pharmacognostic studies of selected medicinal plants...

ETHNOPHARMACOLOGICAL VALIDATION OF MEDICINAL PLANTS TREATING SKIN DISEASES IN HYDERABAD KARNATAKA REGION 243

CHAPTER-III

Pharmacognostic studies of selected medicinal plants

3.1 Introduction

Standardization of natural products is a complex task due to their heterogeneous

composition, which is in the form of whole plant, plant part or extracts obtained thereo f.

To ensure reproducible quality of herbal products, proper control of starting material is

utmost essential. The first step towards ensuring quality of starting material is

authentication. Thus, in recent years there has been a rapid increase in the standardization

of selected medicinal plants of potential therapeutic significance (Reddy and Venkatesh,

2004). Despite the modern techniques, identification of plant drugs by pharmacognostic

studies is more reliable. According to the World Health Organization, (WHO, 1998), the

macroscopic and microscopic description of a medicinal plant is the first step towards

establishing the identity and degree of purity of such materials and should be carried out

before any tests are undertaken.

The term Pharmacognosy was coined by Seydler, a German scientist, which has

root from the green word Pharmakon’ a drug and gignoso, to acquire knowledge is

actually deals with the structural, physical, chemical and biological properties of crude

drugs along with their history, method of cultivation and its preparation (Evans,1989).

During the Vedic age any such person who collected drugs was much reverend and

was known as "parameshti prajapati". In Greece, the same people were known as

"Rhizotomi". Charaka stressed the importance of correct identification of a plant as vital

for treatment. He states that simply confirming the name of the plant does not help in any


way for treatment and that one should be well versed with morphological characters. To

acquire knowledge of plants especially through its name, morphological characters, uses,

etc., one should associate/interact with the hill tribes and cowherds of the forest. The forest

dwellers have a clear idea about the measurements (pramana), colour (varna), physical

characters (akriti) and the specific reproductive characteristic (jatilinga) of each plant.

Dhanvantri nighantu, the ancient master-mind had the unique intuitive and

innovative powers in fixing up the identity of various plants with reference to the latent

properties of the plant on one side and prescribing them for particular ailments all in

scientific objectivity and precision. In addition, the raw material standardization can be

done by correct identification and evaluation of organoleptic characters like colour, odour,

taste, ash and material content, and the fluorescent studies of powdered drug are of utmost

importance to distinguish the different varieties of the plant.

Selected 07 medicinal plants i.e., Annona reticulata L., Annona squamosa L.,

Corchorus olitorius L., Euphorbia tirucalli L., Ficus racemosa L., Pongamia pinnata L.,

Vitex negundo L. parts were having many minor variation creating difficulties in their

proper identification. Hence, the present attempt was made to distinguish the plant parts

through pharmacognostic studies.


3.2 Review of literature

The selected seven ethno medicinal plants for the present investigation is used not

only in this region for curative purposes but also used all over the world. Goel and Aswual

(1990) have made an effort to document the less known medicinal plants which are used in

indigenous folklore from Northern India. Murthi and Sheshadri (1941) have studied the

root characters of Decalepis hamiltoniii and K. indicus and revealed that these two belong

to taxonomically different genera. Chase and Pratt (1949) have investigated the

fluorescence of 151 identified powdered drugs. They also presented a key for the

identification of powdered vegetable drugs based on fluoresence studies. Prasad et al.

(1959) made a comparative study of roots of Withanla somnifera and Withania coagulans.

Dhalla et al. (1961) have studied the pharmcognosy of the roots of both wild Withania

somnifera and cultivated Withania ashwagandha. Prasad and Wahi (1965) have studied

pharmacognosy characteristics of the root of Hemidesmus indicus. Krishnamurthy and Sundaram

(1967) have showed the Morphology of the foliar epidermis of Hemidesmus indicus in which

they have described the type of stomata, cell type and trichomes. Wahi et al. (1971) and

Nayar (1979) made comparative studies of macroscopic and microscopic characters of the

root of four plants. Yoganarasimhan et al. (1981) have given the key characters of some

medicinally important plants like, Tagara, Bharangi, Ishwari, which will help to identify

these plants scientifically to avoid adulterations. Surunge and Deokule (1986) and Mulla

and Datwani (1994) have reported pharmacognstic characters and evaluated the ash values,

soluble and insoluble ash, extractive value of rhizome of Trichodesma indicum R.Br. and

Curculigo recurvata DR. respectively. Fadeyi et al. (1989) have studied four species of the

genus Boerhavia occurring in Nigeria with respect to anatomical and phytochemical

aspects. Dave and Kurjachen (1991) have made an extensive comparative anatomical work

on the fruits of Cryptolepis buchanani and Hemidesmus indicus. Deokule (1991 and 1995)


have also emphasized on morphological, histological and phytochemical studies in his

work on some members of Leguminaceae like Vigna trilobata, Crotalaria return and

Wagatea spicata. Wang et al. (1994) have made thorough investigation of original plants

and commercial products of the traditional Chinese drug Jiuyandu huo. Wang et al. (1995)

have published a review article on recent advances in pharmacognosy research in China.

Lohar et al. (1995) have done standardization of Ptelia trifoliaia Linn by using some

phytochemical and pharmacognostical methods. Fillippini et al. (1995) have done

comparative investigations of finely powdered sample of commercially available capsules of Herba

equiseti with that of authenticated plant material Equisetum arvense L. They concluded that with

the help of scanning electron microscopic technique it is possible to identify the drug even at the

species level. Hamano et al. (1996) have evaluated 48 samples of Cinnamon barks sold in the

markets in Japan. Lim et al. (1996) have done comparative morphological and histological studies

on the Chinese crude drug Huang qi and its adulterants. Dutta and De (1995) have studied

pharmacognostic characters of pods of Cassia fistula and Cassia javanica as the later is often

found as adulterant. Krishnan Nambiar et al. (1998) have published a paper on

pharmacognostic studies on Saraca asoca in comparison with Polyalthia longifolia a

common adulterant of asoca. Madhavan (1999) made an attempt to suggest a few

appropriate standardization techniques to strengthen Ayurvedic system. Krishnan

Nambiar (2000) have studied pharmacognostical characters of Asparagus racemoses

collected from different parts of Kerala. Satakopan (2000) has focused on a few modern

techniques which can be implied to control the quality of Ayurvedic, Siddha and Unani

crude drugs.

