R: Herbal Extract Pre Phytochemical screening:

Concept of standardization, extraction and pre phytochemical screening strategies for
herbal drug
Amita Pandey, Shalini Tripathi

Journal of Pharmacognosy and Phytochemistry 2014; 2 (5): 115-119

http://www.phytojournal.com/vol2Issue5/Issue_jan_2014/11.pdf

Pre Phytochemical screening: Phytochemical examinations were carried out for all the extracts as per the standard methods.

 1. Detection of alkaloids: Extracts were dissolved individually in dilute Hydrochloric acid and filtered.
 Mayer’s Test: Filtrates were treated with Mayer’s reagent (Potassium Mercuric Iodide). Formation of a yellow coloured precipitate indicates the presence of alkaloids.
 Wagner’s Test: Filtrates were treated with Wagner’s reagent (Iodine in Potassium Iodide). Formation of brown/reddish precipitate indicates the presence of alkaloids.
 Dragendroff’s Test: Filtrates were treated with Dragendroff’s reagent (solution of Potassium Bismuth Iodide). Formation of red precipitate indicates the presence of alkaloids.
 Hager’s Test: Filtrates were treated with Hager’s reagent (saturated picric acid solution). Presence of alkaloids confirmed by the formation of yellow coloured precipitate.

2. Detection of carbohydrates: Extracts were dissolved individually in 5 ml distilled water and filtered. The filtrates were used to test for the presence of carbohydrates.
 Molisch’s Test: Filtrates were treated with 2 drops of alcoholic α-naphthol solution in a test tube. Formation of the violet ring at the junction indicates the presence of Carbohydrates.
 Benedict’s test: Filtrates were treated with Benedict’s reagent and heated gently. Orange red precipitate indicates the presence of reducing sugars.
 Fehling’s Test: Filtrates were hydrolysed with dil. HCl, neutralized with alkali and heated with Fehling’s A & B solutions. Formation of red precipitate indicates the presence of reducing sugars.

 3. Detection of glycosides: Extracts were hydrolysed with dil. HCl, and then subjected to test for glycosides.
 Modified Borntrager’s Test: Extracts were treated with Ferric Chloride solution and immersed in boiling water for about 5 minutes. The mixture was cooled and extracted with equal volumes of benzene. The benzene layer was separated and treated with ammonia solution. Formation
of rose-pink colour in the ammoniacal layer indicates the
presence of anthranol glycosides.

4. Legal’s Test:  Extracts were treated with sodium nitroprusside in
pyridine and sodium hydroxide. Formation of pink to blood red
colour indicates the presence of cardiac glycosides.

5. Detection of saponins
 Froth Test: Extracts were diluted with distilled water to
20ml and this was shaken in a graduated cylinder for 15
minutes. Formation of 1 cm layer of foam indicates the
presence of saponins.
 Foam Test: 0.5 gm of extract was shaken with 2 ml of
water. If foam produced persists for ten minutes it
indicates the presence of saponins.

6. Detection of phytosterols
 Salkowski’s Test: Extracts were treated with chloroform
and filtered. The filtrates were treated with few drops of
Conc. Sulphuric acid, shaken and allowed to stand.
Appearance of golden yellow colour indicates the
presence of triterpenes.

 Liebermann Burchard test: Extracts were treated with
chloroform and filtered. The filtrates were treated with
few drops of acetic anhydride, boiled and cooled. Conc.
Sulphuric acid was added. Formation of brown ring at the
junction indicates the presence of phytosterols.

7. Detection of phenols
 Ferric Chloride Test: Extracts were treated with 3-4
drops of ferric chloride solution. Formation of bluish
black colour indicates the presence of phenols.

8. Detection of tannins
 Gelatin Test: To the extract, 1% gelatin solution
containing sodium chloride was added. Formation of
white precipitate indicates the presence of tannins.

9. Detection of flavonoids
 Alkaline Reagent Test: Extracts were treated with few
drops of sodium hydroxide solution. Formation of intense
yellow colour, which becomes colourless on addition of
dilute acid, indicates the presence of flavonoids.
 Lead acetate Test: Extracts were treated with few drops
of lead acetate solution. Formation of yellow colour
precipitate indicates the presence of Flavonoids.

10. Detection of proteins and amino acids
 Xanthoproteic Test: The extracts were treated with few
drops of conc. Nitric acid. Formation of yellow colour
indicates the presence of proteins.
 Ninhydrin Test: To the extract, 0.25% w/v Ninhydrin
reagent was added and boiled for few minutes. Formation
of blue colour indicates the presence of amino acid.

