2H]-,3471288[M-H-H2O]-,
2030935[M-H-glucosyl]-,1790567[glucose-H]-,
1610481[glucosyl-H]-
dihydroharpagide[18]
P7 24748C21H32O14 5531761 -235  5531761[M+HCOO]-,5071677[M-H]-,3451133[M-
H-glucosyl]-,3271006[M-H-glucosyl-H2O]-,1790575
[glucose-H]-
6Oβglucosylaucubinisomer
P8 25407C21H32O14 5531750 -434  5531750[M+HCOO]-,5071746[M-H]-,3230940
[2glucosyl-H]-,1830747[M-H-2glucosyl]-,1790448
[glucose-H]-
 6′Oβglucosylaucubin
isomer
P9 25523C15H24O8 3771427 -689  3551284[M+Na]+,3771427[M+HCOO]-,3311348
[M-H]-,1790543[glucose-H]-
ningpogosideA[19]
P10 25815C21H32O14 5531756 -325  5531756[M+HCOO]-,5071686[M-H]-,3451176
[M-H-glucosyl]-,1830788[M-H-2glucosyl]-,1790702
[glucose-H]-
6Oβglucosylaucubinisomer
P11 26140C21H32O14 5531760 -253  5531760[M+HCOO]-,5071710[M-H]-,3230966
[2glucosyl-H]-
 6′Oβglucosylaucubin
isomer
P12 26315C21H34O15 5711881 018  5492845[M+Na]+,5711881[M+HCOO]-,5251828
[M-H]-,3631358[M-H-glucosyl]-[hargagide-H]-,
3451142[M-H-glucosyl-H2O]-,1830663[M-H-
2glucosyl-H2O]-,1650560[M-H-2glucosyl2H2O]-
6Oβglucosylharpagide[20]
P13 26698C21H32O14 5531749 -452  5531749[M+HCOO]-,5071689[M-H]-,3230959
[2glucosyl-H]-,1610563[glucosyl-H]-
 6′Oβglucosylaucubin
isomer
P14 26982C15H24O10 4091348 -098  4091348[M+HCOO]-,3631304[M-H]-,2010781
[M-H-glucosyl]-,1830669[M-H-glucosyl-H2O]-,
1650604[M-H-glucosyl-2H2O]-,1570574[M-H-glucosyl
-CH3CHO]-,1390425[M-H-glucosyl-H2OCH3CHO]-
harpagide[15]
P15 27215C21H34O15 5711885 088  5491856[M+Na]+,5711885[M+HCOO]-,5251825
[M-H]-,3230938[2glucosyl-H]-,2010740[M-H-
2glucosyl]-,1830714[M-H-2glucosyl-H2O]-,1790593
[glucose-H]-
6′Oβglucosylharpagide[21]
·9521·
2016
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41
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Vol41,No.7 April,2016
,*

