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STANDARDIZATION OF AMRITAPRASHA GHRITA: A HERBAL GHEE BASED

MEDICINAL PREPARATION

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Sivaraj et al., IJPSR, 2018; Vol. 9(11): 4842-4848. E-ISSN: 0975-8232; P-ISSN: 2320-5148

IJPSR (2018), Volume 9, Issue 11 (Research Article)

Received on 26 February, 2018; received in revised form, 03 May, 2018; accepted, 13 May, 2018; published 01 November, 2018

STANDARDIZATION OF AMRITAPRASHA GHRITA: A HERBAL GHEE BASED

MEDICINAL PREPARATION
Saranya Sivaraj, Vijayalaxmi Mallannavar, G. R. Arun Raj * and U. Shailaja
Department of Kaumarabhritya, Sri Dharmasthala Manjunatheshwara College of Ayurveda and Hospital,
Hassan - 573201, Karnataka, India.
Keywords: ABSTRACT: Aim: To standardize Amritaprasha ghrita. Materials
Amritaprasha Ghrita, Under nutrition, and methods: Physico‐chemical studies like refractive index, specific
karshya, Standardization, HPTLC gravity, acid value, saponification value, iodine value, determination
Correspondence to Author: of unsaponifiable matter, peroxide, viscosity, rancidity test and
Dr. G. R. Arun Raj HPTLC were carried out as per the WHO guidelines, Indian
Assistant Professor, Pharmacopoeia and Ayurvedic Pharmacopoeia. Conclusion:
Department of Kaumarabhritya, Standardization tests done on Amritaprasha ghrita helped in
Sri Dharmasthala Manjunatheshwara
College of Ayurveda and Hospital, authenticating and ensuring the quality of the same.
Hassan - 573201, Karnataka, India
E-mail: drdrarunraj26@gmail.com

INTRODUCTION: Standardization is necessary Jivanti (Leptadenia reticulata (Retz.) Wight &

to make sure the availability of a consistent product Arn),10 Shunti (Zingiber officinale Roscoe), 12 Shati
and can assure a reliable product with definite (Hedychium spicatum Sm. in A.Rees),13 Shalaparni
constituents 1 - 2. The standardization of herbal (Desmodium gangeticum (L.) DC.), 14 Prishniparni
medicines is always challenging as it medicines (Uraria picta (Jacq.) DC.), 15 Mudgaparni
contain more than one active principles and the (Phaseolus trilobus Ait.), 16 Mashaparni (Teramnus
active compound is frequently unknown 3 - 4. labialis (L. f.) Spreng.), 16 Meda (Asparagus
Standardization of herbal formulations is essential racemosus Willd.), 10 Mahameda (Asparagus
to assess quality, consistency of active principles racemosus Willd.), 10 Kakoli (Withania somnifera
and therapeutic efficacy of drugs 5 - 6. The quality (L.) Dunal), 17 Ksheerakakoli (Withania somnifera
assessment of herbal formulations is important to (L.) Dunal), 18 - 19 Bruhati (Solanum indicum.), 16
justify their acceptability and safety7 - 8. Kantakari (Solanum xanthocarpum Schrad. & H.
Wendl.), 14 Sweta Punarnava (Boerhavia diffusa
Amritaprasha ghrita 9 comprises of fifty drugs such L.), 10 Rakta Punarnava (T. portulacastrum L.), 20
as Jivaka (Pueraria tuberosa (Willd.) DC), 10 Madhuka (Glycyrrhiza glabra L.), 21 - 24 Kapikachu
Rishabhaka (Pueraria tuberosa (Willd.) DC), 10 (Mucuna pruriens (L.) DC.), 25 Shatavari
Veera (Nardostachys jatamansi (D. Don) DC.), 11 (Asparagus racemosus Willd.), 10 Riddhi (D.
QUICK RESPONSE CODE
bulbifera L.), 10 Vriddhi (Dioscorea bulbifera L.),
10
DOI: Parushaka (Grewia asiatica L.), 26 Bharangi
10.13040/IJPSR.0975-8232.9(11).4842-48 (Clerodendrum serratum. (L)), 27 Mrudvika (Vitis
vinifera L.), 28 Shringhataka (Trapa bispinosa
Article can be accessed online on: Roxb.), 29 Tamalaki (Phyllanthus niruri L.), 10
www.ijpsr.com Vidarikanda (Pueraria tuberosa (Willd.) DC), 30
Pippali (Piper longum L.), 31 Bala (Sida cordifolia
DOI link: http://dx.doi.org/10.13040/IJPSR.0975-8232.9(11).4842-48
L.), 10 Badara (Ziziphus jujube Mill.), 32 Akshotaka

