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Dhiren et al. / Pharma Science Monitor 8(2), Apr-Jun 2017, 583-602

Pharma Science Monitor 8(2), Apr-Jun 2017

PHARMACOGNOSTICAL, PHYSICO-CHEMICAL AND CHROMATOGRAPHIC

EVALUATION OF SEEDS OF ENTIRE HERB OF PORTULACA OLERACEA LINN

Dhiren M. Joshi1*, Ashvin V. Dudhrejiya1, Ravi A. Manek1 and Navin R. Sheth2

1B. K. Mody Government Pharmacy College, Rajkot – 360 003, Gujarat, India2 Hon. Vice-Chancellor, Gujarat Technological University, Ahmedabad, Chandkheda – 382 424, Gujarat, India

ABSTRACTIndia has rich heritage of usage of medicinal plants in its Ayurvedic, Siddha and Unani systemsof medicines besides use of many in folk remedies. The objective of the present investigationwas to evaluate folklore medicine Portulaca oleracea L. for its various pharmacognostic, phyto-chemical studies and chromatographic studies. Portulaca oleracea L. (Common Purslane, alsoknown as Verdolaga, Pigweed, Little Hogweed or Pusley), is an annual succulent in the familyPortulacaceae, which can reach 40 cm in height. It has smooth, reddish, mostly prostrate stemsand alternate leaves clustered at stem joints and ends. The yellow flowers have five regular partsand are up to 6 mm wide. It is called Loni or Ghol in Guajarati, Chhota Lunia in Hindi &Bengali. Portulaca oleracea L. is used as a folk medicine for the treatment of inflammation inIndia. It was found that the plant possesses nor-adrenaline, dopamine, dopa, vitamin C,olearacins – I and II, omega 3 fatty acids, saponins, tannins, flavonoids, saccharides,triterpenoids, α -tocopherol and glutathione, alkaloids. The study of transverse section carriedout using microscopy as well power study of entire herb was performed for the evaluation ofplant. In the present study the methanolic extract of Portulaca oleracea L. was subjected topreliminary phytochemical evaluations for the presence of compounds of different chemicalgroups like carbohydrates, glycosides, saponins, tannins & phenolic compounds, proteins & freeamino acids, flavonoids and gums and mucilage using specific reagents. TLC was performed forthe methanolic extracts of Portulaca oleracea L. using different solvent systems developed for it.Portulaca oleracea L., methanolic extract, Toulene: Ethylacetate : Diethylamine (7:2:1) weredeveloped using Dragendroff’s Reagent. HPTLC fingerprinting of Portulaca oleracea L.methanolic extract were also performed.KEYWORDS: Portulaca oleracea L, HPTLC, Microscopic evaluation.

INTRODUCTION

The major population of the country particularly in rural/tribal areas relies heavily on the use of

herbal medicines for treatment of various diseases. The trust of herbal drug research may be due

to different factors; one may be that no major leads are coming up from research on synthetic

therapeutic agents and antibiotics. Secondly, many of the antibiotic and other synthetic drugs

have shown sensitization reactions and other undesirable side effects and there is a feeling that

herbal drugs are comparatively safer. The direct utilization of plant material as a drug is a feature

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of traditional medicine not only in India but also in the World. Preparations of decoction,

tinctures and total extracts of plants are the part of many pharmacopoeias of the world. Phyto-

pharmaceuticals of proven efficacy which are produced in India are morphine, codeine, thebaine,

papaverine, quinine, quinindine, digoxin, emetin, caffeine, hyoscine, hyoscamine, colchicines,

rutin, berberine, nicotine, strychnine, reserpine, ergot alkaloids, senna glycosides, podophllotoxin

resins, atropine etc.

Portulaca oleracea L. (Common Purslane, also known as Verdolaga, Pigweed, Little Hogweed

or Pusley), is an annual succulent in the family Portulacaceae, which can reach 40 cm in height. .

It has smooth, reddish, mostly prostrate stems and alternate leaves clustered at stem joints and

ends. The yellow flowers have five regular parts and are up to 6 mm wide. It is called Loni or

Ghol in Guajarati, Chhota Lunia in Hindi & Bengali. Portulaca oleracea L. is used as a folk

medicine for the treatment of inflammation in India. It was found that the plant possesses nor-

adrenaline, dopamine, dopa, vitamin C, olearacins – I and II, omega 3 fatty acids, saponins,

tannins, flavonoids, saccharides, triterpenoids, α -tocopherol and glutathione, alkaloids.

