world journal of pharmaceutical researchdosha pitta in the body. in spite of their bland taste,...

www.wjpr.net Vol 6, Issue 5, 2017. 1357

Raja et al. World Journal of Pharmaceutical Research

PHARMACOGNOSTICAL PHYTOCHEMICAL AND LARVICIDAL

ACTIVITY OF ETHANOLIC LEAF EXTRACT OF LUFFA

ACUTANGULA (CUCURBITACEAE)

*R. Ramasubramania Raja1, P.Vijayasanthi

1, G.Mydhili

1, M.Aswini

1, S.Seshadri

1,

V.M.Rajesh1 and M. Sreenivasulu

2

1Departement of Pharmacognosy, Narayana Pharmacy College, Nellore, A.P.

2Principal, Narayana Pharmacy College, Nellore, A.P.

ABSTRACT

Luffa acutangula belongs to the family of cucurbitaceae. The

macroscopical character of the luffa acutangula was colour, odour,

size, shape; taste was performed for the evaluation of morphological

identification character. The microscopical evaluation of luffa

acutangula leaf was performed by using transverse section technique.

The pharmacognostical like extractive value, loss on drying, crude

fibre content was performed. Leaf powder the active constituents by

using colour reaction test. Extraction was performed soxhelet

apparatus by using the solvent ethanol. The ethanolic extract was

analysed by using colour reactions based chemical tests. Thin layer

chromatography used to identify the phytoconstituents. Drug powder with different chemical

reagents used to perform the fluorescence tests by using visible light and UV at 366nm. The

larvicidal activity was performed by using ethanolic extract of luffa acutangula.6 hours, 12

hours, 24 hours, 48 hours, 72 hours were noted the values. In different concentration of

ethanolic extract of luffa acutangula like 10 mg/ml, 20 mg/ml, 40 mg/ml, 80 mg/ml and 160

mg/ml used to analyse the larvicidal activity. Dose depending manner the activity was

responsed. Finally, we concluded the ethanolic extract of luffa acutangula have potential

larvicidal activity in 80 mg/ml and 160 mg/ml.

KEYWORDS: Luffa acutangula, alkaloids, saponins, carbohydrates, larvicidal.

World Journal of Pharmaceutical Research SJIF Impact Factor 7.523

Volume 6, Issue 5, 1357-1378. Research Article ISSN 2277– 7105

*Corresponding Author

R. Ramasubramania Raja

Departement of

Pharmacognosy, Narayana

Pharmacy College, Nellore,

A.P.

Article Received on

19 March 2017,

Revised on 09 April 2017,

Accepted on 29 April 2017

DOI: 10.20959/wjpr20175-8444



INTRODUCTION

Scientific classification

Kingdom : Plantae

Division : Magnoliophyta

Class : Magnoliopsida

Order : Cucurbitales

Family : Cucurbitace

Sub Family : Cucurbitoideae

Tribe : Benincaseae

Sub tribe : Luffinae

Genus : Luffa

Species : acutangula

Biological source : Luffa acutangula( angled Luffa,ridgedLuffa,vegetable gourd)

Synonyms : poppyaNeck.exM.Roem.

Vernacular names

Sanskrit : Gantali, kosataki, ksweda, sutikta.

Bengali : Zinga, Titotorai, Titojhinga, Ghoshalata.

English : Ribbed gourd, silk gourd, Ridge gourd, Angled loofah, Chinese okra, Sinkwa

towel sponge, vegetable sponge.

Gujarat : Turiya, Kadawa.

Hindi : Turai, satputia, Jhimani.

Kannada : Hire- Valli, Kahire, Naagadaaliballi.

Malayalam :PeerKamkai, Athanga.

Marathi : DodkaTuriya, Divali, Kadudodaki, kadushirali, kaduturai, Ranturai.

Punjab : Turiya, Jhinga, Shirola.

Tamil : Peerkku, karniti, Kacappi, Itukari, karnityikkoti, peerkangai.

Telugu : Beera

Urdu : Turai.

Assam : Zika.

Kerala : Peechinga.

Maharashtra : Dodka

Japan : Hechima.

Philippines : Patola.



China : Sigua, Oyong.

Indonesia : Sigua, Oyong.

Korea : Susemi.