There are many more reports on standardization of crude drugs, and to mention a

few, the works of Jaffer et al. (1998), Joshi and Jayshree (1988), Pattanshetty et al. (1989)

Barthwal and Srivastava (1990), Toker et al. (1995) are noteworthy. They found that the


crude drugs were contaminated by Altemaria, Aspergillus, Curvularia, Fusarium, Pennicillium

and Rhizopus. Out of 37 samples screened 30 were found to be aflatoxin positive. The same authors

in the year 1990 found that the active principles have been deteriorated due to fungal association

and stored under different relative humidities. Kulkarni and Lipnis (1992) have made an attempt to

decontaminate some crude drugs by gama radiations and revealed that the irradiation at 10 kg and

15 kg dose of gama radiation could completely decontaminate the samples without altering

their physicochemical parameters and active phytochemical constituents. Fischer et al.

(1993) have analyzed phototherapeutic products of oral dosage forms marketed in Brazil

regarding microbiological quality. Huang and Blume (1994) have tested a few medicinal

plant materials for the microbial contamination. Kivman et al. (1998) have studied the

problems in the standardization of the microbial purity of raw medicinal plant material.

Limyati and Juniar (1998) have assessed Jamu Gendong, a kind of traditional medicine in

Indonesia for microbial contamination of its raw materials and end products. Krishnaveni

and Saaanth Rani have studied pharmacognostic parameters for the leaves of Sapindus

trifollliatus Vahl have studied with the aim of drawing the pharmacopial standard for the

species identification.

Sugumaran and Vetrichelvan have pharmacognostic profile of Bauhinia purpurea.

According them ash value and extractive values are helpful in the differentiation from

other related species. Divyakanth and Vimal Kumar, 2008 have focused on

pharmacognostic characteristic of Neolamarkia cadamba (roxb) in that they have studied

microscopic, macroscopic characteristics of the plant viz., morphological studies, such as

shape, size, apex, surface, base, margin, venation, taste and odour of leaves, as a part of

quantitative microscopy, stomatal number, stomatal index, vein islet number and vein

termination number were determined by using fresh leaves of the plant. The total ash,

water-soluble ash, acid- insoluble ash, sulphated ash, alcohol- and water-soluble extractive


value and foaming index were determined. Satkopan (2000) have focused on a few modern

technique which can be implied to control the quality of Ayurvedic, Siddha and Unani

crude drugs. Nirmaladevi and Periyanayagam (2008) have studied the micro

morphological studies of the plant Plectranthus amboinicus for identification of medicinal

plants and also have studied SEM for better understanding of structural details and to assist

in the solution of taxonomic problem. Nawagish et al., 2007 have studied the preliminary

and pharmacological standardization of Lawsonia inermis Lin. Venkatesh et al. (2008)

have studied the pharmacognostic studies of Dodonaea viscose to overcome the taxonomic

confusion among Dodonaea viscosae varieties. In endemic plant of Jatropa species

pharmacognostic studies were performed by Uthayakumari and Sumathy (2011).

Prasad, Kadam et al., (2012) have studied the pharmacognostic studies of

Mimusops elengi to overcome drug standardisation among Mimusops elengi varieties.

Sumitra Chanda (2014) has given detailed review on importance of pharmacognosy in

medicinal plants.

Kamaruz Zaman and Kalyani Pathak (2013) have studied the

pharmacognostic studies of A. reticulta to overcome drug standardisation among Annona

species, Gorwadiya (2010) drug standardised in F. racemosa, Kumar et al., (2013) have

studied the pharmacognostic studies of P. pinneta, Ahirrao et al., (2011) have

pharmacognostic profile of V. negundo. Seed extract fractions of Corchorus olitorious

pharmacognostic studies were recorded by Maxwell Osaronowen Egua (2014).


3.3 Materials and methods

Organoleptic evaluation

In organoleptic evaluation, various sensory parameters of the plant material, such as

weight, colour, odour, and taste of the leaves were recorded. It includes conclusions drawn

from studies resulted due to impressions on organs of senses.

Florescent studies of powdered drug (Chase, Pratt, 1948)

The dried powder of selected plants parts was sieved through the sieve plate

No.120 and was used for fluorescent studies. A pinch of this powder was taken in a clean

test tube with about 10ml of solvent extract. Likewise, several tubes were made by adding

various solvents like, alcohol, methanol, ethyalacetate, chloroform, benzene, hexane,

acetone, water, hydrochloric acid and conc. sulphuric acid. All the tubes were shaken well

and incubated for about 30min. The colour of the drug solution thus obtained were

observed for their characteristic colour reaction under the visible light(fluorescent tube)

and the ultraviolet light(UV356) and were recorded by comparing with standard colour

charts(Chase and Pratt, 1949).

Determination of extractive value: (Raghunathan, 1982)

Accurately weight 100g of powdered plant materials were extracted successively in

a Sox- let extractor with solvent Petroleum ether, Chloroform, Methanol, Alcohol and

distilled water separately. The obtained extracts were allowed to dry at room temperature.

After complete evaporation of the solvent, weight, colour and nature of the extracts were

noted.

Weight of the residue obtained

Extractive value (%) = x 100

Weight of the plant material taken


Determination of Ash value: (W.H.O. 2002)

Take a known weight of the dried plant material in a pre weighed silica crucible.

Heat it on a Bunsen burner at a low flame, till the plant material gets charred. Then it is

transferred to a muffle furnace and heated strongly at a dull red heat (400-4500C) till a

white ash is obtained. It is cooled in a desiccators for about 15 to 30min, and weighed

using electronic balance and the readings are noted down (Raghunathan, 1982).

