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EXTRACTION AND

PURIFICATION OF PLANT

DERIVED SURFACTANTS

Buse Nur TEKİN, Gülden DÖNMEZ, Merve Deniz KÖSE, Mehmet ÜSTÜNDAŞ, Oğuz

BAYRAKTAR

Ege University Faculty of Engineering, Department of Chemical Engineering

CONTENTS

• Introduction

• Sapindus mukorossi

• Soybean waste water

• Experimental

• Results

• Conclusion

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INTRODUCTION

• Human health has been adversely affected by the

increasing use of synthetic surfactants.

• Hence, studies which are to obtain natural surfactant

from plant and industrial waste are increasing

importance day by day.

• Both Sapindus mukorossi fruit and soybean waste

water rich in saponins were examined in this study.

3

Natural surfactants have gained importance

because realizing economical aspect, health and

environmental effect of usage of synthetic surfactants.

Primary studies in surfactants were concerned with

obtain natural surfactants from plants such as S.

mukorossi, Soybean (Glycine max L. Merrill), Soapwort

(Saponaria officinalis), Bracken (Pteridium aquilinum),

The Horse Chestnut (Aesculus hippocastanum), Soap

Lily (Chlorogalum pomeridianum), Yucca.

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Sapindus mukorossi

It is known as soapberries or soapnut.

Saponin is a type of nonionic surfactant.

Naturally saponins occur from glycosides of steroids,

alkaloids and triterpenoids.

Sapogenin + Monosaccharide = Saponin

5

6

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Soybean waste water

• Soybean waste commonly given to the municipal

water sewer and cause environmental problems like

eutrotification. Soybean waste contains 23%

hemicellulose, 16% cellulose and 28% protein.

Soybean waste water Soybean

8

Food Amount of Saponin (g/kg solid)

Peas (Pisum sativa spp) 11

Soybean (Glycine max L. Merrill) 43

Peanut (Arachis hypogaea L.) 6.3

Lentil 3.7-4.6

Spinach (Spinacia oleracea L.) 47

Asparagus (Asparagus officinalis L.) 15

Oat (Avena sativa L.) 1.0

Garlic (Allium purchased L.) 2.9

Sesame Seeds (Sesame my inducer L.) 3.0

Green beans (P. vulgaris) 13

Chickpea (Cicer arietinum L.) 56

Bean (Vicia faba) 3.5

Beet (Beta vulgaris) 58

Table 1. Natural saponin sources [5].

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Mostly used techniques for purifying saponins;

Ultrafiltration

Column chromatography

Foam fractionation

HPLC analyses .

Among these techniques foam separation is

considered as a simple, energy efficient and

environmentally friendly technique.

10

The leaching liquor from the S. mukorossi has

complex ingredients such as saponin, saccharides,

proteins and other compounds.

The foaming ability of saponins is caused by

the combination of a hydrophobic (fat-soluble)

sapogenin and a hydrophilic (water-soluble) sugar

part.

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S.Mukorossi powder

dissolved in each solvent

(1:10) and (1:20) (w:v)

Extract made a foam

separation

200ml solution at

0.814 L/min at

25℃

UV spectrophotometer

200ml solution

at 0.814 L/min

at 80℃

Soybean waste

water

FT-IR analysis

HPLC analysis

EXPERIMENTAL

For S.mukorossi

For soybean 12

RESULTS

solvent type solid- liquid

ratio (g/ml) yield %

g saponin/10g raw

mat.

Water (1:10) 76 2.4

water@100C (1:10) 71 2.5

ethanol-water (1:10) 78 3.9

Ethanol (1:10) 68 3.5

Acetone (1:10) 65 3.4

Water (1:20) 74 2.8

water@100C (1:20) 70 2.6

ethanol-water (1:20) 66 2.5

Ethanol (1:20) 45 1.2

Table 1. Effects of solvents and solid/liquid ratio on the characteristics of S.mukorossi saponins .

Optimum extraction conditions were found as ethanol water solution at 1:10 ratio. 13

• FTIR spectra of the samples showed –OH, -C=O, C-H, and

C=C absorptions characteristic of oleanane triterpenoid

saponins. The C-O-C absorptions showed glycoside connect to

the sapogenins [3].