11. Detection of diterpenes
 Copper acetate Test: Extracts were dissolved in water
and treated with 3-4 drops of copper acetate solution.
Formation of emerald green colour indicates the presence
of diterpenes [19, 21, 22]

 References
1. Anonymous. The Indian Pharmacopoeia. Govt. of India
publication, New Delhi, 1966, 947-950.
2. Bimakr M. Comparison of different extraction methods
for the extraction of major bioactive flavonoid compounds
from spearmint (Mentha spicata L.) leaves. Food Bioprod
Process 2010; 1-6.
3. Cowan MM. Plant products as antimicrobial agents.
Clinical microbiology reviews 1999; 12(4):564-582.
4. Das K, Tiwari RKS, Shrivastava DK. Techniques for
evaluation of medicinal plant products as antimicrobial
agent: Current methods and future trends. Journal of
Medicinal Plants Research 2010; 4(2):104-111.
5. Eloff JN. Which extractant should be used for the
screening and isolation of antimicrobial components from
plants. Journal of Ethnopharmacology 1998; 60:1-8.
6. Evans WC. Treas and Evans “Pharmacognosy.” Edn 14th
,
WB Saunders company Ltd., 2002, 3-4
7. Handa SS, Khanuja SPS, Longo G, Rakesh DD.
Extraction Technologies for Medicinal and Aromatic
Plants. International centre for science and high
technology, Trieste, 2008, 21-25.
8. Johansen DA. Plant Micro technique. McGraw Hill Book
Co., New York, 1940, 182-197.
9. Kokate CK. Practical Pharmacognosy. Vallabh Prakashan,
New Delhi, 1991, 107-111.
10. Kumar R, Sharma RJ, Bairwa K, Roy RK, Kumar A.
Pharmacological review on natural antidiarrhoel agents.
Der Pharma Chemica 2010; 2(2):66-93.
11. Lapornik B, Prosek M, Wondra AG. Comparison of
extracts prepared from plant by-products using different
solvents and extraction time. Journal of Food Engineering
2005; 71:214-222.
12. Mali RG, Mahajan SG, Mehta AA. In-vitro anthelmintic
activity of stem bark of Mimusops elengi Linn.
Pharmacognosy Magazine 2007; 3(10):73-76.
13. Mute VM. Anthelmintic effect of Tamarind indica linn
leaves juice extract on Pheretima posthuma. International
journal of pharma research and development 2009; 7:1-6.
14. Ncube NS, Afolayan AJ, Okoh AI. Assessment techniques
of antimicrobial properties of natural compounds of plant
origin: current methods and future trends. African Journal
of Biotechnology 2008; 7(12):1797-1806.
15. Parekh J, Karathia N, Chanda S. Evaluation of
antibacterial activity and phytochemical analysis of
Bauhinia variegata L. bark. African Journal of
Biomedical Research 2006; 9:53-56.
16. Remington JP. Remington: The science and practice of
pharmacy. Edn 21, Lippincott Williams & Wilkins, 773-
774.
17. Roopashree TS, Dang R, Rani SRH, Narendra C.
Antibacterial activity of anti- psoriatic herbs: Cassia tora,
Momordica charantia and Calendula officinalis.
International Journal of Applied Research in Natural
Products 2008; 1(3):20-28.
18. Sutar N, Garai R, Sharma US, Sharma UK. Anthelmintic
activity of Platycladus orientalis leaves extract.
International Journal of Parasitology Research 2010;
2(2):1-3.
19. Sharma US, Sharma UK, Singh A, Sutar N, Singh PJ. In
vitro anthelmintic activity of Murraya koenigii linn.
Leaves extracts. International journal of pharma and bio
sciences 2010; 1(3):1-4.
20. Vidyadhar S, Saidulu M, Gopal TK, Chamundeeswari D,
Rao U, Banji D. In vitro anthelmintic activity of the whole
plant of Enicostemma littorale by using various extracts.
International journal of applied biology and
pharmaceutical technology 2010; 1(3):1119-1125.
21. Wallis TE. Text Book of Pharmacognosy. CBS publishers
and Distributors, Delhi, 1989, 356-549.
22. Wang GS, Han J, Zhao LW, Jiang DX, Liu YT, Liu XL.
Anthelmintic activity of steroidal saponins from Paris
polyphylla. Phytomedicine 2010; 17:1102-1105.
23. Wang GX. In vivo anthelmintic activity of five alkaloids
from Macleaya microcarpa (Maxim) Fedde against
Dactylogyrus intermedius in Carassius auratus.
Veterinary Parasitology 2010; 171:305-313.

.







standard strength  pharmacopeial fluid extracts

Pharmacists recognized the destructive influences of heat applied to extracts diminished the efficacy of a number of products that were concentrated through the process of evaporating the solvent (menstruum). This was true in the case of the pharmacopeial fluid extracts. After the drug (marc) was soaked (macerated) with an initial volume of menstruum for 48 hours, it was then allowed to slowly percolate through the marc. This first percolate solution was set aside as the reserve, usually about 4/5 of the final total volume. Further solvent was then percolated through the marc to exhaust it of its active components. However, this second extraction resulted in a dilute solution that required evaporation by heating to concentrate it to a volume that supplied the final 1/5 of the total. This was then added to the reserve in an attempt to achieve a standard strength of solution. Fluid extracts were thus processed so that each milliliter volume represented one gram dry weight of the drug extracted (a 1:1 extract). However, the heating of the second percolate and the mixing of this concentrate with the reserve led to many problems affecting quality.
http://www.freebookcentre.net/

Our Facebook group:
https://www.facebook.com/groups/ADruidHerbal/