Z-X

Total ash value of the sample = x 100

Y

Where, Weight of the empty crucible = Z, Weight of crucible with Ash = X, Weight of the

plant material (g) = Y

Z-X

Weight of the dish + ash = x 100

Y

Determination of acid insoluble ash:

The above obtained total ash was boiled for 15 ml with 25ml of 25% hydrochloric

acid. This was filtered and the insoluble matter was collected on ash less Whatman filter

paper, ignited in sintered crucible, cooled and then kept in a desiccators for 15minute. The

residue was weighed in electronic balance and the acid insoluble ash was calculated.

100

Acid insoluble ash value of the sample = x A

Y

Where, A = Weight of the residue obtained, Y = Weight of the plant material.


3.4 Experimental results

3.4.1 Annona reticulata L.

Organoleptic studies

The detailed study of organoleptic characters of extracts, ash values and the

florescent behaviour of Annona reticulata L. leaf can be easily identified and further

distinguished from their possible adulterants and substituent’s by these parameters.

Organoleptic studies of extracts

The successive Soxhlet extracts viz. petroleum ether. Chloroform ethyalacetate and

98% methanol extracts of A. reticulata L. yields were determined and their differential

values were recorded in the Table. 3.1.4. Among all the extracts obtained, the 98%

methanol extract was having higher percentage of yie ld of compounds then other extracts.

The weight of leaf extracts are 1.50 (pet. ether), 1.80 (chloroform), 1.50 (ethyl acetate) and

2.80 (98% methanol) percent respectively. It is evident from these results, that the 98%

methanol solvent has higher extractive values followed by chloroform, pet-ether and ethyl

acetate.

Table-3.1: Extractive values and organoleptic characters’ of Annona reticulata L. extracts.

Sl no

Plant part

used

Solvents

Weight of

the extracts

Colour of

the extract

Taste of the

extract

Nature of

the extract

Odour of

the extract

1 LEAVES PE 1.50 Yellow Bitter waxy Sweet

2 CHCL3 1.80 Dark Green Bitter sticky Aromatic

3 EtOH 1.50 Light green Bitter sticky Aromatic

4 98% Methanol 2.80 Dark brown Salty Bitter waxy Aromatic

The crude extracts of A. reticulata leaf have shown a wide range of colour. The petroleum

ether extract shows yellow. While, the chloroform extract exhibited dark green colour.

Whereas, the ethyl acetate extract shows light green colour and 98% methanol shows dark

brown colour (Table: 3.1).

A. reticulata leaf extracts taste varies from extract to extract such as tasteless,

sweet. Bitter, light bitter and salty. Of these the petroleum ether, chloroform and ethyl

acetate extract exhibited bitter and 98% methanol show salty bitter in taste.


Further the nature of these extracts varies, pet. ether extract is waxy in nature

whereas chloroform and ethyl acetate extract extracts shows sticky. Whereas the 98%

methanol extract is waxy. Similarly, the extracts were exhibiting variety of odour. The

petroleum ether extract exhibiting sweet and chloroform, ethyl acetate and 98% methanol

extracts exhibited the aromatic odour.

Determination of total ash and acid insoluble ash contents

In the present investigation, the total ash and acid insoluble and water insoluble ash

contents of A. reticulata leaf was studied (Table: 3.2).

Table-3.2: Total ash and acid insoluble ash content in Annona reticulata L. (mg/100g)

Sl no parts Total ash Acid insoluble

ash contents

Water

insoluble

ash

contents

1. Leaf 6.0 3.61 2.42

The total ash value recorded as (07%), acid insoluble ash value determined as 3.61% and

water insoluble ash value recorded 2.42%. This variation in the content of ash is probably

due to the nature of metabolites they possess.

Fluorescent studies of powered drugs

The characteristic colour behaviour of dried and powdered drugs and powder

dissolved in organic solvents like petroleum ether, chloroform, alcohol, water, acetone,

ethyl acetate, Conc. HC1 and H2So4(5%) i.e., leaf of A. reticulata was observed both under

the visible and ultra violet (UV nm) light. The colour reactions of these drugs solutions

thus emitted fluorescence light are tabulated in Table: 3.3.

Table-3.3: Fluorescent studies of powdered drugs Annona reticulata L.

Sl no

Treatment

visible

UV 365 nm

1 Powder Dark green Onion pink

2 Pet. Ether Blackish green Ash

3 Chloroform Yellow green Opaline green

4 Ethanol Light green Light pink

5 Eather Dark green Light pink

6 Acetone Light green Light pink

7 Ethyl acetate Brownish green Light onion


8 Benzene Light green Black

9 Conc. HCl (5%) Light brown Black

10 Conc. H2So4 (5%) Light brown Pale white

The powdered drug solutions have exhibited a wide range of fluorescence colours

under the UV light and visible light. The petroleum ether drug solutions have shown the

fluorescence colours such as blackish green in visible light and ash in UV light, the

chloroform drugs solutions have exhibited a wide range of fluorescence colours as yellow

green in visible in Opaline green UV light . While ethanol (95%v/v) treated drug solutions

exhibited wide range of fluorescence colours light green in visible in UV light pink.

However, the ether drug solutions exhibited dark green in visible light and light pink in

UV light.

In acetone drug solution shows the colours light green in visible light and light pink

in UV light, In ethyl acetate drug solution shows the colours chassis red in visible light and

light pink in UV light, similarly benzene drug solution exhibited light green and black.

Whereas, dilute HC1 (5%) acid drug solution indicating the light brown and black and in

dilute H2So4 (5%) acid drug solution shows light brown and pale white respectively. The

colour reaction varies from non-polar to high polar solvents (Plate-3.1).

