ANALYSIS OF AYURVEDIC

FORMULATIONS

PRESENTED BY:T. LAKSHMI BHAVANI

(2015MPH40023)

Under the guidance of: Dr. B.RAMYA KUBER

SPMVV - TIRUPATI

M.Pharm.,Ph.D Pharmaceutical Analysis

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CONTENTS Introduction to Ayurveda and Ayurvedic

formulations. Analysis of Ayurvedic drugs Analysis of Brahmi hills Analysis of Diabohills juice Analysis of Ashwagandha capsules Analysis of Avaleha Conclusion References

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INTRODUCTION

Ayurvedic medicine is a system of healing that originated

in ancient India.

In sanskrit

Ayur – Life or Living

Veda – Knowledge

So, Ayurveda has been defined as the “Knowledge of

living” or the “Science of longevity”.

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Ayurveda is the science of health and healing practised by

ancient aryans which is based on the Atharvaveda, one of the

oldest scriptures of Hindus, about 3000 years old.

The object of ayurveda is to counteract the imbalance of three

essential elements, vata, pitta, kapha(air,bile and phlegm

respectively),which constitute the Tridosha from which the

body originates.

According to Drugs and Cosmetic Act 1940,ayurvedic

medicine includes all medicines intended for internal or

external use, for or in the diagnosis, treatment, mitigation or

prevention of disease or disorder in human beings or animals.

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According to hypothesis of Ayurveda, the entire universe is

composed of five basic elements called PANCHAMAHABHUTA

and include space, air, energy ,liquid, and solid.

The variable combinations of these principles make tridosha in

human body.

These combinations are vata, pitta and kapha.

They have been interpreted in the following way.

Vata is composed of space and air. It is concerned with the

physical and mental activities which are activating or dynamic in

nature.

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Vata represents nervous system,or nadichakra as per ayurvedic

terminology.

Pitta is composed of liquid and energy, and indicates

biochemical or energy systems.

Kapha is composed of solid and liquid and indicates various

tissues and organs.

Ayurvedic therapeutics is based on five pharmacological

principles (panchsheel) of the drug.

Those are as follows:

1.Rasa 2.Guna 3.Vipaka 4.Virya 5.Prabhava.

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AYURVEDIC FORMULATIONS:

Different solvents (menstrum) used in preparations of

Ayurvedic formulations are water ,oils ,milk ,ghee ,cow’s

urine ,etc.

The use of sweetening agents ,binding agents, colorants,

flavouring principles, and other adjuvents is also very

common in ayurvedic preparations.

With an objective of obtaining maximum therapeutic benefit

and making the formulation palatable, different

pharmaceutical process are prescribed in ayurveda.

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Arka Asava and Arishta Avaleha or Leha and Paka Bhasma Churna Dravaka Fant Ghana-saar Ghrita(snehkalpa) Kalka Kajjali Ksharas Kvatha(Decoction) Kupipakva Rasayana

The Important Ones Are As Follows: Lepa Matras Nassyas Netrabindu and anjana Nikadha Parpati kalpas Pisti Praash Rasa-yoga Sattva Siddha-milks Swarasa Taila (Medicated oils) Vatika and Gutika

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ANALYSIS OF AYURVEDIC DRUGS:

Several Ayurvedic medicines readily available are found

to contain steroids and heavy metals ,both are harmful to

the human body if taken in excess.

Ayurvedic medicines given by the vaidyas –be it in the

form of bhasma or churna which people swallow in with

great faith,could contain steroid and toxic metals such as

lead,mercury and arsenic.

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In order to ensure quality control,quality assurance and safety

for ayurveda and siddha drugs, the quality of herbal products

need to be monitored with creation of awareness amongst all

concerned i.e.;

Physical analysis

Chemical analysis

Microbial analysis

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BRAHMI Synonym: Bacopa monnieri

Biological source:

It consists of dried whole plant, preferably the

fresh leaves and the stem of the plant known

as Bacopa monnieri(Linn.)

Belonging to family Scrophulariaceae and

should contain not less than 2.5% of bacoside

A calculated on the dried basis.

