Winter cherry: Ashwagandha; Withania somnifera

Introduction
Common Name: Ashwagandha Plant
Chemical Constituents
Cultivation
Aeroponic system for cultivation
YIELD
Ashwagandha as Medicinal Herb
Ashwagandha Side Effects
Ashwagandha Benefits
Economics
Recent research
Reference

Introduction
Ashwagandha (Withania somnifera), also known as Indian ginseng, and as Indian Winter Cherry is an important ancient plant, the roots of which have been employed in Indian traditional systems of medicine, Ayurveda and Unani. It grows in dry parts in sub-tropical regions. Rajasthan, Punjab, Haryana, Uttar Pradesh, Gujarat, Maharashtra and Madhya Pradesh are the major Ashwagandha producing states of the country. The estimated production of Ashwagandha roots in India is more than 1500 tonnes and the annual requirement is about 7000 tonnes necessitating the increase in its cultivation and higher production.
Ashwagandha, the Indian ginseng or winter cherry has been used as a quiet valuable herb in the Ayurvedic and indigenous medical system for over 4000 years. The roots, leaves and fruits (berry) possess tremendous medicinal value. A famous Ayurvedic rejuvenative botanical used in many tonics and formulas, Ashwagandha is the best rejuvenation that helps maintain proper nourishment of the tissues, particularly muscle and bones, while supporting the proper function of the adrenals and reproductive system.

Common Name
English - Winter cherry
Latin - WITHANIA somnifera
Sanskrit - Ashwagandha
Hindi - Asgandh
Tamil - Asuragandhi, Amukkira
Kannada - Keramaddinagaddi
Telugu - Vajigandha, Pennerugadda
Malayalam- Amukkuram, Trittavu.
Marathi - Askandha
Marathi - Asgundh, Kanchuki, Askandha
Bengali - Ashvagandh
Punjabi - Asgand
Urdu - Asgandanagaori
Chinese - 睡茄 (Shui Qie)

Ashwagandha Plant
Ashwagandha is an erect branched under shrub up to 1.25 m in height, minutely stellate tomentose. Root fleshy, tapering, whitish brown. Leaves ovate, flower greenish. It grows in dried parts in subtropical regions.
Stem: Ashwagandha stem is terate, branched, cylindrical, solid, clothed with mealy, stellate-hoarytomentun
Root: Ashwagandha roots are straight, um-branched, thickness varying with age. Its roots bear fiber like secondary roots, outer surface buff to gray yellow with longitudinal wrinkles. Stem bases variously thickened; nodes prominent only on the side from where petiole arises, cylindrical, green with longitudinal wrinkles; fracture, short and uneven. Roots odour is characteristic; bitter and acrid. The roots when dry are cylindrical, gradually tapering down with a brownish white surface and pure white inside when broken
Leaves: Ashwagandha leaves are cauline and ramal, simple, exstipulate, petiolate, ovate, acute, entire and up to 10 cm long. Petioles up to 1.25 cm long.
Flowers: Ashwagandha flowers are ebracteate, pedicellate, complete, hermaphrodite, pentamerous, actinomorphic and hypogenous, gamosepalous, 4-6 mm in diameter, lucid-yellow or greenish. Its flowering time is in Winter
Fruits: Ashwagandha fruits a berry enclosed in the green persistent calyx, 5 mm in diameter, smooth, more or less globose, green when unripe, orange-red coloured in ripening stage
Seeds: Ashwagandha seeds are bean shaped, endospermic, yellow and orange-red coloured, somewhat scurfy.

Chemical Constituents
The methanol, hexane and diethyl ether extracts from both leaves and roots of Ashwagandha were found. Alkaloid percentage in roots ranges from 0.13 to 0.31%. The roots of Withania somnifera are alterative, aphrodisiac, deobstruent, diuretic, narcotic, sedative and restorative in nature. The pharmacological activity of the root is attributed to the alkaloids and steroidal lactones. The total alkaloid content in the roots of Indian types has been reported to vary between 0.13 and 0.3, though much high yields (up to 4.3 per cent) have been recorded elsewhere. Many bio-chemical heterogeneous alkaloids, including choline, tropanol, pseudotopanol, cuscokygrene, 3- tigioyloxytropana, isopelletierine and several other steroidal lactories. Twelve alkaloids, 35 withanolides and several sitoindosides have been isolated from the roots of the plant have been studied.
A sitoindoside is a biologically active constituent known as withanolide containing a glucose molecule at carbon 27. Indian ginseng’s pharmacological activity has been attributed to two main withanolides, withaferin A and withanolide D. Withaferin-A is therapeutically active withanolide reported to be present in leaves. In addition to alkaloids, the roots are reported to contain starch, reducing sugars, glycosides, dulcitol, withancil, an acid and a neutral compound. The amino acids reported from the roots include aspartic acid, glycine, tyrosine, alanine, glutamic acid and cysteine.