3.4.2 Annona squamosa L.



fluorescent behaviour of A. squamosa L. leaf can be easily identified and further




98% methanol extracts of A. squamosa L. yields were determined and their differential

values were recorded in the Table. 3.4. Among all the extracts obtained, the 98% methanol

extract was having higher percentage of yield of compounds then other extracts. The

weight of leaf extracts are 0.85 (pet. ether), 1.6 (chloroform), 0.45 (ethyl acetate) and 3.23

(98% methanol) percent respectively. It is evident from these results, that the 98%


methanol solvent has higher extractive values followed by chloroform, pet-ether and ethyl

acetate.

Table-3.4: Extractive values and organoleptic characters’ of Annona squamosa L. extracts

Sl.

no

Plant part used

Solvents

Weight of

the

extracts(%

dry weight

in gm)

Colour of

the extract

Taste of the

extract

Nature of the

extract

Odour of

the extract

1 Leaf PE 0.85 yellow Slight Bitter Light sticky Sweet

2 CHCL3 1.6 Light

green

Sweet bitter sticky Sweet

3 EtOH 0.45 Brownish

green

Bitter Light sticky Aromatic

4 98%

Methanol

3.23 Dark

Brown

Bitter waxy Aromatic

The crude extracts of A. squamosa L. leaf have shown a wide range of colour. The

petroleum ether extract shows yellow. While the chloroform extract exhibited light green

colour. Whereas, the ethyl acetate extract shows brownish green colour and 98% methanol

shows dark brown colour (Table: 3.4).

A. squamosa L. leaf extracts taste varies from extract to extract such as tasteless,

sweet. Bitter, light bitter and salty. Of these the petroleum ether and chloroform extracts

exhibited slight bitter, sweet bitter respectively. The ethyl acetate and 98% methanol

extracts bitter in taste.

Further the nature of these extracts varies, pet. ether and ethyl acetate extracts were

light waxy in nature. Whereas chloroform extract in nature sticky. While the 98%

methanol extract was in waxy.



contents of A. squamosa L. leaf was studied (Table: 3.5.). Table-3.5: Total ash and acid insoluble ash content in Annona squamosa L. (mg/100g)

Sl no parts Total ash Acid insoluble ash

contents

Water insoluble

ash contents

1. Leaf 6.84 0.94 3.54


The total ash value recorded as 6.84%, acid insoluble ash value determined as 0.94% and

water insoluble ash value recorded 3.54% . This variation in the content of ash is probably

due to the nature of metabolites they possess.




ethyl acetate, Conc. HC1 and H2So4 (5%) i.e., leaf of A.squamosa L. was observed both

under the visible and ultra violet (UV nm) light. The colour reactions of these drugs

solutions thus emitted fluorescence light are tabulated in Table: 3.6.

Table-3.6: Fluorescent studies of powdered drugs Annona squamosa L.

Sl no

Treatment

visible

UV 365 nm

1 Powder Green Dark brown

2 Pet. Ether Pastel green Light ash

3 Chloroform Mint green Sky blue

4 Ethanol Light green Slight ash

5 Ether Bus Green Opaline green

6 Acetone Pista green Dark blue

7 Ethyl acetate Slight green Onion

8 Benzene Pastel green Pink

9 Conc. HCl (5%) Pale cream Blackish

10 Conc. H2So4 (5%) Gold brown Light blackish

The powdered drug solutions have exhibited a wide range of fluorescence colo urs


fluorescence colours such as pastel green in visible light and light ash in UV light, the

chloroform drugs solutions have exhibited a wide range of fluorescence co lours as mint

green in visible in sky blue UV light . While ethanol (95%v/v) treated drug solutions

exhibited wide range of fluorescence colours light green in visible in UV light ash.

However, the ether drug solutions exhibited bus green in visible light and opaline green in

UV light.

In acetone drug solution shows the colour pista green in visible light and dark blue

in UV light, In ethyl acetate drug solution shows the colours Slight green in visible light

and Onion in UV light, similarly benzene drug solution exhibited Pastel green and Pink.


Whereas, dilute HC1 (5%) acid drug solution indicating the light Pale cream and Blackish

and in dilute H2So4 (5%) acid drug solution shows Gold brown and Light blackish

respectively. The colour reaction varies from non-polar to high polar solvents (Plate-3.1).

3.4.3 Corchorus olitorius L.



florescent behaviour of Corchorus olitorius L. seed can be easily identified and further




98% methanol extracts of C. olitorius L. yields were determined and their differential

values were recorded in the Table. 3.7. Among all the extracts obtained, the 98% methanol


weight of seed extracts are 0.92 (pet. ether), 1.83 (chloroform), 2.51 (ethyl acetate) and


methanol solvent has higher extractive values followed by ethyl acetate chloroform and

pet-ether.

Table-3.7: Extractive values and organoleptic characters’ of Corchorus olitorius L. seed extracts.

Sl.

no

Plant part used

Solvents

Weight of

the

extracts

Colour of the

extract

Taste of the

extract

Nature of the

extract

Odour of the

extract

1 Seed PE 0.92 Creamy

yellow

Bitter Creamy Sweet

2 CHCL3 1.83 Slight

greenish

Bitter Creamy Sweet

3 EtOH 2.51 Light

Brownish


4 98%

Methanol

4.74 Brown Bitter waxy Aromatic

The crude extracts of C. olitorius L. seed have shown a wide range of colour. The

petroleum ether extract shows creamy yellow. While, the chloroform extract expose the

slight greenish colour. Whereas, the ethyl acetate extract shows light brownish colour and


98% methanol extract shows brownish colour. (Table: 3.7). C. olitorius L. seed all the four

solvent extracts shows bitter in taste.

Further the nature of these extracts varies, pet.ether and chloroform extracts were

creamy in nature. Whereas the ethyl acetate and 98% methanol extracts shows waxy in

nature.


In the present investigation, the total ash, acid insoluble and water insoluble ash

contents of C. olitorius L. seed was studied (Table: 3.8).