The quality and efficiency of the herbal drugs need to be established through systematic

Pharmacognostic, phytochemical and pharmacological evaluation of the drug. In herbal research,

it is also essential to authenticate the plant to establish phytochemical standardization with the

help of reliable instrument like HPTLC and HPLC.

Review of literature:


or Pusley), is an annual succulent in the family Portulacaceae, which can reach 40 cm in height. .

It has smooth, reddish, mostly prostrate stems and alternate leaves clustered at stem joints and

ends. The yellow flowers have five regular parts and are up to 6 mm wide. It is called Loni or

Ghol in Guajarati, Chhota Lunia in Hindi & Bengali. Portulaca oleracea L. is used as a folk

medicine for the treatment of inflammation in India. It was found that the plant possesses nor-

adrenaline, dopamine, dopa, vitamin C, olearacins – I and II, omega 3 fatty acids, saponins,

tannins, flavonoids, saccharides, triterpenoids, α -tocopherol and glutathione, alkaloids (1).

Efforts have been made in last three decades for the development of technical know-how for

extraction of phytochemical on commercial scale by pharmaceutical and chemical industries and

research institutions throughout the world. As a result of modern isolation techniques and

pharmacological testing procedures, new plant drugs usually find their road into the medicines as

purified substances. (2)



As natural products are an integral part of human health care system at present, there is now

popular concern over toxicity and side effects of modern drugs. There is also realization that

natural medicines are safer. Due to these factors over past 10 years, a considerable revival of

interest in the use of herbal medicine in the world has come up.

In present study, the plant Portulaca oleracea L. which is most commonly used plant in world

wide as medicine, also easily available everywhere. The selected plant was screened for it’s all

the connected disciplines like Pharmacognosy and Phytochemistry i.e. microscopy, preliminary

phytochemical screening, TLC fingerprinting, physico-chemical studies, HPLC screening,

HPTLC screening of methanolic extract of Portulaca oleracea Linn.

The whole plant is considered antiphlogistic (takes the heat out) and bactericide in bacillary

dysentery, diarrhoea, haemorrhoids, enterorrhaghia. It has been used in prescriptions as an

antidiabetic. Externally it is used as a cataplasm of fresh leaves for maturing of abscesses. The

whole plant is said to be anaphrodisiac (opposite to aphrodisiac), emollient, calmative, diuretic, a

refreshing agent (3). In Siberia, the herb is used as a gastric sedative. The herb bruised and

applied to the forehead and temple, is said to allay excessive heat and pain, and applied to the

eyes, to remove inflammation (4). Except for the roots, the entire plant is used as an antibacterial,

anti-inflammatory and anthelmintic. It is used in treating bacillary dysentery and dysuria, in a

dose of 250g of fresh plant in the form of a decoction. A combination with equal parts of

Euphorbia thymifolia is also used. The juice extracted from 100g of pounded fresh plant and

diluted with water serves as an anthelmintic against oxyuriasis and ascariasis. It is administered

in the morning for 3-5 days. Poultices of fresh leaves are used to treat mastitis, boils and

impetigo (5) Herb is chiefly valued as a refrigerant and alterative pot herb, particularly useful as

an article of diet in scurvy and liver diseases (6). The native doctors use the plant in

inflammations of the stomach. Bruised and applied to the temples it allays heat, and such pains

as occasion want of rest and sleep. It acts as a refrigerant and alterative in scurvy and liver

diseases (7).

The tasty leaves and stems are stripped from the stalks and used to make nutritious brown-bread

sandwiches; also pickle is prepared from the thick stems. The use of this plant as a vegetable,

spice and medicinal plant has been known since the times of the ancient Egyptians and was

popular in England during the middle Ages (8). The leaf juice is used in spitting of blood. The

fresh leaves bruised are applied to the temples to allay excessive heat and pain; and are also used

as a cooling external application in erysipelas and an infusion of them is given as a diuretic. Sour

leaves are used as a vegetable (6). The bruised fresh leaves are prescribed by Tamil practitioners



as an external application in erysipelas; an infusion of them is also ordered as a diuretic in

dysuria (9). Young stems and leaves are cooked like spinach, with salt and chillies, and are also

used in curries. Juice of the stems may be beneficial in cases of prickly heat and also soothing to

hands and feet whenever a burning sensation is felt.