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Geological source:

It is glabrous, succulent, small, prostrate or creeping

annual herb found in throughout India, in wet, damp and

marshy places, up to 1200m elevation.

Macroscopic characters:

Root - Thin, wiry, small, branched creamish-yellow.

Stem - Thin, green or purplish green, about 1-2 mm thick,

soft, nodes and internodes are prominent,

glabrous; taste, slightly bitter.

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Leaf - Simple, opposite, decussate, green, sessile, 1-2 cm long,

obovate-oblong; taste, slightly bitter.

Flower - Small, axillary and solitary, pedicels 6-30 mm long,

bracteoles shorter than pedicels.

Fruit - Capsules up to 5 mm long, ovoid and glabrous.

CHEMICAL CONSTITUENTS:

Brahmi is found to contain the alkaloids, brahmine, herpestine,

and the mixture of other three alkaloids.

It contains saponins, namely bacosides A and B. Additionally, It

also contains betulicacid, stigmasterol, monnierin and hersaponin.

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.USES:

It is used as nervine tonic,in the treatment of asthma, Epilepsy.

And also used as diuretic and the alcoholic extract of entire

plant is found to have anti-cancer activity.

Important formulations:

Sarasvatarista,

Brahmi Ghrita,

Ratngiri Rasa,

Brahmi vati,etc..

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BRAHMI GHRITA

A polyherbal Ayurvedic formulation.

Preparation: Whole plant of Brahmi was

collected and other ingredients which are

used in the preparation of Brahmi

Ghrita,all are identified and authenticated

in pharmacognosy laboratory.

Brahmi swarasa was extracted by exerting

mechanical pressure on fresh brahmi.

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In a large vessel, Go-Ghrita was poured, when it

liquefies under moderate flame, kalka of vacha, kushtha,

shankhapushpi made in Brahmi swarasa was added,

followed by the addition of Brahmi swarasa.

To get final product, the contents were subjected to heat

till upto the Sneha Siddhi Lakshanas were observed.

PHYSICOCHEMICAL STUDY OF BRAHMI GHRITA:

Brahmi ghrita was analysed with various standard

physico-chemical parameters such as :

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Acid-value – NMT 2

Saponification value – 190 to 230

Refractive index value at 400 – 1.454 to 1.465

Iodine value – 30 to 40

Peroxide value – NMT 4

Specific gravity

HPTLC

Wt/ml at 400 - 0.930g to 0.945g

Congealing point – 210 to 170

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IDENTIFICATIONThin layer chromatography:

Extract 2 g of the sample with 20 ml of alcohol (400 for 3 hrs).

Cool, separate the alcohol layer,

Filter, concentrate to 5 ml

carry out the thin layer chromatography.

Apply 10 μl of the extract on TLC plate

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develop the plate to a distance of 8 cm {Mobile phase: toluene : ethyl acetate : hexane (6 : 3 : 1)}

allow the plate to dry in air

spray with ethanol-sulfuric acid reagent

heating at 1100c for about 10 min.

It shows major spots at Rf 0.15, 0.28, 0.40 and 0.51

(all light grey) under visible light.

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HPTLC

Preparation of sample solution:

The Ghrita sample was adsorbed on silica gel .

The mixture was extracted with hexane.

Hexane fraction was discarded .

The material was extracted with methanol.

Process was repeated for 3 times .

The methanol layer was collected ,filtered , and evaporated off.

The dried material was again dissolved in methanol and used

for TLC identification.

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CHROMATOGRAPHIC CONDITIONS :

1 . Stationary phase :silica gel GF precoated TLC plates.

2 . Mobile phase : Dichloromethane : methanol : water

(4.5 : 1.0 : 0.1 v / v / v).

Sample volume : 5 ml

Sample for HPTLC : Methanol of Brahmi Ghrita.

Spray Reagent:Vaniline-sulphuric acid.

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PROCEDURE :

Before spotting , the plates were prepared with methanol.

Sample solutions were applied to the plates as sharp bands.

The spots were dried in a current of air.

