J
(2015^?°'''*^ **'"'''«"''•'^™''"''*° ""''"''>•''""•'-°"''''"-'^*^™^ • ' • ^ ' " " " ' ' ™ S D...AJ11K D.SimhpPK.
\r,^ ^aemp/ma pan,'iflora (Zingil^eiaeeae) A new record m ttie Flora of Manipyr. Internaiionol Journal of Innovative iW *-™lf i"f™«""S « Technology. 3(7). M l « 5 . 5. Picliearuioonihon P md Koomm S. (200S). Notes on the genus
; I I M T T " ' ^ ' " B I ' " « ' » « ) '" Th-ilmi Joumal cf Thai Tradilional i .ilternatne Medicine UD. 11-51. 6. Sirliugs. P.
. '' tofti r ° u 7 ? , " " J " " * * ™ (Zingiberaceae) in Tliaitod. Thai Fore.t Bullenn. 19. 1-15 7. Tap N (2006).
* c«rric^um%. " " » K.»"rce, Survey. In Nen;en TUnong Dong (lulilors). Sludv o„ Herbal .tiedicine. Gradn.ie
* . N 2 n . i n „ ? , T n " " . , \ u' ' " ' " " ' • " « "•""'• " " ° ' - " ^ «• "">» ><- N. (2007). .tieihod, for Studying Botany.
a c * r of oZ^.„H ; ' " V ° " ^ " " ' ° ' - ' ' • " ' • ^ ™ ^ " " ' ^- Subhadhirasatol S.. Knmmcc S (2008) Anfi-allergfc S ™ r f ' r t ™ f c ; ^ - f » " e l " ™ ~ ' C.. Cheenpracha S (2009). Anti-inHamra.ion effects of compounds Irom
•'* MatsTmura T T l S v °'""'""'''" P-'d'roie. Food Chemistry, 115(2). 534-538. 11. Azuma T. Kava»o S I . i-t ^ S Z e n " fom Z w • « ' ^ ' ' ° ° ^ "" ' ' ' " " • An"»"»S«ii'= " J (.lpha)-glucos,dase inhibiior,. etfecu of ht Z n ™ n p T „ ! Z t o c h T t r . " ' ' " " ' " " ' ' • '-'• ' " - * " l^- >^''«"«k»n. ' • Tong-Un T. MucSmapura S .
!^ subTea™ ra,; 1 " , • ,™'"'™P" * • (^(Xni Aoi-stress effecis oi K^empfcr.a parviilora m immobilization f= i ^ T l ^ L S 71niTt?'nrLi'T''"'°<^ "^ ^"'"'"^ «"'• " - " " • Sa».sdee P., Sabphon C.
•'•« Aniimicrobial mivinTf the e r h Z ; , ; ; i l , . / 1-1. Kummee S. Te.trakul S. Subhadhirasakul S (2008).
k , Jouroal of Seen, OJTMT TmimJ^rtt", T ""f. " T " " "'"••'""' ""'I'"'-'' ^'"g^'-'frin t l'ha™«:otogie.Ucm,iyofZl5-.r«™ 1 . ^"""'f"'«^ V. Tiamxo^en S. Sripanidkulchai BO. (2009).
* Cancer PrJeniioTmllfiSSifiTS'^ T'i ' * " " " """"^ ""'' ' ' " " " " " ' "" '""'• •-'™" ' " C ' •'•>"""' " /
Pbolpramfol C. ,2008), . ^ Z ' c e ^ r ^ f ' i L L ' a c ' ^ r ^ ' m ' l e „?b?etloVe:^^^^^^^^^^^^^ " " T " ' ; i A "iua^iiiciidionoiasirom A.ac/np/enaporMy;oro, Fiioierapia. 7![1). 89-92.