Plant and seeds are used in diseases of the kidney and bladder, as strangury, dysuria, haematuria,

gonorrhoea etc., and also for diseases of the lungs. The plant is also used in haematemesis,

haemoptysis, etc. Externally it is used as an application to burns, scalds, and various forms of

skin diseases. The seeds are described as demulcent, slightly astringent and diuretic; while the

leaves are described as astringent, refrigerant diuretic and emollient. They are beneficial to the

intestinal mucous membrane and therefore relieve tormina, tenesmus and other distressing

symptoms in dysentery and mucous diarrhoea, particularly when combined with other drugs of

similar nature (6). The seeds are said to be used as a vermifuge, and to be useful in dyspnoea

(7).

Fig. 1 : Portulaca oleracea L

In present study, the plant Portulaca oleracea L. which is most commonly used plant in world

wide as medicine, also easily available everywhere. The selected plant was screened for the

connected disciplines like Pharmacognosy and Phyto-chemistry i.e. microscopy, preliminary

phytochemical screening, TLC fingerprinting, physico-chemical studies, HPLC screening,

HPTLC screening of methanolic extract of Portulaca oleracea Linn. These parameters may help

to identify the correct identification of Portulaca oleracea L. plant which have great importance

in traditional medicine.



MATERIAL AND METHODS

Pharmacognostic Studies

Identification and collection of the plants

Fresh plant specimens of Portulaca oleracea L. for the proposed study were collected from

Junagadh District (Gujarat, India). The authenticity of the freshly collected plant was carried out

by sending sample for identification and confirmation by preparing herbarium to National

Institute of Science Communication and Information resource (NISCAIR) and vide its later No

NISCAIR/RHMD/Consult/2011-12/1769/69 dated 07/07/2011 plant was confirmed as Portulaca

oleracea L. and sample was safely stored at Department of Pharmaceutical Science, Saurashtra

University, Rajkot as Sample No DMJ/DPS/SU/Herb/265.

Macroscopic Observations

The drug was subjected to macroscopic studies which comprised of study of organoleptic

characters of the drugs viz., colour, odour appearance, taste, smell and fracture.

Microscopic Studies

Stem

Free hand transverse sections of Portulaca oleracea L. stem were taken separately and cleared

with potassium hydroxide. The sections were treated with phloroglucinol and a drop of

concentrated hydrochloric acid to stain the lignified elements. The sections were also stained

with dilute solution of Iodine to study the starch grains.

Leaf

Free hand transverse sections of Portulaca oleracea L. Leaf were taken separately and cleared

with potassium hydroxide. The sections were treated with phloroglucinol and a drop of

concentrated hydrochloric acid to stain the lignified elements.

Powder Studies of Portulaca oleracea L.

Portulaca oleracea L. whole plant was shade dried for 8 to 10 days to make it moisture free and

by using electric grinder 60# powder was prepared. Powder study is an integrated study of

pharmacognosy. All the herbal origin medicines are studied by this method. The microscopic

examination of powder is used to study the adulteration in herbal drugs. As per the traditional

method, different types of stains are used and qualitative studies are done which reveal the

presence or absence of particular herb. Such crude methods are not enough to standardize the

presence or absence of any herb. Objective of present study is to quantify the microscopic data of

powder studies of plant Portulaca oleracea L. commonly known as luni in local language

respectively. For microscopical examination a slide of powdered drug of Portulaca oleracea L.



was prepared using stains like iodine, methylene blue and saffranine was prepared as per the

standard methods to stain the powder.

Sample preparation

Standard methods as per description in pharmacopeias were used to prepare the slides

Microscopy

Microscopy was done on microscope of Olympus, attached with digital camera. Five different

slides of each stain were prepared and total 25 snaps were clicked to get detail for quantification.

Evaluation of Physical Parameters:

The physical parameter were measured by using entire herb of Portulaca oleracea L. and was

shade dried for 8 to 10 days to make it moisture free and powder was prepared by using electric

grinder. The following physical parameters were measured as per stander procedure listed in The

Ayurvedic Pharmacopoeia of India, 2001.

