chapter 4 trichosanthes dioica trichosanthes...

Chapter 4 Trichosanthes dioica Roxb.

Md. Sarfaraj Hussain, Ph. D, Thesis (2013) Integral University 99

Trichosanthes dioica Roxb.

(Cucurbitaceae)

4.1. Introduction

It is a perennial crop, highly accepted due to its availability for eight months in a year

(February–September). The fruit is edible which is cooked in various ways either alone or

in combination with other vegetables or meats. In India, all parts of this plant have been

traditionally used for various medicinal purposes It contains a variety of trace elements

considered beneficial for the human physiology, such as magnesium, potassium, copper,

sulphur and chlorine. In addition, fruits and other parts such as leaves and tender shoots

have been used in the indigenous system of medicine since ancient times [1, 2].

Vernacular name

Bengali : Patol.

Chinese : Yeshe gua.

English : Pointed gourd, Wild snake gourd.

German : Patol.

Hindi : Palwal, Parval, Parwal, Patola.

Italian : Patole.

Nepalese : Paraval (Parval), Paravara (Parvar)

Distribution

It is a dioecious climber found wild throughout the plains of North and North-East India

from Punjab to Assam and Tripura states of India. It is also commercially cultivated in

India, Pakistan, Bangladesh and Sri Lanka for its fruits, a common culinary vegetable in

the Indian subcontinent [3].

Morphology

The plant is a perennial, dioecious and grows as a vine (Fig. 1). Roots are tuberous with

long taproot system. Vines are pencil thick in size with dark green cordate simple leaves.



Flowers are tubular white with 16–19 days initiation to anthesis time for pistillate flowers

and 10–14 days for staminate flowers. Stigma remains viable for approximately 14 hours

and 40–70% of flowers set fruit. Based on shape, size and striation, fruits can be grouped

into 4 categories: (1) long, dark green with white stripes, 10–13 cm long, (2) thick, dark

green with very pale green stripes, 10–16 cm long, (3) roundish, dark green with white

stripe, 5–8 cm long, and (4) tapering, green and striped, 5–8 cm long [4].

Fig. 4.1 Exomorphic features of the plant

Ethnobotanical uses

The fruits and leaves are the edible parts of the plant which are cooked in various ways

either alone or in combination with other vegetables or meats. Juice of leaves of T. dioica

is used as tonic, febrifuge and in subacute cases of enlargement of liver and spleen. In

Charaka Samhitha the leaves and fruits are used for treating alcoholism and jaundice. The

leaves are used in oedema and alopecia, as antipyretic, diuretic, cardiotonic and laxative.

According to Ayurveda, its root is ingested as a strong purgative, as tonic, febrifuge, in

treatment of jaundice, anasarca and ascites [5-7]. In Ayurvedic medicine, T. dioica is

tridoshic vegetable and is an remedy for balancing of all five fundamental elements. It is

extremely enhancing, easy to digest and assimilate into the physiology does not create

any is effective for all seven layers of the skin.



4.2. Phytoconstituents reported in T.dioica

Previous phytochemical study reveals that in addition to a number of tetra and

pentacyclic triterpenes, the toxic bitter principles cucurbitacins (a group of often highly

oxygenated tetracyclic compounds with a unique carbon skeleton and almost a carbonyl

group in ring C) may be considered as a taxonomic character of Cucurbitaceae. The seeds

of T. dioica contained a large amount of peptides. The seed peptides have the unique

property of being resistant to the action of silver nitrate, a sensitive reagent commonly

used to stain proteins [8]. The seed extract of T. dioica contained 7-

oxidihydrokarounidol-3-benzoate as the most predominant component in the highly polar

fraction of the nonsaponifiable lipid [9]. Two main phytosterols present in T. dioica are

namely, 24α- ethylcholest-7-enol & 24β-ethylcholest-7-enol [10]. The seeds of T. dioica

also contain lectin, a carbohydrate (specifically galactose) binding protein which is

homologous to Type-II ribosome inhibitory proteins (Type-II RIP).

Ali et al., (2004) have studied purification, physicochemical characterization, saccharide

specificity, and chemical modification of a Gal/GalNAc specific lectin from the seeds of

T. dioica [11].

Kabir et al., (2000) evaluated a large amount of peptides. From the seeds peptides have

the unique property of being resistant to the action of silver nitrate, a sensitive reagent

commonly used to stain proteins [12].

Ratnesh et al., (2009) studied that the total phenolic content of T. dioica leaves is about

two times more than that obtained from the fruits and seeds of Moringa olifera and

Embilica officinalis, respectively [13].



HO

H H

HO

H

H H

Stigmast-7-en-3β-ol Stigmasterol

O

O

O

O

HO

OH

HO

Cucurbitacin B

Cucurbitacin D



Cucurbitacin E

Figure 4.2. Structures of previously isolated compounds from T. dioica

4.3. Previously reported bioactivities

Following are the folk and traditional uses of the plant; it has been investigated

scientifically in animal models to validate the potential of the plant in cure of variety of

ailments. The both aqueous and alcoholic extracts have LD50 7.5g/kg [14].

Anti-inflammatory activity

Fulzul et al., (2001) found anti-inflammatory activity of polyherbal formulation “Jatyadi

Ghrita”, the ingredients of Jatyadi Ghrita are Jasmine officinale, Azadirachta indica,

Berberis aristata, Curcuma longa, Picrorrhiza kurroa, Rubia cordifolia, Trichosanthes

dioica, Aristolochia indica, Hemidesmus indicus, Glycyrrhiza glabra and Cow’s ghee

[15].

Antibacterial activity

Hariti and Rathee et al., (1995) showed antibacterial activity of the unsaponifiable

fraction of the fixed oil of T. dioica seeds against Bacilus anthracis and Xanthomonas

malracearum [16].



Rai et al., (2010) reported the in vitro assessment of antimicrobial effect of different

concentration of extract of different part of T. dioica. Five clinical isolates of different

bacterial strains were used and the disc diffusion method was opted. The results revealed

that leaves, fruits and seeds of T. dioica plant may be used as antibacterial agents.

Though the leaves extract was active against all five strains, the highest inhibition was

observed against Mycobacterium smegmatis. Thus the leaves extract could be used for

tuberculosis treatment [17].

Antifungal activity

Hariti and Rathee et al., (1996) stated that the fixed oil of seeds of Trichosanthes species

including T. dioica have antifungal property [18].

