Plectranthus zeylanicus - National Science Foundation

SRI

LANKAN

MEDICINAL

MONOGRAPHS

AND

VOL -

LAKSHMI ARAVINDA

PLANT

ANALYSIS

11

ARAMBEWELA WIJESINGHE

SRI LANKAN MEDICINAL

PLANT

MONOGRAPHS AND ANALYSIS VOL-11

PLECTRANTHUS

£ >om»2

ZEYLANICUS

Lakshmi Arambewela and Aravinda Wijesinghe

Edited by Dilmani Warnasuriya

Industrial Technology Institute (Ceylon Institute of Scientific and Industrial Research) 363, Bauddhaloka Mawatha Colombo 07 Sri Lanka

Published by National Science Foundation 2006

©Industrial Technology Institute & National Science Foundation, 2006. First published in 2006. ISBN 955-590-045-0 Published by National Science Foundation.

Arambewela, Lakshmi Sri Lankan Medicinal plant Monograph and Analysis: Plectranthus zeylanicus / Lakshmi Arambewela and Aravinda Wijesinghe.Colombo: National Science Foundation, 2006. Vol.11, iii,27p. ill.; 30 cm. -(Sri Lankan Medicinal Plants, Vol.11) ISBN 955-590-045-0

Price: Rs. 200.00

i. 615.321 DDC21 iii. Series iv. Arambewela, Lakshmi jt.au. v. Wijesinghe, Aravindajt.au. 1. Medicinal plants

ii. Title

2. Botany, Chemistry, Medical

The information found in this monograph is taken from available scientific literature. The authors accept no liability for any damages arising from any claims contained in this text except the work carried out by the authors.

ii

Preface

Studies on medicinal plants of Sri Lanka have been carried out in the Herbal Technology Division of Industrial Technology Institute (former Ceylon Institute of Scientific and Industrial Research) for almost two decades. This monograph which is the eleventh in this series incorporates information collected from literature surveys, researches and also experiences of the Herbal Technology Division staff. This monograph is intended for a varied reading public, herbal drug manufacturers who need to identify their herbal raw materials, Ayurvedic physicians who need some scientific information on medicinal plants, research workers requiring some quick background information on a plant, industrialists or entrepreneurs pondering on commercial ventures and the inquiring lay readers. W e hope this monograph fulfils some requirements of each of them. The authors wish to thank the members of the Herbal Technology Division for their contribution, the Information Service Center for providing information, Department of Plant Sciences and Department of Zoology of the University of Colombo for assisting in anatomical studies, Food Technology Division of the Industrial Technology Institute for helping in the analysis of powdered plant materials and the Microbiology Laboratory for photographing the slides. They also gratefully acknowledge the sponsor National Science Foundation for the research grant (RG / 2004 / T M / 01).

Herbal Technology Division Industrial Technology Institute P.O.Box 787 Colombo 07 Sri Lanka.

in

Plectranthus zeylanicus Benth Family Lamiaceae

Synonyms 1

Coleus zeylanicus (Benth) Cramer

Selected V e r n a c u l a r Names 1,2

Sinhala - Iriweriya 1,2

Sanskrit-Valakan

3

Tamil- Vettiyar

Pharmacopoeia 4

Ayurveda Pharmacopoeia

Distribution An endemic species to Sri Lanka. It is grown in Kandy, Kadugannawa and Kurunegala. It 1,2,3

can be easily grown in prevailing climatic and soil conditions in Sri Lanka.

Morphology

2

A herbaceous, perennial herb, about 60-90 cm tall, stems branching, somewhat prostrate, bluntly quadrangular, reddish and hairy, leaves simple, opposite, aromatic 5.3-6.5 cm somewhat succulent, dentate, light green on the upper surface, paler below, veins prominent beneath, petioles 2.5-3 cm long, reddish, hairy, faintly grooved above, flowers irregular, bisexual, light blue, 1 cm long and narrow in whorls, each containing about 10 flowers opening at different times along a terminal or branched raceme 15-23 cm long, pedicels 3 m m long reddish and hairy, sepal the largest, broadly ovate, apiculate, others lanceolate acuminate, acute, corolla 2-lipped consisting of 5 segments, upper lip of 4 short segments and a long, boat- shaped lower lip, stamens 4, didynamous, epipetalous, 2 slightly shorter, all included, ovary superior, 2-carpellary, 4-locular with a single ovule in

l

each loculus, style 7 m m long, stigma bifid; fruit of 4, one seeded achenes at the base of a persistent calyx.