Journal of Medicinal Materials, 2016, Vol. 21, No. 5 (pp.297 - 304)
METABOLIC PROFILING OT ARABIDOPSIS THALIANA COL. 0
;' Dao Thi Thanh Hien *
' Hanoi Pharmacy University. Vietnam 'Corresponding aulhor: [email protected]
(Received March, 30^ 2016)"
Summary'
Metabolic Proniing oi Arabidopsis thaliana Col 0
Metabolic proniing of ^r„i,^o^„ „ , „ , „ „ Col 0 . a s m,cs.,g.,cd Mettiaool-d, showed as the best direc, extraction
solvent for ihai purpose. Four navonoids six aminn ir,H- i^,= r,h t -i -, exiraction metabolites weteidemiliedb, using NMR s p e l s c Z P^™>'P™P»9.ds and otber pnmai, .nd seconda,,.
K.r»ord., NMR. Melaholic profiling. Flavonoid Enirocion m.ihod. Arabidop.i, iholiana
A ^ ^ Z a i l a a a .as . e c c . e . „ S S r o ^ r X T t t ^ ^ ^ ' ^ "
extremely popular , „ „ . . s , s t e . Tor stod,i„g of solars, ^ i l t . r i : ™ „ ^ a „ r a ° r j ^ ^ plant biology. Analysis of whole plant alcohols Several himrfr^H . * ntetabolotnes is a difficult task due to the'hi^ robustlv an r i ,y detlTe, P h X t " " H^
number and diversity of primary and secondao^ are ubiquitous constiru mfof h t " 1 ? T t ntetabolites present in plant tissues [1], [2], [3], are seeondaty I t r t o l , / o f t l a ^ i f
Large nuntbers of ntetabolites of Arabidopsis involved inTefen a « „ t u - ^"!i ' have been identified [4], [5], The ntain focus of or aggression b y T a J Z T " ; " ' ' ' ' ' ' ' " "
Utese .ostly gas chrontatography ( G C . n i a s s ^ ^ o ^ " r / S ^ Ire^ Z l ^ ^ ^
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' ' 297
glycosides [6], but quercetin glycosides can also accumulate after exposure to UV radiation [7].
The major anthocyanin in A. timliana has a cyanidin core with four attached sugars [8]. Some fiavonoids from green tissues of ^ . thaliana have been fully structurally characterized as kaempferol 3^3.^[5.I>glucopyranosy l( 1 —»6)D-glucopyranoside]
-7-0-a-L-rhamnopyranoside (1), kaempferol 3- O^-D-glucopyranoside-T-O-a-L-rhamnopyranoside (2), and kaempferol 3-0-a-l-rhamnopyranoside- 7-0-a-L-rhamnopyranoside (3) were identified [9], [10]. Most of studying about flavonoid characterization have done by analytical procedures for identification of the isolated fiavonoids. So an identification of Arabidopsis fiavonoids in plant crude extract still needs to be developed for a metabolic profiling.
The aims of this study are using NMR to identify Arabidopsis thaliana Col. 0 metabolites, focusing on phenolic compounds in plant crude extract. The good extraction method for this purpose also was developed in this article.
2. Methods and Materials Plant materials
Arabidopsis thaliana ecotype Col-0 seeds were supplied by Institute of Biology Leiden (IBL). Seeds were sown on a 2:1:1 (by vol.) mixture of vermiculite, peat moss, and perlite.
The pots were placed at 4 "C for 4 d in the dark and transferred to normal growth conditions.
Plants were grown at 23 °C under long day conditions (16/8 h light/dark cycle). After 4 weeks plants are ready to collect leaves for experiment
General procedures Extraction:
500 ml of methanol was added to 256 g of dried and ground Arabidopsis thaliana leaves and ultrasonicated for 30 minutes and then vacuum filtered. Repealed for 3 times and all the supernatant were pooled and dried using rotary evaporator The dried extract was redissolved in IOO ml of deionized water and partitioned with different solvents like «-hexane, chloroform, and
«-butanol All the fractions were separately dried by rotary evaporator and stored at 4°C until further use.
