6.1 OBJECTIVE - Shodhganga : a reservoir of Indian...

Chapter-6 Herbal formulations

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6.1 OBJECTIVE

To develop simple herbal formulations using the selected plants extract.

6.2 INTRODUCTION

Herbal formulations many times fails to gain the patient acceptability, hence an

attempt was made to increase the same by developing some simple formulations. The

developed formulations were subjected to standardization by using analytical

methods. Standardization of herbal medicine is important and a challenging task, for

this the author selected some marker compounds and quantitative determination

methods were developed to determine the same in final formulation.

6.3 LITERATURE REVIEW

According to World Health Organization (WHO) more than 80% of the world’s

population relies on traditional medicine for their primary healthcare needs. Use of

herbal medicines in Asia indicates a long history of human interactions with the

environment. Plants used as traditional medicine contain a wide range of substances

that can be used to treat chronic as well as infectious diseases. A vast knowledge of

how to use the plants against different illnesses is expected to have accumulated in

areas where the use of plants is still of great importance (Diallo et al, 1999). The

medicinal value of plants lies in some chemical substances that have a definite

physiological action on the human body. The most important of these bioactive

compounds of plants are alkaloids, flavanoids, tannins and phenolic compounds

(Edeoga et al, 2005).

Scoparia dulcis Linn. is reported as a traditional antidiabetic plant (Latha M et

al, 2004). Scoparia acid D isolated from Scoparia dulcis Linn. is reported for

antidiabetic activity (Latha M et al, 2009). Scoparia dulcis Linn. reported to contain

antidiabetic activity (Pari L et al, 2004). Scoparia dulcis Linn. ethanolic extract

reported to contain antidiabetic and antioxidant activity when studied on alloxan

induced albino mice (Abu Hasanat Md. Zulfiker et al, 2010).

The usage of medicinal plants as traditional medicines is well known in rural

areas of many developing countries. Traditional healers claim that their medicine is

cheaper and more effective than the modern medicine. In developing countries,


102

people of small isolated villages and native communities use folk medicine for the

treatment of common infections (Sandhu et al, 2005; Gupta et al, 2005).

Even though pharmaceutical industries have produced a number of new

antibiotics in the last three decades, resistance to these drugs by microorganisms has

increased. In general, bacteria have the genetic ability to transmit and acquire

resistance to drugs, which are utilized as therapeutic agents. Such a fact is cause for

concern, because of the number of patients in hospitals who have suppressed

immunity, due to new bacterial strains which are multi-resistant. Consequently, new

infections can occur in hospitals resulting in high mortality (Cohen, 1992).

Liquid oral formulations: The stability of the active ingredients in the final product

is of prime concern to the formulator. In general drug substance is less stable in

aqueous media than in the solid dosage form and it is important, therefore, to properly

stabilize and preserve, in particular those solutions, suspensions, emulsions that

contain water.

Liquid preparations for oral use are usually solutions, syrups, emulsions or

suspensions containing one or more active substances in a suitable vehicle. Some

preparations for oral use are prepared by dilution of concentrated liquid preparations,

or from powders or granules for the preparation of oral solutions or suspensions, for

oral drops or for syrups, using a suitable vehicle. The vehicle for any preparations for

oral use is chosen based on nature of the active substances and to provide organoleptic

characteristics appropriate to the intended use of the preparation.

Excipients may be defined as the constituents of a pharmaceutical dosage form

that are not the active substance; according to their function they are the stabilizers to

retain the physical and chemical property of dosage form (European Medicines

Agency 2006).

Liquid preparations for oral use may contain suitable excipients such as

antimicrobial preservatives, antioxidants, dispersing agent, suspending agent,

thickening agent, emulsifying agent, buffering agent, wetting agent, solublizing agent,

stabilizing agent, flavoring agent, sweetening agent and coloring matter those are

authorized by the competent authority. Several categories of preparations may be


103

distinguished as oral solutions, emulsions, suspensions, powders and granules for oral

solutions and suspensions, oral drops, powders for oral drops, syrups, powders and

granules for syrups (British pharmacopoeia, 2007).

Syrups: Syrups are aqueous preparations characterized by a sweet taste and a viscous

consistency. They may contain sucrose at a concentration of at least 45 per cent m/m.

The sweet taste can also be obtained by using other polyols or sweetening agents.

Syrups usually contain aromatic or other flavoring agents (British pharmacopoeia,

2007).

In addition to sucrose, certain other polyols such as glycerin or sorbitol may be

added to retard crystallization of sucrose or to increase the solubility of added

ingredients. Alcohol often is included as a preservative and also as a solvent for

flavors; further resistance to microbial attack can be enhanced by incorporating

antimicrobial agents. When the aqueous preparations contain some added medicinal

substance, the syrup is called as medicated syrup. Syrups contain remarkable masking

properties for bitter or saline drugs. Gycerrhiza syrup has been recommended for

disguising the salty taste of bromides, iodides, and chlorides. This has been attributed

to its colloidal character and its double sweetness of the glycyrrhizin. This syrup is

also of value in masking bitterness in preparations containing B complex vitamins

(Alfonso & Gennaro, 1997).

Acacia syrup because of its colloidal character is of great value as a particular

vehicle for masking disagreeable taste of many medicaments. Raspberry syrup BP

1088 is one of the most efficient flavoring agents, and is especially useful in masking

the taste of bitter drugs. A series of papers by Schumacher deals with improving the

palatability of bulk-compounded products using flavoring and sweetening agents

(Schumacher, 1967).

It is important that the concentration of sucrose approach but may not reach

the saturation point. In dilute solutions sucrose provides an excellent nutrient for

molds, yeasts and other microorganisms. In contractions of 65% by weight or more,

the solution will retard the growth of such micro organisms. However, a saturated

solution may lead to crystallization of a part of the sucrose under conditions of

changing temperature (Alfonso & Gennaro, 1997).


104

Sorbitol solution is the sweetening agent and contains 70% w/w of the total

solids, consisting mainly of d-sorbitol. It has about half the sweetening power of

syrup (Alfonso & Gennaro, 1997).

The effect of trace metals can be minimized by using citric acid or EDTA

which are sequestering agents. Antioxidants however, may retard or delay oxidation

by reacting with the free radicals formed in the product. Examples of antioxidants are

the propyl, octyl and dodecyl esters of Gallic acid, butylated hydroxyanisole (BHA)

and the tocopherols or vitamin E (Alfonso & Gennaro, 1997).

Preparation: Syrups are prepared in various ways; the choice of the proper method

depends on the physical and chemical characteristics of the substances entering into

the preparation.