Fig-1. Plectranthus zeylanicus plant 1. Leaf

2. Stem

3. Flower, lateral view

4. Inflorescence

(Source - Jayawcera, D.M.A, Medicinal plants (1982). part V, p 109)

PLANT MATERIAL OF INTEREST

Whole plant'

Official Drug Whole plant

3

Pharmacognostic Features* Anatomy

Fig-2. Cross section of I. Epidermis

Plectranthus zeylanicus

2. Palisade parenchyma

bundles 5. Parenchyma cells

leaf (stained with safranine (10x10))

3. Spongy parenchyma

6. Trichomcs

4. Vascular

Fi«-4. Cross section of Plectrantluts 1. Epidermal hairs 6. Cambium

7. Xylem

2. Epidermis

zeylanicus

stem (stained with safranine (10x10))

3. Collenchyma

8. Pith

4

4. Cortex

5. Phloem

Powder analysis Analyzed part

Aerial parts of the plant

Oruanoleptic properties Colour - Brownish black Odour - Aromatic Taste - No special taste

Microscopic characters

Fig-6. Powder of aerial parts of Plcctranthus 1. Vessel elements

zeylanicus

2. Trichome

under the microscope

3. Parenchyma cells

Fig-7. Schematic diagram of powder of aerial parts 1. Epidermal cells with stomata

2. Stomata with guard cells

4. Vessel segments with different thickenings

3. Trichomes

5. Epidermal cells

Analyzed part - Root Organolcptic properties Colour - Brownish black Odour - No special odour Paste - No special taste M icroseopic characters

Fig-8. Powder of root of'Plectranthus zeylanicus under the microscope. I. Vessel segments

2. Parenchyma cells

ft

Fig-9. Schematic diagram of powder microscopy of roots 1. Root hairs

2. Parenchyma cells

3. Vessel segments with different thickenings

• These analysis were carried out by the authors at Industrial Technology Institute and the Dept. of Plant Sciences and Dept. of Zoology of University of Colombo

6

Physico-chemical A n a l y s i s Extractable

12

matter

Crushed, dried plant material (about 4 g) was weighed to a glass-stoppered conical flask. Solvent (100 mL) was added, weighed, shaken well and allowed to stand for lh. It was then boiled for lh and cooled. The weight was readjusted with specified solvent and filtered. Filtrate (25 mL) was taken, solvent was evaporated and oven dried at 105 °C for 6 h, cooled in a desiccator and weighed.

Total ask

Crushed, air dried plant material (about 4 g) was weighed to a previously ignited crucible. The material was ignited by gradually increasing the temperature to 550 °C until free from carbon. The crucible was cooled and weighed.

Acid insoluble ash

Hydrochloric acid (25 mL, cone. -70 g/L) was added to the crucible containing total ash, covered with a watch glass and boiled gently for 5 min. The insoluble matter was collected on an ashless filter paper and washed with hot water until the filtrate was neutral. Thefilterpaper containing the insoluble matter was transferred to the original crucible and ignited to a constant weight.

Water soluble ash

Water (25 mL) was added to the crucible containing total ash, covered with a watch glass and boiled gently for 5min. The insoluble matter was collected on an ashlessfilterpaper and washed with hot water. Thefilterpaper containing the insoluble matter was transferred to the original crucible and ignited for 15 min. at a temperature not exceeding 450 °C. Water soluble ash is the calculated difference in weight between the total ash and the residue remaining after treatment of the total ash with water.

Moisture content of the samples was estimated and all the calculations were done on dry weight basis.

