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CHAPTER - II
THE CONSTITUENTS OF THE LEAVES OF MURRAYA KOENIGII (RUTACEAE).
Introduction 26-33
Results and Discussion 34-42
Experimental 43-47
References 48-51
26
INTRODUCTION
1. CARBAZOLE ALKALOIDS
(a) Carbazole alkaloids containing secondary nitrogen :
Tricyclic : Tricyclic carbazole alkaloids isolated from stem bark of M. koenigii
were mukoeic acid (la) -2, mukonal (lb)^ 1-hydroxy-3-methyl carbazole (Ic)" and
me-2-methoxy carbazole-S-carboxylateCId)". Koenoline (le) isolated from root
bark of M. koenigii exhibited cytotoxic activity against the KB cell-culture test
system^. 2-Methoxy-3-methyl carbazole (If)^ from stem bark and murrayanine
(ig) ' from leaves and stem bark of M. koenigii yNere also isolated. Other tricy
clic carbazole alkaloids isolated from M. koenigiiwere mukonidine (Ih)^, mukoline
(li)^°, mukolidine (Ij)'", mukonine (lk)^\ 2-hydroxy-3-methyl carbazole (ll)^^ 3-
methyl carbazole (Im)", glycozoline^^ mukoenine-A, -B and -D^^ The tricyclic
carbazole alkaloids having branched side chain, isolated from the stem bark
and seeds of M. koenigii, were grinimbilol (li)^^ mahanimbinol = mahanimbilol
(liia)«' ^ and murrayanol (lllb)^^
Tetracyclic : Murrayacine (iVa)"'^^ and koenidine = koenigicine = koenimbidine
(IVb) -i9-2i from leaves and roots, grinimbine (IVc) ^^^ from stem bark, leaves
and roots, koenimbin = koenimbine (IVd^^'^from fruits, mukonicine {IVe) '*from
leaves along with koenine (IVf) ^^^ and koenigine (IVg) ' ^ were isolated from
M. koenigii having no side chain. However tetracyclic carbazole alkaloids
containing branched side chain included mahanimbine {Way'*^-^^-^^^^ irom fruits
and leaves, mahanimbicine = isomahanimbin (Vb) - - from roots and leaves,
isomahanine (Vc)'^ from seeds, mahanine (Vd)^^2^3° from leaves and
27
I
a
b
c
d
e
f
9
h
i
i
k
1
m
R2 = R''
R^ = R^
R2 = R4
R^ = R^
R2 = R''
R^ = R''
R2 = R'
R^ = R^
R2=R3
R2 = R3
R2 = R''
R^ = R^
Ri = R2
H, R = OCH3, R3 = CO^H
H, R2 = 0 H , R3 = C H 0
H, R = OH, R3 = CH3
H, R2 = OCH3, R3 = CO2CH3
H, R = OCH3, R3 = CH^OH
H, R2 = OCH3, R ' = CH3
H, R = OCH3, R^ = CHO
H, R2 = OH, R3 = CO2CH3
: H, R = OCH3, R" = CH^OH
: H , R = OCH3, R^ = CHO
: H, R'= OCH3, R3 = CO2CH3
: H, R2 = OH, R3 = CH3
: R'' = H, R3 = CH,
II
28
a R2 = R3 = H, Ri = C K
b Ri = H, R2 = CH„ R ' = OH
^ ^ ^ ^ % , ^
IV
a Ri = R3 = R" = H, R2 = CHO
b R = H, R = CH,, R2 = R3 = OCK
c R2 = R3 = R4 = H, R = CH,
d R3 = R" = H, R =CH,, R2 = OCH,
e R3 = H, Ri = CH„ R2 = R* = OCH,
f R3 = R4 = H, R = CH,, R2 = OH
R'' = H, R = CH„ R = OCH,, R = OH
29
a Ri = R2 = R3 = H
b Ri = R3 = H, R2 = CH3
C Ri = H, R2 = CH3, R3 = OH
d R2 = H, R = CH3, R3 = OH
(CHJXH(OH)
C(CH3):CH,
VI
Vil
30
VIII
a R = C(CH3)pH
b R = C(CHJCH,
a
b
R = H
R = CH,
31
mahanimboline (Vl)^^ were also isolated from M. koenigii.