The mobile phase (20ml) was poured into a twin trough glass

chamber, whole assembly was left to equilibrate for 30 min and

the plate was placed in the chamber.

The plate was then developed until the solvent front had

travelled at a distance 80mm above the base of the plate.

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The plate was then removed from chamber

and dried in a current of air.

Detection and quantification was performed

with camag TLC scanner 3 at a wavelength

of 254 and 366 nm .

It shows major spots at Rf 0.78 (pink color)

which indicates the presence of Bacoside A.

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BRAHMI HILLS CAPSULES:

BRAIN TONIC:

COMPOSITION:

Each 300mg capsule contains:

Brahmi extract (Bacopa monnieri) – 150mg

Brahmi powder (Bacopa monnieri) – 150mg

{standardized to contain 25% total Bacosides=75mg}

(27% saponins = 81mg)

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BENEFITS:

In ayurveda brahmi has been historically used to aid

cognitive brain function.

It is cooling and helps in managing of stress.

It has anti-stress properties.

It is a cooling agent.

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ANALYSIS OF BRAHMI CAPSULES BY HPTLC

PREPARATION OF SAMPLE SOLUTION:

Capsule is a modern solid dosage form in which the

powdered crude drug or extract of the herb is filled into the

shells of the capsules.

2 g of capsule content was transferred to 100 ml volumetric

flask containing 50 ml of methanol and the mixture was

macerated on a shaker for 24 hrs at room temperature.

Then 1.0 ml of this extract was diluted to a 10 ml with

methanol.

PREPARATION OF STANDARD SOLUTION:

It is prepared by dissolving 5.0 mg of bacoside A in 10 ml

methanol, yielding stock solution of concentration = 0.5mg

ml-1.

THE MOBILE PHASE: Toluene: ethyl acetate: methanol: Gla.

acetic acid (3:4:2.5:1) v/v

PROCEDURE

The sample solutions were spotted on the precoated silica gel

aluminum plate 60F254.

The plates were pre-washed by methanol and activated at

60°C for 5 min prior to chromatography.

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15 ml of mobile phase was used per chromatography. Linear

ascending development was carried out in twin trough glass

chamber saturated with the mobile phase.

The length of chromatogram run was 8 cm.

TLC plates were dried in a current of air with the help of an

air dryer.

Densitometric scanning was performed with Camag TLC

scanner III in the reflectance-absorbance mode at 540 nm

after spraying with 10% Sulfuric acid alcohol reagent (Acid

alcohol) .

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DIABOHILLS JUICE

INTRODUCTION:

It is a quality product containing

EUGENIA JAMBOLANA +MOMODICA CHARANTIA as

its ingredient.

Diabohill juice is potent combination of Bitter Gourd and

Amla.

Synonyms: karela -jamun juice

Phyto constituents: Glucoside, saponin (0.5% charaantin),

aromatic volatile oils,7% bitter principles.

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Fruits and leaves: Momordicine, ascorbigen, aspartic acid,

serine, glutamic acid, threonine, alanine, g-aminobutyric

acid and pipecolic acid.

USES: Diabohills juice is very effective in non-insulin dependent

diabetes.

It is also helps in reducing fasting blood sugar too.

And it is also used as:

Anti diabetic , Hypoglycemic

Antibiotic, Anti mutagenic ,Antioxidant etc..

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HERBAL HILLS : DIABOHILLS (KARELA –JAMUN JUICE)

INGREDIENTS: Jambu, Karela, Guduchi,

Baheda, Harde, Sunthee, Amla, Turmeric,

Neem, Fenugreek, Gurmar, Ashwagandha,

Kadu Chirayati, shatavari.

DIRECTIONS: Take 30ml juice mixed with 100ml

water or fruit juice or have it plain.

PART USED FOR JUICE: Pulp of jamun and fruit of

karela.

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MANUFACTURING PROCESS OF DIABOHILLS JUICE AT HERBAL HILLS

STEP: 1

PROCESSING RAW HERBS:

All the raw material karela, amla fruit ,harde, baheda, neem,

methi, ambehaldi, sunthee, shatavari, Ashwagandha, Kadu

Chirayati, bel leaves, and gudmar are washed, cut and

crushed in to small pieces.

