standard protocols for extractions and formulation development

7/26/2019 Standard Protocols for Extractions and Formulation Development

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Standard Protocols for Extractions and

Formulation Development

Vittal Mallya Scientific Research Foundation

# 94/3 & 94/5, 23rdCross, 29thMain, BTM II Stage, Bengaluru 560 076

Email: [email protected]; Web site: www.vmsrf.org


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1.Extraction of Sweet flag (A c o r u s c a la m u s ) technical

1. Pulverize 50kg of sweet flagrhizome in to fine powder.

2.

Dry the powder at ambienttemparature for 24 hrs.

3.

Charge the powder into 2KLSSreactor.

4.

Charge 490 kg of Ethyl acetate.5.

Stir the mixture at 60 C for 10-

12hrs.6. Slowly cool the mass to RT.7. Filter through celite bed.8.

Collect the clear ethyl acetatelayer in a container.

9. Keep the insoluble for any otherusage.

10. Distil ethyl acetate layer under

vacuum at below 60C.11.

Check the weight of the extract

(brown oil)and record12. Sample for analysis of contents

and record the asarone content

Recovery of solvent

Distil ethyl acetate slowly undervacuum at temperature less than 60C toreuse.


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2.Formulations from Sweet flag (A c o r u s c a l am u s ) technical

1) Production of water soluble powder formulation of sweet flag (-Asarone3% WSP)

1.Charge 2kg of beta methyl cyclodextrin in a 50l vessel with mechanical stirrer.

2.Add 10l of water and stir it for 30min to get a clear solution.

3.

Add 500g sweet flag technical (60-65% asarone).4.

Stir for 4hr at ambient temperature.5.Filter the solution through celite bed.

6.Spray-dry the clear solution to get off-white to white powder.

2) Production of water soluble powder formulation of sweet flag (-Asarone

1% WSP)

1.

Charge 9kg of gum acacia in a 250l vessel having mechanical stirrer2.Add 50l of water3.Stir it for 30min to get clear solution.

4.Add 300g sweet flag technical (60-65% asarone).

5.

Stir for 4hr at ambient temperature.6.

Filter the solution through celite bed.7.Spray-dry the clear solution to get off-white to pale pink powder.

3) Production of emulsifable formulation of sweet flag (-Asarone 1% EC)

1.

Charge 10g of sweet flag technical (60-65% asarone) in a 2l vessel having

mechanical stirrer.2.Add 950ml of water.3.Add 20 g carrier (glycerol/neem oil).4.Add 20 g surfactant (Tween 80).5.

Stir it for 2 hr.

6.

Store the milky white solution for further use and bottling .


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3.Extraction of Shikakai( A c a si a co n c i n n a ) technical

1. Powder 25kg of shikakai fruits.2. Charge the powder into 1KLSS

reactor.

3. Add 250kg of water.4. Stir the mixture at 60 C for 5-6hr.5.

Filter to get clear aqueous layer6. Collect in a container. Draw sample

for analysis

4. Production of emulsifiable formulation of shikakai

1. Charge 100g of shikakai technicalin to 50l vessel.

2.

Add 10l of water.3. Add 50 g of surfactant (Tween 80)4.

Stir at RT for 30min to get clear solution.

Production of WSP formulation of shikakai

1. Charge 1000g of Acacia concinna (shikakai) fruit.2. Add 6 volume of water3. Stir at room temperature for 6hrs4.

Filter to get clear water layer5. Add 1000 g of di-potassium hydrogen phosphate and mono potassium di-hydrogen

phosphate mixture of 1:1 ratio

6. Stir to get clear solution and stir at RT for 1hrs

7.

Lyophilize to get water soluble powder.