Moisture Content

10 grams of accurately weighed drug powder was heated at 105ºC in an oven to constant weight

(10). Weight loss after drying gave the moisture content of the drug.

Determination of Foreign matter

100 gm. of the drug sample to be examined was weighed accurately and spread out in thin layer.

Foreign matter was detected by inspection with the unaided eye or by the use of lens (6x). The

foreign matter was separated and weighed and percentage of the foreign matter was calculated

(10).

Determination of Ash values

Determination of Total Ash

About 2 gm. of the powdered material was accurately weighed in a silica crucible, which was

previously ignited and weighed. The powdered plant was spread as a fine even layer at the

bottom of the crucible. The crucible was incinerated at a temperature not exceeding 450˚C until

free from carbon. The crucible was cooled and weighed and the procedure was repeated to get

the constant weight. The percentage of the total ash was calculated with reference to the air-dried

sample (10). The total ash usually consists of carbonates, phosphates, silicates and silica which

include both physiological ash derived from plant tissue itself and non – physiological ash which

are the residue of adhering materials such sand etc.

Determination of Acid-Insoluble Ash

The ash obtained as described in the total ash was boiled with 25 ml. of 2M hydrochloric acid for

five minutes. The insoluble ash was collected on an ash less filter paper and washed with hot



water. The insoluble ash was transferred into pre-weighed silica crucible, ignited, cooled and

weighed and the procedure was repeated to get the constant weight. The percentage of acid

insoluble ash was determined with reference to the air-dried drug.

Determination of Water Soluble Ash

The ash obtained as described in the total ash was boiled in 25 ml. of chloroform water for five

minutes. The insoluble matter was collected in a Gooch crucible or ash less filter paper and

washed with hot water. The insoluble ash was transferred into pre-weighed silica crucible,

ignited for 15 minutes at a temperature not exceeding 450ºC, cooled and weighed and the

procedure was repeated to get the constant weight. The weight of the insoluble matter was

subtracted from the weight of the total ash. The difference of weight was considered as the

water-soluble ash. The percentage of water-soluble ash was determined with reference to the air-

dried drug (10).

Determination of Extractive Values

Determination of Ethanol-soluble extractive

5 gm. of macerated and air dried coarse powder of drug was mixed with 100 ml of 95% ethyl

alcohol in a closed flask and kept for 24 hours, shaking frequently during the first 6 hours and

then allowed to stand for 18 hours. Thereafter, it was filtered rapidly taking precautions against

loss of the solvent. 25 ml. of the filtrate was evaporated to dryness in a tarred flat-bottomed

shallow dish, dried at 105ºC and weighed. Calculated the percentage of ethanol soluble

extractive with reference to the air-dried drug in %w/w (10).

Determination of Water-soluble extractive

5 gm. of macerated and air dried coarse powder of sample was mixed with 100 ml of chloroform

water (0.25% chloroform in water) and kept in a closed flask for 24 hours, shaking frequently

during the first 6 hours and then allowed to stand for 18 hours. Thereafter, it was filtered rapidly,

taking precautions against loss of the solvent. 25 ml of the filtrate was evaporated to dryness in a

tarred flat-bottomed shallow dish, dried at 105ºC and weighed. Calculated the percentage of

water-soluble extractive with reference to the air-dried drug (10).

Phytochemical Study

Extraction

Air dried & coarsely powdered (50 gm) of Portulaca oleracea L. herb was taken. Extraction was

carried out in a soxhlet extractor using methanol. The extracts were then concentrated to dryness

under reduced pressure and controlled temperature, respectively and they were preserved in a

refrigerator (11, 12). Portulaca oleracea L. was taken in different soxhlet apparatus. The soxhlet



extraction process was carried out until the solvent found to be colorless. Then the solvent was

filtered and distilled off. Final traces of methanol were removed under pressure by using rotary

vacuum flask evaporator.

Preliminary Phytochemical Screening

Plant extracts obtained by soxhlet extraction procedure were subjected to preliminary

phytochemical analysis in order to identify the nature of chemical constituents. The detailed

phytochemical screenings were carried out by the standard procedure (13; 14) for the testing of

presecnce of Alkaloids, Carbohydres, Glycosides, Steroids, Test for Fixed Oils and Fats,

Saponins, Tannins and Phenolic compounds, Proteins and Free Amino acids, Flavanoids and

Gums and mucilages by using various.