STEP: 2

The mixture is boiled at 1100c

The boiled material is then cooled at room

temperature.

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STEP 3:

The pulp of Jamun (seeds of jamun are not used) is

prepared and mixed with all the above ingredients and

crushed and water is added.

After addition of preservative, the homogenized mixture is

mixed thoroughly and in process sample is taken and sent

for testing.

The sample passing the specifications are taken for

packing.

STEP 4:

The bottles are labeled and packed as per guidance.

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FINISHED PRODUCT SPECIFICATION OF HERBAL HILLS DIABOHILLS (KARELA JAMUN) JUICE

S.No. TESTS SPECIFICATIONS

1. Description Fibrous Dark Brown liquid

2. Uniformity of weight 500ml + 7.5 %

HEAVY METALS3. Arsenic(As) NMT 10 ppm

4. Lead(Pb) NMT 10 ppm

5. Mercury(Hg) NMT 1 ppm

6. Cadmium(Cd) NMT 0.3 ppm

MICROBIOLOGICAL ANALYSIS7. Total microbial count NMT 1000 cfu /ml

8. Yeast/mould NMT 100 cfu /ml

9. Coliforms Absent

10. Escherichia coli Absent

11. salmonella Absent

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ASHWAGANDHA CAPSULES Synonym : Withania root , Winter cherry

Biological source:

It consists of dried mature roots of withania

somnifera (linn) Dunal; belonging to the

family Solanaceae and should contain not

less than 0.02% of total withanolide A and

withaferin A on dried basis.

Chemical constituents:

The main constituents are alkaloids and

steroidal lactones.

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The main alkaloid is the withanine and other alkaloids are

somniferine, somnine, somniferinine, withananine,

pseudo-withanine, tropine, pseudo-tropine and anahydrine.

Two acyl steryl glucosides viz.sitoindoside vii and

sitoindoside viii have been isolated from roots.

Therapeutic uses:

Anxiety, stress, depression, Muscle fatigue and neck/back stiffness, arthritis of all types, blood pressure, Insomina, diabetic neuropathy, muscle power and stamina, weakness due to Diabeties.

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Composition of capsule:

Each 500 mg capsule contains:

Concentrated extract of withania …..400mg

Excipients ………q.s

(A) Powder study: The shade dried roots of the plant were

powdered and pass through sieve 60# and used for the observation

of microscopical characters.

The powder drug + Phloroglucinol:HCl(1:1) Lignified cells

The powder drug + Lacto phenol Calcium oxalate crystals

The powder drug + Iodine solution Starch grains

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(B).EVALUATION OF QUALITY CONTROL PARAMETERS FOR RAW MATERIALS:PHYSICO CHEMICAL PARAMETERS:

Foreign organic matter:NMT 2%

Total ash :NMT 7%

Acid-insoluble ash :NMT 1%

Alcohol soluble extractive :NMT 15%

Ethanol soluble extractive :NMT 10%

Water soluble extractive : Nil

Loss on drying : NMT 12% determined on 5g by drying

in an oven at 1050.

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DETERMINATION OF pH: The pH value of a solution was determined

potentiometrically by means of a glass electrode, a

reference electrode and a digital pH meter.

First the apparatus was calibrated using buffer of 4,9 and

7pH.

1gm of powdered extract was taken and dissolved in

100ml of demineralized water.

The electrodes were immersed in the solution and the pH

was measured.

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QUALITATIVE PHYTOCHEMICAL EVALUATION:

TEST FOR ALKALOIDS:

Mayer’s test: The small portions of solvent free chloroform, alcoholic

and water extracts are stirred separately with a few drops

of dil.HCl and filtered.

The filtrate may be tested carefully with various alkaloidal

reagents such as Mayer's reagent, cream ppt formed.

It indicates the presence of alkaloids.

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Drug extract + Dragendorffs reagent orange brown ppt.

Drug extract + Hager's reagent yellow ppt.