Sample fractionation:
The n-butanol extract (1.2 g) was selected for fractionation as high fiavonoids content was expected in this fraction. Sephadex column LH- 20 (145 cm length x 16 mm diameter) was used for sample fractionation with 100% methanol asa mobile phase. Total 84 fractions were collected of 5 ml each. TLC indexing was performed fw every fourth fraction and observed under 254 nm and 366 nm. The solvent system for TLC indexing composed of ethyl acetate, fonnic acid, acetic acid, and water, in the ratio of 100:11:11:27 (v/v/v/v), respectively. The fractions showed same pattems under UV were pooled together and seven combined fractions (from A to G) were obtained Fraction A contained fractions from 1-19, B from 20-30, C from 31-34, D from 35-38, E ftom 394S, F ftom 49-71, and G from 72-84. ' H NMR analyses were performed for all the groups and on the basis of high flavanoid signals, fraction C, D, E, and F were selected for further purifications,
HPLC analysis:
The selected fractions were separated using an Agilent 1100 series HPLC with Variable Wavelength Detector (VWD, Agilent, Waldbronn, Germany). A semi-preparative reversed phase column (Phenomenex Luna 5n CIS; 250 x 10 mm, 5[i) was used for separations, with a gradient solvent of O.I % formic acid with water and O.I % formic acid with methanol. The gradient starts from water-methanol (60:40) for the first 30 minutes, then shifted towards 20:80 for two minutes and then again shifted to 60:40 for the final eight minutes wilh the flow rate of 2 ml/min. After the HPLC analyzing, different sub- fractions were combined. For fraction C, sub- fractions 5-7 were combined as C l , 10-13 as 02, and 14-16 as C3. For fraction D, sub-fractions 5- 7, 10-16, and 22-28 were combined as Dl, D2, and D 3 , respectively. Similarly, for fraction E, sub-fractions 5-7, 13-14, 22-26, and 28-32, were combined as E l , E2, E3, and E4. Likewise for fraction F, sub-fractions 24-29, 33-36, 40-43, and 46-48, were pooled and designated as F l , F2, F3, and F4 correspondingly. All the pooled sub- fractions were dried using rotary evaporator and then analyzed by ' H NMR spectroscopy.
298 Journal of Medicinal Materiab, 2016, Vol. 21, No. 5
Sample preparation for NMR measurements:
• 25 mg of freeze-dried material were transferred to a micro-centrifiige tube before adding 600 jil
; of methanol-rf,. Vortexes for 2 minutes and sonicated for 20 minutes then eentrifiigation at 13,000 rpm for 5 mmutes. 500 ^i.l of the
; supematant were transferred into 2ml micro- centrifuge tubes and were added 250 jiI of KHjPO^ buffer, pH 6.0, containing 0.1%
trimethyl silyl propionic acid sodium salt (w/v).
700 nl of the supematant were then transferred into NMR tubes for analysis.
NMR measuremenis:
'H NMR, 2D J-resolved and 2D-NMR COSY spectra were recorded at 25°C on a Bruker DMX- 500 MHz NMR spectrometer (Bmker, Karlsmhe, Germany).
3. Results and discussions Extraction method development:
In order to optimize a good extraction solvent system, difference ratio of water was mixed with methanol, from 0% to 100% (0%, 25%, 50%, 75%, 100%). We found that the solvent MeOH/HjG ratio of I/I (v/v) give divest signal of primary and secondary metabolites in 'H-NMR spectra but the solvent 100% MeOH showed a higher efficiency of phenolic compounds. Thus CD3OD was chosen as direct extraction solvent and D3O was added (30% in total volume) in primary cmde CD3OD extract to get ride of chlorophyll.
Isolation and characterization of Arabidopsis thaliana Col. 0fiavonoids:
Fourteen sub-fractions (Cl-3, Dl-3, El-4, and FM) were analyzed with NMR The results showed that fiavonoids are mainly in sub-fraction F2 and F4. F4 sub-fi-action contains more than one flavonoid so we applied one more HPLC to fractionation F4 and four factions (F4.I, 4.2, 4.3, 4.4) were collected which are contained mainly a single compound in each fraction.
The two-dimensional J-resolved spectmm of the F2 sub-fraction shows four characteristic signals .5 6.84 (H-6, d, J = 2.0 Hz); 3 6.82 (H-8, d,J= 2.0 Hz); S 6.99 (H-3'& 5', d, 7 = 8,4 Hz); S 8.11 (H-2' &6\d,J=9 Hz). This compound was assigned as kaempferol 3-0-[rhamnopyranosyl (|-f2)-glucopyranoside]-7-0-rhamnopyranoside.
