ANALYSIS OF AYURVEDIC FORMULATIONS

1. 1
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

2. 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
2

3. 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”.
3

4.  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.
4

5.  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.
5

6.  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. 6

7. 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.
7

8.  Arka
 Asava and Arishta
 Avaleha or Leha and Paka
 Bhasma
 Churna
 Dravaka
 Fant
 Ghana-saar
 Ghrita(snehkalpa)
 Kalka
 Kajjali
 Ksharas
 Kvatha(Decoction)
 Kupipakva Rasayana
8
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

9. 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.
9

10.  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
10

11. 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.
11

12. 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.
12

13. 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. 13

14. .
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..
14

15. 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.
15

16.  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 :
16

17.  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
17

18. IDENTIFICATION
Thin 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
18

19. 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. 19

20. 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.
20

21. 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.
21

22. 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. 22

23.  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.
23

24. 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)
24

25. 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.
25

26. 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. 26

27. 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.
27

28.  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) .
28

29. 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.
29

30.  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.. 30

31. 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.
31

32. 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.
32

33. 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. 33

34. FINISHED PRODUCT SPECIFICATION OF HERBAL
HILLS DIABOHILLS (KARELA JAMUN) JUICE
34
S.No. TESTS SPECIFICATIONS
1. Description Fibrous Dark Brown liquid
2. Uniformity of weight 500ml + 7.5 %
HEAVY METALS
3. Arsenic(As) NMT 10 ppm
4. Lead(Pb) NMT 10 ppm
5. Mercury(Hg) NMT 1 ppm
6. Cadmium(Cd) NMT 0.3 ppm
MICROBIOLOGICALANALYSIS
7. Total microbial count NMT 1000 cfu /ml
8. Yeast/mould NMT 100 cfu /ml
9. Coliforms Absent
10. Escherichia coli Absent
11. salmonella Absent

35. 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.
35

36.  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. 36

37.  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 37

38. (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.
38

39. 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. 39

40. 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.
40

41.  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 .
41

42.  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.
42

43. 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.
43

44. 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.
44

45. 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.
45

46.  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.
46

47. 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.
47

48.  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%. 48

49. 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.
49

50. 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
50

51.  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.
51

52.  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.
52

53.  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.
53

54. 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
54

55.  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.
55

56. 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. 56

57.  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).
57

58. 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).
58

59. 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%
59

60. 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. 60

61. 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)
61

62. 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.. 62

63. 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.
63

64. 64
Thank you