Manipur : Sebot.

Growth and distribution

The Luffa acutangula Linn. Var. Amara Roxb is a large monoeious annual climber. It is

indigenous to western, Central and Southern regions of India, and regarded as Wild variety of

cultivated species. It resembles to Luffa acutangula in every ascepts, except that it has

smaller leaves, flowers, fruits, and seeds.

Distribution : Luffa acutangula is pantropical and cultivated throughout India.

Habit : Herb.

Propagation: propagation of Luffa acutangula by seeds.

Native range : India and naturalised tropic and Sub tropics.

Cultivation : Luffa acutangula can grow in all type of soils and can be grown in summer

(or) rainy Season.

Seeds can accordingly be down either in February- march (or) june- July.

MORPHOLOGY

Leaf : Petiole is brownish yellow coloured, 3-8cm in length; somewhat twisted, wrinkled

and angular. While lamina having Pale (or) light- green colour, 6-9cm long, crumpled and

broad.

Uses of Luffa acutangula

1. Luffa acutangula has diuretic properties

2. It is used as an expectorant and hypoglycemic.

3. It is used as a bitter tonic.

4. It is used for to reduce hyper acidity.

5. The leaves of Luffa acutangula are useful in the treatment of dysentery conditions.

6. The leaves (or) juice of Luffa acutangula are used as dressing in the diseases such as

Inflammation of spleen, Ring worms, Piles, Leprosy.

7. Pounded leaves mixed with garlic are applied locally for relief in Leprosy.

8. The roots of Luffa acutangula added to milk (or) water is helpful in the removal of kidney

stones.



9. Roots of Luffa acutangula are added to cooled water. Luffa acutangula boiled in hot

water and applied on skin is helpful in the swelling of the lymph glands.

10. Oil is extracted from the seeds of Luffa acutangula which is used in the treatment of skin

diseases.

11. Luffa acutangula is also an effective home remedy for the prevention of premature

greying of hair.

12. Luffa acutangula is chopped in small pieces along with the ribbed skin and completely

dried in the Sun. Once, the Luffa acutangula is fully dry, it is made into a powder and

used to prevent the premature greying of hair.

Luffa acutangula in ayurveda

Ayurveda has attributed luffa acutangla with a number of health benefits which current

clinical research is supporting as well. Theluffa acutangula are rich in mineral and are

very alkaline for the body and hence they have acooling effect on the body.

From ayurveda point of view, increases vata and kapha, but it cools down and pacifies the

dosha pitta in the body. In spite of their bland taste, luffa acutangula have many health

benefits;

Nutrition: luffa acutangula hasmany nutritional benefits as it loaded with fibers, vitamins

and minerals. it also has low calories and fats which make them an integral part a healthy

diet.

Luffa Acutangula and weigh loss: Since the luffa acutangula is low in saturated fat and

cholesterol. it is the ideal diet for those who are looking for weigh loss. luffa acutangula

has a high water content which makes it a food with very less calories. Luffa acutangula

and treatment of jaundice; the luffa acuangula juice is very good natural remedy for the

treatment of jaundice. The juice which is prepared by pounding the luffa acutangula or

the powder which made from the bitter luffa acutangula seeds and crust is helpful in the

cure of jaundice. The dried fruits are powdered and used as snuff in the treatment of

jaundice.

Luffa acutangula and blood purification: Ithelps in the purification, restoration and

nourishment of the liver and is also helpful in the liver detoxification resulting from

alcohol intoxication.



Luffa acutangula and hypoglycaemia: luffa acutangula has certain peptides which are

exactly like insulin and cherantin chemicals which help in reducing and urine sugar

levels.

Luffa acutangula in constipation: The cellulose fibres present in the luffa acutangula are

help full in the treatment of constipation and also effect in the treatment of piles.

Luffa acutangulaand skin care: Luffa acutangula allowed to dry and mature on the

vine, it can be harvested has a sponge. Thisloofah sponge has been used traditionally

exfoliating product while bathing. They are considered helpful in removing dead cells

from the skin thus making the skin smooth and conditioned. The blood purifying

properties the luffa acutangula are helpful against pimples and acne problems. Loofah

sponge is also effective in fighting off foot and body odor.

