Vinca rosa.doc

Minia J. Agric. Res & Dev. Vol. 13 No. 2 June, (1991)

INSECT GROWTH INHIBITORY FLAVAN GLYCOSIDES FROM VINCA RQSAE
Salah M. Abdel-Kader
Agriculture Chemisttry Dept.
Salah M. Hussein
Plant Protection Dept.

Faculty of Agriculture, Minia University

ABSTRACT

Chemical and biological studies were carried out on methanol extract of the
herb of Vinca rosa_e L..These studies led to isolation and identification
of one active flavan glycosides (4, 2, 6 -trihydroxy chalcone 4,( J3, D-
glucurono-side). This compound showed a high biological activity as feed
deterrent, retarded, and reduced the larval growth rate of Sp_o_dop_tera
littor_alis at concentration of 0.01%.
INTRODUCTION
Much efforts to develop non-toxic, safe and biodegradable alternatives to
chemical pesticides has been concerned with the use of natural products.
Periwinkle ( Vdjncja nosae_ L.) , known also as Caijtharantlvus jrosea. G.
is grown as an ornamental plant in tropical and subtropical countries. It
has many uses in folk medicine, and was also used for treatment of
diabetes,cancer chemotherapy (Taylor and Fransworth 1975; Morton 1977 and
Trease and Evans (1978). The chemical constituents in this plant have been
studied by many authors notably (Heal e_t aJL.1950; Chapman 1974 ; and
Meisner et a1.1981).



Methanol extract of the leaves showed insect growth inhibitory against
larvae of cotton leafworm Spodop_t.ei:a litjto£alis (Boisd). This
observation gave an evidence to undertake chemical and biological study to
clarify the major constituent responsible for such activity

MATERIALS and METHODS
a~MaJtejri.a.ls: Plants of VjLrica jrpsae, (L.) were harvested after 120
days growth at the experimental farm of the Faculty of Agriculture at Minia
University and the herb were immediately prepared by cutting into pieces
and extracted as shown below, b- General methods: Melting points were
determined by the micro hot-plate method.
Instrumentation: IR and UV spectra were recorded on Unicam
1 SP200 G and Beckman Model DKI istruments , respectively. H-NMR
13 spectrum was determined on a JEOL C-90 Spectrometer. C-NMR spectrum was
obtained on JEOL FX-400 Spectrometer operating at 100 MHz. Gas-liquid
chromatography (GLC) was performed with a Shimadzu model GC-GAM gas
chromatography apparatus as described in previous paper ( Abdel-Kader e_t
al . 1990 }. Analytical TLC and PTLC separation were carried out on Merck
pre-coated Silica gel 60 plates ( F254 , layer thickness 0.25 mm or 0.5 mm
).
Flavonoid moiety was detected by inspecting developed TLC/ PTLC plates
under (365 nm) and (254nm) UV light, and by the characteristic colours
formed with Gibbs reagent.The compounds of interest were eluted from the
silica gel with EtOAc. Gibbs and Shinoda tests were carried out according
to Tahara et al. (1987).