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Sivaraj et al., IJPSR, 2018; Vol. 9(11): 4842-4848. E-ISSN: 0975-8232; P-ISSN: 2320-5148

(Juglans regia L.), 33 Kharjura (Phoenix dactylifera (L.) Maton), 44 Patra (Cinnamomum zeylanicum
L.), 34 Vatama (Prunus amygdalus Batsch), 35 Blume), 45 Nagakesara (Mesua ferrea L.) 46.
Abhishuka (Pistacia vera L.), 36 Dhatri Amritaprasha Ghrita is made use in the
(Phyllanthus emblica L), 37 Ikshu (Saccharum management of Karshya (Grade 1 & 2 under
officinarum L.), 38 Chaaga Mamsarasa (meat soup nutrition) in children. Literature survey did not
of goat fried with ghee), 39 Go ksheera, 40 Go reveal any standards for Amritaprasha ghrita and
ghrita, 41 Madhu (honey), 42 Sarkara (sugar), hence the current study was undertaken to
Maricha (Piper nigrum L.), 43 Twak (Cinnamomum standardize the same. The ingredients of
zeylanicum Blume), 10 Ela (Elettaria cardamomum Amritaprasha ghrita is detailed in Table 1.
TABLE 1: SHOWING INGREDIENTS OF AMRITAPRAASHA GHRITA
S. no. Sanskrit name Substitute used Botanical name Part used
1 Jivaka Vidarikanda Pueraria tuberosa (Willd.) DC Tuber
2 Rishabhaka Vidarikanda Pueraria tuberosa (Willd.) DC Tuber
3 Veera - Nardostachys jatamansi (D.Don) DC. Root
4 Jivanti - Leptadenia reticulata (Retz.) Wight & Arn Root
5 Shunti - Zingiber officinale Roscoe Rhizome
6 Shati - Hedychium spicatum Sm. in A. Rees Rhizome
7 Shalaparni - Desmodium gangeticum (L.) DC. Root
8 Prushniparni - Uraria picta (Jacq.) DC. Root
9 Mudgaparni - Phaseolus trilobus Ait. Root
10 Mashaparni - Teramnus labialis (L. f.) Spreng. Root
11 Meda Shatavari Asparagus racemosus Willd. Root tuber
12 Mahameda Shatavari Asparagus racemosus Willd. Root tuber
13 Kakoli Aswagandha Withania somnifera (L.) Dunal Root
14 Ksheerakakoli Aswagandha Withania somnifera (L.) Dunal Root
15 Brihati - Solanum indicum Root
16 Kantakari - Solanum xanthocarpum Schrad. & H. Wendl. Root
17 SwetaPunarnava - Boerhavia diffusa L. Root
18 RaktaPunarnava - Trianthema portulacastrum L. Root
19 Madhuka - Glycyrrhiza glabra L. Root
20 Kapikachu - Mucuna pruriens (L.) DC. Seed
21 Shatvari - Asparagus racemosusWilld. Root tuber
22 Riddhi Varahikanda Dioscorea bulbifera L. Tuber
23 Vriddhi Varahikanda Dioscorea bulbifera L. Tuber
24 Parushaka - Grewia asiatica L. Fruit
25 Bharangi - Clerodendrum serratum .(L) Root
26 Mrudvika - Vitis vinifera L. Fruit
27 Brihati - Solanum indicum Root
28 Shringataka - Trapabi spinosa Roxb. Fruit
29 Tamalaki - Phyllanthus niruri L. Whole plant
30 Vidarikanda - Pueraria tuberosa (Willd.) DC. Tuber
31 Pippali - Piper longum L. Fruit
32 Bala - Sida cordifolia L. Root
33 Badara - Ziziphus jujube Mill. Fruit
34 Akshotaka - Juglans regia L. Seed
35 Kharjura - Phoenix dactylifera L. Fruit
36 Vatama - Prunus amygdalus Batsch Seed
37 Abhishuka - Pistacia vera L. Seed
38 Dhatri - Phyllanthus emblica L Fruit
39 Vidarikanda - Pueraria tuberosa (Willd.) DC. Tuber
40 Ikshu - Saccharum officinarum L. Stem
41 Chaaga Mamsarasa - Goat’s meat
42 Go Ksheera - Cow’s milk
43 Go Ghrita - Cow’s ghee
44 Madhu - Honey
45 Sarkara - Sugar
46 Maricha - Piper nigrum L. Fruit
47 Twak - Cinnamomum zeylanicum Blume Bark
48 Ela - Elettaria cardamomum (L.) Maton Fruit
49 Patra - Cinnamomum zeylanicum Blume Leaf
50 Nagakesara - Mesua ferrea L. Flower