No tR/min —ïó [M+HCOO]-/m/zÄÔ/ppm –+æ-žï/m/z _V˜®¼
P16 27598C21H34O15 5711869 -193  5491792[M+Na]+,5711869[M+HCOO]-,5251792
[M-H]-,3631648[M-H-glucosyl]-,3230946[2gluco
syl-H]-,2010760[M-H-2glucosyl]-,1830714[M-H-
2glucosyl-H2O]-
  6′Oβglucosylharpagide
isomer
P17 28290C21H34O15 5711862 -315  5491669[M+Na]+,5711862[M+HCOO]-,5251796
[M-H]-,3631246[M-H-glucosyl]-,3230954[2gluco
syl-H]-,2010800[M-H-2glucosyl]-,1830686[M-H-
2glucosyl-H2O]-
  6′Oβglucosylharpagide
isomer
P18 28790C16H24O10 4211335 -404  3991205[M+Na]+,4211335[M+HCOO]-,3751286
[M-H]-,2130757[M-H-glucosyl]-,1950673[M-H-
glucosyl-H2O]-,1830677[M-H-glucosyl-HCHO]-,
1630433[M-H-glucosyl-H2OCH3OH]-,1510461[M-
H-glucosyl-HCHOCH3OH]-
6Omethylcatalpol[15]
P19 29207C16H26O10 4231490 -425  4011377[M+Na]+,4231490[M+HCOO]-,3771424
[M-H]-,2150921[M-H-glucosyl]-,1970828[M-H-
glucosyl-H2O]-,1790696[glucose-H]-,1610484
[glucosyl-H]-
3,4dihydromethylcatalpol[22]
P20 30373C21H28O13 48714351) -452  4871435[M-H]-,1790376[cafeicacid-H]-,1350577
[cafeicacidHCO2]-
cistanosideF[15]
P21 30848C16H24O9 4051381 -518  3831260[M+Na]+,4051381[M+HCOO]-,3591317
[M-H]-,1970837[M-H-glucosyl]-,1650488[M-H-
glucosyl-CH3OH]-,1350489[M-H-glucosyl-CH3OH
HCHO]-
6Omethylaucubin[23,24]
P22 32373C21H28O14 50313771) -576  5031377[M-H]-,3411034[sucrosylH+H2O]-,
3230779[sucrosylH+H2O]-,1790374[cafeicacid-H]-,
1610343[cafeoyl-H]-
  6OcafeoylβDfructo
furanosyl2αDglucopyrano
side[25]
P23 35132C26H40O16 60722621) 296  6072262[M-H]-,4611635[M-H-rhamnosyl]-,
4431561[M-H-rhamnosyl-H2O]-,3111071[M-H-rh
amnosylarabinosyl-H2O]-,1613469[glucosyl-H]-
deferuloylangorosideC[26]
P24 35415C21H28O13 48714761) 390  5111261[M+Na]+,4871476[M-H]-,1630375[M-H-
sucrosyl]-,[cumaricacid-H]-,1451054[cumaroyl-H]-
6Opcoumaroylsucrose[25]
P25 35840C18H26O10 40114471) -150  4471525[M+HCOO]-,4011447[M-H]-,2691014[M-
H-arabinosyl]-,1610523[glucosyl-H]-
 phenylmethyl6OαLara
binopyranosylβDglucopyr
anoside[27]
P26 36823C22H30O14 5631604 -249  5411503[M+Na]+,5171560[M-H]-,5631604
[M+HCOO]-,1930560[ferulicacid-H]-、[M-H-sucros
yl]-,1750414[feruloyl-H]-
6′Oferuloylsucrose[28]
P27 40065C30H38O16 65320921) 077  6532092[M-H]-,4911566[M-H-glucosyl]-,2910847
[M-H-catalpol]-
premnacorymbosideB[29]
P28 40273C19H28O10 4611660 -108  4611660[M+HCOO]-,4151591[M-H]-,1490397[ara
binosylH+H2O]-[M-H-phenethylglucosyl]-
  phenethyl βprimevero
side[30]
P29 40648C24H30O13 52516251) 209  5491499[M+Na]+,5251625[M-H]-,5071513
[M-H-H2O]-,3230771[cafeoyl+glucosyl-H]-,1610219
[cafeoyl-H]-
6″Ocafeoylharpagide[31]
P30 41265C27H38O17 6792089 -029  6572172[M+Na]+,6792089[M+HCOO]-,6331985
[M-H]-,4851475[M-H-cinnamoyl-H2O]-,3230864
[sucrosyl-H]-[M-H-cinnamicacidglucosyl]-
  6OcinnamoylDfructo
furanosylOglucopyrano
side[31]
P31 41523C21H28O12 5171537 -503  5171537[M+HCOO]-,4711471[M-H]-,3230935[su
crosyl-H]-,[M-H-cinnamicacidglucosyl]-,1610557[glu
cosyl-H]-
sibiriosideAisomer
P32 44123C24H30O12 50916521) -255  5331607[M+Na]+,5091652[M-H]-,3070789[cour
maroyl+glucosyl-H]-,1630425[courmaricacid-H]-,
1610612[glucosyl-H]-
  8Ocourmaroylharpagide
isomer
·0621·
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GÝ_–—±
HPLCESIITTOFMS
—à
,*

