Polysciosides J and K, two new oleanane-type triterpenoid saponins from the leaves of Polyscias fruticosa (L.) harms. cultivating in An Giang Province, Viet Nam

Article abstract of DOI:10.1080/14786419.2018.1557657

For the first time, the phytochemical constituents of the leaves of Polyscias fruticosa (L.) Harms. cultivating in An Giang Province, Viet Nam were investigated and led to purify two new oleanane-type triterpenoid saponins, named polyscioside J (1) and polyscioside K (2) together with two known saponins, ladyginoside A (3) and chikusetsusaponin IVa (4) using variously chromatographic methods. Saponin (4) was reported for the first time from this species. Their structures were verified by IR, UV, HR-ESI-MS, NMR 1D and 2D experiments and compared with previous literatures.

Full text of DOI:10.1080/14786419.2018.1557657

Natural Product Research
Formerly Natural Product Letters
ISSN: 1478-6419 (Print) 1478-6427 (Online) Journal homepage: http://www.tandfonline.com/loi/gnpl20
Polysciosides J and K, two new oleanane-type
triterpenoid saponins from the leaves of Polyscias
fruticosa (L.) harms. cultivating in An Giang
Province, Viet Nam
Van Mai Do, Cong Luan Tran & Tan Phat Nguyen
To cite this article: Van Mai Do, Cong Luan Tran & Tan Phat Nguyen (2019): Polysciosides
J and K, two new oleanane-type triterpenoid saponins from the leaves of Polysciasꢀfruticosa
(L.) harms. cultivating in An Giang Province, Viet Nam, Natural Product Research, DOI:
10.1080/14786419.2018.1557657
To link to this article: https://doi.org/10.1080/14786419.2018.1557657
View supplementary material
Published online: 19 Jan 2019.
Submit your article to this journal
View Crossmark data
Full Terms & Conditions of access and use can be found at
http://www.tandfonline.com/action/journalInformation?journalCode=gnpl20

NATURAL PRODUCT RESEARCH
https://doi.org/10.1080/14786419.2018.1557657
Polysciosides J and K, two new oleanane-type
triterpenoid saponins from the leaves of Polyscias
fruticosa (L.) harms. cultivating in An Giang
Province, Viet Nam
Van Mai Doa,b, Cong Luan Trana and Tan Phat Nguyenc,d
b
aNursery and Pharmaceutical Faculty, Tay Do University, Can Tho City, Viet Nam; Faculty of
c
Pharmacy, University of Medicine and Pharmacy, Ho Chi Minh City, Viet Nam; Graduate University
of Science and Technology Vietnam Academy of Science and Technology, Hanoi, Viet Nam;
dInstitute of Chemical Technology, Vietnam Academy of Science and Technology, Ho Chi Minh City,
Viet Nam
ABSTRACT
ARTICLE HISTORY
Received 8 November 2018
Accepted 6 December 2018
For the first time, the phytochemical constituents of the leaves of
Polyscias fruticosa (L.) Harms. cultivating in An Giang Province,
Viet Nam were investigated and led to purify two new oleanane-
type triterpenoid saponins, named polyscioside J (1) and polyscio-
side K (2) together with two known saponins, ladyginoside A (3)
and chikusetsusaponin IVa (4) using variously chromatographic
methods. Saponin (4) was reported for the first time from this
species. Their structures were verified by IR, UV, HR-ESI-MS, NMR
1D and 2D experiments and compared with previous literatures.
KEYWORDS
Polyscias fruticosa;
araliaceae; triterpenoid;
polyscioside J;
polyscioside K
1. Introduction
Polyscias is the second largest genus in the family Araliaceae, with 159 species and is
distributed from tropical Africa to the islands of the eastern Pacific Ocean (Bean 2015),
which 7 species and 1 variety are found in Vietnam (La et al. 2013). Polyscias fruticosa
CONTACT Tan Phat Nguyen
ntphat@ict.vast.vn
Supplemental data for this article can be accessed at https://doi.org/10.1080/14786419.2018.1557657.