The F4.1 fraction shows characteristic signals at S 6.52 (H-6, d, y = 2.0 Hz); S 6.82 (H-8, d, J = 2.0 Hz); 6 7.0 (H-3'& 5', d, 7 = 8.4 Hz); S 8.09 (H-2' & 6', d, J = 9 Hz) which was assigned as kaempferol glycoside i (3-0-glucopyranoside-7- rhamnopyranoside) and the J-resoIve NMR signals of the F4.3 fraction place at d 6.43 (H-6, d, / = 2.0 Hz); S 6.81 (H-8, d. J = 2.0 Hz); S 7.83(H-2'& 6', d,J= 9.0 Hz) 5 7.04 (H-5', d, J = 9 Hz) was elucidated as kaempferol glycoside 2 (3,7-0- dirhamnopyranoside) [5], [1 i]. The NMR spectmm of F4.4 fraction shows typical querctine NMR spectra but we could not identify the attached sugar. For the confirmation of the stmctures, LC-ESI-MS in negative mode was applied to the extract. Major [M-HJ" signals of F2, F4 1. F4.2, F4.4 respectively are m/z 739, 577, 593 and 447 which correspond with the expected molecular weights of kaempferol gljtoside 1 (3-a^ucop>Tanoside-7-ihamnopyranoside), kaempferol 3,7-0- dirhamnopyranoside, kaempfenai 3-0-[rhamnopyranosyI( 1 ->2)-glucopyranoside]- 7-0-rhamnopyranoside and quercetin 3-0- rhamnoside (Fig.l)
NMR signal assignments of Arabidopsis in MeOH crude extract:
Figure 2 shows the 'H-NMR spectrum of the Arabidopsis Col.O. The combined information gathered from 'H-NMR, COSY and ^-resovle spectra and Ihe use of a library of 'H-NMR spectra of reference compounds have allowed an almost complete assignment. Table 1 summanzes the chemical shift information from the 2D-NMR spectra of the identified compounds.
Sugars, organic acid and amino acid present in high field region NMR spectra, between 0.5 to 6.0 ppm (Figure 2 b) In the amino acid region (S 0 8-,5 4.0), alanine, glutamic acid (glutamine), threonine, valine and asparagines (aspartic acid) were identified (Table 1). In organic acids region NMR signals only formic and malic acid can be detectable because the other organic acids have very poor solubility in MeOH. The signals of the temiinal CH3 of choline was identified at ,5 3 23 (s). For sugars, the anomeric proton of fl-glucose at 6 4.58 (d, J = 7.8 Hz), «-g!ucose at ^ 5 i 8 (d y ' 3.7 Hz), sucrose at <5 5 4 (d, y = 4.0 Hz),
Journal of Medicinal Materials, 2016, Vol. 21, No. 5
299
rhamnose at 5 5.62 (d, J - 8.0 Hz), and fmctose at ^4.17 (d, J = 9,0 Hz) were assigned.
Most of signal of fiavonoids and phenylpropanoids are presented in low filed region (6.8-8.2 ppm) has been analyzed. In the aromatic region, the presence of five major doublets with the same coupling constants (d, J = 16.0 Hz) in the range of ^ 6.'i\-d 6.50 indicated the presence of the trans olefinic protons of the phenylpropanoids (Figure 3). This was confirmed that H-8' of the phenylpropanoids correlated with the H-7' protons at S 1.S9-6 1.11 in the COSY spectmm (Figure 4). Five trans- phenylpropanoids forms were elucidated by two dimensional NMR. Those are cafferoyi malate {5 6.32), hydroxyfemloyi malate {S 6.34), coumaroyl malate {S 6.37), synapoyi malate (i5 6.48), synapoyl
glucose {S 6.49) (Figure 3).