NotR/min —ïó [M+HCOO]-/m/zÄÔ/ppm –+æ-žï/m/z _V˜®¼
P3344782C29H36O15 62319591) -353  6471903[M+Na]+,6231959[M-H]-,4611606[M-
H-cafeoyl]-,3151036[M -H -cafeoylrhamnosyl]-,
2970946[M-H-cafeoylrhamnosyl-H2O]-
acteoside[15]
P3445848C21H28O12 5171543 -387  5171543[M+HCOO]-,4711503[M-H]-,3230953[su
crosyl-H]-、[M-H-cinnamoylglucosyl-H2O]-,1610557
[glucosyl-H]-
sibiriosideA[15]
P3546198C35H46O19 76925501) -143  7692550[M-H]-,5932011[M-H-ferulicacid+H2
O]-,4471468[M-H-ferulicacid+H2Oarabinosyl]-,
4611559[M-H-feruloylrhamnosyl]-,4291368[M-H-fer
uloylrhamnosyl-H2O]-
scrophulosideB1[32]
P3648323C29H36O15 62319451) -578  6231945[M-H]-,4611588[M-H-cafeoyl]-,3150980
[M-H-cafeoylrhamnosyl]-
isoacteoside[33]
P3748473C22H30O13 5471645 -420  5251321[M+Na]+,5471645[M+HCOO]-,5011619
[M-H]-,3230915[sucrosyl-H]-[M-H-methylcinnamic
acid]-
sibiriosideB[34]
P3848915C35H46O19 76925331) -364  7692533[M-H]-,5932015[M-H-feruloyl]-,4471398
[M-H-feruloylarabinosyl]-
scrophulosideB1isomer
P3949615C24H30O13 5711679 193  5491589[M+Na]+,5251624[M-H]-,3812213,
4431637,3230795[cafeoyl+glucosyl-H]-,2010800[M-
H-cafeoylglucosyl]-,5711679[M+HCOO]-,1829433
[M-H-cafeoylglucosyl]-,1790356[cafeicacid-H]-,
1610223[cafeoyl-H]-
 6″Ocafeoylharpagideiso
mer
P4051857C30H40O16 7012266 -456  7012266[M+HCOO]-,6552184[M-H]-,5071673[M-
H-cinnamoyl-H2O]-,3271006[harpagideH-H2O]-[M-
H-cinnamoylglucosyl2H2O]-
  6OαDgalactopyranosyl
harpagosideisomer
P4152548C36H48O19 78326971) -255  8072765[M+Na]+,7832697[M-H]-,6072179[M-
H-feruloyl]-,5892677[M-H-feruloyl-H2O]-,4611592
[M-H-feruloylrhamnosyl]-,3291231[M-H-feruloylrham
nosylarabinosyl]-
angorosideC[15]
P4252890C30H38O15 63721211) -267  6372121[M-H]-,4751766[M-H-cinnamoyl]-,
4611537[M-H-cinnamoylCH2]-,3291247[M-H-cin
namoylrhamnosyl]-
cistanosideC[35]
P4353115C36H48O19 78327011) -204  7832701[M-H]-,6072177[M-H-feruloylrhamnosyl]-,
5892087[M-H-feruloyl-H2O]-,4611591[M-H-feruloyl
rhamnosyl]-,4431566[M-H-feruloylrhamnosyl-H2O]-,
3291149[M-H-feruloylrhamnosylarabinosyl]-
angorosideCisomer
P4453290C24H30O12 50916591) -118  5331662[M+Na]+,5091659[M-H]-,3470936[M-
H-glucosyl]-,3290949[M-H-glucosyl-H2O]-,1830714
[harpagideHglucosyl-H2O]-,1630420[courmaricacid-
H]-
8Ocourmaroylharpagide[15]
P4553440C32H40O17 69521901) -043  6952190[M-H]-,5331622[M-H-glucosyl]-,3330949
[M-H-glucosyl-catalpol]-,1630414[courmaricacid-
H]-,1449913[courmaroyl-H]-
scrophulosideA2[32]
P4653765C32H40O17 69521951) 029  6952195[M-H]-,5331667[M-H-glucosyl]-,3331035
[M-H-glucosyl-catalpol]-,1450289[courmaroyl-H]-
scrophulosideA2isomer
P4753823C25H32O13 53917611) -167  5631732[M+Na]+,5391761[M-H]-,3451247[M-
H-feruloyl-H2O]-[harpagideH-H2O]-,1930518[ferulic
acid-H]-,1830714[harpagideHglucosyl-H2O]-,1750458
[feruloyl-H]-
6″Oferuloylharpagide[31]
P4853998C30H38O15 63721271) -173  6372127[M-H]-,4611601[M-H-cinnamoylCH2]-,
3151036[M-H-cinnamoylrhamnosylCH2]-
cistanosideCisomer
P4954057C30H40O16 7012271 -385  6792184[M+Na]+,7012271[M+HCOO]-,6552213
[M-H]-,5071651[M-H-cinnamoyl-H2O]-,3451165
[M-H-cinnamoyl-H2Oglucosyl]-,[harpagideH-H2O]-
 6OαDgalactopyranosyl
harpagoside[15]
·1621·
2016
y