Luffa acutangula as immune system booster: Juice of luffa acutangula mixed with

other healthy vegetables taken daily helps in strengthening of immune system and helps

the body fighting off infections effectively.

Luffa acutangula for eyes: The high β carotin content of Luffa acutangula iut in eyes to

treat granular conjunctivitis and styes.

Luffa acutangula is also useful as emetic, expectorant and demulcent.

Luffa acutangulaayurvedic remedies

1. For jaundice: One cup Luffa acutangula juice mixed with 2 spoons of sugar, if taken

twicedaily cures jaundice.

2. For bleeding from wound: The Pulp of the Luffa acutangula is ground and applied on the

wound. This will stop bleeding from the wounds.

3. For stomach worms: 1 Luffa acutangula boiled in two glasses of water and then adequate

Salt is added to it. If this mixture is killed twice daily, stomach worms will be killed.

4. Forasthma: Grind the Luffa acutangula to extract ½ cup Luffa acutangula juice mixed

with sugar and taken twice a day will cure cancer.

MATERIAL AND METHODS

PLANT MATERIAL

The crude drug was collected from podalakuru road (NELLORE). The month of november

2016, intially the plant was identified by local people and then authorisied by Dr. Bhaskar,



Botanist VR college, Nellore. The specimen of plant material was kept in the college for

further work.

chemicals

All the chemicals procured from rankem, aventor Performance material India limited,

Harayana.

Preparation of exract

The plant leaf was dried under shadow (5 days) and then powdered by using grinder and then

sieved. The powdered material of the plant extracted by using the solvent of ethanol. 15 gm

of powder was taken in soxhelet apparatus and then extracted by using ethanol for the

duration of 6hrs. Finally, the ethanolic crude extract was found to be 4 gm.

Pharmacognostical studies

The pharmacognostical studies on extractive value, moisture content, crude fibre content, loss

on drying, foreign organic matter was performed and reported on table.

Microscopical evaluation of leaf

The leaf was examined the microscopical characterised by using transverse section technique

by using the staining reagent like safranin, bromothymol blue and iodine solution. Observe

the characters reported on result.

Powder microscopical examination by using eyepiece and stagemicrometer:

The leaf powder length and width was determined by using the stage and eyepiece

micrometer.

Procedure

1. Using stage micrometre, calibrate the eyepiece micrometer. Calculate the factor (average

distance between two lines in microns).

2. Take a little quantity of powdered drug (Luffa acutangula leaf) in a test tube and boil with

clearing agent, chloral hydrate solution.

3. Transfer cleared powder in a watch glass.

4. Stain the lignified trichome with the staining reagent (phloroglucinol and concentrated

hydrochloric acid).



5. Mount this treated powder in glycerine water and observe the slide under low power.

(Powder should be thinly, uniformly scattered, without overlapping of particles).

6. Focus a stained trichome (intact trichome). By rotating the scale of eyepiece micrometer,

note the numbers of divisions of the eyepiece micrometer covered by the width of the

same trichome.

7. Again rotate the eyepiece micrometer without disturbing the slide and find the numbers of

divisions of the eyepiece micrometer covered by the width of the same trichome.

8. Similarly, calculate the length and width of about 25 trichome, and write the readings in

two separate columns.

9. Multiply each value by the factor calculated in the first step to get the value in microns.

Then calculate the average value and write the range for the length and width of trichome.

Extractive value

1. Determination of methanol- soluble extractives

1. Weigh about 5 g of the powdered drug in a weighing bottle and transfer it to a dry 250ml

Conical flask.

2. Fill a 100ml graduated flask to the delivery mark with the solvent (90% methanol). Wash

out the weighing bottle and pour the washings, together with the remainder of the solvent

into the conical flask.

3. Cork the flask and set aside for 24 hours, shaking frequently(Maceration).

4. Filter into a 50ml cylinder. When sufficient filtrate has collected, transfer 25 ml of the

filtrate to a weighed, thin porcelain dish, as used for the ash values determinations.

5. Evaporate to dryness on a water -bath and complete the drying in an oven at 100°C.

6. Cool in a dessicator and weigh.

7. Calculate the percentage w/W of Extractive with reference to the air- dried.

2. Determination of water- soluble extractives

1. Weigh about 5 g of the powdered drug in a weighing bottle and transfer it to a dry 250 ml.

Conical flask.