Solution with heating: This is usual method for making syrups when the valuable

constituent is neither volatile nor injured by heat, and when it is desirable to make the

syrup rapidly. The sucrose usually is added to the purified water or aqueous solution

and heated until solution is affected, then it is strained and sufficient purified water

added to make the desired weight or volume (Alfonso & Gennaro, 1997).

Agitation without heating: This process is used in those cases where heat would

cause the loss of valuable, volatile constituents. Glass-lined tanks with mechanical

agitators, especially adapted to dissolving of sucrose, are used for making syrups in

large quantities (Alfonso & Gennaro, 1997).

In the manufacturing, packaging, storage and distribution of liquid

preparations for oral use, suitable measures are taken to ensure their microbial quality;

recommendations on this aspect are provided in the text on Microbiological quality of

pharmaceutical preparations (Alfonso & Gennaro, 1997).

Herbal drugs and herbal drug preparations present in the dosage form are not subject

to uniformity of dosage units (British pharmacopoeia, 2007).

Preservation: Syrups should be made in quantities that can be consumed within few

months, except in those cases where special facilities can be employed for their

preservation; a low temperature is the best method. Concentration with out

supersaturating is also a condition favorable to preservation. The USP states that the


105

syrups may contain preservatives, glycerin, methyl paraben, benzoic acid and sodium

benzoate which may prevent bacterial and mold growth. Combination of alkyl esters

of p-hydroxy benzoic acid are effective inhibitors of yeast that have been implicated

in the contamination of commercial syrups (United States Pharmacopeia, 2007).

Syrups can be preserved by a) storage at low temperature, b) adding preservative such

as glycerin, benzoic acid, sodium benzoate, methyl paraben or alcohol in the

formulation. High sucrose concentrations will usually protect an oral liquid dosage

form from growth of most microorganisms. A problem arises, however, when

pharmacist must add other ingredients to syrups that can result in a decrease in the

sucrose concentration. This may cause loss of the preservative effectiveness of the

sucrose. This can be overcome however by calculating the quantity of a preservative

(such as alcohol) to be added to the formula to maintain the preservative effectiveness

of the final product (Loyd V Allen, 2004).

General preservatives used:

Alcohols: Ethanol is useful as a preservative when it is used as a solvent, however, it

does need a relatively high concentration, some what greater than 10%, to be

effective. Too high a concentration may result incompatibilities in suspension and

emulsion systems. Propylene glycol is also used as a solvent in oral solutions and

topical preparations, and it can function as a preservative in the range of 15% to 30%.

It is not volatile like ethanol and is used frequently not only in solution but also in

suspensions and emulsions.

Acids: Benzoic acid has low solubility in water, about 0.34% at 25ºC, the

concentration range used for inhibitory action varies from 0.01 to 0.5%. Only the

ionized form is effective, and therefore its use is restricted to preparations with a pH

below 4.5. Sorbic acids also have a low solubility in water, 0.3% at 30ºC. Suitable

concentrations for preservative action are in the range of 0.05 to 2%. Its preservative

action is due to the non ionized form; consequently, it is only effective in acid media,

because of double bonds in its structure, it is subjected to oxidation.

Esters: Parabens are the esters of Para hydroxyl benzoic acid and include the methyl,

ethyl, propyl and butyl derivatives. The solubility in the water decreases as the

molecular weight increases from 0.25% for the methyl esters to 0.02% for the butyl


106

ester. These compounds are effective and stable over a pH range of 4-8; they are

applied at a concentration up to about 0.2%. Frequently two esters are used in

combinations.

Other excipients: Hydroxyethylcellulose (HEC) is nonionic cellulose ether that

readily dissolves in water to form viscous solutions. Chemically it is high purity

cellulose that has been etherified to form hydroxyethyl groups to give the desired

solubility in water. In solution the product acts as a thickener and rheology control

agent, as a protective colloid, binder, stabilizer and suspending agent (British

pharmacopoeia 2007).

Polymers such as hydroxypropylcellulose, hydroxyethylcellulose, sodium

carboxymethyl cellulose, hydroxypropyl methylcellulose, polyvinylpyrrolidone,

pectin, carrageenan and guar gum have wide application in the pharmaceutical

industry (Jian-Hwa Guo, 1998).

Sorbitol may have a mild laxative effect, Calorific value 2.6 kcal/g (European

commission; 2003), Sorbitol is a sugar which can be used in diabetic patients because

of its special properties. The slow absorption of sorbitol, and its conversion to fructose

and glycogen prior to its availability as glucose, has led to consideration of this

substance for use as an adjunct in diabetic diets (Charles R Shuman, 1956; Brian A.

Burt, 2006). Several reporters reported sorbitol also show side effects like diarrhoeia

(Chusid, 1981; Jain, 1985; Badiga, 1990).

Glycerol is used as humactant, it is a chemical compound that is also known as

glycerin. It is a neutral, sweet tasting, colorless, thick liquid. It can easily dissolve in

water or alcohol but not in oils. But, there are many substances which are more

soluble in glycerin than in water or alcohol. So, it is said that glycerin is a good

solvent. A very interesting property of glycerin is that it is highly hygroscopic, which

means that it absorbs water even from the air (Jain, 2006).

Saccharin sodium is used as sweetening agent (Cooney, 1977).

Comprehensive reviews had been done by WHO in 1993 on biological risk

assessment of sodium saccharin. The usage of saccharin sodium had no effect on


107

bladder cancer (Chappel, 1992; Elcock and Morgan, 1993). Samuel M. Cohen et al,

1991 reported sodium saccharin having tumor promoting activity.

6.4 MATERIALS AND METHODS

Scoparia dulcis Linn. plant powder extract, Achyranthes aspera Linn. plant powder

extract, hydroxy ethyl cellulose (Natrosol 250 HX PHARM), sorbitol 70% solution,

Glycerol, Saccharin sodium, Sodium benzoate, Sorbic acid, Propylene glycol, Apricot

168153 flavour and peppermint troomint flavor.

Scoparia dulcis Linn. plant powder extract herbal formulation:

Sorbitol based liquid oral syrups: Finished product specification includes

description, identification, weight per ml, pH of the oral liquid dosage form, assay of

some active ingredients and microbiological purity. Specification for microbiological

purity is included and the specification covers total bacterial, fungi & yeast count and

some pathogens.

Table 6.1. Specifications for Scoparia dulcis Linn. syrup.