Anthelmintic activity

The in vitro activities of defatted methanol (MeOH) extract of the leaves from T. dioica

and its ethyl acetate (EtOAc) and n-butanol (n-BuOH) fractions were evaluated against

Pheretima posthuma (Annelida) and Ascaridia galli (Nematoda). All the extracts

demonstrated concentration dependent paralytic and lethal effects on P. posthuma and

lethal effects on A. galli. The EtOAc fraction was found to be the most potent followed

by the defatted MeOH extract and its n-BuOH fraction. A. galli was found to be more

sensitive than P. posthuma against all tests extracts indicating T. dioica as an effective

nematocide [19].

Anti-oxidant activity

Shivhare et al., (2010) studied anti-oxidant activity of aqueous extract of T. dioica fruits

for its free radical scavenging property in different in vitro methods as 1, 1 diphenyl-2-

picryl hydrazyl, nitric oxide, reducing power assay and hydrogen peroxide radical

method. The findings could justify the inclusion of this plant in the management of

antioxidant activity [20].

Cholesterol-lowering activity

Sharmila et al., (2007) observed cholesterol lowering activity of the aqueous fruit

extract of T. dioica in normal and streptozotocin diabetic rats [21].



Sharma and Pant et al., (1992) showed influence of alcoholic extract of whole

fruit of T. dioica on blood sugar, serum lipids, lipoproteins and faecal sterols in

normal albino rabbits. Effect of oral administration of 2 ml per day of suspension

(in water) of alcoholic extract of whole fruit of T. dioica (2%) with basal diet for

four weeks was studied in the normal albino rabbits. It was observed that this

extract lowered the blood sugar, total cholesterol, low density lipoprotein

cholesterol and triglyceride levels, and increased the high density lipoprotein

cholesterol, phospholipid and faecal sterol levels [22].

Hepatopotective activity

Ghaisas et al., (2008) showed hepatoprotective activity of aqueous and ethanolic extracts

of T. dioica (whole plant) in ferrous sulphate-induced liver injury. Ethanolic and aqueous

extracts of T. dioica at different doses (100, 200 and 400 mg/kg) and silymarin (100

mg/kg) were administered orally for 10 days. The groups treated with 400 mg/kg aqueous

and ethanolic extract showed significant reduction in AST, ALT, ALP level. The

pretreatment with T. dioica extracts showed profound histopathological protection to

liver cells as evident from histopathological studies. Hence, it can be concluded that T.

dioica has significant hepatoprotective activity [23].

Hypoglycemic activity

The anti-diabetic activity was examined to study the effects of single and repeated

oral administration of the aqueous fruit extract of T. dioica at a dose of 50 ml/kg

b.w in normal and streptozotocin-induced diabetic rats. The aqueous fruit extracts

of T. dioica (50 ml/kg) were administered orally for 15 days, to normal and

diabetic rats. The effect of the fruit extracts on cholesterol and triglycerides, were

studied. The body weights of the rats were observed. In normal rats, the aqueous

fruit extract of T. dioica induced significant decrease of plasma cholesterol and

triglyceride concentrations 6 hrs after a single oral administration, and also in 2

weeks after repeated oral administrations. One week after repeated oral

administration of aqueous extract of T. dioica, the plasma triglyceride levels were

significantly decreased. The decreasing trend continued even after 2 week. Study



indicates that the aqueous fruit extract of T. dioica exhibits cholesterol and body

weight-lowering activities in both normal and hyperglycemic rats [21].

The hypoglycaemic activity of aqueous extracts of T. dioica fruits at a dose of

1000 mg/kg body weight daily once for 28 days reduced the levels of fasting

blood glucose, postprandial glucose, asparate amino transferase, alanine amino

transferase, alkaline phosphatase, creatinine, urine sugar and urine protein where

as total protein and body weight was increased. The study showed that aqueous

extract of Trichosanthes dioica possessed significant hypoglycemic activity [14].

The extract significantly lowers the fasting blood glucose level and markedly

improves the glucose tolerance of rats [24].

In normal, sub-diabetic, and mild diabetic animal models the graded doses of the

extract, viz., 500, 750, 1000, and 1250 mg/kg b. w. were administered orally. It

was found that the blood glucose concentration decreased in a dose-dependent

manner. The dose of 1000 mg/kg b.w. was found to be most effective with a

maximum fall of 30.4% at 6 h during FBG studies in normal rats. However, the

GTT studies showed the maximum reduction of 26.6% at 5 h in normal rats.

Moreover, in case of sub-diabetic and mild diabetic rats, the observed reduction in

blood glucose levels was 32.8% and 35.9%, respectively, at 3 h during GTT. [25].

This study was to screen the glycemic attributes of an aqueous extract of T. dioica

leaves in normal as well as various diabetic models. The variable doses of 250,

500, and 750 mg kg−1 body weight (bw) of the extract were administered orally

to normal and streptozotocin (STZ)-induced sub- and mild-diabetic rats in order

to define its glycemic potential. The dose of 500 mg kg−1

b.wt. was identified as

the most effective dose which brings down the blood glucose level (BGL) by

32.9% (P<0.001) at 6 h during fasting blood glucose (FBG) studies in normal

rats. However, glucose tolerance test (GTT) showed the maximum reduction of

30.9% (P<0.001) in BGL at 5 h in normal rats with the same dose, whereas the

reduction observed was by 40.3% and 88.6% (P<0.001) in sub- and mild-diabetic

rats, respectively, at 3h of glucose administration only [26].



Wound healing activity

Shivhare et al., 2010 studied methanolic extract of the plant T. dioica for assessment of

healing potential in the form of simple ointment using full thickness burn wound model

in rats. The effect produced by the extract ointment provides significant healing when

compared with the control and standard groups [27, 28].

Clinical Investigation

T. dioica is known to have antiulcerous effect in polyherbal preparation. Two

formulations have been clinically investigated as given below:

Rai and Tripathi, (1968) showed that Patoladi kasaya, a polyherbal formulation,

consisted of 11 herbs viz., Patola, Haritaki, Bibhitaka, Amalaki, Kutaki, Cirayata,

Amrta, Pittapapada, Sunthi, and Bhrngaraja exhibited complete improvement in

50% cases and partial improvement in 40% cases with peptic ulcer (10 patients

case study) [29].

Tripathi and Pathak, (1975) evaluated another Patoladi kasaya which consisted of

only four herbs namely Sunthi, Patola, Amrta, Kutaki in the 33 case study of

duodenal ulcer. It kept the patients symptoms/complication free when given in

dose of 40 ml/day in two divided doses. It normalized both hyper and hypoacidity

of these patient [30].

Aryavansha et al., (1981) studied the efficacy of single herb patola in 20 patients

with duodenal ulcer. Effectively of patola in duodenal ulcer was found 45%

excellent response out of 20 cases [31].