Figure 1. FTIR spectra of the foam fractination product and oleanolic

acid [2]. 14

Figure 2. FT-IR spectra of S. mukorossi Extract. (W:Water, E:Ethanol,

EW:Ethanol-water, BW:Water at 100℃)

15

Exp. No Initial Solution Final Foam Residual Solution

Air Flow Rate (L/min) Time (min) Volume (ml) 200 28 172

pH 5,58 6,2 6,6

Colour Slightly yellow White Slightly yellow

Volume (ml) 200 30 170

pH 6,86 7,54 7,47

Colour Slightly yellow White Slightly yellow

B

A

Parameters

Temperature 25℃

Temperature 80℃

0,814 75

Table 2. Results of foam separation

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Figure 3. FT-IR Spectra of samples: RW: Raw material, EW:Ethanol-

water solution, FA:Foam phase A, FB:Foam phase B.

Foam phase was rich in saponin compared with

ethanol – water extract and raw material.

With increasing the temperature, the adsorption is

enhanced as in the case of foam fraction experiment

carried out at relatively higher temperature. 17

-500

0

500

1000

1500

2000

2500

3000

0 0,5 1 1,5 2 2,5 3 3,5 4 4,5 5

mA

U

min

FP

IS

BP

IS

Figure 4. HPLC chromatogram analysis of saponins in soybean

waste water. (FP:Foam phase, IS:Initial solution).

Calculated enrichment ratio was found as 57% using spectrophotometric analysis, and 61% using HPLC analysis.

By using foam separation method it was possible to obtain value added natural surfactants from soybean waste waters. 18

CONCLUSION

• With foam separation technique higher amount of

saponins can be obtained from S.mukorossi and soy

bean waste. The obtained product is non-toxic and

eco-friendly. This product could be used in baby

shampoo, detergents and pesticide residue as a

replacement ingredient.

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CONCLUSION

• By using foam separation technique it is possible to

obtain value added natural surfactants from waste

water. Valorization of these wastes are highly

important for the national economy.

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REFERENCES [1] Yan, J., Wu, Z.L., Zhao, Y.L., Jiang, C.S., 2011. Seperation of tea saponin by two stage foam fractionation. Seperation and Purification Technology. 80, 300-305.

[2] Li, R., Wu, Z. L., Wang, Y.J., Li.L.L., 2013. Separation of total saponins from the pericarp of S. mukorossi Gaerten. by foam fractionation. Industrial Crops and Products. 51, 163– 170.

[3] Kareru, P. G1. Keriko, J. M1., Gachanja, A. N1., Kenji, G. M., 2008. Direct Detection of Triterpenoid Saponins in Medicinal Plants. Afr. J. Trad. CAM 5 (1): 56 – 60.

[4] Akbel, E., Karadağ, Funda., 2012. Saponin ve Reproduktif Etkileri,Türk Bilimsel Derlemeler Dergisi 5, 25-29.

[5] Dinda, B., Debnath, S., Mohanta B. C., Harigaya, Y.,2010. Naturally occurring triterpenoid saponins. Chemistry and Biodiversity. 7, 2327-2580.

[6] Cheok, C.Y., Salman, H. A. K., Sulaiman, R., 2014. Food Research International. 59, 16-40.

[7] Du, M., Huang, S., Zhang, J., Wang, J., Hu, L., Jıang, J., 2013. Isolation of Total Saponins from Sapindus mukorossi Gaerth. Vol.4, No.1, 24-27.

[8] Heng, W., Ling, Z., Na, W., Youzhi, G., Zhen, W., Zhiyong, S., Deping, X., Yunfei, X., Weirong, Y., 2015. Extraction and Fermantation-Based Purification of Saponins from Sapindus mukorossi Gaertn. 18, 429-438.

[9] Ibrahim, M., Khan, A.A., 2006. Antimicrobial Activity of Sapindus Mukorossi and Rheum Emodi Extracts Against H pylori: In itro and in vivo studies. World J Gastroenterol , 12(44), 7136-7142.

[10] Sparg, S.G. , Light, M.E. , van Stade, J., 2004. Biological activities and distribution of plant saponins. Journal of Ethnopharmacology 94 (2004) 219–243.

[11] Takeuchi, T. M., Pereira, C. G., Braga, M. E. M., Marostica, M. R., Leal, P. F., & Meireles, M. A. A., 2009. Low pressure solvent extraction (Solid liquid extraction, microwave assisted, and ultrasound assisted) from condimentary plants. In M. A. A. Meireles (Ed.), Extracting bioactive compounds for food products-Theory and applications (pp.140-144). Boca Raton: CRC press, 151-158.

[12] Bhargava, D., Shivapuri, J.N., Kar, S., Pandit, B.R., Sidhiqie, A., Upadhyay, A., Thakur, S., Mondal, K.C., 2012. Evaluation of Antigonorrhoeal Activity of Saponins Extract of Sapindus Mukorossi Gaertn. Vol.3, No.2, 459.

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Thank you very much for your kind Attention!

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