Cultivation
W.somnifera grows well in sandy loam or light red soil, having pH 7.5-8.0 with good drainage. It can be cultivated between 600-1200 m altitudes. The semi-tropical areas receiving 500-750 mm rainfall are suitable for cultivation of this rained crop. The crop requires dry season during its growing period. Temperature between 200C to 350C is most suitable for cultivation. Late winter rains are conducive for the proper development of the plant roots

LAND PREPARATION
Ashwagandha is usually grown in fields which are not well covered by the irrigation systems. The field on which food crops cannot be taken profitably for the above reason may be used for Ashwangandha cultivation. The soil of the field selected for Ashwagandha cultivation is well pulverized by ploughing, disking and/or harrowing. The field may be then levelled by the application pata.

PLANTING
The crop can be sown either by broad casting or in lines. Live to line method is preferred as it increases root production and also helps in performing intercultural practices properly. The seeds are usually sown about 1-3 cm deep in June- July in nursery. A light shower after shower after sowing ensures good germination. About 500-750 gm seeds are sufficient for 1 ha. Field. Seeds can be treated, with Thiram or Indofil or Dithane medicinal plants - 45 (@ 3 gm/kg seed), before sowing to protect seedlings from seed borne diseases. The seedling after 25-35 days after sowing can be transplanted in the field marinating 60 x 60 cm. Spacing between the plants & the rows. It may be noted that since 'Ashwagandha' is a rainy season Kharif crop, the time of sowing is decided by date of arrival of monsoon in that area.

THINNING AND WEEDING
The seeds sown by broadcasting or in the line in furrows should be thinned out by hand at 25-30 days after sowing to maintain a plant population of about 30-60 plants per square meter (about 3.5 to 6 lakh plants/hectare). The plant density to be used may depend on the nature and fertility of the soil. On the marginal land the population is kept high. If some fertiliser (N: P: K: 20:20:0) is applied then the population should preferably be kept at a lower level. One hand weeding at an early stage is sufficient to enable the Ashwagandha plants to take over the growth of weed which get suppressed by its smothering effect.

MANURES, FERTILISERS AND PESTICIDES
The medicinal plants have to be grown without chemical fertilizers and use of pesticides. Organic manures like, Farm Yard Manure (FYM), Vermi-Compost, Green Manure etc. may be used as per requirement of the species. To prevent diseases, bio-pesticides could be prepared (either single or mixture) from Neem (kernel, seeds & leaves), Chitrakmool, Dhatura, Cow's urine etc.
Withania somnifera is prone to several pests and diseases. Leaf spot disease caused by Alternaria alternata is the most prevalent disease, which is most severe in the plains of Punjab, Haryana, and Himachal Pradesh. In some cases insects or mite infestations are noticed. To spray pest repellents such as roger or nuvan 3 per cent diluted in one litre of water three times a week to control this infestation.
Damping off is a major disease in Withania somnifera at seedling stage and results in heavy seedling mortality under field condition. However, it can be controlled by application of Dithane M-45 (0.3%) as foliar spray.

IRRIGATION
Light shower after transplantation ensures establishment of seedlings. There is no need of irrigation if rainfall is at regular intervals. Excessive rainfall/water is harmful to the crop. Life saving irrigations may be applied, if required.

HARVESTING/ POST HARVESTING
The plants start flowering and bearing fruits from December onwards. The crop is ready for harvest in January- March at 150 to 180 days after sowing. The maturity of crop is judged by drying out of leaves and yellow red berries. The entire plant is uprooted for roots which are separated from aerial parts by cutting the stem 1-2 cm above the crown. The roots are then either cut transversely into small pieces (7 to 10 cm) or dried as it is in the sun. About 650-800 kg roots can be obtained from 1 ha on drying it comes to 350-435 kg. Berries are hand plucked separately. They are dried and crushed to take out the seeds.
The dried roots, entire or transversely cut into smaller pieces, have to be further cleaned, trimmed and graded. The roots are beaten with a club which removes adhering soil and breaks off the thin, brittle lateral rootlets. Lateral branches, root crown and stem remains on roots are carefully trimmed with the help of knife. Roots are collected 6 months after the sowing of crop.

Aeroponic system for cultivation
Ashwagandha traditionally grown outdoors in soil, the UA team decided to use an entirely non-traditional method - Aeroponic - to produce bulk amounts of withaferin A needed for biological evaluation. In Aeroponic, plants are set over enclosed chambers where their suspended roots are misted with water and nutrients, instead of growing in soil. The Withania plants grew about five times larger using this method than if they had been grown in soil.