4.4. EXPERIMENTAL

MATERIALS & METHODS

Phytochemical Investigation

Collection of plant material

The fresh leaves of T. dioica were collected from the basin area of Koshi river, Saharsa

District Bihar, India, in May 2009. The plant specimen was authenticated by Prof (Dr.)

Anjani Kumar Sinha, Principal, M L T Saharsa College Saharsa, Saharsa, Bihar. A

voucher specimen no SHC 57/05/2009 has been deposited at the herbarium, Department

of Botany, M L T Saharsa College Saharsa- 852201.

Preparation of extract

The leaves of T. dioica (3.5 kg) was shade dried, coarsely powdered and extracted

exhaustively with methanol in a Soxhlet apparatus. The methanolic extract of the plant

was then concentrated on a steam bath and dried under reduced pressure to get 87.9 g of

dark brown mass.

Preparation of slurry

The concentrated extract (80 g) of the drug was dissolved in minimum amount of

methanol with constant stirring, till desired consistency was obtained. A weighed

quantity of silica gel for column chromatography (60-120 mesh) was then added slowly

with continuous mixing until the whole methanolic solution of plant extract adsorbed on

the silica gel particles. It was dried in the air; the larger lumps were broken and finally

passed through a sieve (No. 8) to get uniform particle size.

Packing of column & Isolation of phytoconstituents

A column of 3.0 feet, height and 16 mm internal diameter was taken, cleaned properly

and dried. The lower end of the column was plugged with non-absorbent cotton wool.

The column was clamped and fitted in a vertical position on a stand. The column was half

filled with petroleum ether (b.p. 60-80 ºC). Silica gel (for column, 60-120 mesh) was then

poured in small portions and allowed to settle down and the dried plant extract slurry was



loaded over the column. The developments and elution of the column were carried out

with successive series of different solvents in various combinations, such as petroleum

ether (100), petroleum ether: chloroform (3:1, 1:1, 1:3), chloroform (100), chloroform:

methanol (99:1, 98:2, 97:3, 19:1, 9:1, 4:1) and methanol to isolate the compounds.

Homogeneity of the fractions

The fractions collected were subjected to thin layer chromatography (TLC) to check

homogeneity of various fractions. Chromatographically identical fractions (having same

Rf values) were combined together and concentrated. They were then crystallized with

suitable solvent system.

Observation

Following compounds have been isolated from the T. dioica:

Heneicosanyl oleate (TD-01)

Elution of the column with petroleum ether: chloroform (9:1) gave afforded colourless

crystal of TD-01, recrystallized from chloroform: methanol (1:1), 180 mg, (0.36%,

yield).

M.P.: 91-92°C

UV λmax (MeOH): 308 nm

IR max (KBr): 2921, 2851, 1735, 1458, 1384, 1261, 1096, 803 cm-1

1H NMR (DMSO-d6): δ 5.19 (1H, m, H-9), 5.09 (1H, m, H-10), 4.40 (2H, t, J=9.5 Hz,

H2-1ꞌ), 2.27 (2H, t, J=7.2 Hz, H2-2), 2.18 (1H, m, H-8), 1.96 (2H, m, H-11), 1.48 (4H, m,

2×CH2), 1.23 (52H, brs, 2×CH2), 1.13 (4H, m, 2×CH2), 0.85 (2H, t, J=6.3 Hz, Me-18)

,0.81 (3H, t, J=6.1 Hz, Me-21ꞌ)

13C NMR (CDCL3): δ 172.61 (C-1), 129.87 (C-9), 118.25 (C-10), 62.03 (C-1ꞌ), 32.54

(CH2), 29.92 (30×CH2), 27.16 (CH2), 22.19 (CH2), 14.33 (Me-18, Me-21ꞌ)

+ve ESI MS m/z (rel:int): 576 [M]+(C39H76O2) (32.6), 282 (18.2), 265 (14.9).



Farnasonoic acid α-L-glucoside (TD-02)

Further elution of the column with chloroform: methanol (19:1) furnished pale yellow

crystalline mass of TD-02, recrystallized fromchloroform: methanol (1:1), 205 mg,

(0.43% yield)

Rf value: 0.72;

Solvent: Chloroform: methanol (19:1)

m.p.: 105- 106°C


IR max (KBr): 3399, 3251, 2931, 2842, 1682, 1458, 1384, 1047, 822 cm-1

1H NMR (DMSO-d6): δ 6.16 (1H, brs, H-1ꞌ), 4.47 (1H, m, H-5ꞌ), 4.22 (1H, dd, J=6.3,6.5

Hz, H-2ꞌ), 3.62 (1H, m, H-3ꞌ), 3.53 (1H, m, H-4ꞌ), 3.05 (2H,brs, H2-6ꞌ), 2.32 (1H, d,

J=15.6 Hz, H2- 2a), 2.23 (1H, d, J=9.9 Hz, H2- 2b), 2.18 (1H, m, H-18), 1.99 (1H, m, H-

11), 1.91 (1H, m, H2-6), 1.86 (1H, m, H2-8), 1.82 (2H, m, H2-5), 1.78 (2H, m, H2-9),

1.64 (2H, m, H2-4), 1.42 (2H, m, H2-10), 1.23 (3H, brs, Me-14), 1.13 (3H, d, J=5.7 Hz,

Me-12), 0.97 (3H, d, J=6.0 Hz, Me-15), 0.85 (3H, d, J=7.3 Hz, Me-13).

13C NMR (DMSO-d6): δ 177.03 (C-1), 42.69 (C-2), 36.84 (C-3), 28.86 (C-4), 28.83 (C-

5), 27.91 (C-6), 74.01 (C-7), 26.06 (C-8), 25.86 (C-9), 24.93 (C-10), 36.89 (C-11), 19.20

(C-12), 20.46 (C-13), 23.01 (C-14), 18.99 (C-15), 104.26 (C-1ꞌ), 80.06 (C-2ꞌ), 64.86 (C-

3ꞌ), 84.50 (C-4ꞌ), 77.76 (C-5ꞌ), 63.11 (C-6ꞌ).

+ve ESI MS m/z (rel:int): 420 [M]+(C21H40O8) (18.2), 377 (21.7), 335 (15.3), 291 (6.5),

240 (7.3), 214 (9.1), 179 (19.4), 172 (6.6), 163 (7.8), 129 (20.5).

Lanosten-5-en-3β-ol-26-oic acid glucosyl capriate (TD-03)

Further elution of the column with chloroform: methanol (19:1) gave colourless crystal of

TD-03, recrystallized from chloroform: methanol (1:1), 205 mg (0.38% yield).

