Ijrtem 110103

Invention Journal of Research Technology in Engineering & Management (IJRTEM) www.ijrtem.com ǁ Volume 1 ǁ Issue 10 ǁ

ISSN: 2455-3689

Fractionation of Crude Dye Extracted From Cucurbita Pepo Leaves by Cold Extraction Method Shradhanjali Mohapatra1 & Pragnyashree Mishra2 1

Assistant Professor, Department of Textile and Apparel designing, College of Home Science, OUAT. 2 Assistant Professor, Department of Floriculture and Landscaping, College of Horticulture, OUAT.

ABSTRACT: Natural dyes are those dyes obtained from natural sources. The majority of natural dyes are usually collected from roots, berries, bark, leaves, wood, fungi and lichens. Usually in ancient days people have dyed their textiles by using locally available materials. Cold extraction for crude dyes extraction from Cucurbita pepo leaves. Theextract obtained quantitatively from cold extraction method was 6.81g and 2.27g respectively from 100g and 50g of C. pepo dry mass taken in 750ml and 500ml of ethanol solvent.6 components/functional groups were confirmed in crude dye fractioned with n hexane, chloroform and ethyl acetate but only 4 components/functional groups were confirmed in crude dye fractioned with acetone.

INTRODUCTION Natural dyes are those dyes obtained from natural sources. The majority of naturaldyes are usually collected from roots, berries, bark, leaves, wood, fungi and lichens.Usually in ancient days people have dyed their textiles by using locally available materials. Woad, indigo, saffron, and madder were raised commercially and were important trade goods in the economics of Asia and Europe.A large number of investigations on extraction of colorants from natural sources like plants, microbes, insect and animals and their different kinds of applications have been done till date. In functional finishing oftextiles, coloration of food and dyesensitized solar cells natural dyehas already appliedShahid& Islam (2013). According toGrifoniet al. (2011)with naturaldye fabrics made of vegetable fibres has role in UV protection. Textiles offered a safestprotection from (UV) radiation exposer with the use of natural dyesincrease the UV protection of fabrics madeof vegetable fibre.A pumpkin is a gourd-like squash of thegenus Cucurbita belongs to the familyCucurbitaceaeCucurbita pepo or pumpkins are widely grown for commercial use and having good values in both food and recreation. This family consists of 125 generaand 960 species, mainly in tropicaland subtropical regions. Since long time efforts have been made by variousscientists to develop an efficient extractiontechnique for natural dye extraction.Baliarsinghet al.(2012) has isolated principal colour compounds such as flavonoids and tannin moietiesfrom two plant species (SaracaasocaandAlbizialebbeck), using solvent extraction method and standardised dyeing techniques, identified colourcomponents and checked their potentialantimicrobial properties against commonhuman pathogens. Hee-OckBooaet al (2012). extracted and characterized somenatural plant pigments and their use for cosmetics and determined antioxidantactivities, total polyphenols and flavonoids, and antimicrobial effects of some plant pigments.Natural pigment from the petals of the Flame of forest(Buteamonosperma) flower under differentoperating conditions such as extraction time,temperature and mass of the petals conventional extraction technique were extracted by by Sinha et al (2012). In the present study crude dyes are extracted from C. pepo leaves by cold extraction method. The fractionation of the dye was done with various solvents andanalysed by simple chromatographic technique.

MATERIAL AND METHODS This experiment was conducted at OUAT, Bhubaneswar All commercially available solvents such as were carried out. The curves were visualized under ultra violet at 254 nm in TLC. 2.1. Collection of C. pepo leaves The leaves of Cucurbita pepo was collected from nearby areas of OUAT in November 2016.

2.2. Preparation of sample The leaves of C. pepo were washed with tap water followed by distilled water toremove impurities/dust from the surface. The leaves were then dried in shade and powdered by blender.

