Isolation of Curcuminoids from Turmeric Using Non Chlorinated Solvents

Journal of Critical Reviews

ISSN- 2394-5125 Vol 7, Issue 14, 2020 ISOLATION OF CURCUMINOIDS FROM TURMERIC USING NON CHLORINATED SOLVENTS

S.I.Davis Presley,1*V.G.Karthik Ramanan,1 S.M.Prasanth1

Department of Chemistry, SSN College of Engineering, Chennai, India. Email: [email protected] *

Received:13.04.2020 Revised: 18.05.2020 Accepted: 17.06.2020

Abstract For centuries turmeric is used as a spice and medicinal herb in India. Turmeric contains a natural phenolic compound called curcuminoids and curcumin being its principal composition. Curcuminoids are used widely as aconventional medicine and in food for their coloring and aromatic properties. Curcumin has received a lot of research interest because of its anti-inflammatory, anti- carcinogenic and anti-tumour properties .T urmeric is commonly available in grocery stores. Turmeric contains two percent of curcumin by weight and also contain smaller amount of other curcuminoids. Curcumin from turmeric is extacted by column chromatography or by soxhlet extraction.From literatures we find chlorinated solvents are widely used for extraction.So we want to try a suitable mobile phase free from chlorinated solvents and also to get maximum yield of curcuminoids.

Keywords: Turmeric, curcumin, methanol, column chromatography, ethylacetate, hexane

INTRODUCTION The biological activity of the turmeric is due to curcumin and Turmeric (Curcuma longa) is used as an spice for a very long its two analogues.[4] Variety of turmeric rhizomes are found in time, a native of southeast Asia, used from ancient times as a different geographical zones which contains about 2-9% of dye and a condiment. It is a flowering plant and it belongs to curcuminoids, but in some very special species the content of the ginger family, Zingiberaceae. It is a herbaceous plant which curcuminoids may go up to 15% .Highest curcuminoid content is perennial. Highly branched, yellow to orange, cylindrical, is found in curcuma zedoaria(>100 µg g-1)whereas in curcuma aromatic rhizomes are found. In many Asian dishes turmeric is longa the content is (1–2 µg g-1)and curcuma aromatic the a key ingredient which imparts a mustard-like, earthy aroma content is (0.1 µg g-1) [5]. and pungent odour which gives a slightly bitter flavor to foods. Curcuma longa is found to possess many properties as There are experimental evidences to show the effects of per medical literature like anti-cancer, anti-coagulative and curcumin for the treatment of variety of diseases[6].One of the anti-hepatotoxic properties [1] . Turmeric rhizome powder important property of curcumin is anti-inflammatory initiates immune response and aspartate aminotransferase properties which is useful in making drugs.Curcumin also (AST) and lactate dehydrogenase (LDH) activities in the serum inhibits the formation of plaques, fibrils and amyloid b of caged chickens [2]. oligomers, and rescues amyloid in vivo[7].Also, as per [8].curcumin also reduces induced cytotoxicity in parkinson’s Turmeric mainly consists of three constituents, which are disease. commonly called curcuminoids. They are curcumin, demethoxycurcumin and bis-demethoxycurcumin. Curcumin Curcumin another important function is that it reverses (1,7-bis(4-hydroxy 3-methoxyphenyl)-1,6-heptadione-3,5- impaired hippocampal neurogenesis and increases serotonin dione) is a polyphenolic compound obtained from turmeric receptor 1A m-RNA and brain-derived neurotrophic factor which contains about 2 to 5% of the rhizome content and also expression in chronically stressed rats[9]. Curcuminoids it is a more valuable compound of turmeric[3]. improves the life of patients with solid tumours by suppressing their systemic inflammation.

Curcumin an orange-yellow crystalline powder, was first microwave assisted extraction[11], high performance sub- extracted in 1895. Many extraction methods were followed in critical water extraction, soxhlet extraction, column recent decades to extract curcumin. Some of them include chromatography[12], and ionic liquid extraction assisted by