Rf value: 0.72



Solvent: Chloroform: methanol (19:1)

m.p.: 135- 136°C

UV λmax (CHCl3): 243 nm

IR max (KBr): 3510, 3417, 2923, 2852, 1737, 1688, 1635, 1463, 1383, 1262, 1075,

811cm-1

1H NMR:(DMSO-d6):δ 5.42 (1H, d, J=5.7 Hz, H-6), 3.73 (1H, dd, J=5.5, 8.8 Hz, H-

3α), 1.20 (3H, brs, Me-29), 1.13 (3H, d, J=6.6 Hz, Me-27), 1.02 (3H, brs, Me-19), 0.96

(3H, brs, Me-28), 0.92 (3H, d, J=6.3 Hz, Me-21), 0.85 (3H, brs, Me-30), 0.81 (3H, brs,

Me-18), 5.07 (1H, d, J=7.3 Hz, H-1ꞌ), 4.22 (1H, m, H-2ꞌ), 3.85 (1H, m, H-3ꞌ), 3.63 (1H,

m, H-4ꞌ), 3.05 (2H, brs, H2-6ꞌ), 2.26 (2H, t, J=7.2 Hz, H2-2ꞌꞌ), 1.62 (2H, m, H2-3ꞌ), 1.23

(8H, brs,4×CH2), 0.83 (1H, t , J=6.5 Hz, Me-8ꞌꞌ),

+ve ESI MS m/z (rel:int): 746 [M]+(C44H74O9) (4.1), 602 (5.2), 457 (22.8), 440 (19.7),

179 (12.6), 144 (15.7), 127 (23.1).

Lanosten-3β-ol-26-oic acid tetraglucoside (TD-04)

Elution of the column with chloroform: methanol (19:1) gave colourless crystals of TD-

04, recrystallized from methanol, 103 mg (0.23% yield).

Rf value: 0.39

Solvent: chloroform: methanol (9:1);

m.p.: 190-192°C.


IR max (KBr): 3515, 3406, 3245, 3406, 3245, 2925, 2843, 1688, 1455, 1384, 1264,

1074, 811cm-1

.

1H NMR:(DMSO-d6): δ 5.41 (1H, dd, J=6.8, 6.5 Hz, H-6), 3.84 (1H, dd, J=5.3, 8.8 Hz,

H-3α), 1.22 (3H, brs, Me-29), 1.15 (3H, d, J=6.5 Hz, Me-27), 1.02 (3H, brs, Me-27), 0.96




Me-18), 5.41 (1H, d, J=6.6 Hz, Me-1ꞌ), 4.39 (1H, dd, J=6.3 Hz, Me-2ꞌ), 3.70 (1H, m, H-

3ꞌ), 3.41 (1H, m, H-4ꞌ), 4.59 (1H, m, H-5ꞌ), 3.16 (2H, d, J=8.6 Hz, H2-6ꞌ), 5.06 (1H, d,

J=9.0 Hz, H-1ꞌꞌ), 4.26 (1H, m, H-2ꞌꞌ), 3.62 (1H, m, H-3ꞌꞌꞌ), 3.69 (1H, m, H-5ꞌꞌ), 3.22 (4H,

brs, H2-6ꞌꞌ, H2-6ꞌꞌꞌ), 4.98 (1H, d, J=7.5 Hz, H-1ꞌꞌꞌ), 4.22 (1H, m, H-2ꞌꞌꞌ), 3.66 (2H, m, H-3ꞌꞌꞌ,

H-3ꞌꞌꞌꞌ), 3.34 (1H, m, H-4ꞌꞌꞌ), 4.56 (1H, m, H-5ꞌꞌꞌ), 4.89 (1H, d, J=9.0 Hz, H-1ꞌꞌꞌꞌ), 4.30 (1H,

m, H-4ꞌꞌꞌꞌ), 3.36(1H, m, H-4ꞌꞌꞌꞌ), 4.44 (1H, m, H-5ꞌꞌꞌꞌ), 3.05 (2H, brs, H2-6ꞌꞌꞌꞌ),

13C NMR (MeOD): δ 36.79 (C-1), 30.84 (C-2), 80.19 (C-3), 42.36 (C-4), 139.24 (C-5),

119.71 (C-6), 30.55 (C-7), 40.71 (C-8), 48.29 (C-9), 36.68 (C-10), 22.95 (C-11), 28.91

(C-12), 44.52 (C-13), 54.25 (C-14), 34.99 (C-15), 42.83 (C-16), 51.18 (C-17), 18.49 (C-

18), 20.46 (C-19), 35.13 (C-20), 19.17 (C-21), 33.15 (C-22), 25.74 (C-23), 45.16 (C-24),

30.23 (C-25), 181.26 (C-26), 24.87 (C-27), 26.14 (C-28), 26.77 (C-29), 19.11 (C-30),

104.86 (C-1ꞌ), 74.84 (C-2ꞌ), 72.41 (C-3ꞌ), 71.65 (C-4ꞌ), 79.70 (C-5ꞌ), 64.74 (C-6ꞌ), 103.89

(C-1ꞌꞌ), 73.90 (C-2ꞌꞌ), 72.36 (C-3ꞌꞌ), 71.20 (C-4ꞌꞌ), 78.28 (C-5ꞌꞌ), 64.47 (C-6ꞌꞌ), 100.94 (C-

1ꞌꞌꞌ), 73.88 (C-2ꞌꞌꞌ), 72.11 (C-3ꞌꞌꞌ), 69.86 (C-4ꞌꞌꞌ), 77.64 (C-5ꞌꞌꞌ), 62.87 (C-6ꞌꞌꞌ), 94.56 (C-

1ꞌꞌꞌꞌ), 73.86 (C-2ꞌꞌꞌꞌ), 72.11 (C-3ꞌꞌꞌꞌ), 69.19 (C-4ꞌꞌꞌꞌ), 75.19 (C-5ꞌꞌꞌꞌ), 61.65 (C-6ꞌꞌꞌꞌ),

+veESI MS m/z (rel:int): 1106 [M]+

(C54H90O23) (2-1), 592 (11.5), 504 (8.6), 459 (42.8),

442 (33.6), 341 (10.2), 179 (18.1).

Lanastan-3β-ol-26-oic acid tetraglucoside (TD-05)

Further elution of the column with chloroform: methanol (9:1) gave afforded colorless

crystal of TD-05, recrystallized from methanol, 155mg, (0.34% yield).