2. Fill a 100ml graduated flask to the delivery mark with the solvent (90% water). Wash out

the weighing bottle and pour the washings, together with the remainder of the solvent into

the conical flask.






5. Evaporate to dryness on a water -bath and complete the drying in an oven a100°C.



3. Determination of acetone - soluble extractives


Conical flask.

2. Fill a 100ml graduated flask to the delivery mark with the solvent (90% acetone). Wash

out the weighing bottle and pour the washings, together with the remainder of the solvent

into the conical flask.

3. Cork the flask and set aside for 24 hours, shaking frequently. (Maceration)





7. Calculate the percentage w/w of Extractive with reference to the air- dried.

4. Determination of chloroform- soluble extractives

1. Fill a 100ml graduated flask to the delivery mark with the solvent (90% chloroform).

2. Wash out the weighing bottle and pour the washings, together with the remainder of the

solvent into the conical flask.

3. Cork the flask and set aside for 24 hours, shaking frequently (Maceration).






5. Determination of pet ether- soluble extractives


Conical flask.


3. Fill a 100ml graduated flask to the delivery mark with the solvent (90% pet



4. ether). Wash out the weighing bottle and pour the washings, together with the remainder

of the solvent into the conical flask.






6. Determination of acetic acid- soluble extractives

1. Weigh about 5 g of the powdered drug in a weighing bottle and transfer it to a dry 250 ml

conical flask.

2. Fill a 100ml graduated flask to the delivery mark with the solvent (90% acetic acid).

Wash out the weighing bottle and pour the washings, together with the remainder of the

solvent into the conical flask.

3. Cork the flask and set aside for 24 hours, shaking frequently (Maceration).



5. Evaporate to dryness on a water-bath and complete the drying in an oven a100°C.



7. Determination of moisture (Loss on drying)

1. weigh about 1.5g of powered drug into a weighed flat and thin porcelain dish.

2. Dry in the oven at 1000 C and 105

0.

3. Cool in the desiccator and watch. The loss in weight is usually recorded as moisture.

Note

A very use form of dish for the determination of moisture and of ash is a thin flat

porcelaindish. If a platinum dish is available it may be used. The burning of the powder

should proceed slowly and the material must not be allowed to catch fire or to give off smoke

as dense fumes. The most common method for the determination of moisture is to heat the

drug till one gets constant weight at 1000C as is done for digitalis. However many substance

loose other volatile constituents or some of their constituents undergo change with

consequent loss of weight at a temperature of 1000C other methods are used for material of

these types.



Determination of foreign organic matter

Foreign organic matter means the material consisting of any one or all of the following

substances:

1. Material not coming from the original plant sources.

2. Insects, moles or other animal contamination.

3. Parts of the organ or organs from which the drugs is delivered other than the parts named

in the definition and description.

4. Any other organ than the those named in the definition and description.

Procedure

1. Weigh 100 to 500g of the sample (or the quantity specified in the monograph of the

drug).

2. Spread the sample on a white tile or a glass plate uniformly without overlapping.

3. Inspect the sample with naked eyes or by means of a lens (5x or above).

4. Separate the foreign organic matter (mentioned above) manually.

5. After complete separation, weigh the matter and determine % w/w present in the sample.

Determination of crude drug fiber by the dutch method

Crude fiber is the residue of resistant tissues which can be obtained giving treatment to the

vegetable powder with dilute acids and alkali.

If crude drug contains appreciable amount of fat or oil, it must be removed first by extraction

with suitable lipid solvent, before processing.

(a) Weigh 2g of powdered drug in a beaker.

(b) Add 50ml 0f 10% v/v nitric acid.

(c) Heat to boil with constant stirring (Till about 30 seconds after boiling starts).

(d) Strain through fine cotton cloth on a buchner funnel.

(e) Give washing to the residue with boiling water (suction may be used).

(f) Transfer residue from the cloth to a beaker.

(g) Add 50ml of 2.5% v/v sodium hydroxide solution.

(h) Heat to boil. Maintain at boiling point for 30 seconds, stirring constantly.

(i) For quantitative determination, transfer the residue in a cleaned and dried crucible.

(j) Weigh the residue and determine % crude fibers.

