Indian Journal of Chemistry
Vol. 49B, March 2010, pp. 356-359
Note
Chemical constituents isolated from
Andrographis paniculata
Poonam Kulyal*1, U K Tiwari1, A Shukla2 &
A K Gaur3
1
Department of Chemistry, A.N.D.N.N.M. College,
C.S.J.M. University, Harsh Nagar, Kanpur 208 012, India
2
Department of Genetics & Plant Breeding,
College of Agriculture, G.B. Pant University of Agriculture &
Technology, Pantnagar, 263 145, India
3
Department of Molecular Biology and Genetic Engineering,
College of Basic Science & Humanities, G.B. Pant University Of
Agriculture & Technology, Pantnagar, 263 145, India
E-mail: chem_poonam11@yahoo.com
Received 18 August 2008; accepted (revised) 17 August 2009
Phytochemical investigation of the aerial parts of Andro-
graphis paniculata give diterpenic constituents andrographolide,
14-deoxy-11,12-didehydroandrographolide, 14-deoxyandrogra-
pholide, 3,14-dideoxyandrographolide, 14-deoxy-11-oxoandro-
grapholide, 14-deoxy-12-hydroxyandrographolide, neoandrogra-
pholide, andrographiside and 14-deoxyandrographiside. The
structures of these compounds have been established on the basis
of spectral data analysis.
Keywords: Andrographis paniculata, Kalmegh, Acanthaceae,
diterpenes, diterpenoidal glycoside
Andrographis paniculata (Acanthaceae), commonly
known as Kalmegh is found in the plains of India,
Pakistan, Sri Lanka and West Indies. About 26
different poly herbal formulations of this plant are
mentioned in Ayurveda as a popular remedy for the
treatment of various liver disorders1,2. Aerial parts of
the plant are used for jaundice, colic dysentery and
dyspepsia3,4, as a bitter tonic, stomachic, anthelmintic
and anti plasmodial5. It is widely used in traditional
medicine as an antidote against poisons of snakes and
insect, and as an antimalarial agents6-8. It is beneficial
in general debility, asthma, bronchitis, filariasis8 and
in hepatitis during clinical studies10. It has also shown
immunomodulating/immunostimulatory and anti
cancerous activity11-15. One of the active compound
neoandrographolide has been reported to show anti-
HIV activity16. The present paper describes the
isolation and characterization of six diterpenoides and
three diterpenoidal glycosides from the aerial parts of
A. paniculata grown in Pantnagar.
Results and Discussion
The compounds 1-5 were identified as andro-
grapholide, 14-deoxy-11,12-didehydroandrographo-
lide, 14-deoxyandrographolide, 3,14-dideoxyandro-
grapholide, 14-deoxy-11-oxoandrographolide and
compounds 7-9 were identified as neoandrographolide,
andrographiside (Figure 1), 14-deoxyandrographiside.
Compound 6 was obtained as amorphous powder. The
molecular formula, C20H30O5 was established by mass
spectrum giving a molecular ion at m/z 350(M+) and
named as 14-deoxy-12-hydroxyandrographolide. The
IR spectrum showed a broad peak at 3420 cm-1 due to
hydroxyl group. Peaks at 1730 and 1656 cm-1 were due
to lactone ring. Peak at 904 cm-1 was due to presence of
exocyclic methylene group. The 1H NMR spectrum
exhibited a singlet at  0.69 for methyl groups present
at C18 and C20. Peak at  3.01 (singlet) and 2.20 (triplet)
was obtained for a hydroxyl proton and a proton
present at C3. Peak at  3.98 gave a doublet for two
protons present at C15. Broad singlet was obtained at 
5.20 and 5.66 for hydroxyl group present at C19 and
C12, respectively. Mass spectra gave peaks at m/z 350
(M+), 332, 223, 299 and 335. Characterization of
phytoconstituents of Dendrobium gratiosissium17 and
Hyatella cribrifomis18 has been reported recently for
their cytotoxic as well as antimicrobial activities.
Experimental Section
All the melting points were uncorrected. The UV
spectra were determined on a Perkin-Elmer Lambda
15 UV/VIS spectrophotometer. The IR spectra were
determined on a Perkin Elmer spectrum RX1 (4000-
450 cm-1). The 1H NMR spectra were measured on a
Bruker DRX-300 spectrometer using CDCl3 as
solvent and TMS as internal standard in ppm. The
mass spectra were measured on Jeol SX-102 at 70 eV.
Plant Material
The whole plant except root was collected from
G.B. Pant University of Agriculture and Technology,
Pantnagar, UK, India in July 2003 and identified as
Andrographis paniculata (Acanthaceae).
 NOTES 357