Rf value: 0.43 (Chloroform: methanol- 9:1)

m.p.: 125-127 °C


IR max (KBr): 3510, 3406, 3345, 3225, 2930, 2845, 1685, 1462, 1384, 1074 cm-1

1H NMR (DMSO-d6):δ 3.89 (1H, dd, J=5.5,8.8 Hz, H-3α), 2.27 (1H, m, H-25), 1.23


Me-28), 0.93 (3H, d, J=6.6 Hz, Me-21), 0.84 ( 3H, brs, Me-30), 0.81 (3H, brs, Me-18),



5.49 (1H, d, J=7.3 Hz, H-1ꞌ), 4.71 (1H, m, H-5ꞌ), 4.43 (1H, dd, J=7.3, 6.5 Hz, H-2ꞌ), 4.12

(1H, m, H-3ꞌ), 3.74 (1H, m, H-4ꞌ), 3.22 (2H, brs, H2-6ꞌ), 5.37 (1H, d, J=7.5 Hz, H-1ꞌꞌ),

4.63 (1H, m, H-5ꞌꞌ), 4.39 (1H, m, H-2ꞌꞌ), 4.02 (1H, m, H-3ꞌꞌ), 3.72 (1H, m, H-4ꞌꞌ), 3.16

(2H, brs, H2-6ꞌꞌ), 5.09 (1H, d, J=7.2 Hz, H-1ꞌꞌꞌ), 4.58 (1H, m, H-5ꞌꞌꞌ), 4.33(1H, m, H-2ꞌꞌꞌ),

3.96 (1H, m, H-4ꞌꞌꞌ), 3.13 (2H, brs, H2-6ꞌꞌꞌ), 4.93 (1H, d, J=7.1 Hz, H-1ꞌꞌꞌꞌ), 4.52 (1H, m,

H-5ꞌꞌꞌꞌ), 4.24 (1H, m, H-2ꞌꞌꞌꞌ), 3.79 (1H, m, H-3ꞌꞌꞌꞌ), 3.53 (1H, m, H-4ꞌꞌꞌꞌ), 3.03 (2H, brs, H2-

6ꞌꞌꞌꞌ).

13CNMR (MeOD): δ 35.03 (C-1), 30.22 (C-2), 80.20 (C-3), 42.68 (C-4), 50.94 (C-5),

18.50 (C-6), 28.83 (C-7), 38.32 (C-8), 48.30 (C-9), 36.82 (C-10), 22.66 (C-11), 26.80

(C-12), 44.67 (C-13), 52.63 (C-14), 34.36 (C-15), 44.71 (C-16), 51.28 (C-17), 17.37 (C-

18), 20.46 (C-19), 36.84 (C-20), 19.20 (C-21), 33.54 (C-22), 25.52 (C-23), 45.03 (C-

24), 27.89 (C-25), 182.61 (C-26), 24.90 (C-27), 25.84 (C-28), 26.19 (C-29), 18.56 (C-

30), 105.41 (C-1ꞌ), 74.04 (C-2ꞌ), 72.16 (C-3ꞌ), 71.01 (C-4ꞌ), 78.35 (C-5ꞌ), 64.84 (C-

6ꞌ),101.35 (C-1ꞌꞌ), 73.99 (C-2ꞌꞌ), 71.79 (C-3ꞌꞌ), 69.55 (C-4ꞌꞌ), 77.74 (C-5ꞌꞌ), 64.53 (C-

6ꞌꞌ),100.99 (C-1ꞌꞌꞌ), 73.97 (C-2ꞌꞌꞌ), 71.81 (C-3ꞌꞌꞌ), 69.55 (C-4ꞌꞌꞌ), 76.44 (C-5ꞌꞌꞌ), 63.09 (C-

6ꞌꞌꞌ), 95.78 (C-1ꞌꞌꞌꞌ), 73.06 (C-2ꞌꞌꞌꞌ), 71.31 (C-3ꞌꞌꞌꞌ), 69.38 (C-4ꞌꞌꞌꞌ), 77.01 (C-5ꞌꞌꞌꞌ), 62.81 (C-

6ꞌꞌꞌꞌ).

+ve ESI MS m/z (rel:int): 1108 [M]+(C54H92O23) (1.8), 504 (10.2), 459 ((15.6), 442

(16.7), 397 (6.3) , 342 (7.2), 179 (10.8).

4.5. RESULT & DISCUSSION

Compound TD-01 heneicosanyl oleate, was obtained as a colourless crystalline mass

from chloroform: methanol (9:1) eluant. Its IR spectrum showed absorption band for

ester group at 1735 cm-1.The mass spectrum exhibited a molecular ion peak at m/z 576

corresponding to an alkenyl ester, C39H76O2. The ion peaks arising at m/z 265

[CH3(CH2)7CH=CH(CH2)7CO]+ and 282 [CH3(CH2)7CH=CH(CH2)7COOH]

+ indicated

that oleic acid was esterified with a C21 alcohol. The 1H NMR spectrum displayed

two one –proton multiplets at δ 5.19 and 5.09 assigned to vinylic H-9 and H-10

protons , respectively , a two–proton triplet at δ 4.40 (J=9.5Hz) ascribed to oxygenated

methylene H2-1ꞌ protons, other methyene protons between δ 2.27-1.13 and two three–



proton triplets at δ 0.85 (J=6.3Hz) and 0.81 (J=6.1 Hz) accounted to terminal C-18 and

C-21ꞌ primary methyl protons , respectively. The 13

C NMR spectrum of TD-01

displayed signals for ester carbon at δ 172.61 (C-1), vinylic carbons at δ 129.87 (C-9)

and 118.25 (C-10), oxygenated methylene carbons at δ 62.03(C-1ꞌ) and methyl carbons

atδ14.33 (C-18,C-21ꞌ). On the basis of above results the structure of TD-01 has been

identified as heneicosanyl n-octadec-9-enoate.

Compound TD-02 named farnasanoic acid α-L- glucoside, was obtain as a pale yellow

brown crystalline mass from chloroform: methanol (19:1) eluant. Its responded

positively to test for glycosides and showed IR absorption bands for hydroxyl groups

(3399, 3251cm-1

) and carboxyl function (1682 cm-1

). On the basis of mass and 13

C

NMR spectra, the molecular ion peak of TD-02 was determined at m/z 420

corresponding to a sesquiterpenic acid glycoside C21H40O8. The ion fragments arising at

m/z 129 [C6-C7 fission, (CH2)3 CH(CH3)CH2COOH]+, 291 [M-129]

+ and 335 [C7-C8

fission, (C6H11O5) OCH(CH3)(CH2)3 CH (CH3)CH2COOH]+ indicated the existence of

the sugar unit at C-7. The ion peaks generating at m/z 377 [M-C3H7]+, 214 [377-

C6H11O5]+, 240 [M-C6H11O6]

+, 172 [335- C6H11O5]

+ and 290 [335-COOH]

+ supported

the presence of the glycosidic unit at C-7 and carboxylic function in the molecule

(Scheme-1). The 1H NMR spectrum of TD-02 exhibited a one-proton broad singlet at δ

6.16 assigned to anomeric H-1ꞌ. The other sugar protons appeared from δ 4.47 to 3.05.