Pentacyclic : The only pentacyolic carbazole alkaloid without any side chain
isolated from stem bark of M. koenigii was murrayazolidine (Vn)^^ while
murrazolinine (VlllaP and cyclomahanimbin = currayanine (Vlllb)"*' isolated
from leaves, possess branched side chain.
Hexacyclic : Bicyclomahanimbicine (IXa)^from leaves and bicyclomahanimbin
(rXb) '2^ from both leaves and stem bark of M. koenigii v^ere isolated.
Carbazole alkaloids murrayanine, grinimbine and mahanimbine
containing secondary nitrogen isolated from stem bark of M. koenigiia\so showed
antifungal activity against some human pathogenic fungii^^.
(b) Carbazole alkaloids containing tertiary nitrogen :
Isomurrayazoline (Xa)^^, currayangine = mahanimbidine = murrazoline
(Xb)^2S2^** from leaves and stem bark while murrazolinol (Xc)^^ from stem bark
of M. koenigiiyNere isolated. The crystallographic parameters for murrayazoline^^
are a 11.19, b 11.05, c 15.65 e, p 115.27°, d. (exptl.) = 1.24, z = 4, d. (calcd.) =
1.258, space group P21/c, R = 0.074.
(c) Binary carbazole alkaloids and quinones :
Binary carbazole alkaloids and quinones isolated from M. koenigii
included murrastifojine - F, bis-2-hydroxy-3-methyl carbazole, bismahanine,
biskoeniquinone-A and bis murrayaquinone-A^".
2. ESSENTIALOILS
a- pinene^^"", l-caryophylene^^, sabinene^^ P-caryophyllene'*\ p-
elemene'*\ p-phellandrene^V terpenen-4-ol' '° from leaves while 8-
HsC
a R = H, R' = H
b R = CH3, R' = H
c R = CH„ R' = OH
H3ca
GluO
XI Glu = glucopyranosyl
/SD
XII
CH„OH r^/T^O' 2 ' OCMGg
. / ^ O
H H
XIII
33
geranyloxypsoralene''2, imperatorin''^, heraclenin''^, isosaxalin''^ and
heraclenoFwere isolated from the seeds of M. koenigii.
3. MISCELLANEOUS
Glycoside scopoline (Xl)^ glucose^ from leaves and 3-(1,2-dimethyl
allyl) xanthyletin (Xll)"^ from stem bark of M. koenigii were isolated. Marmesin-
1"-0-p-D-galactopyranoside (XIII) and umbelliferone isolated from stem bark of
M. koenigii showed antiinflammatory and antimicrobial activities'".
34
RESULTS AND DISCUSSION
The dried leaves of Murraya koenigii were extracted with methanol and
methanol extract was concentrated and treated with petrol, benzene, ethyl ac
etate and acetone. The petrol fraction on column chromatography using silica
gel as adsorbent and eluting with petrol-benzene (1:1), yielded a crystalline
product MK-1.
MK-1, crystallized from benzene, m.p. 205°C, gave brown fluorescence
in UV light after developing on thin layer chromatography. In the infra red spec
trum bands appeared at 3400 (N-H stretching), 1640, 1610 and 1580 cm-
(unsaturation and aromatic system). It analysed for C^gH jgNOg (M* 293). In the
mass spectrum the molecular ion appeared at m/z 293 and base peak appeared
at m/z 278 (M-CH3). Other prominent peaks appeared at m/z 235 (M-58) and
m/z 139. The odd mass number for molecular ion is indicative of the presence
of nitrogen.
In the 'H NMR spectrum (600 MHz, CDCI3) given in Table 1 and Fig.1, a
singlet integrating for six protons at 8 1.485 can be assigned to gem dimethyl
protons. The singlet at 5 2.340 is attributable to benzylic methyl group while a
singlet at 5 3.903 can be assigned to methoxy protons. A pair of doublet
appearing at 5 5.640 (J=10 Hz) and 6.536 (J=10 Hz) were assigned to 3'-H and
4'-H, respectively, of chromene ring. The spectrum shows signals for aromatic
protons at 5 7.627 (s, 4-H), 6.940 (dd, J=8.4. 2.2 Hz, 7-H), 7.214 (d. J=8.4 Hz,
8-H) and 7.421 (d, J=2.2 Hz, 5-H). The broad singlet at 6 7.686 was assigned to
N-H proton. The assignments were supported by DEPT experiments (Fig. 2)
and HHCOSY correlations (Fig. 3).