Code Test Specifications

T-01 Description Clear Green colored syrup

T-02 Identification

( By HPLC)

In the Assay test, the retention time of the peaks

corresponding to Scopadulcic acid B, Alpha amyrin in

the Sample solution matches with the retention time of the peaks corresponding to Scopadulcic acid B, Alpha

amyrin obtained with Standard solution

T-03 Weight per ml 1.10 to 1.20 gram per ml

T-04 pH 2.50 to 3.50

T-05 Viscosity Between 100 and 210 cps

T-06

Assay 90.0 % to 110.0 % of label claim

Assay of Scopadulcic Acid B,

Sod. Benzoate & Sorbic acid

90.0 % to 110.0 % of label claim

T-07 Heavy Metals (By AAS) Arsenic : Not more than 2 ppm

Mercury : Not more than 1 ppm

Lead : Not more than 10 ppm

Cadmium : Not more than 0.3ppm

T-08 Microbiological quality TAMC : NMT 105 (Maximum acceptable count: 500

000) CFU/g or CFU/mL

TYMC : NMT 104 (Maximum acceptable count: 50 000)

CFU/g or CFU/mL

Bile tolerant gram-negative bacteria : 104 CFU/g or

CFU/mL

Escherichia coli : Absence (1g or mL)

Salmonella : Absence (25g or 25mL)


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Pharmaceutical development:

Preparation of ethanol extract by sox let apparatus: 10 gram of Scoparia dulcis Linn.

plant powder was accurately weighed and packed in vertical tube, which is connected

to the round bottom flask (500ml volume) containing 100 ml of 90% ethanol. The

round bottom flask was connected on the heating mantle having auto control for the

temperature selected. The temperature was fixed as 90ºC and heated for 1 hour for

proper extraction of active constituents. After completion of one hour the apparatus

allowed to cool and the extract was collected aside. Again 50ml of fresh solvent

added to the round bottom flask and subjected for same process. The two extractives

were mixed and subjected for Rota evaporation until the residue remained 50ml.

The extract obtained was subjected to analysis of the Scopadulcic acid B and Alpha

amyrin content.

Development objective: The development objective was to formulate a liquid oral

preparation of plant extract with acceptable taste and flavor.

Developmental pharmaceutics:

Conventional vehicle for medicated syrup is sugar syrup. However it is associated with

crystallization and cap locking problem. It can be replaced with other vehicles either

alone or in combination to give properties similar to sugar syrup. Such preparations

can be called as sugar free syrups and can be administered by diabetic patients

especially.

In developing antidiabetic syrup this approached was used, combination of

35% w/v sorbitol solution (70% Non crystallizing) with 15% w/v glycerol produced

excellent vehicle. It also imparted sweetness to the syrup. However this vehicle did not

have suitable viscosity. Hence, in order to impart a viscosity to the formulation,

hydroxyethylcellulose which is commonly used as a viscosity-builder was used.

Natrosol 250 HX PHARM is the commercially available grade of

hydroxyethylcellulose. It readily dissolves in cold or hot water and gives clear, smooth

suspension. When used in 0.2% w/v concentration it gave a clear solution without

formation of lumps.

Some of the components are sparingly soluble in water. However it possesses

adequate solubility in propylene glycol, warm water and in acidic environment. Hence


109

the pH of formulation was aimed at 2.5-3.5 where it possesses optimum stability

(Loyd V Allen, 2004).

Liquid oral dosage forms such as syrups and suspensions are susceptible to

microbiological growth during its shelf life. Addition of antimicrobial preservative is

essential for such dosage forms. But this liquid oral dosage form consisting of plant

extract will assign antimicrobial activity to the formulation and hence preservatives are

not necessary. Ethanol present in the formulation will also apart the same and prevents

the growth of microbes during its stability (Guidance on Development Pharmaceutics

1996).

The ethanol may tend to evaporate and can leads to the growth of microbes; hence

sorbic acid and sodium benzoate are used as preservatives in the present study.

For medicated syrup organoleptic properties such as taste and flavor are important to

make it palatable to patients. Plant extract possesses bitter taste which remains

persistent after administration. To mask this bitter taste saccharin sodium was used as

a sweetener since it is approximately 300 times sweeter than sugar and possesses

excellent solubility in water. The concentration of saccharin sodium was optimized to

0.03 %w/v. Apricot flavor was used as flavoring agent. When only apricot flavor was

used in the concentration of 0.264%v/v it did not mask the bitter taste. Hence, a

combination of Apricot flavor and menthol was tried. However, the bitter taste still

persisted. On replacing menthol with peppermint troomint flavor in the concentration

of 0.1325%v/v along with Apricot, the bitter taste was successfully masked and a

pleasantly flavored syrup formulation was obtained.


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Table 6.2. Formulae for Scoparia dulcis Linn. syrup

SN Ingredient Per 5 ml Function Manufacturer

1 Scoparia dulcis Linn. Liquid

extract 500 mg

Active

ingredient ----

2 Hydroxy ethyl cellulose

(Natrosol 250 HX PHARM) 10.0 mg

Viscosity

building agent Hercules

3 Sorbitol 70% solution 1750.0 mg Vehicle Noble Drugs

Ltd.

4 Glycerol 750.0 mg Vehicle Godrej

Industries

5 Saccharin sodium 1.5 mg Sweetener Tiansin

Changhe

6 Propylene glycol 150.0 mg Solvent for

preservative

Regent

Chemicals

7 Apricot 168153 flavour 0.0132 ml flavor Symrise

8 Peppermint troomint flavour 0.0066 ml flavor Quest

international

9 Sodium Benzoate 1.125 mg Preservative Across

10 Sorbic acid 1.25 mg Preservative S.D fine

11 Purified water q.s. 5 ml Solvent In house

Table 6.3. Manufacturing Formulae for Scoparia dulcis Linn. Syrup (Batch 1

liter)

Ingredient Typical Quantities

Plant extract 100ml

Hydroxy ethyl cellulose ( Natrosol 250 HX PHARM) 2g

Sorbitol 70% solution 350g

Glycerol 150g

Saccharin sodium 0.3g

Propylene glycol 30g

Apricot 168153 Flavor 0.25 ml

Peppermint Troomint Flavor 0.15 ml

Sodium Benzoate 0.225g

Sorbic acid 0.25g

Purified water q.s. 1000 ml


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Manufacturing process:

Stage 1) Preparation of active solution:

a) Dispersed 2.0 g Hydroxyethyl cellulose in 300 ml purified water and allowed

to swell at room temperature for 30 minutes. Added Sodium benzoate.

b) Heated the solution to 800C under stirring for 1 hour.

c) Allowed the solution to cool to room temperature and added 100ml of plant

extract and Sorbic acid under stirring.

Step 2) Preparation of flavor solution:

Mixture of 0.25 ml Apricot 168153 Flavor and 5 g propylene glycol, 0.15 ml

Peppermint troomint flavor in additional 5 g propylene glycol were prepared

separately.

Stage 3) Preparation of vehicle solution:

a) To a mixing vessel added 350 grams of Sorbitol 70% solution.

b) Further added 150 grams glycerol to mixing vessel under stirring, rinsed the

container with 50 ml purified water and added it to mixing vessel under

stirring.

c) Dissolved 0.3 grams saccharin sodium in 10 ml purified water and added to

mixing vessel, rinsed the containers with 10 ml purified water and added to

mixing vessel under stirring.