Determination of Total Phenolic content

The total phenolic content in the methanolic extract of Portulaca oleracea L. was determined as

per the reported method.

Preparation of test sample

Stock solutions of samples were prepared by dissolving 10 mg of dried methanolic

extract in 10 ml of methanol.

Preparation of reagent used

Folin ciocalteu reagent, 20% sodium carbonate solution: 20gms of anhydrous sodium

carbonate was dissolved in 100ml of distilled water

Protocol for total phenolic content

A method described by Singleton and Rossi was used. In this assay 500 µl of the sample was

taken in 25 ml volumetric flask. To this, 10 ml of water and 1.5 ml of Folin ciocalteu reagent

were added. The above mixture was kept for 5 min. and then 4 ml of 20 sodium carbonate

solution was added and the volume was made up to 25 ml with distilled water. The mixture was

kept for 30 min. and the absorbance of the blue color developed was recorded at 765nm, in UV-

visible spectrophotometer. The total Phenolic content was calculated from calibration curve of

Gallic acid plotted by using the above procedure. The total Phenolic content was expressed in

terms of µg/mg of dry extract equivalent to Gallic acid. The entire assay was performed in

triplicate.

Thin Layer Chromatography:

Preparation of various Extract

Thin layer chromatography is an important analytical tool in the separation, identification and

estimation of different components which are separated by the differential migration of solute



between two phases – a stationary phase and a mobile phase. Here the principles of separation

are adsorption and the stationary phase acts as an adsorbent. Depending on the particular type of

stationary phase, its preparation and use with different solvent can be achieved on the basis of

partition or a combination of partition and adsorption. The methanolic extracts of Portulaca

Oleracea L. was prepared as per the procedure given in chapter 4.2.1; showed good resolution in

solvent system by trial and error method.

TLC Studies

The TLC plate prepared with silica gel-G (activated) was the stationary phase having a thickness

of about 0.5mm. 20μl each of test solution was applied on silica gel-G plate (5x15 cm.). The

TLC plate was developed in the saturated chromatographic chamber containing the following

solvent systems: Toluene : Ethylacetate : Diethylamine (7:2:1)

HPTLC studies

Commercially available precoated aluminium sheets silica gel G60 F 254 [E. Merck], 10 x 10

plate size, were used for this study. Methanolic plant extract of Portulaca oleracea L. herb was

used for analysis

Chromatogram Development

The plates were developed in Camag Twin Trough Chamber using the solvent system n-butanol:

acetic acid: water (4:3:1) for the methanolic extracts. After developing the plates were air dried

and observed under UV chamber (Camag TLC Scanner)

Densitometric Scanning

The developed plates were scanned using densitometric at 254 nm, 366 nm. [range 200 – 400

nm.]. The development chromatograms of these extracts are shown in the HPTLC

Chromatogram.

Chromatographic Development System for HPTLC

Plate Aluchrosep silica Gel 60 F 254

Thickness 0.2mm

Plate size 10 x10 cm

Sample application 4,6,8,10,12,14µl

Solvent system n-butanol: acetic acid: water (4:3:1)

Activation Temperature 110o C for 10 min

Detecting UV chamber at 254nm

Instrument CAMAG LINOMET –V AND CAMAG TLC SCANNER



HPLC Finger printing Profile:

CHROMATOGRAPHIC CONDITIONS

Chromatographic mode : Reverse Phase

Mobile delivery mode : Gradient mode

Tray Temperature : 5° C

Injection Volume : 10 µl

Column : C18

Mobile Phase : Toluene: acetic acid (4:1)

Column Temperature : 40° C

Flow Rate : 0.5 ml/min

Back pressure : 55 bar (Approximately)

Retention Times (in min)

Methanolic extract of Portulaca oleracea L. 4.086 min,5.256min, 6.127 min, 7.108 min, 7.723

min, 8.253 min, 9.991min,11.153 min, 12.273min, 17.681 min. Run Time

RESULT AND DISCUSSION

Pharmacognostic Studies

Macroscopical study of Portulaca oleracea L.


or Pusley), is an annual succulent in the family Portulacaceae, which can reach 40 cm in height..

It is a native of India and the Middle East, but is naturalised elsewhere and in some regions is

considered an invasive weed.