2.3. Cold Extraction 100g of sample wassoaked in 500ml of ethanol solvent for twoweeks. 250 ml of ethanol was added to the soaked sample material in two batches for one week after removal of previous batch ethanol. Then all the layerswere separated and then mixed and filtered.In Rotary evaporator at reduced pressure, the ethanol filtrate wasevaporated to get dark green residue. Then Residue wasplaced in the water bath at the temperature 70oC for 4 hours. By this method ethanol content got evaporated and the ethanol extract which was collected in a vialand stored for fractionation.

2.4. Concentration of C. pepo leaves coldextract with rotary evaporator

| Volume 1 | Issue 10 |

www.ijrtem.com

|1|

Fractionation of Crude Dye Extracted From Cucurbita Pepo Leaves by Cold Extraction Method C. pepo (leaves) extract was kept in a cleaned flask. Temperature of rotary evaporatorwas set at 78 oC for removal of solvent. The concentrated extract was stored for Fractionation and other analyticaltest.

2.5. Fractionation of C. pepo (leaves) coldextract For fractionation the concentrated coldextract was taken. The solvent used were n-hexane,chloroform, ethyl acetate and acetone. All the solvents were added to the conc. cold extract of C. pepo leaves till the extract were dissolved completely in the solvent.Fractions were stored in vials for fractionation in TLC.

RESULTS AND DISCUSSIONS 3.1. Extraction of crude dye from C. pepoleaves Two methods of dye extraction, cold extractionand hot extraction were evaluated. Thequantitative amount of extract obtained from coldextraction method is 6.81 g and 2.27grespectively while from hot extraction method is2.48g as in Table 1 and Table 2. From the tables it can be concluded thaton the basis of quantitative yield of dye hot extractionmethod is better than cold extractionmethod due tothe influenceof temperature on the extraction process.

It is noted from the literature that the extractconcentration increases due to increasing temperature. Hot extractionis better than cold extraction because it`s arapid process of extraction, extraction isgood due to the involvement of temperature and due to the use offilter paper in hot extraction no filtration is required. Usually a lowboiling solvent like n-hexane are used in hotextraction methods. Though hot extraction is better than cold extraction when quantitative yield is considered but in the sense of cost efficacy and availability cold extraction is better than hot extraction as cold extraction canbe carried out in a simple pot but for hotextraction a heating source, condenser,round bottom flask and the most importantelectricity is required.

S.no 1 2

.

Table1.Quantitative production of crude extract of the leaves of C. pepo using ethanol solvent.

Sample Solvent volume Weight of Weight of beaker Weight of extract weight (g) (ml) beaker (g) +extract (g) 100 750 161.83 168.64 6.81 50 500 161.83 164.10 2.27

Table 2. Quantitative production of crude extract of the leaves of C. pepo using n-hexane solvent Sample weight Solvent volume Weight of beaker Weight of beaker Weight of extract (g) (ml) (g) +extract (g) 50 500 35.18 37.66 2.48 No. of spots 1 2 3 4 5 6

No. of spots 1 2 3 4 5 6

No. of spots 1

| Volume 1 | Issue 10 |

Table 3. Fractionation with n-hexane Distance trveled by Distance traveled by solvent front spots 3.6 cm 3.42cm 3.6 cm 3.3cm 3.6 cm 3.0cm 3.6 cm 2.3cm 3.6 cm 1.2cm 3.6 cm 0.8cm Table 4.Fractionation with Chloroform Distance trveled by Distance traveled by solvent front spots 3.3 cm 3.2cm 3.3 cm 2.6cm 3.3 cm 2.3cm 3.3 cm 1.7cm 3.3 cm 1.3cm 3.3 cm 0.9cm Table 5. Fractionation with Ethyl acetate fraction Distance trveled by Distance traveled by solvent front spots 3.9cm 3.3cm

www.ijrtem.com

Rf value Rf1 = 0.98 Rf2 = 0.92 Rf3 = 0.83 Rf4 = 0.63 Rf6 = 0.21 Rf5 = 0.32

Rf value Rf1 = 0.92 Rf2 = 0.79 Rf3 = 0.69 Rf4 = 0.51 Rf6 = 0.38 Rf5 = 0.26

Rf value Rf1 = 0.85

|2|

Fractionation of Crude Dye Extracted From Cucurbita Pepo Leaves by Cold Extraction Method 2 3 4 5 6