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Neem Seeds - 1000 kg

CakeOil

Water - 2000 litres +

Stirring for 6 hrs

Filtration through Filter Press

Extract Filtration

Water Extract

+ Ethyl Acetate 250 kg

Stirring & Settling for 6 hrs

Extract I

+ Ethyl Acetate 150 kg

Stirring & Settling for 6 hrs

Extract IIWater Extract + Extract I

Distilled at 30 to 35 C under

vacuum 450 to 500 mm

Crude Extract - 10 kg

Hexane - 100 litres +Add extract slowly to Hexane

while stirring

White Powder Precipitation

Filteration

Air dried - White Powder

TECHNICAL AZA

NeutscheFilter

Vacuum Distillation Unit

through Buchner under vacuum

Oil Expeller Unit

FLOW CHART INSTRUMENTSPROCESS

Water Extract

5.Extraction of Neem( A z a d i r a c h t a i n d i c a ) technical

1. Expel oil from 1000kg of Neem seedand collect the cake

2.

Extract neem seed cake with 8vol

water at room temp under stirringfor 6hr

3.

Filter to get clear water layer4. Extract with Ethyl acetate by adding

1/6 volume of water layer at roomtemp for 6hr

5. Settle for 2-3 hrs and separate ethyl

acetate layer6. Extract once again with Ethyl

acetate by adding 1/6 volume ofwater layer and separate the layer

7. Concentrate ethyl acetate extractunder vacuum by distillation to 90-95%

8.

Add 10 times of hexane understirring

9. Filter and air dry to get the technicalwith about 40% purity of

Azadirachtin Technical

6.Formulations from Azadirachtin technical

1. Process of making water soluble powder formulation (Azadirachtin 3% WSP)

1. Charge 5kg of beta methyl cyclodextrin in a 50l vessel with mechanical stirrer.2. Add 30l of water and stir it for 30min to get a clear solution.

3. Add 1kg Azadirachtin technical (40% Azadirachtin).4.

Stir for 4hr at ambient temperature.5.

Filter the solution through celite bed.

6. Spray-dry the clear solution to get the water soluble powder.

2. Process of making water soluble powder formulation (Azadirachtin 0.3%WSP)

1.

Charge 1.5kg of neem seed kernel in to 50lts reactor.2.

Add 6vol water3. Stir at room temperature for 6hrs4. Filter to get clear water layer

5. Charge above water layer back in to the clean reactor6. Add 2kg of di-potassium hydrogen phosphate and mono potassium di-hydrogen

phosphate mixture of 1:1 ratio

7. Stir to get clear solution and stir at RT for 1hrs8. Spray dry to get 1.7-1.8kg of water soluble powder (light brown solid) of about

3200-3500ppm azadirachtin content


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3. Process of making emulsifiable formulation (Azadirachtin 0.1% EC)

1.

Charge 50kg Neem oil (~2000ppm Azadirachtin) in 50l SS reactor

2. Add 16kg of carrier (Glycerol)3. Add 10l of water4. Add 24kg of surfactant (TWEEN 80)5. Stir for 1hr to get uniform EC formulation


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6.Extraction of Lantana (Lantana camara) technical

1. Powder 5kg of Lantana leaves and put

into 250l SS reactor.

2.

Charge 50l of water.3. Stir the mixture at 60 C for 5-6 hrs.4.

Filter to get clear aqueous layer.

5. Charge clear aqueous layer in toanother clean 250l SS reactor.

6. Distil water almost completely under

vacuum at temperature below 80C.7.

Collect the extract and weigh to record

7.Formulations from Lantana technical

1. Charge 1 kg of Lantana technical in to 20l vessel

2. Add 10l of carrier (IPA)

3. Add 10l of water

4. Add 100g of surfactant (TWEEN 80)

5. Add 0.4kg of stabilizer (glycerol)

6.

Stir at ambient temperature for 30min to get uniform EC formulation


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Pongammia Seeds - 100 kg

Oil Expeller

Seed cake

Hexane(250Kgs)

Ethyl acetate/water

Stirring for 6 hrs at RT

Pongamia Technical (Thick

syrupy mass )

Filtration

Ethyl acetate layer

FLOW CHART INSTRUMENTSPROCESS

Filter

Hexane layer Insoluble mass

Distil at

standard protocols for extractions and formulation development

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