INSECT GROWTH INHIBITORY FROM Vinca rosae 845
Extraction and Separation Qf Yincaiinrl :
The herb ( 1.2 Kg.) of Vinca ro_sae, (L.) was extracted with methanol (10
liters) for three weeks at room temperature (Abd El-Kader 1991). Removal of
the solvent ( at 30 C ) and complete drying of the methanol extract gave a
brown powder (70.3 gm. ) , which was chromatographed on silica gel ( 400
gm, Silica gel 60, Merck ) by successive elutions with 1:1 ( 2 liter ) and
7:3 (2 liter ) mixture of n-hexane and ethyl acetate (EtOAc). The activity
was observed in the latter fraction (5.7gm). Silica gel (400 gm)
chromatography of the active fraction was eluted with (13 : 7 : 4; v/v )
solvent mixture of CHCl^- MeOH-H^O, and was collected in fractions of 15 ml
each to give the active fractions ( 450 ml ). The concentrate of the active
fractions afforded a crude crystalline material, which was recrystalllized
from the solvent mixture (n- hexane- EtOAc, 4 : 1 ) to give pure crystals
o r
(1.04 gm), mp. 138-140 C, ( = 11.52 ppm
^ attributable to H- bonded C-2-OH. Similarly, no shift was given
upon the addition of NaOAc, In addition obvious bathochromic shifts
occurred in the presence of sodium methoxide ( 39 nm, aromatic OH group).
The flavone skeleton for Vincalin-1 was deduced from the positive result of
a Shinoda test (Tahara, et. aJL.,1974), which gave an anthocyanidin product
with a decisive reddish colour if the tested compound did not have an
isoflavoni-dal but flavonoidal structure. IR of Vincalin-1 ( nujol ) 3360-
3500 (-OH), 1680 (C=0), 1725 (C=0 acid), 2650 (strectching OH
1
acid) and 1430, 1310 cm (COOH). The H-NMR (100 MHz) spectrum of

Vincalin-1 (DMSO-d ) showed one hydrogen-bonded hydroxyl proton
s at 11.52 (lH,s,H-2) indicating the presence of Chalcone moiety
(Hedin and Waagea, 1986 ). Moreover, the presence of the signal


Table 1: Carbon chemical shift of Vincalin-1.


Chalcone nuclei
Glucuronic moiety


H
H
"c- "1 "1"27 ".70 "s "c- "1 "101."51 "
"C-7 "1 "4".43 "d "c- "6' "168. "28 "s "
" " "1" " " " " " " "
"P "9 "200 ".3 "s " " " " " "
"— " " " " " " " " " "
"c- "1 "
" "i. "2 " "3 "4 "5"6"7 " "


accumulative mort.%
"20.00
"25
".00
"27.97
"50.00
"70.83
"79.16
"90
".47
"100
"0.0
" "No. survival larvae 30
"30
"28
"
"25
"24
"24
"24
"24
"
"21 1
"00
" "survival %
"100
"93
".33
"83.33
"80
"en
"80
"80
"
"70
"-
" "Aveg. wt ./larvae 43.1
"132.1
"208
".3
"288.7
"387.2
"423.1
"712
"516
".6
"413.2
"
" "Untreated standard error i4.3 +
"13.2
"±6
".5
"±29.07
"±20.68
"±13.6
"±21.6
"±16
".3
"±7.3
"
" "Aveg food consum.
"62.29
"126
".3
"144.2
"215.3
"313.1
"310.9
"420
".6
"286.1
"-
" "standard error (mg/L
"t 3.6
"+ 13
". 2
"±20.1
"+ 17.3
"±21.2
"+16.22
"±16
".7
"±3.6
"-
" "RGR
"0.128
"0
".101
"0.093
"0.093
"0.081
"0.114
"0
".071
"0.0499
"-
" "accumolitive % Mort.
"0.0
"6
".66
"16.66
"20.00
"20.00
"20.00
"20
".00
"30.00
"~
" "RGR = (Wf / W0)/Tf Where W = weight ; Tf = time since start ; f =
final and 0 = start.














natural compounds are known as potent phagodeterrent for jous insect (
Chapmar !Q74- Wada and Munkata, 1986 )*
findings have been reported with other insects and alka-3ill, 1972 and Ma,
1972 ). It could be concluded that -1 inhibited the feeding activity and
retarded the larval it the concentration used. Further work is needed on
the value of this material as insectstatics to better iding the role and
function of this natural product as an ; tool in pest control managment.

ACKNOWLEDGEMENT
.uthors wish to thank Prof. Dr. F.S.Badran, Department of ure, Faculty of
Agriculture, Minia University for us with plant samples used in this
investigation. S. der is indebted to Prof. M.A. Selim, Director of & Devel.
H. J. Gruy and associates, Inc., University of a for their encouragement
and for C- NMR measurement.

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INSECT GROWTH INHIBITORY FROM Vinca rosae

ATCC- 842. Can.J. chem.68,323.


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