Drug extract + Wagers reagent reddish-brown ppt.

TEST FOR GLYCOSIDES:

GENERAL TEST: Extract 200mg of the drug using 5ml of dilute

(10%) sulphuric acid and boil on waterbath.

After boiling add equal volume of water to the volume of NaOH .

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Add 0.1ml of Fehlings reagent A and B alkaline (red

litmus changes to blue ) and heat on water bath for 2 min.

Note the red ppt formed represents the glycoside after

acid hydrolysis.

TEST FOR SAPONINS: Frothing test:The sample drug was vigorously shaken

with distilled water and was allowed to stand for 10 min.

Stable froth more than 1.5cm indicates the presence of

saponins.

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TEST FOR TANNINS:

Ferric chloride test: The water extract of the sample drug was

treated alcoholic ferric chloride.

Blue colour indicates the presence of tannins.

Lead acetate test: With 5% lead acetate solution tannins give

ppt which turns to red colour on addition of KOH

solution ,on excess addition ppt is dissolved.

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TEST FOR CARBOHYDRATES:

Molisch’s test: Small quantities (200mg) of alcoholic and aqueous extract are

dissolved seperately in 5ml of distiiled water and filtered.

The filtrate is treat with Molisch’s reagent.

It gives red or purple colored compound.

It indicates the presence of carbohydrates.

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EVALUATION OF QUALITY CONTROL PARAMETERS FOR FINISHED PRODUCT(CAPSULES):

DISINTEGRATION TEST FOR CAPSULES:

Disintegration test was performed using the digital

microprocessor based disintegration test apparatus by VEEGO.

One capsule was introduced into each tube and added a disc to

each tube.

The assembly was suspended in the water in a 1000ml beaker.

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The volume of water was such that the wire mesh at its highest

point is atleast 25mm below the surface of the water,and at its

lower point was at least 25mm above the bottom of the beaker.

The apparatus was operated and maintained with temperature

at 37 + 20 c.

Noted down the time require to all capsules to disintegrate and

pass through wire mesh.

LIMITS: disintegration time must not be more than 30mins.

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DISSOLUTION TEST FOR CAPSULES:

The dissolution test was performed for capsule using USP

dissolution apparatus 2 by VEEGO.

The 900 ml of the 5N HCl as dissolution medium was

introduced into the vessel of the apparatus.

The dissolution medium was warmed to 36.5 -37.50c.

For the capsules basket type dissolution apparatus was

used.

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The apparatus was operated immediately at the speed of 50

rpm for 2 hrs.

After 2 hrs a 25ml specimen was withdrawn from a zone

midway between the surface of the dissolution medium and

top of the rotating blade or basket,in evaporating dish.

Filter and evaporate the specimen.

For the each of the capsule tested, the amount of dissolved

active ingredients in the solution was calculated as a

percentage dissolved in 2 hrs.

LIMITS: Dissolution of capsules should not be less than 70%.

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MICROBIAL ANALYSIS: The test included total bacterial count, total yeast, and mould

count.

And also the identification of specified organisms such as

Escherichia coli, Salmonella species, Staphylococcus aureus,

and pseudomonas aeuruginosa.

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AVALEHA Avaleha or Lehya is a semisolid preparation of drugs,

prepared with addition of jaggery,sugar or sugar-candy and

boiled with prescribed juices or decoction.

These preparations generally have

1) Kasaya or other liquids

2) Jaggery ,sugar or sugar-candy

3) Powders or pulps of certain drugs

4) Ghee or oil

5) Honey

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Jaggery, sugar or sugar-candy is dissolved in the liquid and

strained to remove the foreign particles.

This solution is boiled over a moderate fire.

When pressed between two fingers if paka becomes

thready, or when it sinks in water without getting easily

dissolved, it should be removed from the fire.

Fine powders of drugs are then added in small quantities

and stirred continuosly to form a homogeneous mixture.

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Ghee or oil, if mentioned, is added while the preparation is

still hot and mixed well.

Honey, if mentioned is added when the preparation becomes

cool and mixed well.