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35
The "C NMR spectrum (150.8 MHz, CDCg given in Table I and Fig.
4,shows signals for all the 19 carbons. The signal at 516.022 was assigned to
methyl carbon C-3 of carbazole nucleus which was supported by DEPT experi
ments. The geminal methyl group carbons at C-2' absorb at 6 27.596 while
methoxy carbon attached to C-6 gives signal at 556.038 and C-2' absorb at 5
75.849 which was confirmed by DEPT experiments. The ^ C NMR signals for
sp^ carbons in the carbazole skeleton are as follows 5102.680 (C-5 ), 104.505
(C-1), 111.048 (C-8), 113.021 (C-7), 116.868 (C-4b), 117.197 (C-4'), 118.299
(C-3). 121.028 (C-4), 124.448 (C-4a). 129.199 (C-3'), 134.410 (C-8a), 135.759
(C-9a), 149.832 (C-2), 153.975 (C-6). The assignments were further confirmed
by DEPT experiments, HMBC (Fig. 5), COLOC (Fig.6), and CHCOSY (Fig.7)
correlations. On the basis of above data MK-1 was confirmed as koenimbine
(IVd) m.p. 205° C (lit. 194-95° C) ^^^ which was characterized earlier on the
basis of ^H NMR, IR, UV and MS only. The ^^c NMR and 2D NMR is discussed
for the first time.
IVd
Further elution of the column with petrol - benzene and crystallization
from benzene gave colourless crystals of MK-2.
MK-2, m.p. 139-40°C was UV inactive and it was detected by charring of
TLC plate (SiOg) with perchloric acid. In the IR spectrum a broad band at 3400
36
Table I. 'H NMR and '^C NMR data of MK-1 (IVd)
Assignment
C/H
Chemical Shift (S.ppm)
13 C NMR (150.8 MHz, CDCL) 'H NMR (600MHz, CDCL)
1
2
3
4
4a
4b
5
6
7
8
8a
9
9a
3-Me
6-OMe
1'
2'
3'
4"
2'-Me.
104.505
149.832
118.299
121.028
124.448
116.868
102.680
153.975
113.021
111.048
134.410
-
135.759
16.022
56.038
-
75.849
129.199
117.197
27.596
7.627 (s)
7.421 (d, J=2.2 Hz)
6.940(cld,J=8.4,2.2 Hz)
7.214(d, J=8.4Hz)
7.686 (br,s)
2.340 (s)
3.903 (s)
5.640 (d,J=10Hz)
6.536 (d,J=10 Hz)
1.485 (s)
37
cm^ indicated the presence of -OH group and the other bands were compa
rable with the IR spectrum of authentic sample of p-sitosterol"^. The occur
rence of p-sitosterol from M. koenigii has not been reported so far.
Elution of the column with benzene followed by preparative thin layer
chromatography yielded MK-3 and MK-4.
MK-3 was crystallized as colourless granular compound from alcohol -
benzene (7:3) melting at 94.5°C. It was UV inactive and its homogeniety was
checked on TLC plate (SiOg) with iodine as well as perchloric acid. In the mass
spectrum, molecular ion appeared at m/z 476 with successive loss of 14 show
ing peaks at m/z 462, 448, 434, 406 etc. and base peak appearing at 57, indi
cating the presence of long chain hydrocarbon.
The ^H NMR spectrum (600 MHz, CDCy is given in Fig. 8. It shows
signals at 8 0.866 (t, J=6.6 Hz), 1.243 (s), 1.542 (d, J=5.88) and 3.618 integrat
ing in the ratio 3 : 66 : 3 : 2 respectively. The consideration of ^H NMR integra
tion of 74 protons and the signal at 5 3.618 for two protons can lead to a
molecular ion C^^H^JD (M^522), but the molecular ion peak at m/z 522 is not
indicated in MS. In MS, a fairly unstable peak appeared at m/z 476 (3.5) as
molecular ion peak.