7

Table 1. Physico-chemical parameters of Plectranthus zeylanicus**

Physico-chemical parameter Ethanol extract of aerial parts Ethanol extract of root Water extract of aerial parts Water extract of root

A mo unt % 14.4-15.1 3.5-3.8 33.3-35.4 7.0-7.5

Total ash content of aerial parts

13.6-15.9

Total ash content of root

13.2-13.6

Acid insoluble ash content of aerial parts

0.9-1.0

Acid insoluble ash content of root

6.8-6.9

Water soluble ash content of aerial parts Water soluble ash content of root

11.1-14.9 0.2-0.3

(Results arc expressed as percentages on dry weight basis)

T H I N L A Y E R C H R O M A T O G R A P H I C PROFILE

Plectranthus zeylanicus water extract of aerial parts Sample preparation

: P. zeylanicus

aerial parts (4 g) were boiled for one hour with

water (100 mL) and the extract was filtered and evaporated to dryness. Ten microliters (10 uL) of the diluted extract (0.1 g in 5 mL) was spotted on TLC plate. Adsorbent

: Silica g e l - G F 5 4

Solvent system

: Ethyl acetate : Acetic acid : Water (4.6 : 0.2 : 0.2)

2

Detection Direct evaluation

:

Scanning

: Densitometer at 254 nm (before spraying) and 450 nm (after

UV

2 5

4nm,

Rf values - 0.39. 0.52. 0.93

spraying) Spray reagent

Fig-10.

T

L

: Vanillin sulphate

C

f i n g e r

p r i n t

p r o f i l e

o f

w a t e r

e x t r a c t

9

o f

Plectranthus zeylanicus

a e r i a l

p a r t s

a. ?.

oo

l> . 250

i"

0 . 20 j

•:7*

"ii;!

a -15 c

•

i!'.2

•

•

•) f n

.... t.c. :

i \ . osa

3

'

!

Table 2. Description of densitogram (Fig-V1)

:

Peak no.

Y(mm)

1

8.10

Relative area % 20.87

2

12.12

5.31

3

20.05

3.45

4

65.45

31.21

r

; ^ T \ ; V

. ooo
.

c

o

Table 3. Description of densitogram (Fig-12)

O

Peak no.

Y(mm)

1

9.61


2

20.94

5.10

3

28.25

7.44

4

34.18

7.02

5

66.80

32.67

1;!J0 - •

O.01C.i)iO.(.'.".O.O-y. Or'.'.OiO.r,?}.

,

,

Fig-12. Densitogram of TLC finger print profile of water extract of Plectranthus zeylanicus aerial parts at 450 nm

10

Plectranthus

zeylanicus ethanol extract of aerial parts

Sample preparation

: P. zeylanicus aerial parts ( 4 g) were boiled for one hour with 9 5 % ethanol ( 1 0 0 mL) and the extract was filtered and evaporated to dryness. Ten microliters ( 1 0 uL) of the diluted extract ( 0 . 9 g in 5 mL) was spotted on TLC plate.

Adsorbent

: Silica g e l - G F 5 4

Solvent system

: Chloroform : Methanol ( 4 . 9 : 0 . 1 )

2

Detection Direct valuation

:

Scanning

: Densitometer at 2 5 4 nm (before spraying) and 4 5 0 nm (after

UV

2 5

4nm,

K t

values - 0

. 1 9 , 0 . 4 8 , 0 . 6 7 , 0 . 8 0 ,

0 . 9 6


: Vanillin sulphate

Fig-13. TLC finger print profile of ethanol extract of Plectranthus zeylanicus aerial parts

11


-it .

1 O" Stage

Y(mnO

Peak no.

Y(ram)

1

10.06


2

23.73

10.35

3

30.00

4.24

4

71.49

25.12

Fig-14. Densitogram of TLC finger print profile of ethanol extracl of Plectranthus zeylanicus aerial parts at 254 nm

Table 5. Description of densitogram (Fig-15) Peak no.

Y (mm)

1

10.29


2

23.44

7.93

3

36.04

5.28

4

55.59

3.73

5

63.98

3.39

6

71.58

9.21

2.

• .". . • ;i i 3 -

4

.v/

5 ! '• • (i

jt.iijc V : inn

Fig-15. Densitogram of TLC finger prim profile of ethanol extract of Plectranthus zeylanicus aerial parts at 450 nm

12

Plectranthus zeylanicus Sample preparation

w a t e r

e x t r a c t

o f

r o o t s

: P. zeylanicus roots (4 g) were boiled for one hour with water (100 mL) and the extract was filtered and evaporated to dryness. Nine microliters (9 uL) of the diluted extract (0.9 g in 5 ml.) was spotted on TLC plate.

