Pharmacognostical Standardization of the Plant Santolina chamaecyparissus

 

R. Vijaya Bharathi*, P. Shanthi, N. Jayshree and K.T. Kasthuri

Department of Pharmacognosy, College of Pharmacy, Madras Medical College, Chennai, Tamilnadu, India

*Corresponding Author E-mail: rvbharathi2003@yahoo.com

 

ABSTRACT:

Standardization was carried out on the basis of detailed organoleptic and microscopical evaluation of the plant which includes morphology and microscopy as well as WHO recommended physicochemical studies. The results of the standardization may throw immense light on the botanical identity of the plant Santolina chamaecyparissus which may furnish a basis of judging the authenticity of the plant and also to differentiate the drug from its allied species and detect adulterants.

 

KEYWORDS: Santolina chamaecyparissus, Micromorphology, Physicochemical

 


 

INTRODUCTION:

India has a rich cultural heritage of traditional medicines. The crude drugs have the advantage of being always easily available in abundance, comparatively cheaper, with negligible side effects and have frequently been prescribed to the people of all ages. Santolina chamaecyparissus (Cotton lavender) is an evergreen perennial herb belonging to the family Asteraceae. The plant has been used as medicine in many countries. Evidences reveal that the Greeks use Santolina as a dewormer and moth repellant and Arabs as an eye wash and topical anti-inflammatory. Romans use Santolina to treat snake bites and other insect bites. The plant was used in folk medicine because of its antispasmodic, digestive, anti-inflammatory, emmenagogue, vermifuge, worm infestation stimulant, antiseptic, astringent and antimicrobial properties1. As there is no record on pharmacognostical work on this plant, the present study has been undertaken to study its morphology, microscopy and physico-chemical constants. The study parameters includes organoleptic features, macroscopy, microscopy, fluorescence analysis, ash values, extractive values, crude fibre content, loss on drying, foaming index, swelling index, estimation of heavy metal and inorganic elements and preliminary phytochemical studies.

 

MATERIALS AND METHODS:

Plant Material:

The plant Santolina chamaecyparissus Linn. was collected from Nilgris hills, Tamil nadu, India during the month of July 2009.

 

The plant was identified, confirmed and authenticated by Botanical survey of India, Coimbatore and a voucher specimen has been deposited in the Department of Pharmacognosy (BSI/SC/5/23/09-10/Tech-620).

 

Macroscopical and microscopical studies:

The macroscopy and microscopy of the plant was studied according to the method of Brain and Turner2. For the microscopical studies, the cross sections were prepared and stained as per the procedure of Johansen3 and the quantitative microscopy was studied as per the procedure given by Wallis 4. The powder analysis has been carried out according to the method of Brain and Turner5.

 

Physico-chemical constants:

The ash values, extractive values, crude fibre content, loss on drying, foaming index, swelling index and volatile oil content were performed according to the official methods prescribed in Indian Pharmacopoeia6. Fluorescence analysis was carried out according to the method of Chase and Pratt7 and Kokoski et al8.

 

Preliminary phytochemical Studies:

Preliminary phytochemical Screening was carried out by using standard procedures described by Harborne9.

 

RESULTS AND DISCUSSION:

Macrosopy:

The plant is an evergreen sub shrub around 30-45 cm in height with woolly silvery hairs on its camphor scented leaves and upright stems having it a whitish grey appearance. Leaves are short oblong with acute apex, de-current base, crenate margin and bumpy rubbery texture. The upper surface is smooth dark grey and lower surface is rough pale grey with strong odour and aromatic taste. Stems usually erect branched mostly from bases, often tomentose to lanate.

Microscopical studies:

T.S of leaf:

In the cross sectional view, the leaves appear circular with undulated outline, circular with deep lobes or somewhat flat with lobed marginal part of the lamina (Fig. 1a). The circular leaf lacks dorsiventral differentiation. It has a central vascular strand and radiating small air chambers separated by thin, uniseriate partition filaments. The outer part of the arenchyma has elongated cylindrical palisade cells .The epidermal layer has continuous wavy outline, the cells are circular or spindle shaped with thick walls. The central vascular strand has a layer of parenchymatous bundle sheath enclosing fairly prominent mass of xylem and small nests of phloem. Xylem elements are thick walled and angular. The circular leaf is 450 µm in diameter.