z ³
41
ų

}
Vol41,No.7 April,2016
,*

NotR/min —ïó [M+HCOO]-/m/zÄÔ/ppm –+æ-žï/m/z _V˜®¼
P5054557C30H40O16 7012317 271  7012317[M+HCOO]-,6552191[M-H]-,5071663[M-
H-cinnamoyl-H2O]-
 6OαDgalactopyranosyl
harpagosideisomer
P5155223C38H50O20 8712848 -333  8492618[M+Na]+,8712848[M+HCOO]-,8252728
[M-H]-,7832651[M-H-acetyl]-,7652528[M-H-ace
tyl-H2O]-,6492128[M-H-feruloyl]-,6072319[M-H-
feruloylacetyl]-,5892035[M-H-feruloylacetyl-H2O]-,
4611553[M-H-feruloylacetylrhamnosyl]-,4431438[M-
H-feruloylacetylrhamnosyl-H2O]-,3291247[M-H-feru
loylacetylrhamnosylarabinosyl]-
acetylangorosideC
P5256282C31H40O15 6972365 229  6752321[M+Na]+,6972365[M+HCOO]-,6512288
[M-H]-,5051616[M-H-rhamnosyl]-,4751781[M-H-
feruloyl]-,4571662[M-H-feruloyl-H2O]-,3291171[M-
H-feruloylrhamnosyl]-
cistanosideD[15]
P5357828C24H30O11 5391745 -464  5171632[M+Na]+,5391745[M+HCOO]-,4931693
[M-H]-,3451138[M-H-cinnamoyl-H2O]-,2010733[M-
H-cinnamoyl-H2Oglucosyl]-,1830726[M-H-cinnamoylglu
cosyl2H2O]-,1790765[glucose-H]-,1650540[M-H-cin
namoylglucosyl3H2O]-,1610583[glucosyl-H]-
harpagosideisomer[36]
P5458403C25H32O12 5691865 -193  5471826[M+Na]+,5691865[M+HCOO]-,5231836
[M-H]-,3451132[M-H-methoxycinnamoyl-H2O]-
 8Omethoxycinnamoyl
harpagideisomer
P5558878C30H40O16 7012284 -200  7012284[M+HCOO]-,6792350[M+Na]+,6552165
[M-H]-,5071682[M-H-cinnamoyl-H2O]-,3451175
[M-H-cinnamoyl-H2Oglucosyl]-,[harpagideH-H2O]-
 6OαDgalactopyranosyl
harpagosideisomer
P5659387C31H40O15 6972377 402  6972377[M+HCOO]-,6512279[M-H]-,4731681[M-
H-methoxylcinnamoyl-H2O]-,3271063[M-H-methoxylcin
namoyl-H2Orhamnosyl]-[aucubinH-H2O]-
undulosideⅢ[37]
P5760853C30H40O16 7012277 -299  6792027[M+Na]+,7012277[M+HCOO]-,6552208
[M-H]-,5071671[M-H-cinnamoyl-H2O]-,3411092
[2glucosylH+H2O]-,3231031[2glucosyl-H]-,1790702
[glucose-H]-
 6″OαDgalactopyrano
sylharpagoside[38]
P5861845C24H30O11 5391769 -019  5171644[M+Na]+,5391769[M+HCOO]-,4931712
[M-H]-,3451148[M-H-cinnamoyl-H2O]-,1830971
[M-H-cinnamoyl-H2Oglucosyl]-,1650599[M-H-cin
namoyl2H2Oglucosyl]-,1610571[glucosyl-H]-
harpagoside[15]
P5964693C25H32O12 5691844 -562  5471735[M+Na]+,5691844[M+HCOO]-,5231772
[M-H]-,3451138[M-H-methoxylcinnamoyl-H2O]-
1790503[glucose-H]-,1650586[M-H-methoxylcinnamoyl
2H2Oglucosyl]-
 8Omethoxycinnamoyl
harpagide[15]
P6070110C33H42O17 7552370 -450  7552370[M+HCOO]-,7092307[M-H]-,6491994[M-
H-acetyl-H2O]-,5471845[M-H-glucosyl]-,5171683
[M-H-glucosyl-CH2O]-
scrophulosideA5[32]
P6172058C35H44O17 73525501) 598  7592325[M+Na]+,7352550[M-H]-,6932347[M-
H-acetyl]-,6512325[M-H-2acetyl]-,5051685[M-H-
2acetyl-rhamnosyl]-,3291179[M-H-2acetyl-rhamnosyl
feruloyl]-
diacetylmartynoside[39]
P6278758C43H50O19 9152879 -535  9152879[M+HCOO]-,8692947[M-H]-,7072281[M-
H-glucosyl]-,6472078[M-H-glucosyl-acetyl-H2O]-,
5931957[M-H-acetyl-H2Omethoxylcinnamoyl-H2O-
C3H4O]-
scrophulosideA4[40]
P6379458C42H48O18 8852795 -316  8852795[M+HCOO]-,8392803[M-H]-,6772158[M-
H-glucosyl]-,6171974[M-H-glucosyl-acetyl-H2O]-,
5631883[M-H-acetyl-H2Omethoxylcinnamoyl-H2O-
C3H4O]-
scrophulosideB4[41]
·2621·
I:Šè