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MATERIALS AND METHODS: Physico‐ shaken well. Added One ml of Phenolphthalein

chemical studies like refractive index, specific solution and titrated against 0.1M Potassium
gravity, acid value, saponification value, iodine hydroxide solution. End point is the appearance of
value, determination of unsaponifiable matter, pale pink colour. Repeated the experiment twice to
peroxide, viscosity, rancidity test and HPTLC were get concordant values.
carried out as per the WHO guidelines, Ayurvedic
Pharmacopoeia and Indian Pharmacopoeia. Saponification Value: Weighed 2 g of the
Amritaprasha ghritha into a 250 ml RB flask fitted
Plant Material: The constituents of Amritaprasha with a reflux condenser. Added 25ml of 0.5M
Ghrita were collected from the local market of alcoholic potash. Refluxed on a water bath for 30
Hassan District, Karnataka State, India in the minutes. Cooled and added 1 ml of phenolphthalein
month of March 2017. The collected drug was solution and titrated immediately with 0.5 M
identified and authenticated (no: SDMCAH- Hydrochloric acid (a ml). Repeated the operation
DG/2017/16) at the teaching pharmacy of omitting the substance being examined (blank) (b
Department of Dravyaguna (Ayurveda ml). Repeated the experiment twice to get
Pharmacology), SDM College of Ayurveda and concordant values.
Hospital, Hassan, Karnataka State, India.
Iodine Value: The sample was accurately weighed
Methodology: The studies were done at SDM in a dry iodine flask. Dissolved with 10 ml of CCl4,
Centre for Research in Ayurveda and Allied 20 ml of iodine monochloride solution was added.
Sciences, Kuthpady, Udupi, Karnataka State, India Stopper was inserted, which was previously
as per standard procedure. moistened with solution of potassium iodide and
flask was kept in a dark place at a temperature of
Refractive Index: Placed a drop of water on the about 17 ºC for 30 min. 15 ml of potassium iodide
prism and adjusted the drive knob in such a way and 100 ml of water was added and shaken well.
that the boundry line intersects the separatrix This was titrated with 0.1N Sodium thiosulphate,
exactly at the centre. Noted the reading. Distilled starch was used as indicator. The number of ml of
water has a refractive index of 1.33217 at 28˚C. 0.1N sodium thiosulphate required (a) was noted.
The difference between the reading and 1.3325 The experiment was repeated with the same
gives the error of the instrument. If the reading is quantities of reagents in the same manner omitting
less than 1.3320, the error is minus (-) then the the substance. The number of ml of 0.1N sodium
correction is plus (+) if the reading is more, the thiosulphate required (b) was noted. The
error is plus (+) and the correction is minus (-). experiment was repeated twice to get concordant
Refractive index of oil is determined using 1 drop values.
of the sample. The correction if any should be
applied to the measured reading to get the accurate Determination of Unsaponifiable Matter:
refractive index. Refractive index of the test Weighed 5 g of the Amritaprasha ghritha into the
samples were measured at 28˚C. flask. Added 50 ml alcoholic KOH into the sample.
Boiled gently but steadly under reflux condenser
Specific Gravity: Cleaned a specific gravity bottle for one hour. The condensor was washed with 10ml
by shaking with acetone and then with ether. Dried of ethyl alcohol and the mixture was collected and
the bottle and noted the weight. Cooled the sample transferred to a separating funnel. The transfer was
solution to room temperature. Carefully filled the completed by washing the sample with ethyl
specific gravity bottle with the test liquid, inserted alcohol and cold water. Altogether, 50 ml of water
the stopper and removed the surplus liquid. Noted was added to the separating funnel followed by an
the weight. Repeated the procedure using distilled addition of 50 ml petroleum ether. The stopper was
water in place of sample solution. inserted and shaken vigorously for 1 min and
allowed it to settle until both the layers were clear.
Acid Value: Weighed 2-10 g of ghritha in a
The lower layer containing the soap solution was
conical flask. Added 50 ml of acid free alcohol-
transferred to another separating funnel and
ether mixture (25 +25ml) previously neutralised
repeated the ether extraction six times more using
with the 0.1M potassium hydroxide solution and

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Sivaraj et al., IJPSR, 2018; Vol. 9(11): 4842-4848. E-ISSN: 0975-8232; P-ISSN: 2320-5148