 

Fig. 1 a. T.S of leaf entire view

 

Fig. 1 b. T.S of leaf a sector enlarged

 

 

Fig. 1 c. T.S of leaf –vascular bundle magnified

EP-Epidermis; VB-Vascular bundle; Ph-Phloem; PM-Palisade mesophyll; X-Xylem; MT-Mesophyll tissue.

The circular lobed leaf has a similar structure as the circular leaf. It has deeply indented epidermal layer of small spindle shaped cells and wide air chambers separated by thin partition filaments. A flat collateral vascular bundle occurs in the middle of the leaf which has horizontal short rows of thick walled xylem elements and abaxial wide zone of phloem elements (Fig. 1 b and 1c).

 

Epidermal trichomes are non glandular, covering type and curiously lobed. Some of the trichomes are long, tubular and wide with less prominent lobes at the tip. Some of the trichomes are long, narrow and have lateral, short branch. There are also trichomes which are wide, short with short spring outgrowth and thick terminal branches. More commonly the trichomes are long, wide with short or long lateral branches as well as terminal branches.

 

T.S. of Thick stem:

The thick stem is nearly 2mm in diameter. There is a narrow but distinct superficial periderm, equally developed all around the stem, there are narrow fissures at certain places. The periderm consists of four or five layers of phellem cells with straight radial walls and curved tangential walls. The periderm is 120µm wide. Inner to the periderm is a wide zone that includes secondary phloem, secretory ducts and isolated patches of fibres. The secondary phloem occurs in several successive thin layers with parenchyma tissue in between. The secretory ducts are 70µm in diameter. They occur in the middle of the phloem, the duct is surrounded by four or five layers of rectangular thick walled cells.

 

The distribution pattern of the vessels is unique. They occur in thin bands which are tangential in orientation and undulate in outline. They are in successive waves till the outer part of the cylinder. The vessels are angular, thick walled and are 20 µm wide. They are mostly in multiples of radially oblique or tangential wavy bands (Fig. 2 a).

 

Fig. 2 a. Anatomy of the thick stem

 

Young stem:

Young stem measuring nearly 1mm thick was studied. It has an intact epidermal layer bearing dense trichomes. The epidermal cells are thick walled papillate. The cortex consists of outer zone of six layers of compact parenchyma cells and inner zone of three or more wide tangentially stretched air chambers separated by partitions comprising of cylindrical cells. Phloem is a narrow continuous zone ensheathing the xylem. Along the outer portion of the phloem, there are thick masses of cortical fibres.

 

Xylem is in the form of thick, hollow cylinder around the pith. Xylem has fibres and vessels. The vessels are narrow, circular or angular thick walled and are distributed in straight or oblique radial multiplies or in short clusters. The pith is wide and consists of angular, thick walled and lignified compact cells (Fig. 2 b and 2 c).

 

Fig. 2 b. Structure of the young stem

 

Fig. 2 c. T.S of stem –half portion enlarged

Pe-Periderm; Co-Cortex; Pi-Pith; AC-Aerenchyma; CF-Cortical fibre; Ph-Phloem; SPh-Secondary Phloem; SX-Secondary Xylem

 

Physicochemical parameters:

The physiochemical parameters are mainly used in judging the purity and the quality of the drug (Table 1). Ash values give an idea of the earthy matter or inorganic composition or other impurities present along with the drug. The ash value of the powdered plant revealed a high percentage of sulphated ash as compared to other ash values. Extractive values give an idea about the chemical constituents present in the drug as well as useful in the detection of exhausted or adulterated drugs. The result suggests that the powdered plant have high alcohol soluble extractive value as compared to other extractive values. The loss on drying reveals the percentage of moisture present in the drug. The crude fibre content determines the excessive woody material criteria for judging the purity. The foaming index and swelling index also studied. The fluorescence analysis of powdered drug and extracts was studied in both UV and daylight. The powdered plant showed green fluorescence with 1N alcoholic NaOH, 1N NaOH, 1N KOH and 1N alcoholic KOH in UV light at 254 nm, which indicates the presence of chromophore in the drug (Table 2).