ß 2019 Informa UK Limited, trading as Taylor & Francis Group

2
V. DO ET AL.
has been used traditionally in Vietnam for the treatment of ischemia, inflammation
and increase blood in the brain (Do 2004; Vo 2012). Pharmacological investigation of
extracts from P. fruticosa possessed anti-asthmatic, anti-inflammatory (Bernard et al.
1998; Koffuor et al. 2014, 2016); anti-histaminic, mast cell stabilization effect (Koffuor
et al. 2016); anti-pyretic, anagesic, molluscicidal properties (Bernard et al. 1998), diur-
etic effect (Varadharajan and Rajalingam, 2011), anti-diabetic activity (Divakar and
Bensita, 1998; Hanh et al. 2016). Phytochemical study of P. fruticosa illustrated triter-
penoid saponins (Chaboud et al. 1995; Proliac et al. 1996; Hanh et al. 2016), and polya-
cetylens (Lutomski et al. 1992) as the main ingredients. Moreover, steroids (Tram et al.
2017) and sesquiterpenoids (Brophy et al. 1990) were reported. Continuing a study of
the bioactive triterpenoids (Nguyen et al. 2015, 2016; Ngo et al. 2017, 2018), this paper
concentrates the isolation and structural elucidation of two new oleanane-type triter-
penoid saponins (1, 2) and two known saponins from the leaves of P. fruticosa culti-
vating in An Giang Province, Viet Nam.
2. Results and discussion
The n-butanol extract from the leaves of P. fruticosa cultivating in An Giang Province,
Viet Nam was subjected to column chromatography over silica gel normal-phase
and reversed-phase RP-18 to give two new oleanane-type triterpenoid saponins,
polyscioside J (1) (3-O-[b-D-glucopyranosyl-(1!4)]-b-D-(6-O-methyl)glucuronopyranosyl
oleanolic acid 28-O-b-D-glucopyranosyl ester) and polyscioside K (2) (3-O-[b-D-
glucopyranosyl-(1!4), b-D-glucopyranosyl-(1!2)]-b-D-(6-O-methyl)glucuronopyranosyl
oleanolic acid) together with two known saponins, ladyginoside A (3-O-[b-D-glucopyra-
nosyl-(1!4)]-b-D-glucuronopyranosyl oleanolic acid) (3) (Huan et al. 1998) and chiku-
setsusaponin IVa (3-O-b-D-glucuronopyranosyl oleanolic acid 28-O-b-D-glucopyranosyl
ester) (4) (Nguyen et al. 2015) (Figure 1).
Figure 1. Chemical structures of compounds 1–4.

NATURAL PRODUCT RESEARCH
3
Compound (1) was obtained as a white amorphous powder. The molecular formula
was established as C49H78O19 by HR-ESI-MS data ([M ꢀ H]ꢀ m/z 969.5011, calcd.
969.5059). The IR spectrum of 1 showed absorptions of hydroxyl group (3316 cmꢀ1
)
and the C–O stretching (1023 cmꢀ1). The 13C-NMR and DEPT spectrum of 1 (Table S1)
showed forty-nine carbons including: two carbonyl carbons, two olefinic carbons, three
anomeric carbons, thirteen oxygenated methine carbons, two oxygenated methylene
carbons, one oxygenated methyl carbon, six quaternary carbons, three methine car-
bons, ten methylene carbons, seven methyl carbons. The presences of seven tertiary
methyl groups at dC 15.5–33.1, one oxygenated methine carbon at dC 89.3 (C-3) corre-
sponding to proton at dH 3.28 (dd, 4.0 and 11.5, H-3), two olefinic carbons at dC 144.5
(C-13) and 122.0 (C-12) corresponding to proton at dH 5.40 (brs, H-12), and one car-
bonyl carbon at dC 176.5 (C-28) were confirmed a bidesmosidic saponin of oleanolic
1
acid in 13C and H-NMR (Nguyen et al. 2015, 2016; Ghislain et al. 2018). Moreover, the
1
13C and H-NMR of 1 revealed three anomeric carbons at dC 106.8 (C-1’), 105.0 (C-1’’)
and 95.7 (C-1’’’) corresponded to three anomeric protons at dH 4.94 (d, 8.0, H-1’), 4.96
(overlap, H-1’’) and 6.28 (d, 8.0, H-1’’’), respectively; one carbonyl carbon at dC 170.0
(C-6’), one oxygenated methyl carbon at dC 52.4 (-OMe); two oxygenated methylene
carbons at dC 62.5 (C-6’’) and 62.2 (C-6’’’) were assigned to methyl b-D-glucuronate
(Me-GlcA), b-D-glucose (Glc-1) and b-D-glucose (Glc-2), respectively (Nguyen et al.