The main fiavonoids can be detected in the NMR spectmm of the Arabidopsis Col.O were kaempferol 3 - O - rhamnopyranoside -7-0- riiamnopyranoside, kaempfenDl 3-0-[rhamnopyranosyl ( l - » 2 ) - glucopyranoside]-7-0-rhamnopyranoside, kaempferol 3,7-0-dirhamnopyranoside. Quercetin derivatives are very minor in cmde extract and difficult to detect in NMR spectmm. The H-2' and H-6' chemical shifts of kaempferol 3-0- [rhamnopyranosyl (l-»2)-glucopyranoside]-7-0- rhamnopyranoside differ by approximately 0.2 ppm which can be detected at 8.09 and 8.11 ppm (d, J = 9.0 Hz). The doublet presented at 7.83 ppm (d, y = 9.0 Hz) that were assigned as H-2' and H-6' signal of the kaempferol 3,7-0-di rhamnoside.
Table 1. 'H chemical shifts (J) and couplmg constants (Hz) af Arabidopsis thaliana Col.O metabolites identified by references and usmg ID and 2D NMR spectra (CDjOD-KHjPO., in DjO (pH 6 0)
Chemical shifts (ppm) and coupling conslants (Hz) Amino/oreanic acids
S 1 32(H-5.d.^=6.6Hz)
^l.48(H-3.d.J-7.0Hz) Glutamine (glutamic acid) J2.l2(H-2,m)62.48(H-3,m)
S 1.03 (H-, d. J= 7.8 Hz) 5 1.07(H-. d. J= 7.8 Hzl 6 0.96 (H- ,d,J= 8,0 Hz) S 0.98 (H- . d, J = 8 Q Hz) Asparagines or aspartic acid S 2 8 (m). 2.97 (m)
5 4.32(H2, dd,J=4.0Hz, II Hz) S 2 80 (H3, dd, J = 8 8 Hz, 16.0 Hz) 5 2.96 (H2, dd. J = 3.6 Hz, 16.0 Hz)
Formic acid (formate)
mtm^^
J4.58(H-l,d.J-92Hz)•j^mmmm^^m^^^i
^5.18(H-l,d,y=4 0Hz)
J 5.62 (H-l. d. 7= 8 0 Hz) ^ 4 (H-l.d.J^Hz)
Kaempferol 3-0-glucopyranoside-7- rhamnopyranoside
Kaempferol 3,7-0- dirhamnopyranoside
g4.17(H-l.d.J^9.0H
d 6.52 {H-6, d. J= 2.0 Hz) 3 6.82 (H-8, d. J= 2.0 Hz) S 7.0 (H-3'& 5', d, J = 8 4 Hz) S 8.09 (H-2' & 6'. d,J=9 9 Hz)
Kaempferol 3-0 -rhamnosyl (12) glucoside-7-0- rhamno pyranos ide
3 6,43 (H-6, d, J = 2.0 Hz) 6 6.81 (H-8, d, J = 2.0 Hz) S 7.83(H-2'&
6', d. J= 9.0 Hz) 5 7.04 (H-5'. d. J^ 9 Hz)
iraas -S-hydroxyfemoyl malate
S 6.84 (H-6, d,J= 2.0 Hz) S 6.82 (H-8, d.J = 2.0 Hz) S 6.99 (H-3'&
5'. d, J = 8.4 Hz) S 8.11 (H-2' & 6'. d, J= 9 Hz) Irons caffeoyl malate S 6.36 (H-8, d 16 Hz) S 7.66 (H-7, d, ^ 1 6 Hz)
S 6.32 (H-8, d 16 Hz) 6 7.66 (H-7. t
d 4.1 {H-2, dd, J= 2.0 Hz, 13Hz) <5 3.62(H-( and 6, dd, J= 8.8 Hz, 16.2 Hz) 5 3.46 (H-l and 3. dd,y=6.5H2.13.9Hz] confirm by cosy.