O O
15

14 16

13 O
O
HO
12

11

20
17
1

9
2
10 8

3 5 7
4 6 HO
H
OH
HO H
18 19
OR
1R=H 2
8 R = glu
O O

O O

HO H
H OR1
OR

3 R = H, R1 = H 4R=H
6 R = OH, R1 = H 7 R = glu
9 R = H, R1 = glu
O

HO H
OH

5
Figure 1 ― Diterpenes and diterpenoidal glycosides isolated from Andrographis
paniculata

extracts were concentrated under reduced pressure,

Isolation Procedure affording the residue (70 g). The crude residue (70 g)
The aerial part (1 kg) of A. paniculata was was defatted with hexane and then fractionated into
percolated with 95% ethanol (5 × 1L). The combined chloroform (10 g) and methanol (20 g). Methanol
fraction (5 g) was subjected to silica gel column
358 INDIAN J. CHEM., SEC B, MARCH 2010
chromatography eluted with chloroform-methanol
(1.5-21% methanol in chloroform) to yield four
fractions (A-D). All fractions were further subjected
to repeated column chromatography on silica gel
eluted with chloroform-methanol (1-35% methnol in
chloroform). Repeated column chromatography of
fraction A on silica gel (chloroform-methanol; 1-2%,
2.5-3.5%) afforded compounds 1 (20 mg) and 2 (13
mg). Compounds 3 (18 mg) and 8 (16 mg) were
obtained from the fraction B after repeated silica gel
chromatography (chloroform-methanol; 5-7% and 8-
12% respectively). The fraction C on repeated silica
gel chromatography gave compounds 4 (14 mg) and 5
(18 mg) (chloroform-methanol; 14-18% respectively).
Compounds 6 (13 mg) and 9 (15 mg) obtained from
fraction D after repeated silica gel chromatography
(chloroform-methanol; 22-35% and 17-20%,
respectively). The structure for each compound
isolated was determined by IR, 1H NMR and MS
studies carried out at Central Drug Research Institute,
Lucknow, India.
Andrographolide 1
Colourless cubes, m.p. 218-22°C, UV(methanol):
235 nm; IR (KBr): 3400, 3299, 1727, 1673, 908 cm-1;
1
H NMR (300 MHz, CDCl3):  6.91 (1H, t, C12), 4.90
(1H, d C14), 4.21- 4.50 (2H, m, C15), 1.81-1.96 (5H,
m, C2, C7, C9), 0.72 (2 × CH3, s, C18, C20); MS: m/z
350(M+), 332, 314 and 237 for C20H30O5.
14-Deoxy-11,12-didehydroandrographolide 2
White crystal, m.p. 204-205°C; UV(methanol): 252
nm; IR (KBr): 3328, 2930, 2849, 1727, 1669 and 901
cm-1; 1H NMR (300 MHz, CDCl3):  0.71 (3H each, s,
C18, C20), 2.51(1H, s (br), C3-OH), 2.39 (1H, t, C3),
4.51(2H, d, C15), 4.87 (1H, t, C14), 5.16 (1H, s (br),
C19-OH), 5.68 (1H, d, C12); MS: m/z 332(M+), 317,
314, 296, 281 and 223 for C20H28O4 .
14-Deoxyandrographolide 3
Fine needles, m.p. 170°C; UV(methanol): 238 nm;
IR (KBr): 3399, 3280, 2930, 2850, 1728, 1632, 1458,
1369, 908 cm-1; 1H NMR (300 MHz, CDCl3):  0.66