For microscopial examination, residue is suspended in water for alcohol (70%) until required



for use.

Vein islet number

1. Clear a piece of the leaf by boiling in chloral hydrate solution for about thirty minutes.

2. Arrange camera lucida and drawing board for making drawings to scale.

3. Place stage micrometer on the microscope and using 16 mm objective, draw a line

equivalent to 1 mm as seen through the microscope.

4. Construct a square on this line.

5. Move the paper so that the square is seen in the eye piece, in the centre of the field.

6. Place the slide with the cleared leaf (epidermis on the stage)

7. Trace off the veins which are included with in the square, completing the outlines of those

islets which overlap two adjecent sides of the square.

8. Count the number of vein islets in the square millimeter. Where the islets are intersected

by the sides of the square include those on two adjacent sides and exclude those islets on

the other sides. (To obtain a critical result for a leaf, 4sq. mm. Should be used, Preferably

in one large area of 4sq. mm.

9. Find the average number of vein islets from the four adjoining square to get the value for

one sq.mm.

Veinlet termination

1. Clear a piece of the leaf by boiling in chloral hydrate solution for about thirty minutes.

2. Arrange camera lucida and drawing board for making drawings to scale.

3. Place stage micrometer on the microscope and using 16 mm objective, draw a line

equivalent to 1 mm as seen through the microscope.

4. Construct a square on this line.

5. Move the paper so that the square is seen in the eye piece, in the centre of the field.

6. Place the slide with the cleared leaf (epidermis on the stage)

7. Trace off the veins which are included with in the square, completing the outlines of those

islets which overlap two adjecent sides of the square.

8. Count the number of vein islets in the square millimeter. Where the islets are intersected

by the sides of the square include those on two adjacent sides and exclude those islets on

the other sides. (To obtain a critical result for a leaf, 4sq. mm. Should be used, Preferably

in one large area of 4sq. mm.



Count the numer of veinlet termination present within the square. Find the average number of

veinlet termination number from the four adjoining square to get the value for one sq.mm.

vein termination number of luffa acutangula is between.

Stomatal number

1. Clear the piece of the leaf (middle part) by boiling with chloral hydrate solution or

alternatively with chlorinated soda. peel out upper and lower epidermis separately by

means of forceps. Keep it on slide and mount in glycerin water.

2. Arrange a camera lucida and drawing board for making drawings to scale.ι

3. Draw a square of 1mm by means of stage micrometer.

4. Place the slide with cleared leaf (epidermis) on the stage. Trace the epidermis cell and

stomata.

5. Count the number of stomata present in the area of 1 sq.mm. Include the cell if at least

half of its area lies within the square.

6. Record the result for each of the ten fileds and calculate the average number of stomata

sq.mm.

Stomatal index

Stomatal index is the percentage which the number of stomata form to the total number of

epidermal cells, each stomata being counted as one cell. Stomatal index can be calculated by

using following equation.

I = S/E+S 100

Where, I = Stomatal index

S = No. of stomata per unit area

Procedure

1. Clear the piece of the leaf (middle part) by boiling with chloral hydrate solution or

alternatively with chlorinated soda. peel out upper and lower epidermis separately by

means of forceps. Keep it on slide and mount in glycerin water.

2. Arrange a camera lucida and drawing board for making drawings to scale.

3. Draw a square of 1mm by means of stage micrometer.

4. Place the slide with cleared leaf (epidermis) on the stage. Trace the epidermis cell and

stomata.

5. Count the number of stomata, also the number of epidermal cells in each field.



6. Calculate the Stomatal index using the above formula.

7. Determine the values for upper and lower surface (epidermis) separately.

Chemical test

1. Detection of alkaloids

The small portions of solvent free chloroform, alcoholic and water extracts are stirred

separately with a few drops of dilute hydrochloric acid and filtered. The filtrate may be tested

carefully with various alkaloidal reagents; Mayer's reagent (cream precipitate), Dragendorff's

reagent (orange brown precipitate), Hager's reagent (yellow precipitate) and Wagner's reagent

(reddish-brown precipitate).

2. Detection of carbohydrates and glycosides

(a) Small quantities(200mg) of alcoholic and aqueous extracts are dissolved separately in 5ml

of distilled water and filtered. The filtrate may be subjected to Molisch's to detect the

presence of carbohydrates.