300 Journal of Medicinal Materials, 2016, VoL 21, No. S
0 3(4°H 0
(r) Kaempferol 3-0-glucopyranoside-7-rhamnopyraiioside
(2) K»mpft,o) 3-0-,h™„os,l (I-.2) Elucop,™,„side-7-0- rl,™„„py™„s,de 0 ) Kaempferol 3,7.0- d„l,am„op,™os,de, (4) Querce.inc 3-0-rt,™„op,T.oo»de (5) Phenylpropanoids: R, - OCH,. R. - OH, hydroxyfemloyi ™Iat=
Ri - OH, R, . H, cafferoyi malate R| ^ H, R2 = H, coumaroyl malate , , , . , R | - O C H J , R ; . OCH,, synapoyl malate (6) Synapoyl glucose
Flg»r. I. Chemical structures of fl.vouotds »,d phenylpropanoids in Arabidop.a thaliana Coi 0
Figure 2. 'H-NMR speclt. of A thaliana Col 0 (a), extended high fled re.ion 0 0 5 ! „ ,M
5 6-8.2 ppm (e) 1 Leucine., 2. Valine. 3 Tl,ren„ine,4 Alanine 5 Gim™ f S * ' " ' ' " " ' ' " " ° " " " " "»»»
glucose. 10. u.glucose. 11. Sucrose. r2 Rhamnose, 13. Phenyprop™! 4 ' ' ^ " T " ' ' ' """= " " " • ' ' '^"""' ' • " • Kaempferol 3-<>glucopyr.noside-7.rham„„p,™„,,derKTem„fcol 3 O h " ' " ^ " ''"""" « ' « » - • '^
. « n „ o p y ™ o s i d e , . . K . m p f e r « 1 3 . , - 0 - d ; r h ™ ^ : Z ™ i : t r p r r : : : ' d e ; ; L t : " - = - - -
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301
_ I - J L „ LuiiiJ-iALLo^AL' -J»L_
-^'Ll---iii'**^--Jk^^
Figure 3.2D-NMR J-resolved spectra of A. thaliana Col.O in aromatic region ftom 6,2 - 7.0 ppm (a), and from 7.0 - 8.2 ppm (b). 1. rrnn, 5-hydroxyferuoyl malate, 2. trans caffeoyl malate, 3. /rii»» coumaroyl malate.
4. Syui^yl malate. 5. Synapoyl glucose. 6. Kaempferal 3,7-0- dirhamnopyranoside, 7. Kaempferol 3-0-rhamnosyl (1-2) glucosLde-7-O-riiamnopyranoside, 8. Kaempferol 3-0-glucopyranoside-7-rhanmopyranoside.
9. Quercetin derivatives, 10. els phcnylpropanoid.
302 Journal of Meilicinal Materials, 2016, VoL 21, No. S
^-...i=_..iMu..^
87
tl '
• f i l ll 1
, i .|(,, )
)
1
(
! M 1
" f 1!"
=, synapoyl glucose. 6. Kaempferol 3.7-0- d t r h a m n ^ ^ T , " ' r m p Z l T o T " ' ' t ' r " ' " " " ' " ' ' rh^nopymnctde. 8. Kaempferol J - O - g i u c i p y ^ I S ^ Z ^ p T l i r ' ' ^ ' " " " = - ' - ' ' - 4. Conclusion
This study shows ihat methanol-d^ can be used as the best direct solvent to extract phenolic compounds and divert other metabolites from A. thaliana Col.O. The plant crude extract can be applied directly to NMR
Ihfee keampferol glycosides anti o n , quercettn glycoside were isolated ,„H .demtned. By using N M R s p e i r f e c h e
Journal of Meilicinal Materials, 2016, Vol. 21, No. S
References
1. DivLon R A.. Smick D (2003). Phuoc he mistiy meets genome analysis, and beyond. Phytochemistry 62, 815-816.1 Sumner L W. Mendes P , Dixon R A. (2003), Plant metabolomics' large-scale phytochemistry in the fiinctional genomics era, Phviochemisirv. 62. 817-836 3. Stobieeki M , Kachlicki P (2005), Metabolomics and metabolite profiling - can w achieve the %oa.P. Ada Pliysiologiae Planianim, 27, 109-116. 4. Roessner U., Luedemann A., Brasi D., Fiehn O.. LinkeT Wiilmilzer L, Femie A R (2001), Metabolic profiling and phenoiyping of genetically and environmentally modified planl si^icms, Planl Cell. 13, 11-29- 5. Roessner U , Wagner C , Kopka J., Trethewey R. N , Willmitzer L. (2000), Simultaneous anal) sis of metabolites in potato tuber by gas chromatography-mass spectrometry, Planl Journal, 23, 131-142. 6. RohdeA, Morreel K, Ralph J. (2004), Molecular phenoiypmg of the pall and pal2 mutants of Arabidopsis thaliana reveals far- reaching consequences on phenylpropanoid, amino acid, and carbohydrate metabolism. Plant Cell, 16, 2749-2771 7.