A three-proton broad singlet at δ1.23 and three doublets at δ 1.13 (J=5.7Hz), 0.97

(J=7.3Hz), and 0.85 (J=7.3Hz) integrating for three-protons each were attributed to

tertiary C-14 and secondary C-12, C-15 and C-13 methyl protons, respectively. The

remaining methine and methylene proton resonated between δ 2.32 - 1.42. The 13

C

NMR spectrum displayed signals for carboxylic carbon at δ 177.03 (C-1), anomeric

carbon at δ 104.26 (C-1ꞌ), other sugar carbons between δ 84.50-63.11, oxygenated

quaternary carbon at δ 74.01 and methyl carbons from δ 23.01 to 19.20. The absence

of any signal between δ 6.16-4.47 in the 1H NMR spectrum and from δ 177.03 to

104.26 in the 13

C NMR spectrum supported saturated nature of the molecules. The

existence of anomeric proton at δ 6.16 as singlet and sugar carbons at δ 80.06 (C-2ꞌ)

and 84.50 (C-4ꞌ) indicated furanose nature of the sugar unit in L-form. The presence of

two one –proton doublets at δ 2.32 (J=15.6Hz) and 2.23 (J=9.9Hz), accounted to



methylene H2-2 protons, suggested the location of the carboxylic function at C-1

carbon. Acid hydrolysis of TD-02 yield L-glucose. On the basis of above results the

structure of TD-01 has been established as 3, 7, 11-trimethyl-7α-L-glucofuranosyl-oxy-

n-dodecanoic acid. This is a new sesquiterpenic glucoside.

Compound TD-03 A lanostenoic acid glucosidic ester, was obtained as a colourless

crystalline mass from chloroform: methanol (19:1) eluants. It produced effervescence

with sodium bicarbonate solution, responded positively to glycoside tests and had

distinct IR absorption bands for hydroxyl groups (3510, 3417 cm-1

), ester group (1737

cm-1

), carboxyl function (1688 cm-1

), and unsaturation (1635cm-1

). Its mass spectrum

showed a molecular ion peak at m/z 746 consistent to the molecular formula of a

triterpenic glycosidic ester C44H74O9. The ion peaks arising at m/z 127 [CH3(CH2)CO]+,

144 [CH3(CH2)6COOH]+ and 602 [M-144]

+ indicated that capric acid was attached to

the sugar unit. The ion peaks generating at m/z 457 [M-C6H10O5-CO(CH2)6CH3]+, 440

[M-C6H10O6-CO(CH2)6CH3]+ and 179 [C6H11O6]

+ suggested attachment of the C6 sugar

unit to the tetracylic triterpenic acid. The 1H NMR spectrum of TD-03 displayed a one-

proton doublet at δ 5.42 (J=5.7 Hz) assigned to vinylic H-6 proton. A one–proton

double doublet at δ 3.73 (J=5.5, 8.9 Hz) was ascribed to oxygenated methine H-3α

proton. Four broad singlet’s at δ 1.20, 1.02, 0.96, and 0.85 and two doublets at δ 1.13

(J=6.6 Hz), 0.92 (J=6.3 Hz), all integrated for three-proton each, were attributed to

tertiary C-29, C-19, C-28 and C-30 and secondary C-27 and C-21 methyl protons,

respectively. A one- proton doublet at δ 5.07 (J=7.3 Hz) was due to anomeric H-1ꞌ

proton. The other sugar protons appeared as one-proton multiplets at δ 4.22, 3.85 and

3.63 and as a two-proton broad singlet at δ3.05. A two-proton triplet at δ 2.26(J=7.2

Hz) was accounted to methylene H2-2ꞌꞌ adjacent to the ester function .A three proton

triplet at δ 0.83 (J=6.5 Hz) was accounted to primary C-8ꞌꞌ methyl protons. The 1H and

13C NMR spectral data of the lanostene unit were compared with related triterpenoids

(Ching et al., 2005, Ching et al., 2006 & Ali, 2001). Acid hydrolysis of TD-03 yielded

D-glucose and capric acid (Co- TLC) comparable. On the basis of this discussion the

structure of TD-03 has been elucidated as lanosten-5-en-3β-ol-26-oic acid -3β-D-

glucopyranosyl-2ꞌ-octanoate. This is a new triterpenic glycosidic ester.



Compound TD-04, named lansotein-3β-ol-26-oic acid tetraglucoside, was obtained as

colourless crystals from chloroform: methanol (19:1) eluants. It gave positive tests for

glycosides and showed IR absorption bands for hydroxyl groups (3515, 3406, 3245 cm-

1), carboxylic function (1688 cm

-1) and unsaturation (1641cm

-1).On the basis of mass

and 13

C NMR spectra the molecular ion peak of TD-04 was determined at m/z 1106

consistent to the molecular formula of triterpenic tetraglycoside C54H90O23.The ion

peaks arising at m/z 179 [C6H11O6]+, 342 [C12H22O11]

+ and 504 [C12H22O11-C6H10O5]

+

indicated chain of hexose sugar in the glycone chain. The ion fragment generating at

m/z 459 [M-tetraglycoside, C30H51O3]+ and 442 [459-OH]

+ supported that the triterpene

contained one each vinylic linkage and carboxylic function. The 1H NMR spectrum of

TD-04 showed two one–proton double doublets at δ5.41(J=6.9, 6.5 Hz) and 3.84

(J=5.3, 8.8 Hz) assigned to vinylic H-6 and oxygenated methine H-3α protons,

respectively, five three-proton broad singlet’s at δ 1.22, 1.02, 0.96, 0.84 and 0.81

ascribed tertiary C-29 , C-19, C-28, C-30 and C-18 methyl protons, respectively and

two three-proton doublets at δ 1.15 (J=6.5 Hz) and 0.90(J=6.2 Hz) attributed

correspondingly to secondary C-27 and C-21 methyl protons and the all methyl

functionalities were attached to the saturated carbons. Four one–proton doublets at δ