3.9 cm 3.9 cm 3.9 cm 3.9 cm 3.9 cm

3.0cm 2.2cm 1.6cm 1.1cm 0.7cm

No. of spots

Table 6.Fractionation with Acetone Distance travelled by Distance travelled by solvent front spots

Rf value

1 2 3 4

3.5 cm 3.5 cm 3.5 cm 3.5 cm

Rf1 = 0.56 Rf1 = 0.42 Rf1 = 0.26 Rf1 = 0.18

2.0 cm 1.5 cm 1.0 cm 0.7 cm

Rf2 = 0.77 Rf3 = 0.56 Rf4 = 0.40 Rf6 = 0.27 Rf5 = 0.16

Table 7.Percent yield/ Percent recovery of cold extract of C. pepo Mass of plant leaves: 100g S. No 1 2 3 4

Fractionation solvent Ethanolic extract . n-hexane Chloroform Acetone

Percent Yield (w/w) 6.81% 3.42% 0.77% 2.48%

TLC and UV-Vis study of variousfractions of C. pepo cold extract At ambienttemperature and pressure in 30% of mobile phase (n-hexane 7ml + ethyl acetate 3ml) system a good TLC was developed for various fraction of C. pepo cold extract.The solvents used for fractionation were nhexane, chloroform and acetone. The extracted dye consist of six components while the TLC was developed with n hexane, chloroform and acetone fractions but the extracted dye consist of four components while TLC was developed with acetone fraction. All components have distinct Rf values which are shown in the Tables 4-7. Due to variation of solvent used in fractionation deviation are seen in all cases.

6components/functional groups were confirmed in crude dye fractioned with n hexane, chloroform and ethyl acetate but only 4 components/functional groups were confirmed in crude dye fractioned with acetone. Maximum Rfvalues were observed forchloroform, like Rf1 for chloroform is 0.92 cm. For n-hexane, ethyl acetate and acetone were 0.98cm, 0.85cm and 0.56cmrespectively. All fractions except acetone, 6spots were observed on developed TLCwhile acetone has 5 spots on developedTLC. Similar results have been observed by Shah et al. 2013.

Conclusion From the experiment it can be concluded that the cold extraction method is more suitable while cost and availability is concerned but hot water extraction method is more rapid. Among all samples while 100 gm sample is dissolved in 750 ml the crude extract will be more.The crude dye fractionation with different solvents in TLC showedinmajor thedye has six distinct components.

REFERENCES [1] [2] [3]

[4] [5] [6]

Shahid, M.&Ul-Islam, S. (2013). Recent advancements in natural dye applications. Journal ofCleaner Production, 53: 310–331.

Grifoni D.,Bacci, L.,Zipoli, G., Albanese, L. and Sabatini, F.(2011). The role of natural dyes inthe UV protection of fabrics made of vegetable fibers. Dyes and Pigments, 91: 279–285.

Baliarsingh, S., Panda, A. K., Jena, J., Das, T. & Das, N. B. (2012). Exploring sustainable techniqueon natural dye extraction from native plants for textile: identification of colourants, colourimetric analysis of dyed yarns and their antimicrobial evaluation. Journal of Cleaner Production, 37: 257–264.

Booa H.O., Hwangb, S. J.,Baec, C. S., Parkc, S. H. ,Buk-GuHeod&Gorinsteine, S. (2012).Extraction and characterization of some natural plant pigments. Industrial Crops and Products, 40: 129–135.

Sinha K., SahaaP.D. ,Datta B. S. (2012). Extraction of natural dye from petals of Flame of forest(Buteamonosperma) flower: Process optimization using response surface methodology(RSM). Dyes and Pigments, 94: 212–216.

Shah, N., Farman, S., Hussain,Z., Arain,M. B. &Sulaiman Sham, S. (2013). Pakhtunkhwa J. Life Sci. 1 (03):130-144

| Volume 1 | Issue 10 |

www.ijrtem.com

|3|