Leaves :

Alternate, sessile or very sort petiolate glabrous, oblanceolate to obovate, obtuse to truncate at

apex, tapering slightly to base, entire, glabrous, succulent, to 3cm. long, 1.2cm. broad.

Stem :

From stout taperoot, multiple from base, prostate greenish with some red tinge, branching

succulent, herbaceous.

Flower :

Petals 5, yellow glabrous, 3-4mm. long, 2.5-3mm broad, distinct. Stamen 6-10. Filsment 1 mm

long yellow translucent glabrous. Anthers yellow 2-3mm. broad. Style 5 lobed, 1.1mm. long

glabrous.Calyx tube 2mm. long, glabrous, green, 2-lobed with transverse groove. Lobes subequal



to unequal, to 4mm. long, glabrous acute. Capsule circumissle to +5mm long. Seeds many.

Plancentation free central.

Microscopical Study Transverse section of Portulaca oleracea L. stem

Transverse section of the stem shows (Fig. 5.1 to 5.3) single cuticularised epidermis and wide

zone of thick and irregular walled parenchymatous cells (cortex region). Pericycle layer is

present. It also shows 3-4 layers of phloem. Xylem is present. Vascular bundle is endarch. In the

pith region, few calcium oxalate rosette crystals were seen.

Fig. 2 : T.S. of Portulaca oleracea L. stem

(Epidermal cells)Fig. 3 : T.S. of Portulaca oleracea L. stem

(Xylem vessel & Phloem)

1: Epidermis ; 2: Rosette of Calcium Oxalates ; 3: Pith ; 4: Xylem Vessel 5: Phloem

Fig. 4: T.S. of Portulaca oleracea L. stem



Transverse section of Portulaca oleracea L. leaf

Fig. 5 : T.S. of Portulaca oleracea L. leaf

(Upper Epidermis, Vascular bundles and

Collenchyma)

Fig. 6 : T.S. of Portulaca oleracea L.

leaf(Collenchyma and Lower Epidermis)


(Vascular bundle and Palliside cells)


(Xylem vessels)

TS passing through midrib show deep groove on the upper side and broad ridges and furrow on

lower side, lamina narrow gets sometimes curved at its extremities, single layered upper

epidermis followed by single layered palisade cells. The mesophyll cells are parenchymatous,

vascular bundle bicollateral, lying in the midrib region are bigger, lower portion of the midrib is

occupied by collenchymatous cells.



Powder characteristics of Portulaca oleracea L. herb

Fig. 9 : Trichomes Fig. 10 : Xylem Fibre

Fig. 11 : Sclerenchymatous cells Fig. 12 : Stone Cells

Fig. 13 : Xylem vessels Fig. 14 : Stone Cell & Scelerenchyma

Powder characteristics of Portulaca oleracea L. herb (Fig. 9 to 14)



Quantitative standards for Ipomoea reniformis and Portulaca oleracea

The herbs were preliminary evaluated by determining the several physical constants like foreign

matter, ash value, extractive values and moisture content. All this data are presented in Table No.

1.

Physico-Chemical Standards Portulaca oleracea

A Moisture Content 8.90 %B Foreign matter 1.38 %

C Ash Value % w/w

1. Total Ash 8.282. Water Soluble Ash 2.853. Acid Insoluble Ash 2.56D Extractive Value % w/w1. Alcohol Soluble Extractive 4.942. Water Soluble Extractive 6.26

Table No. 1: Physico-Chemical Standards

Phytochemical Study

Preliminary Phytochemical Analysis

The powder of Portulaca oleracea L.was extracted with methanol as procedure given in the

previous chapter. Extract was subjected to preliminary phytochemical evaluation for the presence

of compound of different chemical groups as shown in Table No. 2.

Phytochemical Analysis Portulaca oleracea

Extractive value of methanolicextracts

7.14% w/w

Alkaloids +

Carbohydrates +

Glycosides +

Steroids --

Furanoids --

Fixed oils & fats --Saponins +

Tannins & Phenolic compounds +

Proteins & Free amino acids +

Flavonoids +

Gums & Mucilage ++ = indicates presence of that chemical constituent group

- = indicates absence of that chemical constituent group

Table No. 2 : Preliminary Phytochemical Analysis of extracts



Thin Layer chromatography

In the present experiment, different solvent systems were tried to resolve the components of

methanolic extract of Portulaca oleracea L. TLC plates for each extract for each solvent system

were developed, for visualization with dragendroff’s reagent and fluorescent wavelength (UV

366nm.).