GÝ_–—±
HPLCESIITTOFMS
—à
,*

NotR/min —ïó [M+HCOO]-/m/zÄÔ/ppm –+æ-žï/m/z _V˜®¼
P6490817C20H28O2 29920011) -535  3012119[M+H]+,2992001[M-H]-,2271082[M-H-
C4H8O]-
sugiol[40]
  
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rã˜
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2 
M‰ÏQMBCèêĖ—Ý7‹š

a
harpagide
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Fig2 MSfragmentationruleofcyclopentanetypeiridoidglycosides(egharpagide)
  
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m‘¬£ê
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š

ì
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·3621·
2016
y

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Vol41,No.7 April,2016
P4(6Oβglucosylaucubin)ÊÓ¦—Õ÷°(P7¬
P10),P5(6′Oβglucosylaucubin)ÊÓ¦—Õ÷°
(P8,P11,P13)、P21(6Omethylaucubin)
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aucubin
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Fig3 MSfragmentationruleofcyclopentenetypeiridoidglycosides(egaucubin)
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·4621·
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Table2 Distributionofthefirstidentifiedconstituentsfrom
ScropharariaeRadixinplants
No
_V˜ —‹
P4 6Oβglucosylaucubin
GÝ&[tÐÇ
[16]
P5 6′Oβglucosylaucubin
GÝ&¡Ç
[17]
P6 dihydroharpagide
GÝ&GÝÇ
[18]
P12 6Oβglucosylharpagide
Œ_&
[20]
P15 6′Oβglucosylharpagide
Œ_&
[21]
P19 3,4dihydromethylcatalpol
GÝ&GÝÇ
[22]
P21 6Omethylaucubin
GÝ&FÍÇ
[2324]
P25  phenylmethyl6OαLarabinopyranosyl
βDglucopyranoside
Ÿ©&
[27]
P26 6′Oferuloylsucrose
Xí&
[28]
P27 premnacorymbosideB
GÝ&GÝÇ
[29]
P28 phenethylβprimeveroside
GÝ&FÀÇ
[30]
P35 scrophulosideB1 GÝ&GÝÇ[32]
P70 sibiriosideB
GÝ&ý¢Ç
[34]
P37 scrophulosideA2 GÝ&GÝÇ[32]
P56 undulosideⅢ
GÝ&[tÐÇ
[37]
P60 scrophulosideA5 GÝ&GÝÇ[32]
P61 diacetylmartynoside
GÝ&GÝÇ
[39]
3 
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

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βglucosylharpagide), P15 (6′Oβglucosylharp
agide),P25(phenylmethyl6OαLarabinopyranosyl
βDglucopyranoside),P26(6′Oferuloylsucrose),
P51(acetylangorosideC),
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P2
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P51
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àïGÝ¿mˆ,ÉO

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„q=

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