Stage 4) Preparation of final syrup:

a) Added herbal extract to mixing vessel under stirring. Rinsed the containers

with 10 ml purified water and added the rinsing to the mixing vessel under

stirring. Give such 5 consecutive rinsing.

b) Cooled the solution to 35-400

C.

c) Added solution of Apricot 168153 flavor to mixing vessel under stirring.

Rinsed the container with 20 ml purified water and added to mixing vessel

under stirring.

d) Added solution of peppermint troomint flavour to mixing vessel under stirring,

rinsed each container separately with 10 ml purified water each and added to

the mixing vessel.


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Figure: 6.1. Process flow chart for Scoparia dulcis Linn. herbal formulation

Disperse

Hydroxyethylcellulose

(Natrosol 250HX PHARM)

in water and allow swelling

for 30 minutes. Add sod.

Benzoate, Heat to 800C for

60 minutes under stirring.

Cool to room temperature

and add Plant extract

,Sorbic acid, maintain at 40-

45 0 C for 30 minutes

Dissolve Apricot 168153

flavour in part of propylene

glycol under stirring.

Mix Peppermint troomint

flavour separately in part %

of propylene glycol.

1

Mixing Vessel

Sorbitol 70%

solution and

Glycerol under

stirring

Appearance

(Uniform

solution free

Dilute to volume with purified

water and mix for 10 minutes

Appearance,

pH, Specific

gravity, assay

Filtration

Machine /Manual Filling

Bottle filling

Labeling and Packaging

Fill

Volume

2

3

4

5

6

7

8

9

10

200 mesh.

Dissolve Saccharin sodium

in purified water.


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e) Made up the volume of syrup with purified water and continued stirring for 15

minutes to get syrup free from lumps, adjusted pH with ortho-phosphoric acid

to get required pH.

Stage 5) Filtration and packing of final syrup:

a) Filtered the syrup through 200 mesh.

b) Filled the syrup into bottles.

Proposed dosage for administration:

Table 6.4 Dosage of Administration for Scoparia dulcis Linn. herbal formulation

Dosage

Adult 30ml / day in 3 divided doses

Children 15ml / day in 3 divided doses

Table 6.5. Dosage of administration for formulation of Scoparia.

Number of teaspoonfuls (5 ml) of suspension

Adult 2 Teaspoonfuls of liquid syrup 3 times daily

Children 1/2-1 Teaspoonful of liquid 1-3 times daily

Proposed usage: Anti diabetic and anti viral syrup

Adverse effects (Undesirable effects): Not studied

Contra-indications: Not studied

Interactions: Not studied

Pregnancy and lactation: Not recommended

Side effects: Not studied

Pharmacokinetics: Currently not available


114

Table 6.6. Pharmaceutical label for Scoparia dulcis Linn. sugar free syrup

Scoparia dulcis Linn. sugar free syrup

Label claim:

Each 10ml. contains

Scopadulcic acid B ……………………5.8 mg

Scoparia dulcis Linn. plant ethanol extract….994 mg

Sorbitol (70%).…………………………3.5 ml

Sodium Benzoate……………………….2.25 mg

Sorbic acid……………………………….2.5 mg

Dosage: 30ml per day in 3 divided doses.

Batch No: Scop003 Mfg. Lic. No: xxxxxx

Manufacturing date:Dec.2006 Expiry date: xxxxxx

Mfg.by: xxxxx

Achyranthes aspera Linn. plant powder extracts herbal formulation:

Table 6.7. Specification for Achyranthes aspera Linn. Syrup.

Code Test Specifications

T-01 Description Clear green colored syrup

T-02 Identification

(By HPLC)

In the Assay test, the retention time of the peaks

corresponding to Oleanolic acid solution matches with

the retention time of the peaks corresponding to

Oleanolic acid with Standard solution.

T-03 Weight per ml 1.10 to 1.20 gram per ml

T-04 pH 3.50 to 4.50

T-05 Viscosity Between 100 and 210 cps

T-06

Assay 95.0 % to 105.0 % of label claim

Assay of

Oleanolic acid

Sod. Benzoate & Sorbic

acid

95.0 % to 105.0 % of label claim

T-07 Heavy Metals

(By AAS)

Arsenic : Not more than 2 ppm




T-08 Microbiological quality TAMC : NMT 105 (Maximum acceptable count: 500

000) CFU/g or CFU/mL

TYMC : NMT 104 (Maximum acceptable count: 50 000)

CFU/g or CFU/mL

Bile tolerant gram-negative bacteria : 104 CFU/g or

CFU/mL




115

Pharmaceutical development: Preparation of ethanol extract by Sox let apparatus:

10 gram of Achyranthes aspera Linn. plant powder was accurately weighed and

packed in vertical tube, which is connected to the round bottom flask (500ml volume)

containing 100 ml of 90% ethanol. The round bottom flask was connected on the

heating mantle having auto control for the temperature selection. The temperature was

fixed 90ºC and allowed for 1 hour for proper extraction of active constituents. After

completion of one hour, allowed the apparatus to cool and the extract was collected

aside. Again added 50ml of fresh solvent to the round bottom flask and subjected for

same process. The two extractives were mixed and subjected for rota evaporation

until the residue remained 50ml.

The extract obtained was subjected to analysis for the content oleanolic acid.

Formulation development:

Development objective: The development objective was to formulate a liquid oral

preparation of plant extract with acceptable taste and flavor.

Developmental pharmaceutics (IP, BP, USP): Sugar free syrups can be administered

by diabetic patients especially, hence the advantage applied in developing antidiabetic

syrup. Combination of 35% w/v sorbitol solution 70% (non crystallizing) with 15%

w/v glycerol produced excellent vehicle. It also imparted sweetness to the syrup.

However this vehicle did not have suitable viscosity. Hence, in order to impart a

viscosity to the formulation, Hydroxyethylcellulose which is commonly used as a

viscosity-builder was used. Natrosol 250 HX PHARM is the commercially available

grade of Hydroxyethylcellulose. It readily dissolves in cold or hot water and gives

clear, smooth suspension. When used in 0.2% w/v concentration it gave a clear

solution without formation of lumps.

Some of the components are sparingly soluble in water. However it possesses

adequate solubility in propylene glycol, warm water and in acidic environment.

Hence the pH of formulation was aimed at 2.5-3.5 where it possesses optimum

stability.

The ethanol may tend to evaporate and lead to the growth of microbes, hence

sorbic acid and sodium benzoate are being used as preservatives in the present study.