Table-3.8: Total ash and acid insoluble ash content in Corchorus olitorius L. seed (mg/100g)

Sl. no part Total ash Acid insoluble ash

contents

Water insoluble

ash contents

1. Seed 6.18 2.43 1.14

The total ash value recorded as (6.18%), acid insoluble ash value determined as

2.43% and water insoluble ash value recorded 1.14% . This variation in the content of ash

is probably due to the nature of metabolites they possess.




ethyl acetate, Conc. HC1 and H2So4(5%) i.e., seeds of Corchorus olitorius L. was

observed both under the visible and ultra violet (UV nm) light. The colour reactions of

these drugs solutions thus emitted fluorescence light are tabulated in Table: 3.9.


Table-3.9: Fluorescent studies of powdered drugs in Corchorus olitorius seed

Sl. no

Treatment

visible

UV 365 nm

1 Powder Pale cream with Blackish Light yellowish

2 Pet. Ether Transparent Green Pale Yellow precipitate

3 Chloroform Light green Light greenish

4 Ethanol Transparent Transparent

5 Ether Light brownish Light yellowish

6 Acetone Transparent Transparent

7 Ethyl acetate Blackish Blue

8 Benzene Transparent Transparent

9 Conc. HCl (5%) Highly Transparent White creamy

10 Conc. H2So4 (5%) Slight Pinkish Transparent Creamy



fluorescence colours such as transparent green in visible light and pale yellow precipitate

in UV light, the chloroform drugs solutions have exhibited a wide range of fluorescence

colours as light green in visible in light greenish UV light . While ethanol (95%v/v)

treated drug solutions exhibited wide range of fluorescence colours transparent in visible

in UV light Transparent. However, the eather drug solutions exhibited light brownish in

visible light and light yellowish in UV light.

In acetone drug solution shows the colours transparent in visible light and

transparent in UV light, In ethyl acetate drug solution shows the colour blackish in visible

light and blue in UV light, similarly benzene drug solution exhibited transparent and

transparent. Whereas, dilute HCl (5%) acid drug solution indicating the light brown and

black and in dilute H2So4 (5%) acid drug solution shows highly transparent and white

creamy respectively. The colour reaction varies from non-polar to high polar solvents

(Plate-3.1).

3.4.4 Euphorbia tirucalli L.



98% methanol extracts of E. tirucalli L. yields were determined and their differential

values were recorded in the Table. 3.4.4. Among all the extracts obtained, the ethyl acetate



weight of leaf extracts are 0.97 (pet. ether), 1.21 (chloroform), 2.94 (ethyl acetate) and

2.31 (98% methanol) percent respectively. It is evident from these results, that the ethyl

acetate solvent has higher extractive values followed by 98% methanol, chloroform and

pet-ether.

Table-3.10: Extractive values and organoleptic characters of E. tirucalli L. extracts.

Sl no

Plant part used

Solvents

Weight of

the extracts

Colour of

the extract

Taste of the

extract

Nature of the

extract

Odour of the

extract

1 Leaf PE 0.97 Creamy

yellow

Bitter waxy Sweet

2 CHCL3 1.21 Greenish Bitter Creamy Sweet

3 EtOH 2.94 Light

Brownish


4 98%

Methanol

2.31 Brown Bitter waxy Aromatic

The crude extracts of E. tirucalli L. leaf have shown a wide range of colour. The

petroleum ether extract shows creamy yellow. While, the chloroform extract exhibited

greenish colour. Whereas, the ethyl acetate extract shows light brownish colour and 98%

methanolic extract shows brown colour (Table: 3.10).

The four solvent extracts of E. tirucalli L. leaf shows bitter in taste.

Further the nature of these extracts varies, pet. ether, ethyl acetate and 98%

methanol extracts were waxy in nature whereas chloroform extract shows creamy in

nature. Similarly, the extracts were exhibiting variety of odour. The petroleum ether and

chloroform extracts exhibiting sweet odour. Whereas the ethyl acetate and 98% methanolic




contents of E. tirucalli L. leaf was studied (Table: 3.11).

Table-3.11: Total ash and acid insoluble ash content in E. tirucalli L. (mg/100g)


contents

Water

insoluble ash

contents

1. Leaf 5.39 2.28 1.08



2.28% and water insoluble ash value recorded 1.08 %. This variation in the content of ash





ethyl acetate, Conc. HC1 and H2So4(5%) i.e., leaf of E. tirucalli L. was observed both



Table-3.12: Fluorescent studies of powdered drugs in E. tirucalli L.

Sl. no

Treatment

visible

UV 365 nm

1 Powder Light green Light pink

2 Pet. Ether Pista green Ash

3 Chloroform yellow Opaline green

4 Ethanol Brownish Light pink

5 Eather Yellow Light ash

6 Acetone Greenish Sky blue

7 Ethyl acetate Light yellowish Light pink

8 Benzene Pastal green Onion pink

9 Conc. HCl (5%) Brownish Dark brownish

10 Conc. H2So4 (5%) Brownish Light black



fluorescence colours such as pista green in visible light and ash in UV light, the

chloroform drugs solutions have exhibited a wide range of fluorescence colours as yellow

in visible in opaline green UV light. While ethanol (95%v/v) treated drug solutions

exhibited wide range of fluorescence colours brownish in visible in UV light pink.

However, the ether drug solutions exhibited yellow in visible light and light ash in UV

light.

In acetone drug solution shows the colours greenish in visible light and sky blue in

UV light, In ethyl acetate drug solution shows the colours light yellowish in visible light

and light pink in UV light, similarly benzene drug solution exhibited pastal green and


onion pink. Whereas, dilute HC1 (5%) acid drug solution indicating the dark brownish and

black and in dilute H2So4 (5%) acid drug solution shows brownish and light black

respectively. The colour reaction varies from non-polar to high polar solvents (Plate-3.1)

3.4.5 Ficus racemosa L.



fluorescent behaviour of F. racemosa L. leaf can be easily identified and further




98% methanol extracts of F. racemosa L. yields were determined and their differential

values were recorded in the Table. 3.5.4. Among all the extracts obtained, the 98%

methanolic extract was having higher percentage of yield of compounds then other

extracts. The weight of leaf extracts are 1.76 (pet. ether), 1.60 (chloroform), 0.67 (ethyl

acetate) and 2.58 (98% methanol) percent respectively. It is evident from these results, that

the 98% methanolic solvent has higher extractive values followed by, pet-ether,

chloroform and ethyl acetate.