2015, 2016 ). Moreover, acidic hydrolysis and TLC comparision of the hydrolysate with
authentic sugar verified sugar moiety as D-glucose (Rf 0.40). The HMBC spectrum of 1
(Figure S1) showed correlations between oxygenated methyl protons at dH 3.85 (s)
and carbon carbonyl of glucuronic unit at dC 170.0, which identified a methyl b-D-glu-
curonate moiety (Me-GlcA). On the orther hands, three anomeric protons at dH 4.94
(H-1’) of Me-GlcA, 4.96 (H-1’’) of Glc-1 and 6.28 (H-1’’’) of Glc-2 corresponded to car-
bons at dC 89.3 (C-3) of aglycone, 82.7 (C-4’) of Me-GlcA and 176.5 (C-28) of aglycone,
respectively. Based on data of HR-ESI-MS, 1D, 2D-NMR and compared with previous
published data (Huan et al. 1998); the structure of 1 was evidenced as 3-O-[b-D-gluco-
pyranosyl-(1!4)]-b-D-(6-O-methyl)glucuronopyranosyl oleanolic acid 28-O-b-D-gluco-
pyranosyl ester, named Polyscioside J.
Compound (2) was obtained as a white amorphous powder. The molecular formula
was established as C49H78O19 by HR-ESI-MS data ([M-H]- m/z 969.5013, calcd.
969.5059). The 1H and 13C-NMR data (Table S1) demonstrated that 2 has the same
aglycone and sugar moieties as 1, except for the down field shift of carbon due to
C-2’ of Me-GlcA (dC 80.2), indicating one b-D-glucose moiety (Glc-2) linked at this car-
bon. Further, the HMBC spectrum of 2 (Figure S1) showed correlations between three
anomeric protons at dH 4.88 (d, 7.5, H-1’) of Me-GlcA, 4.90 (d, 8.0, H-1’’) of Glc-1, 5.37
(d, 7.5, H-1’’’) of Glc-2 and carbons at dC 89.5 (C-3) of aglycone, 81.7 (C-4’) of Me-GlcA,
80.2 (C-2’) of Me-GlcA, respectively. Thus, the structure of 2 was recognized as 3-O-
[b-D-glucopyranosyl-(1!4), b-D-glucopyranosyl-(1!2)]-b-D-(6-O-methyl)glucuronopyra-
nosyl oleanolic acid, named Polyscioside K.
The genus Polyscias were acknowledged as a rich source of triterpenoid saponins
which revealed mainly oleanane-type aglycones. Oleanolic acid framework was
described from P. scutellaria (Paphassarang et al. 1988; 1989a, 1989b, 1990; Mitaine-
Offer et al. 2004). Whereas, hederagenin genin was reported from P. dichroostachya

4
V. DO ET AL.
(Gopalsamy et al. 1990) and P. scutellaria (Mitaine-Offer et al. 2004). Moreover,
some rare skeletones, such as echinocystic acid and collinsogenin were detailed from
P. scutellaria (Mitaine-Offer et al. 2004), 12-oxo-3b,16b,20(S)-trihydroxydammar-24-ene
was illustrated from P. fulva (Bedir et al. 2001). In our study, four oleanane-type
triterpenoid saponins were indicated as well as the previous investigations (Chaboud
et al. 1995, 1996; Proliac et al. 1996; Huan et al. 1998; Hanh et al. 2016). However,
it is the first time that the methyl glucuronate saponins were characterized from
the species P. fruticosa.