"Using the Aeroponic system for cultivation, we were able to produce more than 20 grams of withaferin A in a single greenhouse. It normally costs around $195 for just 10 milligrams," Gunatilaka said. "Also, it usually takes two to three years to mature to sizeable roots to be commercially viable, but here it takes just six to nine months." Not only did the Aeroponic method yield bigger plants faster, with more withaferin A than usual, it also unexpectedly stimulated the plants to produce large amounts of the new natural product - a water-soluble sulphate form of withaferin A.

Upon testing, this new form demonstrated the same bioactivity as withaferin A. It was able to inhibit the proliferation and survival of tumour cells, disrupt tumour formation and induce the healthy cells' heat-shock response to reduce stress and increase survival, according to the researchers.

The difference is that the sulphate form of withaferin A is slower acting and water-soluble; it can be converted to withaferin A in cell culture media. The researchers, expecting that this withaferin A form will convert to its active form when metabolized in the body, are pursuing further testing in animal models. The patent will be held by the UA and the Massachusetts Institute of Technology.

YIELD
On an average, the yield from 1 hectare of commercial cultivation is approximately 3 to 5 q. / of dry roots and 50 to 75 kg of seeds. A maximum yield can be procured up to 6.5 to 7.0 q/ha. There are instances where farmers have achieved root yields as high as 1 tonne. Commercially, 6 to 15 mm diameter and 7 to 10 cm length root species are better. Alkaloid percentage in roots ranges from 0.13 to 0.31%. And an average yield from one hectare land under commercial cultivation is approx 3-5 quintals of dried roots and 50-75 kg seeds.

Ashwagandha as Medicinal Herb:-Ashwagandha is considered to be one of the best rejuvenating agents in Ayurveda. Its roots, seeds and leaves are used in Ayurvedic and Unani medicines. Ashwagandha root drug finds an important place in treatment of rheumatic pain, inflammation of joints, nervous disorders and epilepsy. Dried roots are used as tonic for hiccup, cold, cough, female disorders, as a sedative, in care of senile debility, ulcers, etc. Leaves are applied for carbuncles, inflammation and swellings. Leaf juice is useful in conjunctivitis. Bark decoction is taken for asthma and applied locally to bed sores. Ashwagandha and its extracts are used in preparation of herbal tea, powders, tablets and syrups.

Ashwagandha has anti-inflammatory, anti-tumour, anti-stress, antioxidant, mind-boosting, immune-enhancing, and rejuvenating properties. Ashwagandha root has also been noted to have sex-enhancing properties. Ashwagandha is mentioned in the ancient Kama Sutra as an herb to be used for heightening sexual experience. Ashwagandha has the ability to restore sexual health and improve overall vitality while promoting a calm state of mind. A 2002 laboratory study indicates Ashwagandha stimulates the growth of axons and dendrites. A 2001 study in rodents showed Ashwagandha had memory boosting ability. A 2000 study with rodents showed Ashwagandha to have anti-anxiety and anti-depression effects.

The plant has been used as an aphrodisiac, liver tonic, anti-inflammatory agent, and more recently to treat asthma, ulcers, insomnia, and senile dementia. Clinical trials and animal research support the use of Ashwagandha for anxiety, cognitive and neurological disorders, inflammation, and Parkinson's disease. Incorporation of Ashwagandha in the diet may prevent or decrease the growth of tumours in human.

It helps in providing progressive, long lasting results for various health concerns like aging, anaemia and slow growth, arthritis, fatigue, waning memory, sports fitness and stress-disorders. Pharmacological studies and research so far have indicated that Ashwagandha has anti-tumour, anti-stress, antioxidant boosting, haemopoeitic and rejuvenating properties. It is also an exceptional nerve tonic and nourishes the nerves and improves nerve function to maintain calm during stressful conditions. It also nourishes crucial mind and body connection and psychological immune response.

Ashwagandha Side Effects:-Ashwagandha does not have any significant side effects reported in the medical literature. Safety in pregnancy has not been fully established for Ashwagandha.

Ashwagandha Benefits
Ashwagandha benefits all parts of the body and can be used as a tonic or in oral form. Several studies have shown that Ashwagandha is useful in addressing the following health problems:
1) Osteoarthritis: A study in 2008, scientists tested Ashwagandha effects on human cartilage and found that the herb may help protect against inflammation and cartilage damage associated with osteoarthritis.

2) Anxiety: In an animal-based study published in 2000, researchers found that Ashwagandha had an anti-anxiety effect similar to that of lorazepam (a medication used to treat anxiety disorders). The herb also appeared to ease depression.

3) Type 2 Diabetes: Ashwagandha may help normalize high blood sugar and improve insulin sensitivity, according to preliminary, animal-based research published in 2008.