Table-3.13: Extractive values and organoleptic characters’ of Ficus racemosa L. extracts

Sl.

no

Plant part

used

Solvents

Weight of the

extracts

(% dry weight

in gm)

Colour of

the extract

Taste of the

extract

Nature of the

extract

Odour of the

extract

1 Leaf PE 1.76 Pale yellow Tasteless waxy Sweet

2 CHCL3 1.6 Dark

brown

Tasteless sticky Sweet

3 EtOH 0.67 Dark green Bitter sticky Aromatic

4 98% Methanol 2.58 Dark

brown

Salty waxy Aromatic

The crude extracts of F. racemosa L. leaf have shown a wide range of colour. The

petroleum ether extract shows pale yellow. While, the chloroform extract exhibited dark

brown colour. Whereas, the ethyl acetate extract shows dark green colour and 98%

methanolic extract shows dark brown colour. (Table: 3.13). F. racemosa L. leaf extracts

taste varies from extract to extract such as tasteless, Bitter and salty. Of these the

petroleum ether and chloroform exhibited tasteless. The ethyl acetate extract having bitter


in taste. Whereas the 98% methanolic extract shown salty in taste. Further the nature of

these extracts varies, pet. ether and 98% methanolic extracts were waxy in nature whereas

chloroform and ethyl acetate extracts shown sticky in nature. Similarly, the extracts were

exhibiting variety of odour. The petroleum ether and chloroform extracts exhibiting sweet

odour. Whereas the ethyl acetate and 98% methanolic extracts exhibited the aromatic

odour.



contents of F. racemosa L. leaf was studied (Table: 3.14).

Table-3.14: Total ash and acid insoluble ash content in Ficus racemosa L. (mg/100g)


contents

Water insoluble

ash contents

1. Leaf 8.43 3.33 7.8


3.33% and water insoluble ash value recorded 7.8% . This variation in the content of ash





ethyl acetate, Conc. HCl and H2So4(5%) i.e., leaf of F. racemosa L. was observed both


solutions thus emitted fluorescence light are tabulated in 3.15.

Table-3.15: Fluorescent studies of powdered drugs Ficus racemosa L.

Sl. no

Treatment

visible

UV 365 nm

1 Powder Light green Light pink

2 Pet. Ether Pista green Ash

3 Chloroform Bus green Opaline green

4 Ethanol Light green Light pink

5 Eather Mint green Light ash

6 Acetone Pista green Sky blue

7 Ethyl acetate Light green Light pink


9 Conc. HCl (5%) Sandal wood Dark brownish

10 Conc. H2So4 (5%) Pale cream Light black




fluorescence colours such as pista green in visible light and ash in UV light, the

chloroform drugs solutions have exhibited a wide range of fluorescence colours as bus

green in visible in Opaline green UV light . While ethanol (95%v/v) treated drug solutions

exhibited wide range of fluorescence colours light green in visible in UV light ash.

However, the ether drug solutions exhibited mint green in visible light and light ash in UV

light.

In acetone drug solution shows the colours pista green in visible light and sky blue

in UV light, in ethyl acetate drug solution shows the colours light green in visible light and

light pink in UV light, similarly benzene drug solution exhibited pastal green and onion

pink. Whereas, dilute HCl (5%) acid drug solution indicating the dark sandal wood and

dark brownish and in dilute H2So4 (5%) acid drug solution shows pale cream and light

black respectively. The colour reaction varies from non-polar to high polar solvents

(Plate 3.1).

3.4.6 Pongamia pinnata L.


The detailed study of organoleptic characters of extracts, ash values a nd the

fluorescent behaviour of Pongamia pinnata L. seed can be easily identified and further




98% methanol extracts of P. pinnata L. yields were determined and their differential

values were recorded in the Table. 3.6.4. Among all the extracts obtained, the pet-ether

extracts are having higher percentage of yield of compounds then other extracts. The

weight of seed extracts are 5.21 (pet. ether), 1.42 (chloroform), 1.00 (ethyl acetate) and

0.73(98% methanol) percent respectively. It is evident from these results, that the pet-ether

solvent has higher extractive values followed by chloroform, ethyl acetate and 98%

methanol.


Table-3.16: Extractive values and organoleptic characters’ of Pongamia pinnata L. extracts.

Sl.

no

Plant part used

Solvents

Weight of

the

extracts

Colour of

the extract

Taste of the

extract

Nature of

the extract

Odour of

the extract

1 Seed PE 5.21 yellow Tasteless Oily Sweet

2 CHCL3 1.42 Slight

white


3 EtOH 1.00 Ash Bitter Creamy Aromatic

4 98%

Methanol

0.73 White Bitter Amorphous Aromatic

The crude extracts of P. pinnata L. seed have shown a wide range of colour. The

petroleum ether extract shows yellow. While the chloroform extract exhibited slight white

colour. Whereas, the ethyl acetate extract shows ash colour and 98% methanol s hows

white colour (Table: 3.16).

P. pinnata L. extracts taste varies from extract to extract such as tasteless, sweet.

Bitter, light bitter and salty. Of these the petroleum ether extract exhibited tasteless and

chloroform, ethyl acetate, 98% methanol show bitter in taste.

Further the nature of these extracts varies, pet. ether extract is oily in nature and

chloroform extract shows waxy. Similarly, the ethyl acetate extracts exhibiting creamy in

nature. Whereas the 98% methanol extract is amorphous.



contents of seed of P. pinnata L. was studied (Table: 3.17).