(b) Another small portion of extract is hydrolyzed with dilute hydrochloric acid for few hours

in water-bath and is subjected to Liebermann-Burchard's, legal's and borntrager's tests to

detect presence of different glycosides.

(c) A smallportions of extractnis dissolved in water and treated with fehlings's Barfoed's and

Benedict's reagents to detect presence of different sugars.

3. Detection of phytosterols

The petroleum ether, acetone and alcoholic extracts are refluxed separately with solution of

alcoholic potassium hydroxide till complete saponification takes place. The saponification

mixture is diluted with distilled water and extracted with ether. The ethereal extract is

evaporated and is evaporated and the residue (unsaponifiable matter) is subjected to

Lieberman’s and Burchard's tests.

4. Detection of fixed oils and fats

A small quantity of petroleum ether and benzene extracts is pressed separately between two

filter papers. Oil stains on the paper indicate the presence of fixed oil.

A few drops of 0.5N alcoholic potassium hydroxide is added to a small quantity of petroleum

ether or Benzene extract along with a drop of phenolphthalein. The mixture is heated on



water bath for 1-2hours. Formation of soap or partial neutralisation of alkali indicates the

presence of fixed oils and fats.

5. Detection of saponins

About 1ml of alcoholic and aqueous extract s is diluted separately with distilled water to 20

ml and shaken in a graduated cylinder for 15 minutes. One cm layer of foam indicates

presence of saponins. The test solution may be subjected to test for haemolysis.

6. Detection of phenolic compounds and tannins

Small quantities of alcoholic and aqueous extracts in water are tested for the presence of

phenolic compounds and tannins with dilute ferric chloride solution (5 percent), 1 percent

solution of gelatin containing 10 percent sodium chloride, 10%lead acetate and aqueous

bromine solutions.

Fluorescence analysis of crude powder

The fluorescence analysis was performed by crude powder with various chemical reagents

under day light and UV light and reported on the table.

Thin layer chromatography

1. Amino acids

Stationary phase : silica gel-G

Mobile phase : Butanol: acetic acid: water (4: 5: 1)

Detection : Iodine chamber

2. Glycosides


Mobile phase : chloroform: Benzene:Ethanol: Acetate(11 : 4: 2 : 1 : 2 )

Detection : Iodine chamber.

3. Saponins


Mobile phase : Chloroform: Methanol: Water (7: 4: 1)


4. Sterols




Mobile phase : Hexane: Acetone (9: 1)


5. Steroids


Mobile phase : Ethyl acetate: Acetone: pet ether (4: 2: 2)


Larvicidal bioassay

Bioassay for the larvicidal activity was carried out using WHO [11] procedure with minor

modifications. Twenty larvae, each were introduced into treatment trays containing 250 ml of

their natural growth medium (Tap water - untreated - added with dog biscuits and yeast in the

ratio 3:1). To the treatment set, respective concentrations of the plant extracts (0.5, 1.0, 2.0,

4.0 and 8.0 ml) were added from the stock solution; maintaining a relative concentration of

the plant extract as 10, 20, 40, 80 and 160 mg/ml respectively. A control was maintained,

containing only larvae and natural growth medium. Mortality counts of larvae were

monitored at regular intervals i.e. 6, 12, 24, 48, 72 and 96 Hours after Treatment. Larvae

were considered dead if they settle and remain motionless in the bottom of the test beaker

with no response to light or mechanical stimulus or not recovering life functions even after

being transferred to their growth medium.

Macroscopy

Fig: 1 Luffa acutangula plant

S.no Macroscopy Petiole Lamina

1. Colour Brownish yellow Light green

2. Odour Characteristic

4. Size 3-8 cm 6-8cm

5. Shape Angular,Twisted Curled, corrugated



Microscopy character of luffa acutangula

Fig: 2 midrib, and lamina (Transverse section of luffa acutangula leaf)

Fig: 3Mid rib (Transverse section of luffa acutangula leaf)

Fig: 4 Transverse section of luffa acutangula leaf (lamina)



Epidermal layer is surrounded by thin cuticle, upper and lower epidermis is present. The

lower epidermis having numerous trichomes (Uniseriate multicellular, uniseriate unicellular).