4) Cancer: In a 2003 study, tests on human tumour cell lines revealed that Ashwagandha may slow the growth of lung, breast, and colon cancer cells. Published in 2007, another study on human cells shows that Ashwagandha may inhibit tumour growth without harming normal cells.

5) Anti-Oxidant: Ashwagandha used as an anti-oxidant, as studies have shown that it can eliminate free radicals from your immune system. Free radicals are the agents that cause the breakdown of your body’s tissue, alternatively known as aging.

6).Provide energy: Studies show that supplementing with Ashwagandha can provide the energy needed to get through long workouts while also allowing for maximum recovery and cell re-growth.

7) General tonic: Ashwagandha is a tonic, which increases sperm count and sexual potency. In the rural areas vegetable made out of this plant is given to tuberculosis patients. It also increases the iron content in the blood.

Economics
Unit cost: The unit cost for development of 1 ha transplanted Ashwagandha cultivation is Rs.22,400/-. The details of the income assumed are also presented in the given table.
Income (Rs. /ha)
From roots 35000
From seeds 3600
Total gross income 38600
Cost 22400
Net income 16200

The estimated production of Ashwagandha roots in India is more than 1,500 tonnes and the annual requirement is about 7,000 tonnes necessitating the increase in its cultivation and higher production. The most important trade centre of the country for availability of Ashwagandha is Nimach Mandi in Madhya Pradesh and other trade centres
are Bombay, Calcutta, Delhi and Amritsar. Market for medicinal plants is volatile and the economics may vary.

HELP: For details on cultivation and marketing details some of the following institutes can be contacted:
* National Medicinal Plants Board, Department of AYUSH, Ministry of Health & Family Welfare, Government of India, Chandralok Building, 36, Janpath New Delhi - 110001, Phone: 011 - 23319255, email: nmpbindia1@indiatimes.com
* Central Institute of Medicinal and Aromatic Plants, P.O. - CIMAP, Near Kukrail Picnic Spot, Lucknow - 226 015, Indiaphone: 0522-2359623 E-mail: director@cimap.res.in
* Medicinal Plants Unit, Horticultural College and Research Institute, Tamil Nadu Agricultural University (TNAU), Coimbatore: 641 003, phone: 0422-5511365, email: herbs@tnau.ac.in
* Aromatic and Medicinal Plants Research Station, Odakkali, Asamanoor post-683549, Ernakulam district, phone: 0484-2658221, email: amprs@satyam.net.in
* Gandhi Krishi Vigyan Kendra, University of Agricultural Science, Bangalore-560065, phone: 080-55315598, email: vasunuthan@rediffmail.com
* Central Institute of Medicinal and Aromatic plants, Allalasandra, GKVK post, Bangalore- 560 065, phone: 080-28460563.
* National Research Centre for Medicinal and Aromatic plants, Boriavi-387 310, Anand, Gujarat, Phone: 0268-2578602, or 0268-2578644, email: nrcmap@wilnetonline.net
* College of Agriculture, Medicinal and Aromatic plants division, Mandsaur-458001, Madhya Pradesh., Phone: 07422- 242289.

Recent research
New anti-cancer drug from Ashwagandha University of Arizona scientists have used a new quick-growing technique to produce a water-soluble form of withaferin A compound that combats cancer and encourages the survival of healthy cells. Research trials are under way on this sulphate form of withaferin A, which could develop into a new anti-cancer drug. Scientifically studied since the 1960s, withaferin A reduces tumour mass by preventing the growth of blood vessels that make a tumour malignant.

The compound is derived from the roots of a winter cherry plant, the extracts of which have been used for more than 3,000 years in India as a general tonic to build stamina, improve mental concentration, relieve stress and enhance health. "Finding a water-soluble analog of withaferin A is significant, especially if it turns out to be a clinically useful drug," said Leslie Gunatilaka, director 

Products Research and Commercialization
Recent studies on benefits of Ashwagandha herb reports, how it is effective in the treatment of bone cancer, nerve problems, rheumatism, diabetes, bipolar disease, constipation and impotency. Research of scholars from Banaras Hindu University shows that, elements of Ashwagandha acts as an antioxidant, helps to increase memory power, relieve inflammation, stress and anxiety.
Ashwagandha Cure Alzheimer's: New research has revealed this herb may also fight off the devastating effects of Alzheimer's disease. Researchers at the National Brain Research Centre (NBRC) have conducted studies on mice that suggest Ashwagandha extract may reverse memory loss and improve cognitive abilities in those with the disease. Initially, mice with Alzheimer's were unable to learn or retain what they learned, but after receiving Ashwagandha for 20 days, this improved significantly. After 30 days, the behaviour of the mice returned to normal.