For medicated syrups, organoleptic properties such as taste and flavor are important

to make it palatable to patients. Plant extracts possesses bitter taste which remains


116

persistent even after administration. To mask this bitter taste saccharin sodium was

used as a sweetener since it is approximately 300 times sweeter than sugar and

possesses excellent solubility in water. The concentration of saccharin sodium was

optimized to 0.03 %w/v. Apricot flavor was used as flavoring agent. When only

apricot flavor was used in the concentration of 0.264% v/v it did not mask the bitter

taste successfully. Hence, a combination of Apricot flavor and menthol was tried.

However, the bitter taste still persisted. On replacing menthol with Peppermint

Troomint flavor in the concentration of 0.1325% v/v along with Apricot, the bitter

taste was successfully masked and a pleasantly flavored syrup formulation was

obtained.

Table 6.8. Formulae for Achyranthes aspera Linn. Syrup

SN. Ingredient Per 5 ml Function Manufacturer

1 Achyranthes aspera

Linn. Liquid extract 500mg Active ingredient -----

2

Hydroxy ethyl

cellulose

(Natrosol 250 HX

PHARM)

10.0 mg Viscosity

building agent Hercules

3 Sorbitol 70% solution 1750.0 mg Vehicle Noble Drugs

Ltd.

4 Glycerol 750.0 mg Vehicle Godrej

Industries

5 Saccharin sodium 1.5 mg Sweetener Tiansin

Changhe

6 Propylene glycol 150.0 mg Solvent for

preservative

Regent

Chemicals

7 Apricot 168153

flavour 0.0132 ml flavor Symrise

8 Peppermint troomint

flavor 0.0066 ml flavor

Quest

international

9 Sodium Benzoate 1.125mg Preservative Across

10 Sorbic acid 1.25mg Preservative S.D fine

11 Purified water q.s. 5 ml Solvent In house


117

Table 6.9. Manufacturing Formulae for Achyranthes aspera Linn. Syrup (Batch

Size: 1 Liter)

Ingredient Typical Quantities

Plant extract 100ml

Hydroxy ethyl cellulose ( Natrosol

250 HX PHARM) 2g

Sorbitol 70% solution 350g

Glycerol 150g

Saccharin sodium 0.3g

Propylene glycol 30g

Apricot 168153 Flavor 0.25 ml

Peppermint Troomint Flavor 0.15 ml

Sodium Benzoate 0.225g

Sorbic acid 0.25g

Purified water q.s. 1000 ml

Manufacturing process: The process was same as mentioned above for Scoparia

dulcis Linn. formulation that herbal extract added was Achyranthes aspera Linn..

Table 6.10. Pharmaceutical label for Achyranthes aspera Linn. sugar free syrup

Achyranthes aspera Linn. sugar free syrup

Label claim:

Each 10ml contains

Oleanolic acid ……………………………….2.96 mg

Achyranthes aspera Linn. plant ethanol extract……995 mg

Sorbitol (70%).………………………………3.5 ml

Sodium Benzoate…………………………....2.25 mg

Sorbic acid………………………………..… 2.5 mg


Batch No: Vsp004 Mfg. Lic. No: xxxxxx

Manufacturing date:Nov.2006 Expiry date: xxxxxx

Mfg.by: xxxxx


118


Table 6.11. Dosage for formulation of Achyranthes aspera Linn. sugar free syrup

Dosage


Children 15ml / day in 3 divided doses

Table 6.12. Dosage Administration for formulation of Achyranthes aspera Linn. sugar

free syrup


Adult 2 Teaspoonfuls of liquid syrup 3 times daily


Proposed usage: Hepatoprotective and antiviral








119

Formulation development for poly herbal formulation:

Table 6.13. Specifications & test procedure for strong herbal cough syrup.

TAMC: Total aerobic microbial count

TYMC: Total yeasts and moulds count

Components of drug product:

Extracts

Glycyrrhiza glabra (Yashtimadhu) (Aqueous Extract 5:1)

Adhatoda vasica (Vasaka) (Aqueous Extract 4:1)

Terminalia belerica (Vibhitaka) (Aqueous Extract 5:1)

The above all extracts are ready made, procured from chemiloids & Plantex

Zingiber officinale (Alcohol Extract)

S. No. Tests Specifications

1 Description Dark green colored syrupy liquid with

characteristic flavor of pineapple, taste sweet.

2 pH Between 4.0 and 6.0

3 Weight per ml Between 1.15 and 1.35 g / ml

4 Viscosity Between 100 and 210 cps

5 Assay of Gallic Acid NLT 65.0 mg per 100 ml

6 Assay of

Glycyrrhizic Acid NLT 120.0 mg per 100 ml

7 Assay of Vasicine NLT 2.0 mg per 100 ml

8

Assay of 6-Gingerol

Sod. Benzoate &

Sorbic acid

NLT 2.0 mg per 100 ml

9 Heavy Metals (By

AAS)

Arsenic : Not more than 2 ppm




10

Microbiological

quality

TAMC : NMT 105 (Maximum acceptable count:

500 000) CFU/g or CFU/mL

TYMC : NMT 104 (Maximum acceptable count:

50 000) CFU/g or CFU/mL

Bile tolerant gram-negative bacteria : 104 CFU/g

or CFU/mL




120

Table 6.14. Manufacturing Formulae for poly herbal formulation (Batch Size:1

Liter)

S. No Ingredients Manufacturer Qty. per 1000

ml

1. Glycyrrhiza glabra

(Yashtimadhu)

(Aqueous Extract 5:1)

Chemiloids 20.00g

2. Adhatoda vasica (Vasaka)


Chemiloids 10.00g

3. Terminalia belerica

(Vibhitaka)


Plantex 12.00g

4. Zingiber officinale.

(Alcohol Extract)

Self generated 10.00g

5. Sucrose USP/NF 750.0g

6. Glycerol Noble Drugs

Ltd.

50.00g

7. Propylene Glycol Regent

Chemicals

50.00g

8. Citric Acid Monohydrate S.D fine

chemicals

0.65g

9. Sodium Benzoate Across 0.45g

10. Sorbic Acid S.D fine

chemicals

0.50g

11. Color BQ Supra

Amerind Colors

& Chemicals

0.45g

12. Essence Pineapple Symrise 0.002 ml

13. Purified Water In house Q.s. to

1000 ml


121

Figure: 6.2. Process flow chart for poly herbal formulation

1

Extract Glycyrrhiza glabra,

Adhatoda vasica, Terminalia

belerica, into purified water

Extract the Zingiber

officinale into Ethanol

Add

Colour &Flavour

Filter the

Macerated Herbals

Add glycerol,

citric acid

Dilute to volume with purified

water and mix (10 minutes)

Appearance,

pH, Specific

gravity, assay

Filtration

Machine /Manual Filling

Bottle filling

Labeling and Packaging

Fill Volume

2 3

4

5

6

7

8

9

10

200 mesh

Mixing Vessel

PREPARATION OF

SUGAR SYRUP


122

Step-1: In beaker 100ml of purified water was boiled and added the specified quantity

of sodium benzoate and sorbic acid and mixed well to dissolve the contents. To the

above added the following duly weighed herbal extracts dry powder one by one under

continuous stirring of the contents in suitable capacity.