3. Experimental
3.1. General experimental procedures
€
The optical rotations were determined on a Kruss-optronic GmbH polarimeter
equipped with a sodium lamp (589 nm) (Hamburg, Germany). The IR spectra were
recorded with a Bruker Tensor 27 FT-IR Spectrometer (Bremen, Germany). The UV
spectra were performed with Agilent 1260 HPLC-UV (Waldbronn, Germany). The high
resolution electrospray ionisation mass spectroscopy (HR-ESI-MS) was recorded on
a LC-X500R QTOF spectrometer (Sciex, USA). The 1H-NMR (500 MHz), 13C-NMR
(125 MHz), DEPT, COSY, HSQC and HMBC spectra were recorded on a Bruker
AM500 FT-NMR spectrometer using tetramethylsilane (TMS) as internal standard (MA,
USA). Column chromatography was carried out using Merck Silica gel normal-phase
(230–240 mesh) and reversed-phase C18 Merck (Darmstadt, Germany). Analytical TLC
was carried out in silica gel plates Merck DC-Alufolien 60 F254 (Darmstadt, Germany).
Compounds were visualized by spraying with aqueous 10% H2SO4 and heating for
3–5 min. Camptothecin was purchased from Calbiochem.
3.2. Plant material
The leaves of Polyscias fruticosa were cultivated in An Giang Province, Viet Nam and
identified by Prof. Dr. C.L. Tran, Nursing Pharmaceutical Sciences, Tay Do University.
A voucher specimen (No. Mai-PF-2016) was deposited in Nursing Pharmaceutical
Sciences, Tay Do University.
3.3 Extraction and isolation
Dried powder of leaves P. fruticosa (859 g) were extracted with 96ꢁ EtOH for three
time (3 ꢂ 10 L) at room temperature, residue was filtered, solvents were removed
under low pressure and the crude extract was obtained. The crude extract (156 g) was
applied to liquid-liquid extraction procedures and successively partitioned into diethyl
ether, ethyl acetate, n-butanol and aqueous partition. The n-butanol extract (50 g) was
eluted by Diaion HP-20 column with H2O, 50% MeOH, 80% MeOH, 100% MeOH
and 100% Me2CO to give five major fractions (I–V), respectively. Fraction III (11 g) was
subjected to silica gel normal-phase column chromatography with mobile phase
CHCl3–MeOH gradient (0–100%) to get five subfractions (III.1–III.5).

NATURAL PRODUCT RESEARCH
5
Fraction III.1 (3,0 g) was classified on silica gel column chromatography with eluted
solvent CHCl3–MeOH–H2O (95:5:0 ! 80:20:1, v/v) and reseparated by RP-18 using mix-
tures of MeOH–H2O (60:40, v/v) to obtain 3 (13 mg) and 4 (10 mg). Fraction III.2 (2,5 g)
was chromatographed on silica gel normal-phase with solvent CHCl3–MeOH–H2O
(90:10:0 ! 80:20:1, v/v) and further purified by RP-18 using mixtures of MeOH–H2O
(50:50, v/v) to afford 1 (12 mg). Fraction III.3 (2,0 g) was fractioned on silica gel column
with solvent CHCl3–MeOH–H2O (88:12:0 ! 75:25:2, v/v) and segregated by RP-18 using
mixtures of MeOH–H2O (50:50, v/v) to yield 2 (15 mg).
Polyscioside J (1): Amorphous powder (MeOH); IR (KBr) tmax: 3316, 2943, 2831,
1449, 1416, 1023 cmꢀ1; UV (MeOH) kmax: 207 nm; HR-ESI-MS m/z 969.5011 [M ꢀ H]ꢀ
1
(calcd for C49H78O19, 969.5059); H and 13C-NMR data (pyridine-d5), see Table S1.