Other research work found that
* Free radical scavenging activity of Ashwagandha root powder was found in 15 days of experimental studies on rats.
* Liver and kidney protective roles of Ashwagandha were proved in metal-induced toxicity in experimental studies on mice in Indore.
* Ashwagandha tablets improved the physical and mental health of pre-school children in clinical trials in Chennai.
* In a reported study, this herb was given to 30 mental patients suffering from anxiety neurosis in doses of 40
ml/day. (In two equally divided doses.) For one month. At the end of the month, most of the anxiety disorders, panic attacks and similar mood phobias, had disappeared.

Reference
1. Ashwagandha About Herbs. New York: Memorial Sloan-Kettering Cancer Centre.
2. Ahmad, M. K.; Mahdi, A. A.; Shukla, K. K.; Islam, N.; Rajender, S.; Madhukar, D.; Shankhwar, S. N.; Ahmad, S. (2010). Withania somnifera
3. Mirjalili, M. H.; Moyano, E.;Bonfill, M.; Cusido, R. M.; Palazón, J. (2009). "Steroidal Lactones from Withania somnifera, an Ancient Plant for Novel Medicine".
4. Abraham, A., I. Kirson, E. Glotter and D. Lavie.1968. A chemo taxonomic study of Withania somnifera
5. Ven Murthy, M. R.; Ranjekar, P. K.; Ramassamy, C.; Deshpande, M. (2010). "Scientific Basis for the Use of Indian Ayurvedic Medicinal Plants in the Treatment of Neurodegenerative Disorders: Ashwagandha".
6. Van Der Hooft, C. S.; Hoekstra, A.; Winter, A.;De Smet, P. A.; Stricker, B. H. (2005). "Thyrotoxicosis following
the Use of Ashwagandha"
7. Sandhya Singh, Sushil Kumar: Withania somnifera: the Indian ginseng Ashwagandha, Central Institute of Medicinal and Aromatic Plants, 1998.
8. Dhalla, N.S., K.C. Gupta, N.S. Sastry and C.L. Malhotra: "Comparative studies of Withania somnifera Dunal and Withania Ashwagandha Kaul"
9. Archana, R., Namasivayam, A., 1999. Anti stress effect of Withania somnifera
10. Davis, L., Kuttan, G., 2000, Immunomodulatory activity of Withania somnifera
11.Dhalla N.S., Sastry, M. S. and Malhotra , C. L. 1961. Chemical studies of Withania somnifera -J. Pharm. Sc.

CAPSULES: DEFINITION TYPES AND STANDARD TESTS:


CAPSULES:  A small container, A pill in the form of a small rounded gelatinous container with medicine inside



 DEFINITION TYPES AND STANDARD TESTS:



Capsules are solid dosage forms in which the drug or a mixture of drugs is enclosed in hard gelatin capsule shells in soft soluble shells or gelatin or in hard or soft shell of any other suitable material of various shapes and capacities.



Following are the Types of Capsules:



1. Hard Capsules. 2. Soft Capsules. 3. Modified release Capsules. 4. Enteric Capsules.



Empty Gelatin Capsules Sizes.

HARD GELATIN CAPSULES: - These sizes are designated by numbers.


1. 000(Triple Zero) --- Biggest Size

2. 00(Double Zero) 
3. 0(Zero)
4. 1(One)
5. 2(Two)
6. 3(Three)
7. 4(Four)
8. 5(Five) --- Smallest Size 


SOFT GELATIN CAPSULES: These are classified depending upon the shapes, sizes and capacities.



The numbers represents capacities in minims.



1. Round - 1, 2, 3, 4, 5, 6, 7, 8, 9, 28, 40T, 80T and 90.

2. Oval - 1, 2, 3, 4, 5, 6, 7.5, 10, 12, 16, 20, 40, 60, 80, 85 and 110. 
3. Oblong - 3, 4, 5, 6, 8, 9.5, 11, 14, 16, 20, 90 and 360.
4. Tube - 5, 6, 8, 17.5, 30A, 30B, 35, 45, 55, 65, 90, 160, 250, 320 and 480. 
5. Misc. - 6, 17, 30, 35, 60 and 80. 


CAPSULES: Standards and Limits:



1. Description: It should comply with Specifications of Product.



2. Content of Active Ingredients.

Limit: 90 to 110% of label claim. 


3. Uniformity of Weight.

Average weight of Capsules content Less than 300 mg
Percentage Deviation 10%
Average weight of Capsules content 300 mg and more
Percentage Deviation 7.50%


4. Disintegration Test.

Hard Capsules - Disintegration time shall not be more than 30 minutes.
Soft Capsules - Disintegration time shall not be more than 60 minutes.


Enteric Capsules - Acidic Medium – Shall not disintegrate for 2 hours and in alkaline 

Medium capsules shall disintegrate within 30 minutes.