Glycyrrhiza glabra (Yashtimadhu) extract dry powder [5 : 1]

Adhatoda vasica (Vasaka) extract dry powder [4: 1]

Terminalia belerica (Vibhitaka) extract dry powder [5: 1]

After addition of dry herbal extracts powder the contents stirred for 1 hour and kept

the solution for maceration for a period of not less than 12 hours.

Weighed specified amount of propylene glycol BP in a cleaned suitable capacity of

vessel and heated to 70°C to 80°C.

Step-2: The Zingiber officinale extract was prepared by process of percolation:

100g of Zingiber officinale wet material was weighed and subjected for size reduction

of coarse irregular pieces each of 3 mm using knife and packed in percolator (500ml

volume) as a dense bed. Loaded 100 ml of 90% ethanol; remove the locked air, from

the packed bed by allowing the ethanol to drain out from down side, the herbal drug to

be extracted is always covered with the remaining extraction solvent.

Allowed the whole setup to stand for an appropriate time of 24 hours and collected

the extracted solvent. The residue remained was pressed out and the expressed liquid

combined with percolate. The two extractives were mixed and subjected for rota

evaporation until the residue remained 50ml.

Step-3: Filtration of macerated herbals: The herbals after completion of maceration

were filtered through 200mesh. The collected filtrate was kept for storage under clean

air tight container.

Step-4: Preparation of sucrose syrup: 300ml of purified water was taken in a suitable

capacity of glass vessel & heated up to 55°C temperature, added sodium benzoate and

sorbic acid in sequence, stirred to dissolve for 5 minutes.

Specified amount of Sucrose was added under constant stirring. Boiled the sugar

syrup up to 90°C and maintain the temperature at 90°C for 30 minutes and stirred well


123

until Sucrose gets completely dissolved. Then Glycerol BP was added under

continuous stirring for 10 minutes.

Step-5: Mixing of extracts solution with sucrose syrup: Filtered the supernatant

solution of both the extracts. Stirred the obtained extracts solution for 5 minutes,

boiled the extract solution up to 70°C. Transferred the boiled extract solution to

sucrose syrup. Then added the zingier extract after reaching temperature of 50ºC and

mixed, purified water added up to required volume.

Addition of citric acid: Specified amount of citric acid weighed and dissolved in 10ml

of previously boiled purified water in a separate vessel, citric acid solution slowly

added with constant stirring to syrup.

Addition of color: Specified amount of color BQ supra weighed and dissolved in

20ml of previously boiled purified water in a separate vessel, and added to bulk syrup.

Step-6: Dilution to volume: Measured the volume of syrup obtained, made up to the

required volume with water.

Step-7: Filtration: Filtered the syrup through 200 mesh to remove any unwanted

particles or lumps after attainment to room temperature.

Table 6.15. Pharmaceutical label for poly herbal formulation.

Poly herbal formulation

Label claim:

Each 5ml. contains

Gallic acid …………………… 1.9 mg

Glycyrrhizic acid…………..…. 8.4 mg

Vasicine...……………………… 0.19 mg

6-Gingerol………………………0.35 mg

Sodium Benzoate……………… 2.25 mg

Sorbic acid…………………….. 2.5 mg


Batch No: VSP007 Mfg. Lic. No: xxxxxx

Manufacturing date:Dec.2006 Expiry date: xxxxxx

Mfg.by: xxxxx


124


Table 6.16. Dosage of administration for poly herbal formulation

Dosage


Children 7.5 -15 ml / day in 3 divided doses

Table 6.17. Dosage of administration for poly herbal formulation


Adult 1 Teaspoonful of liquid syrup 3 times daily


Proposed use: Cough suppressant







General reference test procedure section:

1. Description: Placed the Syrup in a clean and dry Petri dish, and observed against

specification.

2. Identification: In the HPLC Assay test, the retention time of the peaks

corresponding to the active in the Sample solution matches with the retention time of

the peaks corresponding to active obtained with standard solution.

3. Weight per ml: Weighed a previously dried 10 ml Pycnometer (W1). Filled the

Pycnometer with sample and adjust the temperature of the sample to 25°C, removed any

excess of sample & weighed (W2).

Determined the weight per ml using the formula given below.

Weight per ml = W2 – W1 / 10


125

4. pH of the solution: Measured the pH with about 30 ml of the Syrup in 50 ml glass

beaker. Allowed the equilibration time of 2 minutes, and noted the pH after

stabilization.

5. Viscosity (using the rotating viscometer):

a. Instrument : Brook field viscometer RVT

b. Spindle No. : 2

c. Factor : 4

d. Speed : 100 RPM

Sample preparation: Transfer 200 ml of the sample to 250 ml beaker and bring the

temperature of the sample to 25°C. Maintain the temperature throughout the test.

Procedure:

i. Fixed the spindle No. 2 on the instrument.

ii. Turned the speed knob to 100.

iii. Switched on the instrument and adjusted the reading to 0.000.

iv. Put off the instrument and placed the sample, so that the spindle dips inside the

sample to the described height.

v. Started the instrument and allowed to running till a constant reading is achieved on

the scale after pressing the switch.

vi. Recorded the reading and calculated the viscosity using the factor.

6. Assay: Individual methodology for the contents mentioned in HPLC chapter.

7. Heavy metals:

Preparation of 2M Sodium Hydroxide Solution: Dissolved 80g of Sodium Hydroxide

in 1000ml water.

Blank Preparations: 0.1M Hydrochloric Acid: Dissolved 8.5ml of Hydrochloric Acid

in 1000ml water.

Standard solution:

A) For Arsenic (As) :

Dissolved about 1.320 g of Arsenic trioxide in 20ml of 2M Sodium hydroxide

(NaOH) and diluted to 1000ml with water (1000 ppm). Diluted 10ml of this solution

to 100ml with 0.1M HCl which contains 100ppm of As.

Above stock diluted 2ml → 100ml (2ppm)


126




B) For Mercury (Hg):

Dissolved about 1.354 gm of Mercuric chloride(HgCl2) in 50ml of 2M nitric acid and

diluted to 1000ml with water (1000 ppm). Diluted 10ml of this solution to 100ml

with 0.1M HCl which contains 100ppm of Hg.