50 ml of petroleum ether for each extraction. All Rancidity Test: 1 ml of melted fat was mixed with
the extracts were collected in a separating funnel. 1ml of conc. HCl and 1 ml of 1% solution of
The combined extracts were washed in the funnel 3 phloroglucinol in diethyl ether and then mixed
times with 25 ml of aqueous alcohol and shaked thoroughly with the fat acid mixture. A pink color
vigorously. And drawing off the alcohol-water indicates that the fat is slightly oxidized while a red
layer after each washing. The ether layer was again color indicates that the fat is definitely oxidized.
washed repeatedly with 25 ml of water until the
water no longer turns pink on addition of a few Sample Preparation for HPTLC: Sample
drops of Phenolphthalein indicator solution. The obtained in the procedure for the determination of
ether layer was transferred to a tarred flask unsaponifiable matter is dissolved in 10 ml of
containing few pieces of pumice stone and chloroform this was followed for all the sample of
evaporated to dryness on a water bath. Placed the Amritaprasha ghritha, and chloroform soluble
flask in an air oven at 85 °C for about 1 h to portion was used for HPTLC.
remove the last traces of ether. A few ml of acetone
HPTLC: 4, 8 and 12 µl of the above sample of
was added and evaporated to dryness on a water
Amritaprasha ghrita was applied on a precoated
bath. Cooled in a desicator to remove last traces of
silica gel F254 on aluminum plates to a band width
moisture and then weighed.
of 8 mm using Linomat 5 TLC applicator. The
Peroxide Value: 5 g of the Amritaprasha ghrita plate was developed in toluene - ethyl acetate (9:1)
was weighed accurately into a conical flask, added and the developed plates were visualized under
30 ml of mixture of 3 volumes of glacial acetic short UV, long UV, and after derivatisation in
acid and 2 volumes of chloroform, added 0.5 ml of vanillin-sulphuric acid spray reagent it was
potassium iodide, allowed it to stand for 1 minute, visualized under white light and scanned under UV
add 30 ml of water titrate gradually with vigorous 254 nm, 366 nm and 620 nm. Rf, colour of the
shaking with 0.1M sodium thiosulphate until the spots and densitometric scan were recorded.
yellow color disappears. Add 0.5 ml of starch TABLE 2: SHOWING RESULTS OF STANDARDIZATION
indicator continued the titration until blue color PARAMETERS
disappears. Parameter Results n = 3 %w/w
Amritaprasha ghrita
Peroxide value= 10(a-b) / W Refractive index 1.45783
Specific gravity 0.9551
Where W= weight in g of the substance Acid value 5.66
Saponification value 124.96
Viscosity: The given sample is filled in a U tube Iodine value 47.71
viscometer in accordance with the expected Unsaponifiable matter (%) 1.03
Peroxide value 0.19
viscosity of the liquid so that the fluid level stands Rancidity Fat is not oxidised
within 0.2 mm of the filling mark of the viscometer
when the capillary is vertical and the specified
temperature is attained by the test liquid. The liquid
is sucked or blown to the specified height of the
viscometer and the time taken for the sample to
pass the two marks is measured. Viscosity is
measured using the formula:
η1= ρ1t1 X η2
ρ2t2
η1 – Viscosity of sample
η2 - Viscosity of water
t1 and t 2- time taken for the sample and water to pass the Short UV Long UV Post derivatisation
meniscus Track 1- Chloroform fraction of Amritaprasha ghrita- 4 µl
ρ1 and ρ2 – Density of sample and water Track 2 - Chloroform fraction of Amritaprasha ghrita - 8 µl
X= Specific gravity of sample x 0.9961/specific gravity of Track 3- Chloroform fraction of Amritaprasha ghrita - 12 µl
water Solvent system- Toluene: Ethyl acetate (9.0:1.0)
= Xx Time for samplex1.004/specific gravity of water FIG. 1: TLC PHOTO DOCUMENTATION OF CHLORO-
x70sec FORM FRACTION OF AMRITAPRASHA GHRITA

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TABLE 3: Rf VALUES OF THE SAMPLE OF AMRITAPRASHA GHRITA

Short UV Long UV Post derivatisation
- 0.10 (FL. blue) -
0.24 (L. green) - -
0.30 (L. green) - -
- - 0.36 (D. purple)
- 0.43 (FL. blue) -
- 0.50 (FL. purple) 0.50 (L. purple)
- 0.74 (F. blue) -
- 0.78 (F. blue) -
- 0.92 (F. blue) -
*F-fluorescent; D-dark; L-light