The ^^C NMR spectrum (150.8 MHz, CDCI3), given in Fig. 9, shows sig
nals at 6 14.065 (-CH3), 22.680 (-CH^-), 25.771 (-CH^-), 29.355 (-CH^-), 29.454
(-CH^-), 29.618 (-CH^-), 29.700 (-CH-)^, 31.936 (-CHg-), 32.873 (-CH^-) and
63.108 (-CH^O-). The appearance of ^ C NMR signal at 5 63.108 and proton
signal at 5 3.618 indicates the presence of CHg group attached to oxygen,
probably an alcohol. The assignment of CH^and CH3 carbons has been
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supported by DEPT experiments (150.8 MHz, CDCy shown in Fig. 10.
MK-4 crystallized from chloroform, m.p. 235-37°C, C gH gNO (M^ 331),
gave brown fluorescence in UV light after developing on TLC plate (silica gel.)
The IR spectrum shows bands at 1640 and 1600 c m \ characteristic of
unsaturation.The ^H NMR spectrum (Fig.11) in CDCI3 at 100 MHz was compa
rable with that of mahanimbidine^ and shows singlets for methyl groups at 6
1.32, 1.48, 1.92 and 2.34 attributable to 8'-Me, 3'-Me, 9'-Me and 3-Me, respec
tively (Table II). The muJtiplets appearing at 8 0.20 and 1.40 were assigned to
5'-ax-H and 5'-eq-H, respectively. The benzylic proton at 1'- position appears
as a broad doublet at 5 3.35. The aromatic proton at C-4 appears as a singlet at
5 7.35. Other aromatic protons absorb between 6 6.30 - 7.90 as double dou
blets or multiplets integrating for four protons at 5, 6,7and 8 carbons. Although
chemical and spectral data of MK-4 (m.p. 234-37°C) are comparable with
mahanimbidine (Xb)^ (scurrayangine = murrazoline)" 7 34 p p 266°C, yet
Table II. 'H NMR data of compound MK-4 (100 MHz, CDCi,)
Assignment Chemical shift ( 5, ppm)
5'-ax-H 0.20 (m,1H)
8'-Me 1.32 (s,3H)
5'-eq-H 1.40 (m,3H)
3'-Me 1.48 (s,3H)
9'-Me 2.34 (s, 3H)
3-Me 2.34 (s, 3H)
1'-H 3.35(brd, 1H)
Ar - CH3 7.35 (s, 3H)
Ar-H 6.30-7.90 (m, 5H)
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39
its structural assignment is not conclusive due to large difference in melting
point. Further studies are required to settle the stereochemistry as well as struc
ture.
The benzene fraction on column chromatography (silica gel), eluting the
column with petrol - benzene (1:1) and benzene - ethyl acetate (8:2) gave
Mk-5andMK-6.
MK-5 was obtained as light yellowish green crystals from benzene - al
cohol. It melts at 225°Cand shows brown fluorescence in UV light after devel
opment on TLC plate (silica gel). The IR spectrum shows bands at 3400 (N-H),
1640, 1620 and 1600 cm" (unsaturation and aromatic system). In the mass
spectrum molecular ion appeared at m/z 323 and base peak at m/z 308.The ^H
NMR spectrum (Table III and Fig. 12) was comparable with that of koenldlne
(IVb)^^ (=Koenigicine = Koenimbldine)^^ 21 n p 224-25°C. The structure was
confirmed by ^^c NMR (Table III and Fig. 13), DEPT experiments, HHCOSY
(Fig. 14), CHCOSY (Fig.15), COLOC and HMBC (Fig. 16) studies which was
not done earlier.