5.41 (J=6.6 Hz), 5.06 (J=9.0 Hz), 4.98 (J=7.5 Hz) and 4.89 (J=9.0 Hz) were accounted

to anomeric H-1ꞌ, H-1ꞌꞌ, H-1ꞌꞌꞌ and H-1ꞌꞌꞌꞌ protons, respectively. The other sugar protons

appeared between δ 4.59-3.05. The 13

C NMR spectrum of TD-04 displayed signals for

vinylic carbons at δ 139.24(C-5) and 119.71(C-6), carboxylic carbons at δ 181.26 (C-

26), oxygenated methine carbon δ 80.19 (C-3), anomeric carbons at δ 104.86 (C-1ꞌ),

103.89 (C -1ꞌꞌ), 100.94 (C-1ꞌꞌꞌ), 94.56 (C-1ꞌꞌꞌꞌ), and other sugar carbons in the range of

78.28-61.65. The presence of oxygenated methylene protons in the slightly deshielded

region at δ 3.16 (H2-6ꞌ) and 3.22 (H2-6ꞌꞌ and H-6ꞌꞌꞌ) and carbons signals at δ 64.74 (C-

6ꞌ), 64-47 (C-6ꞌꞌ) and 62.87 (C-7ꞌꞌ) suggested (1→6) linkage of the sugar unit. Acid

hydrolysis of TD-04 yielded D-glucose, co-TLC comparable. On the basis of these

results the structure of TD-04 has been formulated as lanost-5-en-3β-ol-26-oic acid 3β-

L-glycopyranosyl-(6ꞌ-1ꞌꞌ)-β-D-glucopyranosyl-(6ꞌꞌ→1ꞌꞌꞌ)-β-D-glucopyranosyl-(6ꞌꞌꞌ→1ꞌꞌꞌ)-

β-D-glucopyranoside. This is a new triterpenic tetraglycoside.



Compound TD-05, named lansotanol-26-oic acid tetragluacoside was obtained as

colourless crystals from chloroform: methanol (9:1) eluants. It responded positively to

glycosidic tests and showed IR absorption bands for hydroxyl groups (3510, 3406,

3225 cm-1

) and carboxylic function (1685 cm-1

). On the basis of mass and 13

C NMR

spectra, the molecular ion peak of TD-05 was determined at m/z 1108 corresponding to

the molecular formula of triterpenic acid tetraglycoside C54H92O23. The ion peaks

arising at m/z 179 [C6H11O6]+, 342 [C6H11O6-C6H10O5]

+ and 504 [C6H11O6-C6H10O5-

C6H10O5]+ suggested the presence of hexose units in the glycoside chain. The ion

fragment forming at m/z 459 [M-glycoside chain, C30H51O3]+ and 442 [459-OH]

+

indicated that the triterpenic moiety possessed one each oxygenated methine and

carboxylic group. The 1H NMR Spectrum of TD-05 exhibited a one-proton double

doublets at 3.89(J=5.5, 8.8 Hz) assigned to oxygenated methine H-3α proton. Five

broad singlet’s at δ 1.23, 1.02, 0.97, 0.84, and 0.81 and two doublets of δ1.13 (J=6.1

Hz), and 0.93 (J=6.6 Hz), all integrating three proton each, were attributed to tertiary

C-29, C-19, C-28, C-30, and C-18 and secondary C-27 and C-21 methyl protons, all

attached to saturated carbons. Four one-proton doublets at δ 5.49 (J=7.3 Hz), 5.37

(J=7.5 Hz), 5.09 (J=7.2 Hz), and 4.93 (J=7.1 Hz), were ascribed to anomeric H-1ꞌ, H-1ꞌꞌ,

H-1ꞌꞌꞌ and H-1ꞌꞌꞌꞌ protons, respectively. The other sugar protons appeared from δ 4.71 to

3.03. The 13

C NMR spectrum of TD-05 displayed signals for carboxylic carbons at δ

181.26 (C-26), oxygenated methine carbon at δ 80.20 (C-3), anomeric carbons at δ

105.41 (C-1ꞌ), 103.35(C-1ꞌꞌ), 100.99 (C-1ꞌꞌꞌ), 95.78 (C-1ꞌꞌꞌꞌ), and other sugar carbons

between 78.35-62.81. The presence of oxygenated methylene protons in the deshielded

region in 1H NMR at δ 3.22 (H2-6ꞌ) and 3.16 (H2-6ꞌꞌ) and 3.13 (H-6ꞌꞌꞌ) and carbons

signals at δ 64.84 (C-6 ꞌ), and 64.53 (C-6ꞌꞌ) and 63.09 (C-3ꞌꞌꞌ) suggested (6→1) linkages

of the sugar units. The 1H and

13C NMR spectral data of TD-05 were compared with

the lanostane–type triterpenoids, (Ching et al., 2005; Ching et al., 2006; Ali et al.,

2001). Acid hydrolysis of TD-04 yielded lanoston-3β-ol-26-oic acid and D-glucose. On

the basis three results the structure of TD-04 has been determined Lanastan-3β-ol-26-

oic acid-3β-D-glycopyranosyl-(6ꞌ→1ꞌꞌ)-β-D-glycopyranosyl-(6ꞌꞌ→1ꞌꞌꞌ)- β-D-

glycopyranosyl-(6ꞌꞌꞌ→1ꞌꞌꞌꞌ)-β-D-glycopyranoside. This is new triterpenic

tetraglucoside.



Table 4.1: Phytoconstituents isolated from T. dioica

Code Compound

name

Eluants M. wt.

Mol

.formula

m.p.(C) %

Yield

Nomenclature

TD-01 Heneicosanyl

oleate

PE: C

(90:10)

577

C39H76O2

91-92 °C 0.36% n-Heneicosanyl n-

octadec-9-enoate

TD-02 Farnasonoic

acid α-L-

glucoside

C: M

(19:1)

420.54

C21H40O8

105-106 °C 0.43% 3, 7, 11-Trimethyl-7α-

L-gluco-furanosyloxy-

n-dodecanoic acid

(New)

TD-03 Lanosten-5-en-

3β-ol-26-oic

acid glucosyl

capriate

C: M

(19:1)

746

C44H74O9

135- 136°C

0.38% Lanosten-5-en-3β-ol-

26-oic acid -3β-D-

glucopyranosyl-2ꞌ-

octanoate (New)

TD-04 Lanosten-3β-

ol-26-oic acid

tetraglucoside

C: M

(9:1)

1106

C54H90O23

190-192 °C

0.23% Lanost-5-en-3β-ol-26-

oic acid 3β-L-

glycopyranosyl-(6ꞌ-1)-

β-D-glucopyranosyl-

(6ꞌꞌ-1ꞌꞌꞌ)-β-D-

glucopyranosyl-(6ꞌꞌꞌ-

1ꞌꞌꞌ) -β-D-

glucopyranoside.