Adsorbent

: Silica gel-GFISJ

Solvent system

Detection

: Ethyl acetate : Acetic acid : water (4.6 : 0.2 : 0.2)

Direct evaluation Scanning : Spray reagent

UV

2

5

4nm,

Rf values - 0.32, 0.47. 0.96

: Densitometer at 254 nm (before spraying) and 450 nm (after spraying) : Vanillin sulphate

Fig-16. TLC finger print profile of water extract of Plectranthus zeylanicus roots

iI


C O

2 0 . 0

A 0. C

60.O

Peak no.

Y(mm)

Relative area %

1

8.41

35.70

2

47.41

21.01

3

69.78

34.67

Fig-17. Densitogram of TLC finger print profile of ethanol extract of Plectranthus zeylanicus roots at 254 nm


0

. 'j

0y

Peak no.

Y (mm)

Relative area %

1

9.60

21.18

2

16.93

8.76

3

25.69

11.22

4

66.89

45.14

'

:o . o2n

. c J r>. o .">

a

. o 6

v>.

o" 0 . &


14

Plectranthus

zeylanicus ethanol extract of roots

Sample preparation

: P. zeylanicus

roots (4 g) were boiled for one hour with 9 5 %

ethanol (100 mL) and the extract was filtered and evaporated to dryness. Eight microliters (8 u.L) of the diluted extract (0.8 g in 5 mL) was spotted on TLC plate. Adsorbent

: Silica

Solvent system

rChloroform : Methanol (4.9 : 0.1)

gel-GF254

Detection Detection evaluation

: UV

Scanning

: Densitometer at 254 nm (before spraying) and 450 nm (after

25

4nm,

Revalues - 0.37, 0.49, 0.84


: Vanillin sulphate

Fig-19. TLC finger print profile of ethanol extract of Plectranthus zeylanicus roots

15


' , 2

Y(mm)

Relative area %

1

8.31

56.25

2

23.20

7.50

3

30.01

9.81

'

11,

0 .000

3

Peak no.

I

r-....v^..

S Liige Y ijrmi i



Peak no.

Y (mm)

Relative area %

1

8.42

23.80

?

22.74

37.00

3

49.09

16.07

4

68.49

23.13

i. -31-:,; Y ( :-,-n

Fig-21. Densitogram of TLC finger print profile of ethanol extract a(~ Plectranthus zevlanicus roots at 450 nm

16

High Pressure Liquid Chromatographic Profile** Plectranthus zeylanicus Sample preparation

water extract of aerial parts : P. zeylanicus aerial parts (4 g) were boiled for one hour with water (100 mL) and the extract wasfilteredand evaporated to dryness. The diluted extract (5.2 mg in 5 mL) was purified using Sep-pak CI8 cartridge.

Injection volume

: 20 ixh

Apparatus

: Shimadzu LC - 10 ADvp pumps and Shimadzu SPD - M 10 Avp uv / vis photodiode array detector.

Column

: Inertsil 5U ODS - 2 reverse phase column, (250 m m x 2.6 mm).

Solvent system

: Acetonitrile : Water (50 : 50)

Flow rate

: 1 mL/min

Detection

: 254 nm

Table 10. Retention times of main peaks

Peak no.

Retention

Relative

time

area %

(min) 1

2.32

22.64

2

2.46

16.94

3

2.82

30.00

4

5.08

8.74

5

6.22

21.49

Fig - 22. HPLCfingerprint profile of water extract of Plectranthus zeylanicus aerial parts

17

Plectranthus zeylanicus Sample preparation

ethanol extract of aerial parts

: P. zeylanicus aerial parts (4 g) were boiled for one hour with 95% ethanol (100 mL) and the extract was filtered and evaporated to dryness. The diluted extract (7.9 mg in 5 mL) was purified using Sep-pak CI8 cartridge.

Injection volume

: 20 /xL

Apparatus

; Shimadzu LC - 10 ADvp pumps and Shimadzu SPD - M 10 Avp uv / vis photodiode array detector.

Column

: Inertsil 5U ODS - 2 reverse phase column, (250 m m x 2.6 mm).

Solvent system

: Methanol : Water (85 : 15)

Flow rate

: 0.8 mL/min

Detection

: 254 nm

Table 11. Retention times of main peaks

0

5

1