CH3O
CH3O
IV b
MK-6 was obtained as light green granules from pyridine - benzene -
methanol (1:11:10). It melts at 288 - GO C (lit. 2Q0°CY^ and was detected by
iodine and perchloric acid on TLC plate (silica gel.). In the infra red spectrum it
40
Table III. 'H NMR and ' 'C NMR data of MK-5 (iVb)
Assignment
C/H
Chemical shift ( 5 , ppm)
13 C NMR (150.8 MHz, CDCL) ^H NMR (600 MHz, CDCL)
1
2
3
4
4a
4b
5
6
7
8
8a
9
9a
3-Me
6-OMe
7-oMe
2'
3'
4'
2'-Me,
104.620
148.681
118.282
120.124
124.448
116.868
102.286
148.188
144.390
94.723
134.410
-
135.759
16.022
56.301
56.581
75.668
. 129.429
117.263
27.547
7.532 (s)
7.367 (s)
6.874 (s)
7.726 (s)
2.311 (s)
3.902 (s)
3.957 (s)
5.657 (d,J=10Hz)
6.570 (cl,J=10Hz)
1.463 (s)
41
shows strong absorption at 3350 cm"" indicating the presence of -OH group. Its
melting point and IR spectrum indicated it to be sterol glycoside. The structure
was assigned as sitosterol - 3 - 0 - p - D - glucoside (XlVa) from IR and H
NMR (100 MHz, CDCI3) given in Fig. 17 and Table IV of its acetylated derivative
(XlVb). The occurrence of sitosterol - 3 - O - P - D - glycoside from M. Koenigii
has not been reported so far.
OR'
OR'
XIV
aR' = H
b R' = Ac
42
Table IV. H NMR data of MK-6 tetraacetate (100 MHz, CDCI3)
Assignment Chemical shift ( S , ppm)
18-Me 0.68 (s,3H)
28 - Me 0.76 (d, J=6.8 Hz, 3H)
0.84 (d, J=6.5 Hz. 6H)
0.90 (d, J=6.5 Hz, 3H)
1.00 (s,3H)
1.88-2.32(8, 12H)
3.22 - 3.80 (m, 8H)
5.40 (m, 1H)
1.12 - 1.60 (cyclic & side (chain)
26, 27
21-Me
19-Me
-Me
-OCO Me
-CH,0-
=CH
-CH2, • •CH
43
EXPERIMENTAL
The dried leaves (1.5 kg) of M. koenigii were extracted with methanol.
Repeated extractions were made until the solution become colourless. The
methanol extracts were mixed and dried under reduced pressure. A dark green
gummy mass (100g ) was obtained which was treated with petrol, benzene,
ethyl acetate and acetone several times till solvent become colourless in each
case.
The petrol fraction (40g) was adsorbed on silica gel (30g) and trans
ferred to column of silica gel (150g) set with petroleum ether (60-80°C). The
elution of column with petrol - benzene (1:1) gave crude MK-1 which on disso
lution in benzene - chloroform, crystallization and recrystallization gave pure
MK-1.
MK-1: Crystalline compound (400 mg), crystallized in benzene which melts at
205°C and dissolves readily in chloroform. Rf=0.2958 (silica gel, petrol- ben
zene, 4:6) , in UV light gave brown fluorescent, adsorb iodine, developed on
charring with perchloric acid and with alcoholic FeCl3 gave yellowish green
colouration.
UV, IR, MS and 'H NMR data of MK-1.
UVX^*°'^nm (log e) : 360 (4.20), 340 (4.24), 300 (4.62), 240 (4.69), 230 (4.68). m3x
IRv*^^'cm-^ : 3400, 2950, 2850, 1640, 1610, 1580, 1490, 1450, 1420, 1400, max
1380, 1360, 1330, 1290, 1260, 1240, 1200, 1170, 1130, 1120, 1100,
1050, 1020, 890, 870, 840, 800, 770, 760, 720, 680.
MS m/z (rel. int.) : 293 [M]* (40), 278 [M-15] (100) aromatlzation of chromene
ring by loss of Me group, 262 (10), 235 (22), 191 (10), 139(16), 117(10),
77(10), 53(18).
44
^H NMR (600 MHz ) : 5 1.485 (s, 6H, gem dimethyl protons), 2.340, (s, 3H, 3-
Me), 3.903 (s, 3H, Ar-OCHg), 5.640 (d, J=10 Hz, 3'-H), 6.536 (d, J=10Hz,
4'-H), 6.940 (dd, J=8.4, 2.2 Hz, 7-H), 7.214 (d, J=8.4 Hz, 8-H), 7.421 (d,
J=2.2 Hz, 5-H), 7.627 (s, IH, 4-H), 7.686 (br s, IH, N-H).