 

Table 1. Physico-chemical constants

S. no

Parameters

Values

I

Ash values

 

1

Total ash

7.83 % w/w

2

Acid Insoluble Ash

2.71 % w/w

3

Water Soluble Ash

4.36 % w/w

4

Sulphated Ash

8.51 % w/w

II

Extractive values

 

5

Alcohol soluble Extractive

30 % w/w

6

Water soluble Extractive

18.6 % w/w

7

Non- volatile Ether soluble Extractive

2 % w/w

8

Volatile Ether soluble Extractive

5 % w/w

III

Other parameters

 

9

Crude fibre content

60.07 % w/w

10

Loss on drying

3.2 % w/w

11

Swelling index

10

12

Foaming Index

Nil

13

Volatile oil content

1.33 % v/w

 

Preliminary phytochemical screening:

Preliminary phytochemical screening of the plant extracts showed the presence of terpenoids, alkaloids, carbohydrates, flavanoids, proteins, steroids, tannins and volatile oils (Table 3).


Table 2. Fluorescence analysis of Santolina chamaecyparissus  powder

S. no

Treatment

Day light

Short UV (254 nm)

Long UV (365 nm)

1

Powder

Pale Green

Pale Green

Dark Green

2

Powder + water

Green

Green

Dark Green

3

Powder +1N HCl

Brownish green

Pale Green

Brownish green

4

Powder+1N H2SO4

Brown

Pale Green

Pale Green

5

Powder +1N HNO3

Brown

Brownish green

Brownish green

6

Powder+Acetic acid

Dark Brown

Yellowish brown

Brownish green

7

Powder + 1N NaOH

Dark Brown

Fluorescent green

Brownish green

8

Powder +1N KOH

Dark Brown

Fluorescent green

Brownish green

9

Powder+1NAlc. NaOH

Dark Brown

Fluorescent green

Brownish green

10

Powder +1NAlc. KOH

Dark Brown

Fluorescent green

Pale Green

11

Powder + Ammonia

Brown

Brownish green

Brownish green

12

Powder + Iodine

Pale Brown

Yellowish brown

Brownish green

13

Powder + FeCl3

Reddish Brown

Dark Green

Dark Green

14

Powder + Ethanol

Dark Green

Brownish Green

Brownish Green

 

Table 3. Preliminary Phytochemical Screening of Santolina chamaecyparissus

Test

Hexane

Benzene

Chloroform

Acetone

Ethanol

Water

Carbohydrates

-

-

-

-

+

+

Phytosterols

+

+

-

-

-

-

Terpenes

+

+

-

-

-

-

Volatile oil

+

-

-

-

-

-

Lipids and fats

+

-

-

-

-

-

Saponins

-

-

-

-

-

-

Alkaloids

+

+

-

-

-

-

Phenolic compounds and tannins

-

-

-

+

+

+

Proteins and amino acids

-

-

-

-

+

+

Flavanoids

-

-

-

+

+

-

Gums and Mucilage

-

-

-

-

-

+

+ denotes the presence of the respective class of compounds.

 


 

CONCLUSION:

Developing standards is an integral part of establishing the correct identity and quality of a crude drug. The above studies provide information on the Pharmacognosy, chemical constituents and physicochemical characters which may be useful for standardization of herbal drugs of folk medicine practice.

 

REFERENCES:

1)       Lewis DA. Anti-inflammatory drug from plant and Marine sources, Birkhauser, 1989; pp. 231-283.

2)       Brain KR, Turner TD. The Practical Evaluation of Phytopharmaceuticals. Wright-Scientechnica, Bristol, 1975; pp. 4-9.

3)       Johansen DA.  Plant Microtechnique, McGraw Hill, New York, 1940; pp. 182.

4)       Wallis TE. Text book of Pharmacognosy, CBS publication, 1958; 6th ed: pp. 182.

5)       Brain KR, Turner TD. The Practical Evaluation of Phytopharmaceuticals, Wright-Scientechnica, Bristol, 1975; pp. 36-45.

6)       Indian Pharmacopoeia. Government of India, Ministry of Health and Welfare, Controller of Publications, New Delhi, 1996; vol. II, 4th ed: pp. A53-A54.

7)       Chase CR, Pratt RJ. Fluorescence of powdered vegetable drugs with particular reference to development of a system of identification. J Am Pharmacol Assoc 1949; 38: 32-36.

8)       Kokoshi J, Kokoshi R, Sharma FJ. Fluorescence of Powdered vegetable drug under ultraviolet radiation, J Am Pharmcol Assoc 1958; 47: 715-717.

9)       Harborne JB. Methods of extraction and isolation. In: Phytochemical Methods, Chapman & Hall, London, 1998; pp. 60-66.

 

 

 

 

 

 

Received on 25.04.2010       Modified on 20.05.2010

Accepted on 31.05.2010      © RJPT All right reserved

Research J. Pharm. and Tech. 4 (1): January 2011; Page 85-88

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