TLC of methanolic extracts of Portulaca oleracea L. was carried out in different solvent system

as shown in Table No. 3 Photographs of TLC plates are shown in figure no. 15

Mobile Phase for Methanolic

extract (Portulaca oleracea)

Spraying Reagent Rf

ValueSpot Colour

Toulene:Ethylacetate:

Diethylamine (7:2:1)

Dragendroff’s

Reagent

0.65 Pink

0.73 Purple

Table No. 3: TLC Solvent system

Fig. 15 : Toulene:Ethylacetate:Diethyla-

mine (7:2:1) Methanolic extract

(Portulaca oleracea) Dragendroff’s

Reagent



HPTLC Finger Printing

HPTLC is now-a-days applied to obtain “Finger Print” patterns of herbal formulations,

quantification of active ingredients and also detection of adulteration. The HPTLC

chromatograms were developed for methanolic plant extracts using the same solvent system

utilized for TLC.

Plate : Aluminium plate precoated with Silica Gel GF254

Thickness : 0.2 mm

Plate size : 10 X 10 cm.

Sample Application : 10 µl, 20 µl, 30 µl,

Solvent System : For Portulaca oleracea L. methanolic extract

Toulene: Ethylacetate: Diethylamine(7:2:1)

Detection :For Portulaca oleracea L. methanolic extract: Detection at

366 nm. (Fig. 5.32)

Instrument : CAMAG TLC Scanner 3 and LINOMAT-V

Fig. 16a HPTLC plate at 254nm

UV light

Fig.16b HPTLC plate at

Visible light

Fig. 16c HPTLC plate at

366nm UV light

Fig. 16 HPTLC Finger Printing of Methanolic Extract of Portulaca Oleracea L.



Fig. 17 : Chromatogram of HPTLC fingerprint profile for Methanolic extract of Portulaca

oleracea



HPLC:

HPLC is now-a-days applied to obtain “Finger Print” patterns of herbal formulations,

quantification of active ingredients and also detection of adulteration. The HPLC chromatograms

were developed for methanolic plant extracts using the solvent system Toluene: acetic acid (4:1).

Fig. 18 : Chromatogram of HPTLC fingerprint profile for Methanolic extract of Portulaca

oleracea L.

Peak Time Conc Area Height PeakStart

PeakEnd

Area%

1 4.086 0.76833 26412 2076 3.808 4.384 0.76832 5.256 31.4632 1081553 37513 4.427 5.707 31.46323 6.127 60.33966 2074186 109422 5.707 6.88 60.33974 7.108 4.82366 165814 6646 6.88 7.659 4.82375 7.723 0.28157 9679 805 7.659 7.883 0.28166 8.253 0.77086 26498 857 7.883 8.704 0.77097 9.991 0.40193 13816 732 9.781 10.517 0.40198 11.153 0.66924 23005 825 10.709 11.669 0.66929 12.273 0.25943 8918 442 11.947 12.619 0.259410 17.681 0.22211 7635 279 17.259 18.133 0.2221

Table No.4 HPTLC HPTLC fingerprint profile for Methanolic extract of Portulaca oleracea L.

0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0 27.5 min

-10

0

10

20

30

40

50

60

70

80

90

100

110

mAU254nm,4nm (1.00)

1

2

3

45 6 7 8 9 10



CONCLUSION

The present study on pharmacognostic investigation of Portulaca oleracea L. herb might be

useful to supplement information in regard to its identification parameters assumed significantly

in the way of acceptability of herbal drugs in present scenario of lack of regulatory laws to

control quality of herbal drugs. Portulaca oleracea L. was preliminary evaluated by determining

the several physical constants like foreign matter, ash value, extractive values and moisture

content. Portulaca oleracea L. methanolic extracts was subjected to preliminary phytochemical

evaluation for the presence of compound like carbohydrates, glycosides, saponins, tannins and

phenolic compounds, proteins and free amino acids, flavonoids and gums and mucilage as well

as TLC, HPTLC and HPLC profile of the methanolic extract of Portulaca oleracea L was

developed and these result of research may serve as important tools for the identification of the

plant.

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