Polyscioside K (2): Amorphous powder (MeOH); IR (KBr) tmax: 3262, 2942, 2915,
2831, 1022 cmꢀ1; UV (MeOH) kmax: 204 nm; HR-ESI-MS m/z 969.5013 [M ꢀ H]ꢀ (calcd for
1
C49H78O19, 969.5059); H and 13C-NMR data (pyridine-d5), see Table S1.
3.4. Acid hydrolysis
Two new saponins 1, 2 (2 mg) was refluxed with 2N aq. CH3COOH (5 mL) for 2 h at
100 ꢁC. After extraction with CHCl3 (3x5 mL), the aqueous layer was repeatedly evapo-
rated to dryness with MeOH until neutral, and then analyzed by TLC over silica gel
(MeCOEt–isoPrOH–Me2CO–H2O 20:10:7:6) by comparison with authentic sugar of D-
glucose Rf 0.40 according to Nguyen et al. (2015, 2016, 2017a, 2017b).
Disclosure statement
No potential conflict of interest was reported by the author(s).
Funding
This research is funded by An Giang Department of Science and Technology.
References
Bean AR. 2015. A conspectus of Polyscias J.R.Forst. & G.Forst. (Araliaceae) in Queensland,
Australia. Austrobaileya 9:445–456.
Bedir E, Toyang NJ, Khan IA, Walker LA, Clark AM. 2001. A new dammarane-type triterpene
glycoside from Polyscias fulva. J Nat Prod. 64(1):95–97.
Bernard BM, Pakianathan N, Divakar MC. 1998. On the antipyretic, anti-inflammatory, analgesic
and molluscicidal properties of Polyscias fruticosa (L.). Harms Anc Sci Life. 17:313–319.
Brophy JJ, Lassak EV, Suksamrarn A. 1990. Constituents of the volatile leaf oils of Polyscias fruti-
cosa (L.) Harms. Flavour Fragr J. 5(3):179–182.
Chaboud A, Rougny A, Proliac A, Raynaud J, Cabalion P. 1995. A new triterpenoid saponin from
Polyscias fructicosa. Pharmazie 50:371.
Divakar MC, Bensita MB. 1998. Screening of various leaf extracts of Polyscias fruticosa Harms for
their antidiabetic activity. Indian J Nat Prod. 14:24–28.
Do TL. 2004. Medicinal plants and remedy of Vietnam. Hanoi, Vietnam: Publisher of Medicine.
Ghislain FW, Jean NI, Armelle MT, Patrick AY, Ibrahim D, Victor K, Yeboah S, Bathelemy N,
Thomas E, Bonaventure NT. 2018. Polyacanthoside A, a new oleanane-type triterpenoid

6
V. DO ET AL.
saponin with cytotoxic effects from the leaves of Acacia polyacantha (Fabaceae). Nat Prod
Res. 1–6. doi: 10.1080/14786419.2018.1486312. (Epub ahead of print).
Gopalsamy N, Gueho J, Julien HR, Owadally AW, Hostettmann K. 1990. Molluscicidal saponins of
Polyscias dichroostachya. Phytochemisrty 29(3):793–795.
Hanh TTH, Dang NH, Dat NT. 2016. a-Amylase and a-glucosidase inhibitory saponins from
Polyscias fruticosa leaves. J Chem. 2016:1–5.
Koffuor GA, Boye A, Kyei S, Ofori-Amoah J, Akomanin Asiamah E, Barku A, Acheampong J,
Amegashie E, Kumi Awuku A. 2016. Anti-asthmatic property and possible mode of activity of
an ethanol leaf extract of Polyscias fruticosa. Pharm Biol. 54(8):1354–1363.
Koffuor GA, Boye A, Ofori-Amoah J, Kyei S, Abokyi S, Nyarko RA, Bangfu RN. 2014. Anti- inflam-
matory and safety assessment of Polyscias fruticosa (L.) Harms (Araliaceae) leaf extract in
ovalbumin-induced asthma. BJMMR. 3:337–342.
La DM, Chau VM, Tran VS, Pham QL, Phan VK, Tran HT, Tran MH, Ninh KB, Le MH. 2013.