PHARMACOGNOSY : PHYSICAL METHODS OF EVALUATION OF CRUDE DRUGS

PHYSICAL METHODS OF EVALUATION OF CRUDE DRUGS

1. DETERMINATION OF MOISTURE CONTENT:
The moisture content can be determined by heating the drug in oven at 105°C to a constant weight. Toluene Distillation method is used for determining the Moisture content of drug those contains Volatile active constituents.

2. DETERMINATION OF VISCOSITY: METHOD BY USING OSTWALD VISCOMETER:
Procedure: Fill the Viscometer, Previously washed with chromic acid E and five times with distilled water with the liquid being examined through tube L to slightly above the mark G, using a long pipette to minimize wetting the tube above the mark. Place the tube vertically in water bath which temperature is maintained at the temperature at which viscosity is to be measured. When the temperature of liquid in tube attends, adjust the volume of liquid so that the bottom of the meniscus set at the mark G. Attach the rubber tube to E end of the tube and suck the liquid 5mm above the point E.

After releasing the suction, Measure the time required for the bottom of the meniscus to fall from the top edge of mark E to the top edge of mark F.

Kinematic Viscosity = Kt. (Unit is Sq.mm per second)
Dynamic Viscosity = KPt (Unit is Millipascal second Symbol – mPa S.)

Where, K is constant of instrument, which P determined by using Liquid of known viscosity.

t is time in second for the meniscus to fall from E to F.
P is weight per Ml of liquid in g/Cubic cm.

Viscosity can be measured by using other viscometers like Brookfield Viscometer, which is more accurate.

3. DETERMINATION OF MELTING POINT:
Pure chemicals and Photochemical are having sharp melting points. Crude drugs from animal and plant origin contain the mixed chemicals; they are described with certain range of melting points.

4. DETERMINATION OF ACID VALUES:
Definition: It is defined as the number which expresses in mg the amount of Potassium Hydroxide (KOH) necessary to neutralize the free acids presents in the 1g of the sample. Weight accurately 10g of sample and dissolve it in 50ml of a mixture of Ethanol 95% (25 ML) and Ether (25 ML) previously neutralized with 0.1M Potassium Hydroxide to Phenolphthalein solution. If required dissolve the sample by heating slowly and using reflex condense. To the above sample solution add 1ml of Phenolphthalein indicator and titrate with 0.1M Potassium Hydroxide solution up to the end point pink after shaking for half minute.
Acid value = 5.61n/w

Where n – ml of 0.1 M Potassium Hydroxide solution required.
w – Weight of sample in g.

5. DETERMINATION OF VALUE:
Definition: It is defined as the number of milligrams of Potassium Hydroxide required to saponify the Esters in the 1g of sample substance. It is calculated by subtracting the Saponification value from the Acid value of sample substance.

6. DETERMINATION OF SAPONIFICATION VALUE:
It is defined as the number of milligrams of Potassium Hydroxide required for neutralization of free acid and to saponify the esters present of 1g of sample.

Weight accurately 2g of sample and add it to 200ml glass flask fitted with reflex condenser. Add 25ml of 0.5 M Ethanolic potassium hydroxide and small quantity of pumice powder and boil for 30 min on water bath under reflux. Add 1ml Phenolphthalein solution and titrate immediately with 0.5 M hydrochloric acid (x). Take the blank reading by repeating the same procedure (y). Calculate the saponification values as:
Saponification value = 28.05 (y-x) / w

Where, w - Weight of substance in g.

7. DETERMINATION OF IODINE VALUES:
Iodine value is defined as the number which expresses in grams the quantity of halogen, calculated as iodine, which is absorbed by 100g of the substance under the described condition.
Add accurately weighed quantity of sample in dry 500ml Iodine flask, add 10ml of carbon tetrachloride and dissolve. Add 20 ml of Iodine Monochloride solution fix the stopper and allow it to stand for 30 minute between temperatures 15 to 25°C in dark place. Add 15ml of Potassium Iodide solution in cup top and carefully remove the stopper, rinse the stopper and side of the flask with 100ml of water, mix by shaking and titrate with 0.1 M sodium Thiosulphate solution using Starch solution as indicator towards the end of titration. Note the Burette reading (x) and repeat the same procedure for blank titration and record the Burette reading (y).

Iodine Value = 1.269 (y-x) / w
Where, w is weight of sample in g.

8. DETERMINATION OF ETHANOL SOLUBLE EXTRACTIVE:
Prepare coarse powder of air dried drug. Take 100ml of Ethanol (Specified Strength) in conical flask. Macerate 5g of powdered drug in above conical flask, close and conical flask for 24 hours. Shake the flask frequently during first 6 hours; allow it to stand for 18 hours. Filter rapidly taking precaution against loss of ethanol evaporate 25ml of the filtrate to dryness in a tared flat bottomed shallow dish. Dry at 105°C and weight it. Calculate the % of Ethanol soluble extractive with reference to the air dried drug.