Table-3.17: Total ash and acid insoluble ash content in Pongamia pinnata L. seed (mg/100g)


contents

Water

insoluble ash

contents

1 Seeds 7.00 2.41 1.93

The total ash value recorded as (07%), acid insoluble ash value determined as

(2.41%) and water insoluble ash value recorded 1.93%. This variation in the content of

ash is probably due to the nature of metabolites they possess.





ethyl acetate, Conc. HC1 and H2So4 (5%) i.e., seed of P. pinnata L. was observed both



Table-3.18: Fluorescent studies of powdered drugs in Pongamia pinnata L. Seed.

Sl. no

Treatment

visible

UV 365 nm

1 Powder White Ash

2 Pet. Ether Pale rose Light pink

3 Chloroform White turbidity Ash turbidity

4 Ethanol Turbidity Blue

5 Eather Transparent Transparent

6 Acetone Pale rose Light pink

7 Ethyl acetate Chassis red Onion

8 Benzene Light sard Transparent

9 Conc. HCl (5%) Pace rose Slight blue

10 Conc. H2So4 (5%) Turbidity White



fluorescence colours such as pale yellow in visible light and ash in UV light, the

chloroform drugs solutions have exhibited a wide range of fluorescence colours as white

turbidity in visible ash turbidity in UV light. While ethanol (95%v/v) treated drug

solutions exhibited wide range of fluorescence colours turbidity in visible in UV light blue.

However, the eather drug solutions exhibited transparent in visible light and transparent in

UV light.

In acetone drug solution shows the colours pale rose in visible light and light pink

in UV light, In ethyl acetate drug solution shows the colours chassis red in visible light and

onion in UV light, similarly benzene drug solution exhibited light sard and transparent.

Whereas, dilute Hc1 (5%) acid drug solution indicating the pace rose and slight blue and in

dilute H2So4 (5%) acid drug solution shows turbidity and white respectively. The colour

reaction varies from non-polar to high polar solvents (Plate 3.1).


3.4.7 Vitex negundo L.



florescent behaviour of V. negundo L. leaf can be easily identified and further


Organoleptic studies of V. negundo L. leaf extracts

The successive Soxhlet extracts viz. petroleum ether. Chloroform ethyalacetate and 98%

methanol extracts of V. negundo L. yields were determined and their differential values

were recorded in the Table. 3.19. Among all the extracts obtained, the 98% methanolic


weight of leaf extracts are 1.10 (pet. ether), 1.50 (chloroform), 0.94 (ethyl acetate) and


methanolic solvent has higher extractive values fo llowed by, chloroform, pet-ether and

ethyl acetate.

Table-3.19: Extractive values and organoleptic characters’ of Vitex negundo L. extracts

Sl.

no

Plant part used

Solvents

Weight of

the extracts

Colour of

the extract

Taste of the

extract

Nature of

the extract

Odour of

the extract

1 Leaf PE 1.10 Green Bitter waxy Sweet

2 CHCL3 1.50 Dark

Green

Bitter waxy Sweet

3 EtOH 0.94 Light green Bitter sticky Sweet

4 98%

Methanol

2.05 Dark green Salty Bitter waxy Aromatic

The crude extracts of V. negundo L. leaf have shown a wide range of colour. The

petroleum ether extract shows green. While, the chloroform extract exhibited dark green

colour. Whereas, the ethyl acetate extract shows light green colour and 98% methanolic

extract shows dark green colour. (Table: 3.19).

V. negundo L. leaf extracts taste varies from extract to extract such as tasteless, Bitter

and salty bitter. Of these the petroleum ether, chloroform and ethyl acetate extracts were

exhibited bitter taste. Whereas the 98% methanolic extract was shown salty bitter in taste.


Further the nature of these extracts varies, pet. ether, chloroform and 98%

methanolic extracts were waxy in nature, whereas ethyl acetate extract shown salt bitter in

nature. Similarly, the extracts were exhibiting variety of odour. The petroleum ether,

chloroform and ethyl acetate extracts exhibiting sweet odour. Whereas the 98% methanolic




contents of V. negundo L. leaf was studied (Table: 3.20). The total ash value recorded as

(3.10%), acid insoluble ash value determined as 0.93% and water insoluble ash value

recorded 1.24%. This variation in the content of ash is probably due to the nature of

metabolites they possess.

Table-3.20: Total ash and acid insoluble ash content in Vitex negundo L. (mg/100g)

Sl. no parts Total ash Acid insoluble ash

contents

Water insoluble

ash contents

01. Leaf 3.10 0.93 1.24




ethyl acetate, Conc. HCl and H2So4(5%) i.e., leaf of V. negundo L. was observed both



Table-3.21: Fluorescent studies of powdered drugs in Vitex negundo L.

Sl. no

Treatment

visible

UV 365 nm

1 Powder Green Light Yellow

2 Pet. Ether Fine Green Pale Yellow

3 Chloroform Blackish green Opaline green

4 Ethanol Yellow green Light pink

5 Eather Light green Light pink

6 Acetone Dark green Light pink

7 Ethyl acetate Blackish Blue


9 Conc. HCl (5%) Sandal wood Dark brownish

10 Conc. H2So4 (5%) Pale cream Light black




fluorescence colours such as fine green in visible light and pale yellow in UV light, the

chloroform drugs solutions have exhibited a wide range of fluorescence colours as

blackish green in visible in Opaline green UV light . While ethanol (95%v/v) treated drug

solutions exhibited wide range of fluorescence colours yellow green in visible in UV light

slight pink. However, the eather drug solutions exhibited light green in visible light and

light pink in UV light.

Plate 3.1: Fluorescent studies of powdered drugs of 07 selected plant parts.