^ C NMR data is given in discussion (Table I).
From UV, MS, IR, 'H NMR and ^ C NMR spectral data the compound was
found to be koenimbine = koenimbin.
Further elution of column with petrol - benzene (1:1) and crystallization
in ethanol - chloroform (8:2) crude MK-2 was obtained.
MK-2 : Crude MK-2 on crystallization in benzene gave colourless crystals (180
mg) of Mk-2 (pure), readily dissolved in chloroform and having melting point
139-41 °C. Rf=0.7384 (silica gel, benzene - acetone, 9:1) . UV inactive but ap
peared on absorption with iodine and charring with perchloric acid.
IR, MS and 'H NMR data of MK-2.
IRv l^ax^m ••• : 3400, 3250, 2900, 2850, 1640, 1370, 1050, 950, 880.
MS m/z (rel. int. ) : 414 [M]* (100), 399 (25), 396 (30), 381 (18), 273 (50). 271
(44), 255 (60), 231 (30), 213 (50).
^H NMR (100 MHz) : 5 0.68 (s, 3H, 18-Me), 0.75 (d, J=6.8 Hz, 3H, 28 -Me), 0.84
(d, J=6.5 Hz, 6H, 26, 27-Me), 0.92 (d, J=6.5 Hz, 3H, 21 -Me), 1.01 (s, 3H,
19-Me), 1.07 - 2.34 (-CH^ and -CH protons of cyclic and side chain), 3.52
(m, IH , 3-ax-H), 5.15 (m, IH, -OH), 5.39 (m, IH, oleflnic proton).
From IR, MS, ^H NMR spectral data and on direct comparison with authentic
sample the compound was found to be p- sitosterol.
Elution of petrol fraction column with benzene, applying preparative thin
45
layer chromatography (PTLC) and on crystallization MK-3 was isolated.
MK-3 : Colourless granular compound (120 mg), soluble in chloroform, crystal
lized in alcohol - benzene (7:3) having melting point 94-5°C. Rf= 0.6484 (silica
gel, benzene, PTLC), UV inactive, absorb iodine and developed on charring
with perchloric acid.
Spectral data of MK-3.
IR V *^ '' cm- : 3300, 2900, 2820, 2300, 1460, 1420, 1370, 1330, 1060, max
1010,760,720. •
MS m/z (rel. int.) : 476 [M]* (3.5), 462 (1.2), 448 (7.5), 434 (1.4), 420 (5), 406
(1), 181 (2), 167 (3), 153 (4), 139 (10), 125 (19), 111 (36), 97 (66), 83
(69), 69(56), 57(100).
^H NMR and ^ C NMR is given in discussion.
From IR, MS, ^H NMR and ^ C NMR the compound was found to be a long
chain alcohol.
Further elution of column with benzene, crystallization and recrystalliza-
tion in benzene MK-4 was isolated.
MK-4 : Colourless, crystalline compound (13 mg) which dissolves readily in
chloroform and melts at 235 - 3 7 ^ . Rf = 0.8238 (silica gel, petrol - benzene,
4:6), in UV light gave brown fluorescent, absorb iodine and developed on char
ring with perchloric acid.
Spectral data of MK-4 :
UV>.^^°^(log e) 241 (4.62), 257 (4.37), 307 (4.20), 355 (3.51) iTl3X
IRv'^^'cm-^ : 2950, 2900, 1640, 1600, 1490, 1450, 1370, 1350, 1330, 1300, max
46
1250. 1220, 1200, 1180, 1150, 1120,1080, 1060, 990, 850, 740.
MS 70 eV, m/z : 331 [Mr, 316 [base peak], 248.
H NMR (100 MHz) : 5 0.20 (m, 1H, 5'-ax-H), 1.32 (s, 3H, 8'-Me), 1.40 (m,
1 H,5'-eq-H), 1.48 (s, 3H,3'-Me), 1.92 (s, 3H, 9'-Me), 2.34 (s, 3H, 3-Me),
3.35 (br d, 1H, benzylic proton at 1"), 7.35 (s, 3H, Ph - CH3), 6.30 - 7-90
(m, 5H, Ar-H).