Graham T (1998), Flavonoid and flavonol glycoside metabolism in Arabidopsis, Planl Physiology and Biochemislry, 36, 165-144 8 Bloor S J., Abrahams S. (2002), The structure of the major anthocyanin \n Arabidopsis thaliana, Phytochemisln.
59, 343-346. 9. Kerhoas L., Aouak D., Cingoz A., Routaboui J M , Lepiniec L , Einhom J. and Birlirakis N (20061 Structural characterization of the major fiavonoid glycosides from Arabidopsis thaliana seeds. Journal of Agncullural and Food Chemistry. 54, 6603-6612, 10. Veit M.. Pauli G. F. (1999), Major fiavonoids from Arabidopsis thaliana leaves Jounuil of Natural Products, 62, 1301-1303. 11. Cheynier V. (2005), Polyphenols in foods are more complex than often thought, American Journal of Clinical S'uiniion. 81, 223S-229S
Journal of Medicinal Materials, 2016, Vol. 21, No. 5 (pp. 304-309)
FLAVONOID AND PHENYLPROPANOID GLYCOSIDES ISOLATED FROM ANODENDRONPANICULATUM(ROXB.) A. DC.
Hoang Thi Nhu Hanh, Ho Viet Due, Tran Thi Thuy Linh, Vo Quoc Hung, Nguyen Thi Hoai * Faculty of Pharmacy, Hue Umversity of Medicine and Pharmacy. Hue University, Vietnam
•Corresponding author: hoai77(ggmail.com (Received September, 09"', 2016)
Summary
Flavonoid and Phenylpropanoid Glycosides Isolated from Anodendron paniculaium (Roxb.) A. DC.
Phytochemical sludy on the aerial parts of Anodendron paniculaium led to Ihc isolation of three compounds, including kaemprerol-3-O-rulinoside (1), rutin (2), and sargentol (3). The chemical struclures of the isolated compounds were elucidated on the basis of spectroscopic analyses
Keywords: Anodendron paniculaium, Kaempferol-3-O-rutinoside. Rutin. Sargentol.
1. Introduction still insufficient [5], [6]. In our effort to discover Anodendron is one of about 200 genera anticancerous herbs, the bioactive screening belonging to Apocynaceae family Up to this time, results showed that the methanolic extract from there are approximately 80 compounds which are aerial parts of this plant possessed potent mostly cardenolide glycosides isolated from only inhibitory activity toward LU-1, KB, Hep-G2, 3 species including A. affme, A. formicinum, and MKN-7, and S W ^ 8 0 cancer cell-lines [7].
A. paniculaium [1]. Among those, A. paniculatum Therefore, this study was conducted to clarify the (Roxb.) A. DC. is known to be distributed in the phytochemistry of this plant. Herein we report the woodland or the scrub of Quang Tri, Thua Thien isolation and the structural elucidation of flavonoid Hue, Khanh Hoa, Dong Nai, Ben Tre, and Kien and phenylpropanoid glycoside compounds isolated Giang province, Vietnam [2]. Its root and latex from the aerial parts of ^ . paniculatum collected in have been used as an emetic, cough suppressant Vietnam.
[3] and snakebite treatment [4] Aside from some 2. Material and methods cardenoltdes including anodendroside-A. -E„ -E,, 2.7. Plant material
-h, and -G reported ,n the literature, the knowledge The aerial parts of ^ . paniculatum (Roxb.) A.
about chemical constituents of A. paniculaium is DC.were collected in Dakrong district, Quang Tri
304 Journal of Medicinal Materials, 2016, VoL 21, No. 5