The collenchyma is present in the above the lower epidermis region. The palisade (beam like)

cells present in the below the upper epidermis. The sclernchymatoussheeth mingle with the

xylem and phloem, xylem lignified, phloem unlignified. The remaining space occupied with

parenchymatous cells.

Leaf constant

S.no Leaf constant Result

1. Vein lset number 35

2. Vein termination 25

3. Stomatal number 15

4. Stomatal index 20

Pharmacognostical parameters

The order of polarity wise selected the solvent for extractive values. High extractive values

present in polar solvent of water 30%w/w. The non-polar solvent of Pet ether produced

2%w/w of the extractive only. The crude fibre content and loss on drying also we performed

and noted the value on the table.

Leaf powder chemical test

S.no Chemical test Present/Absent

1. Alkaloids (Dragendroff’s test) Present

2. Glycosides (Bal jet test) present

3. Saponins (Foam test) Present

4. Carbohydrates (Molisch’s test) Present

5. Proteins (Biuret test) Absent

6. Amino acids (Ninhydrin test) Present

7. Lipids(fixed oil) (Filter paper test) Present

8. Volatile oil (distillation method) Absent

S.no Contents Percentage

1. Pet ether soluble extractives 2%

2. Chloroform soluble extractives 2%

3. Acetone soluble extractives 6%

4. Methanol soluble extractives 16%

5. Ethanol soluble extractives 24%

6. Acetic acid soluble extractives 28%

7. Water soluble extractives 30%

8. Crude fibre content 56%

9. Foreign organic matter 12%

10. Loss on drying 15%



Chemical test for ethanolic extract of luffa acutangula

S.no Chemical test Absent /Present

1. Alkaloids (Mayer’s and dragendroff’s test) Present

2. Glycosides (Borntrager’s test) present

3. Saponins (Foam test) Present

4. Carbohydrates (Molisch’s test) Present

5. Proteins (Biuret test) Absent

6. Amino acids (Ninhydrin test) Present

7. Flavonoids (Sinodha test) Absent

8. Steroids (Libermann’sburchard’s test) Present

Fig: 5 Chemical test for ethanolic extract of luffa acutangula

Thin layer chromatography

Fluorescence test

S.no Reagents Visible light Uv light (366nm)

1. Drug + H2So4 Sight brown Brown

2. Drug + HCL Pale green Pale green

3. Drug + NaOH Green Dark green

4. Drug + KOH Pale yellow Pale yellow

5. Drug + Nitric acid Colourless Colourless

6. Drug + Water Colourless Colourless

7. Drug +Acetone Pale green Pale yellow

8. Drug +CHCl3 Dark green Brown

9. Drug + Benzene Brown Black

10. Drug + pet ether Green Green

S.no Contents Rf value

1. Saponins 0.28

2. Amino acids 0.58

3. Glycosides 0.52

4. Sterols 0.44

5. Steroids 0.40



Fig: 6 Fluoresenceanlalysis of leaf powder of luffa acutangula (day light)

Fig: 7 Fluoresenceanlalysis of leaf powder of luffa acutangula (UV light)

Larvicidal activity

Time 6hours 12hours 24hours 48hours 72 hours

10mg/ml 0 0 2 3 4

20mg /ml 0 0 2 7 14

40mg/ml 0 0 3 15 17

80mg/ml 0 0 5 18 20

160mg/ml 0 0 14 20 -

Fig: 8 Larvicidal activity of ethanolic leaf extract of luffa acutangula



Fig: 9 Larvicidal activity in ethanolic leaf of luffa acutangula in 6hrs





Fig: 12 Larvicidal activity in ethanolic leaf of luffa acutangula in 48hrs.

CONCLUSION

The present research work was carried out the standardisation of luffa acutangula leaf, the

pharmacognostical screening work were preformed and reported. This pharmacognostical

screening work including the macroscopy, microscopical character, fluorescent analysis,

physicchemical parameters, extraction, identification of phytoconstituents and larvicidal

activity. This research work on most useful in future research scholars for further studies.

Well developed countries also now-a-days adopting to follow the herbal medicines, for the

reason no risk, side effects less, when compared to allopathic system of medicine. India like

developing countries poor sector of people more, they easily available herbal medicine used

day to day activites.

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