9. DETERMINATION OF WATER SOLUBLE EXTRACTIVE:
Follow the procedure described under Ethanol Soluble Extractive. Use Chloroform water instead of ethanol.

10. DETERMINATION OF ASH FROM CRUDE VEGETABLE DRUGS:
Weight accurately 2 to 3g of air dried crude drug in a treated silica dish and incinerate at a temperature not exceeding 450°C until free from carbon. Cool the Silica dish and weight. If a carbon free ash can not be obtained. Exhaust the charred mass with hot water. Collect the residue on an ash less filter paper, until the ash in white or nearly white, add the filtrate evaporate to dryness and ignite at a temperature not exceeding 450°C. Calculate the % of ash with reference to the air dried drug.

11. DETERMINATION OF REFRACTIVE INDEX:
The instrument used to measure the refractive index if liquids are called refractometer. Refractive index is defined as the ratio of the sin of the angles of incidence and refraction.

n = Sin i
Sin r
Where, n is refractive index,
Sin i – Sine of angle of incidence and
Sin r – Sine of angle of refraction.

RI is useful for measurement of Molecular Formula. It is measured at 20°C ± 5°C temperature.

The Abbe refractometer is used for accurate measurements of RI. It consists of Abbe’s prism, Telescope, Compensation Prism and Sector.

Wash and dry the prisms, place few drops of liquids on surface of lower prism and close the prisms. Focus the eyepiece by sliding it up and down until cross hairs are in sharp focus. Turn the compensator until coloured fringes disappear. Rotate the indicator so that the borderline coincides with the intersection of the cross hairs. Note the reading. Repeat the same experiments for 2 to 3 times. Mean of these repeated readings gives the refractive index.

TYPE OF WATER

TYPE OF WATER

These are several types of waters depending upon their Quality and Purpose of use. Only some types of waters and defined here for understanding the terminologies.

1. Drinking or Potable Water:
Means water that is intended for human consumption and may be sealed in a bottle or other containers with no added ingredients. Drinking water shall comply with the standard specified by Bureau of Indian Standard or WHO Drinking Water Standard. Or US potable Water Standard 40 CFR 141 or such other standards. Indian Pharmacopoeia has given 500 CFU/ml limit for Total Aerobic Microbial Count.

2. Purified Water (IP):
Water prepared by distillation, by means of ion exchange or by any other appropriate means from suitable potable water that complies all relevant statutory regulations. Purified water shall comply with standards given in Monographs of Official Books and Pharmacopoeia – (IP, BP, USP, NF, and EP Etc).10 CFU/ml Limit for microbial Population may be considered for Purified Water.

3. Water for Injection (IP):
It is apyrogenic distilled water intended for use in the preparation of medicines for parenteral administration. When water is used as a vehicle (water for injection in bulk) and for dissolving or diluting substances and preparation for injections.

WFI is obtained by distillation potable water or purified water from a neutral glass, quartz or suitable metal still with an effective device for preventing the entertainment of droplets. This still must be suitably maintained to ensure the production of apyrogenic water. The first portion of distillate is discarded and remainder is collected and stored in conditions designed to prevent the growth of micro-organism and to avoid any other contamination. It should comply with the standards given in the monograph of Official books and Pharmacopoeias.

4. Sterile Water for Injection (IP):
Sterile Water for Injection is water for injection distributed in suitable containers or glass of other material sealed and sterilized by heat under conditions that ensures that the water remains apyrogenic. Each container contains a sufficient amount of WFI to permit the withdrawal of the nominal volume. It should comply with the standards given in the monograph of Official books or Pharmacopoeias.

Recommended microbial limit is not more than 10 organisms per 100 ml and Pathogens should be absent.

STORAGE CONDITIONS AS PER IP Indian Pharmacopoeia,

DEFINITIONS OF STORAGE CONDITIONS AS PER IP 96;
Indian Pharmacopoeia 1996

1. COLD: Any temperature not exceeding 8°C and usually between 2°C to 8°C. A refrigerator is a cold place in which the temperature is maintained thermostatically between 2°C to 8°C.

2. COOL: Any temperature between 8°C and 25°C. An article, for which storage in a cool place is directed may alternately, be stored in a refrigerator unless otherwise specified in the individual monograph.

3. ROOM TEMPERATURE: The temperature prevailing in a working area.

4. WARM: Any temperature between 30°C and 40°C.

5. EXCESSIVE HEAT: Any temperature above 40°C.

6. LIGHT RESISTANT CONTAINERS: A light resistant container protect the content from the effect of actinic light by virtue of the specific properties of the material of which it is made.

7. WELL CLOSED CONTAINER: A well closed container products the contents from contamination by extraneous liquid and from loss of the article under normal condition of handling, shipment, storage and distribution.