and 1.26 (each 3H, s, 2 × CH3,C18,C20), 1.59 (12H, S,
6 × CH2), 3.32 (1H, d, H19), 3.56 (1H, t, 3β-H), 3.78
(H, s (br), C3-OH), 4.17(1H, ABd, C19), 4.58 (2H, s,
C17), 4.73 (2H, m, C15), 5.10 (H, s (br), C19-OH) 6.97
(1H, m, C14); MS: m/z 334 (M+), 316, 298, 297, 237,
255, 219, 215, 201, 199, 185, 157 and 91 for
C20H30O4.
3,14-Dideoxyandrographolide 4
Colourless needles, m.p. 106-107°C; UV (methan -
1 1
; H NMR (300 MHz, CDCl3):  0.88 (3H each, s,
C18, C20), 3.14 (2H, d, C12), 3.88 (2 H, s, C17), 3.99
(1H,t, C14), 1.42 (10H, s, 5 × –CH2), 5.12 (1H, s, C19 -
OH); MS: m/z 318 (M+), 303, 300 and 285 for
C20H30O3.
14-Deoxy-11-oxoandrographolide 5
White crystalline solid, m.p. 101°C; UV
(methanol): 225 nm; IR (KBr): 3340, 2929, 2849,
1726, 1713, 1679, 1458, 1365 and 901 cm-1; 1H NMR
(300 MHz, CDCl3):  0.69 (3H each, s, C18, C20), 2.40
(1H, t, C3), 3.41 (1H, s, (br), C3-OH), 4.33 (2H, d,
C15), 4.42 (1H, t, C14), 4.98(1H, s (br), C19-OH), 5.59
(2H, s, C12); MS: m/z 348(M+), 333, 330, 312, 297
and 223 for C20H28O5.
14-Deoxy-12-hydroxyandrographolide 6
Amorphous power; IR (KBr): 3420, 2932, 2828,
1730, 1656 and 904 cm–1; 1H NMR (300 MHz,
CDCl3):  0.69 (3H each, s, C18, C20), 2.20 (1H, t, C3),
3.01 (1H, s, C3-OH), 3.39 (1H, t, C14), 3.98 (2H, d,
C15), 5.20 (1H, s, C19 - OH), 5.66 (1H, s, C12-OH);
MS: m/z 350 (M+), 335, 332, 317, 299 and 223 for
C20H30O5.
Neoandrographolide 7
Long colourless needles, m.p.160-62°C;
UV(methanol) 212 nm (End absorption); IR (KBr):
3449, 2931, 2852, 1748, 1600, 1447, 1357, 910 cm-1;
1
H NMR (300 MHz, CDCl3):  0.65 and 1.07 (each
6H, s, 2 × CH3, C18,C20), 1.19-2.37 (14H, m,7 × CH2),
3.44 - 4.80 (11H, m, sugar protons, C19) 7.18 (1H,
ABd, J = 4.0Hz, H14); MS: m/z 480 (M+) for
C26H40O8. Acid hydrolysis followed by paper
chromatography confirmed the sugar as glucose.
Andrographiside 8
Amorphous power, m.p. 193°C, UV(methanol) 230
nm; IR (KBr): 3399, 3307, 2929, 2849, 1727, 1674,
1458, 1365, 907 cm-1; 1H NMR (300 MHz, CDCl3): 
0.88 and 1.07 (each 6H, s, 2 × CH3,C18, C20), 1.47-
2.09 (10H, m, 5 × CH2), 3.20 -4.62(11H, m, sugar
protons, C1’-6’); MS: m/z 512 (M+) for C26H40O10. Acid
hydrolysis followed by paper chromatography
confirmed the sugar as glucose.
14-Deoxyandrographiside 9
 NOTES
Colourless crystals, m.p. 200-201°C; IR (KBr):
3369, 1727, 1668 and 900 cm–1; 1H NMR (300 MHz,
CDCl3):  0.78 (3H, 2 × -CH3, C18, C20), 4.58 (2H, d,
C15 ), 1.31- 2.01 (11H, m, sugar protons, C1- 6); MS:
m/z 496 (M+), 478, 386 and 317 for C26H40O9.
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