Prospects of natural bioactive products from Araliaceae Juss. Family in VietNam. The 5-th
national conference on ecology and biological resources, 1152–1158.
Lutomski J, Luan TC, Hoa TT. 1992. Polyacetylenes in the Araliaceae family. Part IV. The antibac-
terial and antifungal activities of two main polyacetylenes from Panax vietnamensis Ha et
Grushv., and Polyscias fruticosa (L.) Harms. Herba Polonica. 38:137–140.
Mitaine-Offer A-C, Tapondjou LA, Lontsi D, Sondengam BL, Choudharyd MI, Atta-Ur-Rahmand,
Lacaille-Dubois M-A. 2004. Constituents isolated from Polyscias fulva. Biochem Syst Ecol. 32:
607–610.
Ngo TN, Phan NM, Bui TD, Dang VS, Tran CL, Mai DT, Nguyen TP. 2017. Cytotoxic cycloartane tri-
terpenoids from the leaves of Markhamia stipulata var. canaense. Phytochem Lett. 22:251–254.
Ngo TN, Vo CT, Pham NKT, Phan NM, Bui TD, Ngo QL, Dang VS, Tran CL, Mai DT, Nguyen TP.
2018. Markhacanasin C, cycloartane triterpenoid from the leaves of Markhamia stipulata var.
canaense V.S. Dang. Nat Prod Res. 1–6.
Nguyen TP, Le TVH, Mai DT, Le TD, Phan NM, Bui TD. 2015. Two new oleanane-type triterpene
saponins from the leaves of Schefflera sessiliflora De. P. V. Phytochem Lett. 11:102–105.
Nguyen TP, Le TD, Phan NM, Bui TD, Mai DT. 2016. Triterpene saponins with a-glucosidase
inhibition and cytotoxic activity from the leaves of Schefflera sessiliflora. J Asian Nat Prod Res.
18(6):542–549.
Nguyen TP, Mai DT, Do THT, Phan NM. 2017a. Flavonoids with hepatoprotective activity from
the leaves of Cleome chelidonii. Nat Prod Comm. 12:1061–1063.
Nguyen TP, Tran CL, Vuong CH, Do THT, Le TD, Mai DT, Phan NM. 2017b. Flavonoids with hepa-
toprotective activity from the leaves of Cleome viscosa L. Nat Prod Res. 31(22):2587–2592.
Paphassarang S, Raynaud J, Lussignol M. 1988. The polysciasaponin P7 of Polyscias scutellaria
(Burm.f.) Fosb. (Araliaceae). Pharmazie. 43:296–297.
Paphassarang S, Raynaud J, Lussignol M, Becchi M. 1989a. Triterpenic glycosides from polyscias
scutellaria. Phytochemistry 28(5):1539–1541.
Paphassarang S, Raynaud J, Lussignol M, Becchi M. 1989b. Triterpenoid saponins from Polyscias
scutellaria. J Nat Prod. 52(2):239–242.
Paphassarang S, Raynaud J, Lussignol M, Cabalion P. 1990. A new oleanolic glycoside from poly-
scias scutellaria. J Nat Prod. 53(1):163–166.
Proliac A, Chaboud A, Rougny A, Gopalsamy N, Raynaud J, Cabalion P. 1996. A oleanolic saponin
from Polyscias fructicosa (L.) Harms var yellow leaves. Pharmazie. 51:611–612.
Tram NTT, Tuyet HD, Minh QN. 2017. One unusual sterol from Polyscias fruticosa (L.) Harms
(Araliaceae). Can Tho Univ J Sci. 7:33–36.
Varadharajan R, Rajalingam D. 2011. Diuretic activity of Polyscias fruticosa (L.) Harms. Int J Innov
Drug Dis. 1:15–18.
Vo VC. 2012. Dictionary of Vietnamese medicinal plants. Hanoi, Viet Nam: Publisher of Medicine,.
Vo DH, Yamamura S, Ohtani K, Kasai R, Yamasaki K, Nguyen TN, Hoang MC. 1998. Oleanane
saponins from Polyscias fruticosa. Phytochemistry. 47(3):451–457.