8. TIGHTLY CLOSED CONTAINERS: A tightly closed container protects the contents from contamination by extraneous liquid and solids or Vapour from loss or deterioration of the article from effervescence, deliquescent or evaporation under normal condition of handling, shipment, storage and distribution

GRADES OF POWDERS


GRADES OF POWDERS: 

Grades (Types) of powders are expressed on degree of Coarseness and Fineness of powder.

1) Coarse Powder: A Powder, all the particles of which pass through a sieve with a nominal mesh aperture of 1,700 micron meter and not more than 40% by weight through a sieve with a nominal mesh aperture of 355 micron meter.

2) Moderately Coarse Powder: A Powder, all the particles of which pass through a sieve with a nominal mesh aperture of 710 micron meter and not more than 40% by weight through a sieve with a nominal mesh aperture of 250 micron meter.

3) Moderately Fine Powder: A Powder, all the particles of which pass through a sieve with a nominal mesh aperture of 355 micron meter and not more than 40% by weight through a sieve with a nominal mesh aperture of 180 micron meter.

4) Fine Powder: A Powder, all the particles of which pass through a sieve with a nominal mesh aperture of 180 micron meter and not more than 40% by weight through a sieve with a nominal mesh aperture of 125 micron meter.

5) Very Fine Powder: A Powder, all the particles of which pass through a sieve with a nominal mesh aperture of 125 micron meter and not more than 40% by weight through a sieve with a nominal mesh aperture of 45 micron meter.

6) Micro Fine Powder: A Powder, of which not less than 90% by weight of the particles pass through a sieve with a nominal mesh aperture of 45 micron meter.

7) Super Fine Powder: A Powder, of which not less than 90% by weight of the particles pass through a sieve with a nominal mesh aperture of 10 micron meter.

CULTIVATION OF MEDICINAL PLANTS


CULTIVATION OF MEDICINAL PLANTS

The cultivation technology of vegetable drugs involves convergence of various factors from agricultural and pharmaceutical sphere such as soil, climate, rain fall, irrigation, altitude, temperature, use of fertilizers and pesticides, genetic manipulation and biochemical aspects of natural drugs.

Cultivation ensures quality and purity of medicinal plants. It gives better yield and therapeutic effects. It ensures regular supply of drug. Cultivation increases industrialization and helps for unemployment problem.

Followings are the methods of cultivation:

1) Sexual Method (Seed Propagation): It this method plants are raised from seeds and such plants are called Seedlings.
Good Quality seeds of high germination rate should be used for cultivation. Seeds should be free from other seeds and impurities. Seeds should be checked for germination. Too old seeds should not be used for cultivation. Some chemical, Hormonal and special treatments should be given to seeds for healthy and accelerated growth of seedling.

2) Asexual Methods: Vegetative Part of plant, such as steam or root is placed in such an environment that develops into a new plant. Vigorous growth and new variety cannot be achieved from this method.

FACTORS AFFECTING THE CULTIVATION:

There are several factors that affect the cultivation directly or indirectly.
1) Altitude, Temperature and Humidity.
2) Rainfall or Irrigation.
3) Soil and Soil Fertility.

Types of Soils:
Depending upon the size of mineral matter following names is given to the soil:
Particle Size (Diameter)/ Types of Soil
Less than 0.002 mm/ Fine Clay
0.002 To 0.02 mm/ Coarse Clay or Silt
0.02 To 0.2 mm/ Fine Sand
0.2 To 2.0 mm/ Coarse Sand
Depending upon the percentage covered by clay, soils are classified as given below:

Type of Soil/ % Covered
1. Clay/ More than 50% of Clay
2. Loamy/ 30 to 50% of Clay
3. Silt Loam/ 20 to 30% of Clay
4. Sandy Loam/ 10 to 20% of Clay
5. Sandy Soil/ More than 70% Sandy Soil
6. Calcareous Soil/ More than 20% of Lime.

1) Fertilisers.
2) Pest and Pest Control: Fungi, Viruses, Insects and Weeds. Non-insects like rats, monkeys, birds, rabbits and hares, squirrels, deer, pig etc.

METHODS OF PEST CONTROL:
1) Mechanical Method: Destruction of pest, manually with the help of different devices.
2) Agricultural methods.
3) Biological methods.
4) Chemical method: Rodentisides, Insecticides, Acaricides, Fungicides, Herbicide etc.

PLANT GROWTH REGULATORS:
Plant growth regulators are organic compounds, other than nutrients which affects the morphological structure and or physiological process of plants in low concentration.
Following are the plant growth regulators:
1) Auxins.
2) Gibberellins.
3) Cytokinins.
4) Ethylene
5) Abscisic Acid (ABA)