1. Annona reticulata L., 2. Annona squamosa L., 3. Corchorus olitorius L., 4. Euphorbia

tirucalli L., 5.Ficus racemosa L., 6. Pongamia pinnata L., 7. Vitex negundo L.,

2. A: Visible, B: UV 365 nm.

1A

al

K

no

wl

ed

ge

Pl

an

ts

Ph

ot

os

1B

al

K

n

o

w

le

d

g

e

Pl

a

nt

s

P

h

ot

o

s

2B

al

K

no

wl

ed

ge

Pl

an

ts

Ph

ot

os

2A

A

Pl

an

ts

Ph

ot

os

3A

Kn

owl

edg

e

Pla

nts

Ph

oto

s

3B

al

K

n

o

w

le

d

g

e

Pl

a

nt

s

P

h

ot

o

s

4B

al

K

no

wl

ed

ge

Pl

an

ts

Ph

ot

os

4A

al

K

no

wl

ed

ge

Pl

an

ts

P

ho

to

s

5A

Kn

owl

edg

e

Pla

nts

Ph

oto

s

6A

Kn

owl

edg

e

Pla

nts

Ph

oto

s

7A Kn

owl

edg

e

Pla

nts

Ph

oto

s

5B

al

K

n

o

w

le

d

g

e

Pl

a

nt

s

P

h

ot

o

s

6B

al

K

n

o

w

le

d

g

e

Pl

a

nt

s

P

h

ot

o

s

7B

al

K

n

o

w

le

d

g

e

Pl

a

nt

s

P

h

ot


In acetone drug solution shows the colours dark green in visible light and

light pink in UV light, In ethyl acetate drug solution shows the colours blackish in visible

light and blue in UV light, similarly benzene drug solution exhibited pastal green and

onion pink. Whereas, dilute HCl (5%) acid drug solution indicating the sandal wood and

dark brownish and in dilute H2So4 (5%) acid drug solution shows pale cream and light

black respectively. The colour reaction varies from non-polar to high polar solvents (Plate-

3.1).

3.5 Discussion

Pharmacognostic studies of selected 07 medicinal plants have been undertaken to

overcome the standardization of drug.

Powder analysis

Powder analysis of 07 selected medicinal plant parts in the present study reveals

the different colours in organoleptic and florescent studies. Yoganarasinhma et al. (1981)

have given the key characters of some medicinally important p lants like, Tagara, Bharangi,

Ishwari, which will help to identify these plants scientifically. Divyakanth and Vimal

Kumar (2008) have studied pharmacognostic characters of the two species and according

to them these characters will helps identify to the species. Nirmaladevi and Periyanayagam

(2008) have also has the opinion that pharmacognostic study will be helpful for the

identification of medicinal plants. Fillippini et al. (1995) have done comparative

investigation of finally powdered sample of Herba equiseti with the help of Scanning

Electron Microscopic technique. Satakopan (2000) has focused on few modern techniques

like, SEM which can be implied to control the quality of Ayurvedic, Siddha and Unani

crude drugs.

Extractive and Ash values

Variation in the content of ash and extractive values is probably due to the nature

of metabolites they posses. The extractive values in different solvents viz., petroleum

ether, chloroform, ethyalacetate and 98% methanol extracts were recorded.

Among the extracts obtained, the 98% methanolic extract showed higher yield.

According to the literature the more polar compound will have high extractive value and

less polar compounds will have less extractive value. But, in the present study, it was


found that less polar solvents also showed high extractive value like E tirucalli in

ethyalacetate, Pongamia pinnata L. in petroleum ether, which indicates the presence of

excessive amount of resinous materials present in these plants. These reports supported to

previous reports of Kamaruz Zaman and Kalyani Pathak (2013).

In the present study A. reticulata L. total ash value 06 mg/100g, A. squamosa L.

(6.84), C. olitorius (06.18), E tirucalli (5.39), F. racemosa (8.43), P. pinnata (07) and V.

negundo L. (3.1) whereas in previous report of Kamaruz Zaman and Kalyani Pathak

(2013) showed high ash value 15 mg/100g (A. reticulata L.), Gorwadiya (2010) recorded

in F. racemosa L., Kumar et al., (2013) recorded in P. pinneta L., Ahirrao et al., (2011)

recorded in V. negundo L.. It may depend climatic conditions. Sugumaran and

Vetrichelvan (2008) have reported pharmacognostic profile of Bauhinia purpurea.

According to them the ash value play important role in the differentiation of other related

species. The ash content which remains after incineration of drug may contain some inorganic

compounds such as phosphates, carbonates, silicates and silica, which occur naturally in drug or

added deliberately in the form of adulterants, this content differ from plant to plant.

Fluorescence characters

The characteristic colour behavior of dried and powdered drug of plant is observed

both under the visible and ultraviolet light by treating with various solvents like, water,

ethanol, methanol, ethyl acetate, hexane, benzene, chloroform, acids like, HCl and H2SO4.

The drug solutions have been exhibiting a wide range of fluorescent colours particularly

under the UV light compared to visible light. These fluorescent colours can be compared

with other plants during identification. Chase and Pratt (1949) have said that the

fluorescence of powdered drugs can be used for the identification of plants. They also

presented the key for the identification of powdered vegetables based on the fluorescent

studies. Kamaruz Zaman and Kalyani Pathak (2013) have carried out pharmacognostic

studies to overcome the Taxonomic confusion among the Annona species. Whereas the

previous similar fluorescence characters were observed Gorwadiya (2010) in F. racemosa

L., Kumar et al., (2013) have recorded in P. pinneta L., Ahirrao et al., (2011) have

recorded in V. negundo L.

Macroscopic as well as microscopical studies of any phyto drug are the primary

steps to establish its botanical quality control before going to other studies. As per WHO

norms, botanical standards are to be proposed as a protocol for the diagnosis of herbal


drug. The above mentioned parameters are helpful for the future identification and

authentification of plant in herbal industry and in factories. The physico-chemical

standards, such as, ash values, extractive values, crude fiber content and fluorescence

analysis will be useful to identify the drug even in the crushed or powdered condition. It

will also serve as a standard data for quality control of the preparations containing this

plant in future. The leaf constants can be included as microscopical standards in Indian

herbal pharmacopoeia. Phytochemical study is also useful to isolate pharmacologically

active principles present in the drug. The information obtained from ash values and

extractive values are useful during the time of collection and also during extraction

process. Using these standards, the plant can be differentiated from other related species

and varieties.

pharmacognostic studies of selected medicinal plants...

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