From UV, IR, MS and ^H NMR spectral data the compound was found to be
mahanimbidine = currayangine = murrazotine.
The benzene fraction (18g) adsorbed on silica gel (15g) and transferred
to the silica gel column set with petroleum ether (60-80''C). Elution of column
with petrol - benzene (1:1) and crystallization in alcohol - benzene (8:2) MK-5
was isolated.
MK-5 : Light yellowish green crystals (600 mg) melting point 225°C and soluble
in chloroform. Rf=0.3584 (SiOj, benzene), in UV light gave brown fluorescent,
adsorb iodine, developed on charring with perchloric acid and with alchoholic
FeClg gave green colouration.
Spectral data of MK-5 .
^j^^EtOH ^i^g gj . 235 (4 5g)_ 240 (4.57) , 288 (4.36), 297 (4.53). 336 (3.97).
IRv^^'cm-^ : 3400, 2900, 2800, 1640, 1620, 1600, 1585, 1490, 1470, 1455, max . I . . . , . I .
1440,1420, 1400,1370,1355, 1340,1290, 1270,1240, 1200, 1185, 1170,
1150, 1120, 1110, 1050, 1030, 990, 970. 940, 920, 860, 820, 760, 740,
720, 670.
MS m/s (rel. int.) : 323 [M]^ (60), 308 [M-15] (100) aromatization of chromene
ring by loss of methyl group, 293 [M-30] (20) further loss of methyl group.
47
264 (10), 250 (16), 154 (14). 133 (10), 71 (16).
^H NMR (600 MHz) : 5 1.463 (s, 6H, 2'-Me2), 2.311 (s, 3H, 3-Me), 3.902 (s, 3H,
6-OMe), 3.957 (s, 3H, 7-OMe), 5.657(d, J=10Hz, 1H, 3'-H), 6.570 (d, J=10Hz,
1H, 4'-H), 6.874 (s, 1H, 8-H), 7.367 (s, 1H, 5-H), 7.532 (s, 1H, 4-H).
"C NMR is given in discussion (Table III). From UV, IR, MS, ^H NMR and ^ C
NMR the compound was found to be koenimbidine s koenigicine = koenidine.
Further elution of column with benzene - ethyl acetate (8:2) and on crys
tallization in pyridine - benzene - methanol (1:11:10) MK-6 was isolated.
MK-6 : Light green granules (105g), soluble in pyridine and boiling methanol,
melt at 290°C. Rt= 0.6421 ( SiO^, ethyl acetate - methanol, 8:2 ), UV inactive,
adsorb iodine and developed on charring with perchloric acid.
IR data of MK-6 :
IR\) 'cm- : 3350, 2950, 2850, 1460, 1430, 1380, 1370, 1160, 1110, 1070. max
1010, 950, 920, 880.
Acetylation of MK-6: A tetra acetate (m.p. 158°C) was prepared by treatment
of MK-6 with A c p and pyridine. Rf=0.900 (SiOg", benzene - acetone, 7:1). de
veloped by adsorption of iodine and charring with perchloric acid.
IR\)'^^'cm-^: 2900. 2850, 1750, 1740, 1460, 1430, 1370, 1210, 1150,1030,900 max
H NMR (100 MHz) : 6 0.68 (s, 3H, 18-Me), 0.76 (d. J=6.8 Hz. 3H, 28-Me), 0.84
(d, J=6.5 Hz, 6H, 26. 27-Me). 0.90 (d, J=6.5 Hz. 3H. 21-Me). 1.00 (s, 3H. 19-
Me). 1.88-2.32 (s, 12H, - OCO CH3), 3.22 - 3.80 (m, 8H.-CHp-). 5.40 (m, 1H.
=CH), 1.12 -1.60 (-CHg, - CH protons of cyclic and side chain).
From IR of MK-6, IR and ^H NMR of its tetraacetate derivative the compound
was found to be sitosterol - 3 - 0 - p - D - glucoslde.
48
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