The Lehya should neither be hard nor a thick fluid .

When pulp of the drugs is added and ghee (or) oil is present in

the preparation, this can be rolled between the fingers.

When metals are mentioned, the bhasmas of the metals are

used.

In case of drugs like Bhallataka,purification process is to be

followed.

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The lehya should be kept in glass or porcelain jars.

It can also be kept in a metal container which does not react

with it .

Normally Lehyas should be used within one year.

Eg. Kalyanavaleha, Draksavaleha, vasavaleha, bilvadileha and

surnavaleha.

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KALYANAVALEHA

Definition: Kalyanavaleha is a semisolid preparation made with

the ingredients of the formulation composition given below…

Haridra API

Vaca API

Kustha API

Pippali API

Sunthi API

Ajaji API

Ajamoda API

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Yasthi madhu API

Saindhava lava API

Sarpi API

METHOD OF PREPARATION: Take all ingredients ,clean and dry and powder the ingredients

numbered 1 to 9 seperately and pass through the sieve number

85.Mix all the ingredients thoroughly.

Add sarpi to the mixture ,stir thoroughly to form a semisolid

mass.

Pack it in tightly closed containers to protect from light and

moisture.

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DESCRIPTION: Semisolid paste, yellowish-brown in color with pungent odour,

astringent and salty taste.

IDENTIFICATION:THIN LAYER CHROMATOGRAPHY: Defat 5mg of kalyanavaleha with 75ml of n-hexane under reflux

on a water-bath for 30 min.

Filter and discard the hexane extract.

Extract the defatted marc with 75ml of chloroform under reflux

for 30 min.

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Filter and concentrate the extract to 10 ml and carry out the TLC.

Apply 10ml of the chloroform extract on TLC plate and develop

the plate to a distance of 8cm using

toulene:ethylacetate:methanol (9:1:1) as Mobile Phase.

After development allow the plate to dry in air and examine

under UV light (366nm).

It shows major spots at Rf 0.22(blue), 0.29,0.45(both yellow),

0.60,0.68(both blue).

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CHEMICAL TESTS:

1.Avaleha + conc.sulphuric acid orange red color

{presence of curcuminoids(Haridra)}.

2. Avaleha + 10% sol. of NaOH /KOH red to violet

color (indicating the presence of curcuminoids).

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PHYSICO CHEMICAL PARAMETERS: Loss on drying :NMT 5.5%

Total ash :NMT12.0%

Acid-insoluble ash :NMT 2.0%

Alcohol soluble extractive :NLT 46.0%

Water soluble extractive :NLT 11.0%

PH (1% aqueous solution ) :5.1 and 5.3

Starch :NLT 42.0%

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OTHER REQUIREMENTS:

MICROBIAL LIMITS: These microbial limit tests are performed for the estimation of

the no. of viable aerobic micro-organisms present and for

detecting the presence of designated microbial species in

pharmaceutical substances.

AFLATOXINS: These are highly dangerous toxic substances,extreme care

should be taken while handling these aflatoxin material.

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This aflatoxin test can be performed to detect the

presence of aflatoxins in any material of plant origin.

STORAGE: Store in a cool place in tightly closed containers,protected from

light and moisture.

THERAPEUTIC USES: Svarabhedha(hoarseness of voice)

Mukata (Aphasia)

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CONCLUSION

By performing the analysis of ayurvedic formulations or

preparations, all the macroscopic, microscopic, physical,

and chemical characteristics of the formulations can be

determined .

If any undesirable effects may be identified during

analysis, they can be reduced to acceptable levels, leads

to get good therapeutic effect without any adverse

reactions..

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REFERENCES

Text book of pharmacognosy by C.K.Kokate.

The Ayurvedic pharmacopoeia of India part-II (formulations);

volume-I

International Journal of Applied Biology and Pharmaceutical

Technology.

Journal of Analytical profile of Ayurvedic Formulations. Anonymous. (1996). Indian Pharmacopoeia, Vol. II,

Government of India, Ministry of Health and Family Welfare, New Delhi, A-81-83, 95, 736.

Thank you