C) For Lead (Pb):

Dissolved about 0.400 g Lead nitrate in water containing 2ml of nitric acid and made

upto 250ml using water (1000 ppm) . Diluted 10ml of this solution to 100ml with

0.1M HCl which contains 100ppm of Pb.



Above stock diluted 15ml → 100ml (15ppm Pb)

Above stock diluted 20ml → 100ml (20ppm Pb)

D) For Cadmium (Cd):

Dissolved about 0.100 g of Cadmium in minimum quantity of mixture of equal

volumes of hydrochloric acid and water and diluted to 100ml with 1% hydrochloric

acid (1000 ppm). Diluted 1ml of this solution to 100ml with 0.1M HCl which contains

10ppm of Cd.

Above stock diluted 3ml → 100ml (0.3ppm)




Sample preparation:

Weighed about 10gm of sample in a suitable crucible. Slowly incinerated on

incinerator to near dryness. Added 2ml of Sulphuric acid to wet the sample. Ignited

carefully at a low temperature until thoroughly charred. Kept charred mass in muffle


127

furnace at 700°C for 2hours. Cooled the crucible and dissolved the ash in 25ml of

0.1N Hydrochloric acid.

Taken the readings of standard on suitable AAS by using respective lamps and plotted

calibration graphs. 0.1N Hydrochloric acid used as blank. The reading of sample

noted and calculated content of respective heavy metals in ppm.

The above sample used for determination of the traces of arsenic, standard readings

noted on suitable AAS by using arsenic lamp at the wavelength 193.7 nm and plotted

calibration graph. Also taken reading of sample and found out arsenic content in ppm.

8.0 Microbiological purity (Ph. Eur. method 2.6.31):

Reagents and solution preparations:

Phosphate buffer stock solution pH 7.2: Dissolve 34g of monobasic potassium

phosphate in 1000ml of water, adjust pH with0.1M Hydrochloric acid or with 0.1M

Sodium hydroxide and sterilize in autoclave at 121°c for 20 minutes. Store at 2°C –

8°C.

Dilute Phosphate buffer solution pH 7.2: 1.25 mL of phosphate buffer stock solution

pH 7.2 and 20 g of polysorbate 80 mixed and dissolved in 500ml of purified water and

make up to 1000ml and sterilize in autoclave at 121°c for 20 minutes.

0.1M Hydrochloric acid: 8.5mL of hydrochloric acid make up to 1000ml with

purified water.

0.1M Sodium hydroxide: 4g of sodium hydroxide dissolved and make up to 1000ml

with purified water.

Culture media preparation:

Soybean–Casein Digest Medium broth pH 7.3±0.2: Dissolve pancreatic digest of

casein 17.0g, papaic digest of soyabean 3.0g, sodium chloride 5.0g, dipotassium

hydrogen phosphate 2.5g and glucose monohydrate 2.5g in 1000ml of purified water,

adjust pH with 0.1M Hydrochloric acid or with 0.1M Sodium hydroxide. Sterilize the

medium in autoclave at 121°c for 20 minutes.

Soybean–Casein Digest Medium agar pH 7.3±0.2: Dissolve pancreatic digest of

casein 15.0g, papaic digest of soyabean 5.0g, sodium chloride 5.0g, and agar 15g in

1000ml of purified water, adjust pH with0.1M Hydrochloric acid or with 0.1M

Sodium hydroxide. Sterilize the medium in autoclave at 121°c for 20 minutes.


128

MacConkey broth pH 7.3±0.2: Dissolve pancreatic digest of gelatin 20.0 g, lactose

monohydrate 10.0g, dehydrated oxybile 5.0g and bromocresol purple 10mg in

1000mL of purified water, adjust pH with0.1M Hydrochloric acid or with 0.1M

Sodium hydroxide. Sterilize the medium in autoclave at 121°c for 20 minutes.

MacConkey agar pH 7.1±0.2: Dissolve pancreatic digest of gelatin 17.0 g, peptones

(Mean and casein) 3.0g, lactose monohydrate 10.0g, sodium chloride 5.0g, bile

salts1.5g, agar 13.5g, neutral red 30mg and crystal violet 1mg in 1000mL of purified

water, adjust pH with0.1M Hydrochloric acid or with 0.1M Sodium hydroxide.

Sterilize the medium in autoclave at 121°c for 20 minutes.

Rapport vassiliadis salmonella enrichment broth 5.2±0.2: Dissolve soya peptone

4.5g, sodium chloride 8.0g, potassium dihydrogen phosphate 0.6g, dipotassium

hydrogen phosphate 0.4g, magnesium chloride hexahydrate 29.0g and malachite green

36mg in 1000mL of purified water, adjust pH with0.1M Hydrochloric acid or with

0.1M Sodium hydroxide. Sterilize the medium in autoclave at 115°c for 15 minutes.

Xylulose – lysine-deoxycholate agar medium pH 7.4±0.2: Dissolve xylose 3.5g, L-

lysine 5.0g, lactose monohydrate 7.5g, sucrose 7.5g, sodium chloride 5.0g, yeast

extract 3.0g, phenol red 80mg, agar 13.5g, sodium deoxycholate 2.5g, sodium

thiosulphate 6.8g and ferric ammonium citrate 0.8g in 1000mL of purified water,

adjust pH with0.1M Hydrochloric acid or with 0.1M Sodium hydroxide. Heat to

boiling does not autoclave.

Enterobacteria enrichment broth-mossel pH 7.2±0.2: Dissolve pancreatic digest of

gelatin 10.0g, glucose monohydrate 5.0g, dehydrated ox bile 20.0g, potassium

dihydrogen phosphate 2.0g, disodium hydrogen phosphate dihydrate 8.0g, and brilliant

green 15mg in 1000ml of purified water, heat at 100°C for 30 minutes and cool

immediately.

Violet red bile glucose agar pH 7.4±0.2: Dissolve yeast extract 3.0g, pancreatic

digest of gelatin 7.0g, bile salts 1.5g, glucose monohydrate 10.0g, lactose monohydrate

10.0g, sodium chloride 5.0g and agar 15.0g in 1000ml of purified water, heat to

dissolve the contents and cool immediately.

Total aerobic microbial count (TAMC):

Diluted 10 ml of the sample to 100ml with sterile phosphate buffer and mixed well.


129

Membrane filtration method: 1 ml of above sample was filtered through 0.45µ sterile

cellulose acetate filter paper under vacuum filtration unit, and then the filter paper

flushed with 4 x 100ml of phosphate buffer solution pH 7.2 containing polysorbate 80

as surface active agent and neutralizer for preservatives. Carefully transferred the filter

paper to surface of casein soyabean digest agar medium plate and incubated the plate

at 30°C – 35°C for 5 days. Then the membrane subjected for counting of colonies.