(New)

TD-05 Lanastan-3β-

ol-26-oic acid

tetraglucoside

C: M

(95:5)

1108

C54H82O23

125-127 °C 0.34% Lanastan-3β-ol-26-oic

acid-3β-D-

glycopyranosyl-(6ꞌ-1ꞌꞌ)-

β-D-glycopyranosyl-

(6ꞌꞌ-1ꞌꞌꞌ)- β-D-

glycopyranosyl-(6ꞌꞌꞌ-

1ꞌꞌꞌꞌ)-β-D-

glycopyranoside.(New)

PE= Petroleum ether, C= Chloroform, M= Methanol



Structure of isolated compounds from T. dioica

Heneicosanyl oleate (TD-01)

Farnasonoic acid α-L-glucoside (TD-02)

OO

O CO CH2 (CH2)5 - CH3

OH

HO

OH

COOH

29 28

30H

1'

2'3'

4'

5'

6'

1''

2'' 8''

1

2

34

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

2122

25

26

27

Lanosten-5-en-3β-ol-26-oic acid glucosyl capriate (TD-03)



O

COOH

H

1

2

34 5

6

7

8

9

10

11

12

1314

15

16

17

18

19

20

21 22

25

26

27O

OOH

HO

OH 1''''

2''''3''''

4''''

5''''

6''''

CH2

OOOH

OH 1'''

2'''3'''

4'''

5'''

CH2

OOOH

OH 1''

3''

4''

5''

CH2

OOH

OH 1'

2'3'

4'

5'

6'

6''

6'''

OH

OH

OH

OH

2829

30

Lanosten-3β-ol-26-oic acid tetraglucoside (TD-04)

Lanastan-3β-ol-26-oic acid tetraglucoside (TD-05).

Mass fragmentation pattern of isolated compounds

265 281

H3C (CH2)7 CH CH (CH2)7 C

O

O CH2 (CH2)9 CH3

Scheme 4.1: Mass fragmentation pattern of heneicosanyl oleate (TD-01)



COOH

124

56

7

89

10

11

12

15

HO

1314

O

OH

HO

HOOH

12

C21H40O8

[M+] m/z 420

12 - C3H7 - C6H11O6

COOH

HO

C6H11O5

COOH

H

O

O5C6H11

C7H13O2

m/z 129C15H27O8

m/z 335

C14H27O6

m/z 291

m/z 377

_ C6H11O5

m/z 214

m/z 241

m/z 172 m/z 290

_ C6H11O5

_ COOH+

++

+

Scheme 4.2: Mass fragmentation pattern of farnasonoic acid α-L-glucoside (TD-02)



TD-03

OO

O CO CH2 (CH2)5 - CH3

OH

HO

OH

COOH

H

619

127

602

144

457

- OH440

C44H74O9= 746

C30H49O3= 457

Scheme 4.3: Mass fragmentation pattern of lanosten-5-en-3β-ol-26-oic acid glucosyl

capriate (TD-03).

TD-05

O

COOH

H

1

2

3 4

6

7

8

9

10

11

12

1314

15

16

17

18

19

20

21 22

25

26

27

OOOH

HO

OH 1''''

2''''3''''

4''''

5''''

6''''

CH2

OOOH

OH 1'''

2'''3'''

4'''

5'''

CH2

OOOH

OH 1''

3''

4''

5''

CH2

OOH

OH 1'

2'3'

4'

5'

6'

6''

6'''

OH

OH

OH

OH

H2829

30504341

179

Scheme 4.4: Mass fragmentation pattern of lanastan-3β-ol-26-oic acid tetraglucoside

(TD-05).



Spectra of isolated compound

Spectrum 5.1. 1H NMR spectrum of heneicosanyl oleate (TD-01)

Spectrum 5.2. 13

C NMR spectrum of heneicosanyl oleate (TD-01)



Spectrum 5.3. Mass spectrum of heneicosanyl oleate (TD-01)

Spectrum 5.4. 1H NMR spectrum of farnasonoic acid α-L-glucoside (TD-02)



Spectrum 5.5. 13

C NMR spectrum of farnasonoic acid α-L-glucoside (TD-02)

Spectrum 5.6. Mass spectrum farnasonoic acid α-L-glucoside (TD-02)



Spectrum 5.7. 1H NMR spectrum of lanosten-5-en-3β-ol-26-oic acid glucosyl capriate

(TD-03)

Spectrum 5.8. 13

C NMRspectrum of lanosten-5-en-3β-ol-26-oic acid glucosyl capriate

(TD-03)



Spectrum 5.9. Mass spectrum of lanosten-5-en-3β-ol-26-oic acid glucosyl capriate (TD-

03)

Spectrum 5.10. 1H NMR spectrum of lanosten-3β-ol-26-oic acid tetraglucoside (TD-04)



Spectrum 5.11. 13

C NMR spectrum of Lanosten-3β-ol-26-oic acid tetraglucoside (TD-

04)

Spectrum 5.12. Mass spectrum of Lanosten-3β-ol-26-oic acid tetraglucoside (TD-04)



Spectrum 5.13. 1H NMR spectrum of lanastan-3β-ol-26-oic acid tetraglucoside (TD-05).

Spectrum 5.14. 13

C NMR spectrum of Lanastan-3β-ol-26-oic acid tetraglucoside (TD-

05).



Spectrum 5.15. Mass spectrum of Lanastan-3β-ol-26-oic acid tetraglucoside (TD-05).



4.6. References

[1] Chakravarthy, H. M. (1982). Fascicles of flora of India-11 Cucurbitaceae.

Botanical Survey of India, 136.

[2] Mythili, J. B. and Pious Thomas. (1999). Micropropagation of pointed gourd

(Trichosanthes dioica Roxb). Scientia Horticulturae, 79, 87-90.

[3] Chadha M. L. (2009). Indigenous Vegetables of India with a Potential for

Improving Livelihoods. International Symposium on Underutilized Plants for

Food Security, Nutrition, Income and Sustainable Development, 2, 1-8.

[4] Singh, K. (1989). Pointed gourd (Trichosanthes dioica Roxb). Indian

Horticulture, 33, 35–38.

[5] Kirtikar, K. R and Basu, B. D. (2001). Indian medicinal plant, 2nd

Edn, Vol 5,

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