Fig 2a. At 254nm Fig 2b. At 366nm

Fig 2c. At 620nm

FIG. 2: DENSITOMETRIC SCAN OF AMRITAPRASHA GHRITA

RESULTS AND DISCUSSION: The values of sample of Amritaprasha ghrita is detailed

standardization parameters of Amritaprasha ghrita in Table 3. The Densitometric Scan of
are detailed in Table 2. The TLC photo Amritaprasha ghrita is shown in Fig. 2. The
documentation of chloroform fraction of physicochemical standards would serve as
Amritaprasha ghrita is shown in Fig. 1. The Rf preliminary test for the standardization of the

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Sivaraj et al., IJPSR, 2018; Vol. 9(11): 4842-4848. E-ISSN: 0975-8232; P-ISSN: 2320-5148

formulation. Tests such as refractive index, specific residual toxicity. Int. J. Res. Ayurveda Pharm 2016; 7(S1):
80-83.
gravity, acid value, saponification value, iodine 9. Das SG: Bhaishajya Ratnvali, Rajayakshma chikitsa,
value, determination of unsaponifiable matter, 14/265-274. 1st ed. Varanasi: Chaukhambha Orientalia;
peroxide, viscosity, rancidity test and HPTLC, 2014: 575.
10. Arun-Raj GR, Shailaja U, Rao PN and Debnath P:
results of TLC photo documentation, the unique Rf Nutraceuticals and Functional foods. 1st ed. Houston-
values, densitometric scan and densitogram Texas, USA: Recent Progress in Medicinal Plants.
obtained at different wavelengths can be used as Studium Press LLC; 2016. Chapter 10, Nutraceuticals and
Functional foods in Ayurvedic perspective 42: 172-199.
fingerprint to identify the herbal drug of 11. Arun-Raj GR, Shailaja U, Sagar K, Viswaroopan D and
Amritaprasha ghrita. Kumar SN: Swarnaprashana to Swarnamritaprashana:
experience inspired modification through ages. Int J Pharm
CONCLUSION: Amritaprasha ghrita has been Sci Res 2017; 8(11): 4546-50.
12. Raj JKT, Mallannavar V, Raj AGR, Shailaja U,
standardized using diverse scientific quality Viswaroopan D and Krishna JG: Exploratory studies on
parameters. The results obtained can be used as the therapeutic effects of Patoladi syrup in the
reference while setting the pharmacopoeial management of chronic tonsillitis in children at a tertiary
care hospital of Southern India. Int. J. Res. Ayurveda
standards for Amritaprasha ghrita for the benefit of Pharm. 2018; 9(2): 62-69.
the end user without any unwarranted 13. Ghildiyal S, Gautam MK, Joshi VK and Goel RK:
complications. Pharmacological evaluation of extracts of Hedychium
spicatum (Ham-ex-Smith) rhizome. Ancient Sci Life 2012;
31: 117-22.
ACKNOWLEDGEMENT: Authors are highly 14. Raj AGR, Shailaja U, Rao PN, Pujar MP, Srilakshmi and
grateful to the constant support and guidance of Dr. Mohan K: Effectiveness of Ayurveda treatment modalities
Prasanna N. Rao, Principal, SDM College of in the management of spasticity in children with Cerebral
Palsy at a tertiary care teaching hospital of Southern India.
Ayurveda, Hassan and Dr. Mallika KJ, Research Int. J. Res. Ayurveda Pharm. 2018; 9(2): 96-100.
Dean, SDM College of Ayurveda, Hassan. The 15. Arun K, Tapas BT, Shivakumar and Raj AGR:
Authors thank Dr. B. Ravishankar, Director, SDM Randomized controlled clinical trial to assess the
effectiveness of Haridradi tablet and Navaka guggulu
Centre for Research in Ayurveda and Allied tablet in the management of obesity. Int. J. Res. Ayurveda
Sciences for providing the facilities and guidance. Pharm. 2016; 7(Suppl 1): 70-75.
16. Raj AGR, Shailaja U, N Rao PN and Ajayan S: Review on
CONFLICT OF INTEREST: Nil the concept of immunomodulation in Ayurveda with
special emphasis on Prakara Yoga. Int J Pharm Sci Res
2014; 5(4): 1116-1123.
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How to cite this article:

Sivaraj S, Mallannavar V, Raj GRA and Shailaja U: Standardization of Amritaprasha ghrita: a herbal ghee based medicinal preparation. Int
J Pharm Sci & Res 2018; 9(11): 4842-48. doi: 10.13040/IJPSR.0975-8232.9(11).4842-48.
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