Total combined yeasts and mold count (TYMC):


Membrane filtration method: 1 ml of above sample was filtered through 0.45µ sterile

cellulose acetate filter paper under vacuum filtration unit, and then the filter paper

flushed with 4 x 100ml of phosphate buffer solution pH 7.2 containing polysorbate 80

as surface active agent and neutralizer for preservatives. Carefully transferred the filter

paper to surface of Saboraouds dextrose agar medium plate and incubated the plate at

20°C – 25°C for 7 days. Then the membrane subjected for counting of colonies.

Controls in the analysis:

Media control: Culture media without sample are kept at same test conditions during

the analysis at each step.

Diluent control1ml of sterile diluent without sample are kept at same test conditions.

Test for Escherichia coli:


Primary incubation: Take 10 ml of above sample Soybean–Casein Digest Medium

broth and mix kept incubator at 30°C – 35°C for 24 hours.

Secondary incubation: Transfer 1 ml of above mixed broth sample in to 100ml

MacConkey broth and kept incubator at 42°C – 44°C for 48 hours.

Plating: Streaked loop full of culture from enriched MacConkey broth on MacConkey

agar medium and incubated at 30°C – 35°C for 72 hours.

Test for Salmonella species:

Primary incubation: Take 10 ml of above sample Soybean–Casein Digest Medium

broth and mix kept incubator at 30°C – 35°C for 24 hours.


130

Secondary incubation: Transfer 0.1 ml of above mixed broth sample in to 10ml of

Rapport vassiliadis salmonella enrichment broth and kept incubator at 30°C – 35°C for

24 hours.

Plating: Streaked loop full of culture from enriched Rapport vassiliadis salmonella

broth on xylulose – lysine-deoxycholate agar medium and incubated at 30°C – 35°C

for 48 hours.

Test for bile-tolerant Gram-negative bacteria


Presence or absence test:

Pre incubation: Incubate the above prepared sample at 20°C – 25°C for 2 hours for

resuscitation.

Primary incubation: Take 10 ml of above 10:100 diluted samples in to 90ml of

Enterobacteria enrichment broth mossel and incubated at 30°C – 35°C for 48 hours.

Plating: Streaked loop full of culture from Enterobacteria enrichment broth mossel on

the violet red bile glucose agar. Incubated the plates at 30°C – 35°C for 24 hours.

Escherichia coli culture kept run simultaneously as reference.

Colony characters were compared with the reference for presence or absence.

Table 6.18. Morphologic characteristics of Escherichia coli and Salmonella species on

selective media.

Selective medium Name of organism Characteristic colonial

morphology

MacConkey agar medium Escherichia coli brick red colonies with or

without surrounding zone

of bile pigments

Rapport vassiliadis

salmonella broth on

xylulose – lysine-

deoxycholate agar medium

Salmonella Red colonies with or

without black centre

Table 6.19. Morphologic characteristics of Enterobacteriace.

Selective medium Name of organism Characteristic colonial

morphology

Violet red bile glucose agar Enterobacteriace

group

Pink color colonies

with bile precipitation.


131

6.5 RESULTS

Table 6.20. Results obtained for Scoparia dulcis Linn. herbal formulation

S. No Name of the test Results

1. Description Clear green colored syrup

2. Identification Complies

3. Weight per ml 1.122

4. pH 3.21

5. Viscosity 130 cps

6. Assay

Scopadulcic acid B

Alpha amyrin

Sod. Benzoate

Sorbic acid

102.22 %

99.02 %

100.62 %

101.84 %

7. Microbial quality

TAMC

TYMC

Escherichia coli

Salmonella

Bile-tolerant Gram-negative bacteria

65 cfu/ml

33 cfu/ml

Absent

Absent

Absent

8. Heavy Metals (By AAS) Arsenic : Not detected

Mercury : Not detected

Lead : Not detected

Cadmium : Not detected

Table 6.21. Results obtained for Achyranthes aspera Linn. herbal formulation


1. Description Clear green colored syrup

2. Identification Complies

3. Weight per ml 1.135

4. pH 3.81


6. Assay

Oleanolic acid

Sod. Benzoate

Sorbic acid

101.25 %

99.91 %

102.24 %


TAMC

TYMC

Escherichia coli

Salmonella


48 cfu/ml

36 cfu/ml

Absent

Absent

Absent


132



Lead : Not detected


Table 6.22. Results obtained for poly herbal formulation

6.6 DISCUSSION OF RESULTS IN LIGHT OF OTHERS WORK

No herbal formulations were reported by using Scoparia dulcis Linn. plant extract

with pharmaceutical excipients.

There are some reports on methanol and ethanol extracts of leaves having anti viral

activity (Hayashi et al, 1988) Analgesic and antipyretic activity (Ahmed et al, 2001),

Anti oxidant properties (Latha & Pari, 2003), anti diabetic activity (Pari &

Venkateswaran, 2002), scavenging activity (Babincova & Sourivong, 2001).


1. Description Dark green colored syrupy liquid

with characteristic flavor of

pineapple, taste sweet.

2. pH 4.53

3. Weight per ml 1.26 g / ml


5. Assay of Gallic acid 101.53 %

6. Assay of Glycyrrhizic acid 100.10 %

7. Assay of Vasicine

6-Gingerol

Sodium Benzoate

Sorbic acid

99.47 %

100.86 %

102.44 %

103.68


TAMC

TYMC

Escherichia coli

Salmonella


92 cfu/ml

80 cfu/ml

Absent

Absent

Absent



Lead : Not detected



133

No herbal formulations were reported by using Achyranthes aspera Linn. plant extract

with pharmaceutical excipients. There are some reports on anti microbial activity

(Naidu et al, 2006), extracts showed moderate larvicidal effects (Bagavan et al, 2008),

antipyretic properties (Gokhale et al, 2002) due to presence of oleanolic acid it also

showed to contain antitumor and hepatoprotective properties (Liu, 1995), as well as

exhibiting antiviral properties, but these all studies were conducted on plant parts but

not as formulation.

Multi herbal formulations were reported for specific activities (Balaraman, 2004), but

there is no herbal formulations reported to contain these Adhathoda vasaka,

Glycyrrhiza glabra, Gingiber officinalis and Terminalia belerica plant extracts.

6.7 CONCLUSION

The herbal formulations developed for Scoparia dulcis Linn. and Achyranthes aspera

Linn. are sugar free hydro alcoholic based sweetened syrups; they can be

administrated by diabetic patients.

The herbal formulations developed for cough suppressant syrup was simple sugar

based syrup. It is contra indicated for diabetic patients because of the presence of

Glycyrrhizic acid and sugar based syrup.

6.8 FUTURE PROSPECTS

Other components need to be identified, quantified and their stability to be monitored

during their shelf life.

Bio assay need to be performed for checking the efficacy of the drug product.

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