Journal of critical reviews 723

ISOLATION OF CURCUMINOIDS FROM TURMERIC USING NON CHLORINATED SOLVENTS

enzyme [13] . Here, the technique of column chromatography Sample preparation is exploited. Here we tried different non chlorinated solvents Five gram of powdered sample is taken in a petri dish dried for based on their polarity. The turmeric we are going to use is 1 hour in a hot air oven. The dried sample is loaded in a wet purchased from a local retailer shop. A mixture of hexane and column chromatography. Mobile phase hexane is allowed to ethyl acetate taken in the ratio of 7:3 (by volume) is used as run on the column for six to seven hours. The fractions were the mobile phase in the column. The main advantage of using collected and spotted on TLC plates. The mobile phase is used this combination of solvents is that, neither hexane nor ethyl as a developing solvent. Both the sample spots and standard acetate constitutes chlorine, as most of the chlorinated spots were matched and the similar fractions were collected. solvents are carcinogenic. The extracts are tested on TLC for The collected fractions were subjected to vacuum distillation the presence of curcumin and (other curcuminoids) and HPLC in a rotary evaporator. After complete removal of the solvent tests are carried out on the powdered extracts to check their crystallization is done by adding 10 ml of isopropyl alcohol. assay. The solids were filtered and washed with isopropyl alcohol.

MATERIALS AND METHODS RESULTS AND DISCUSSION LR grade solvents,Merck TLC plates , Curcumin reference Solvents with different polarity were tried for the isolation of standard (Obtained from VenBiotech Pvt Ltd.India), Column the curcuminoids (Table1). Out of the solvents we find hexane (Diameter: 30 mm, Height: 500 mm ) , Electric hot air oven , and ethyl acetate in the ratio of 7:3 is suitable for isolation. For Rotary evaporator (Model: Evator; Capacity: 2000 mL).The isolation of curcuminoids mostly chlorinated solvents along extracted sample were analysed for its assay by using with methanol is employed. So we need to replace it with new shimadzu HPLC.Separation of curcuminoids by TLC. mobile phase. By using ethyl acetate and hexane in the ratio 7:3 as the mobile phase we collected different fractions and The eluents were tested on merck TLC plates. Spots were checked the spots on TLC.The spots which matches with the placed on TLC plates and kept in a beaker having mobile standard curcumin are collected and the solvents are phase. The plates were dried after the development in a hot air evaporated by using rotary evaporator. Oily residue is oven and the spots were analysed in UV Chamber. obtained. Then the crystallization is done by using isopropyl alcohol. Isopropyl alcohol is added and stirred for 4 hours at 5°C.The solid formed is filtered and washed with isopropyl alcohol. Assay of the extracted sample is checked by HPLC.Weight of the solid formed = 0.05g

Table -I Different solvent combination Dichloromethane : Methanol a) 99:1 Ethyl acetate : Methanol a) 99:1 b) 90:10 c) 75:25 Ethyl acetate : Hexane a) 80:20 b) 60:40 c) 30:70

Table-II HPLC Analysis of isolated curcumin

Parameters Results

Assay Total Curcuminoids(%w/w) 99.869% By HPLC ( as is basis) Bis demethoxy curcumin(%w/w) 3.514% De methoxy curcumin(%w/w) 24.245% Curcumin(%w/w) 72.110%

CONCLUSION Influence of Turmeric Rhizome and Black Pepper on A suitable non chlorinated solvent like ethyl acetate and Blood Constituents and Performance of Broiler Chickens. hexane can be used for the isolation of curcumin the above African J of Biotechnology 2012;34:8606–11. mobile is less hazourdous compared to chlorinated solvents 3. Valizadeh K, Mohammad,Ghasem Najafpour-Darzi, when used in larger quantities, the assay of the isolated Mostafa R, Ali Akbar M,Meisam Valizadeh K, High curcumioids show that a good percentage of curcumin is Performance Curcumin Subcritical Water Extraction from present.So we can conclude the above mobile phase can be Turmeric (Curcuma Longa L.J of Chromatography B: used for the isolation. Analytical Technologies in the Biomedical and Life Sciences,2016;1022: 191–98. ACKNOWLEDGEMENT 4. Ferreira, Izabela M, Rodrigues Uliana M, Marques Costa The authors want to thank SSNCE management for funding S, Massimiliano M, Vianello F, Lin Chau M, Giuseppina this project. Pace Pereira L, Exclusion of Solar UV Radiation Increases the Yield of Curcuminoid in Curcuma Longa L. Industrial CONFLICT OF INTEREST Crops and Products 2016;89:188–94. The authors declare there is no conflict of Interest. 5. Esatbeyoglu, Tuba, Huebbe P, Insa M A, Ernst, Chin D, Anika E,Wagner, Rimbach W, Curcumin-from Molecule to ETHICAL CERTIFICATE Biological Function.Angewandte Chemie - International This project doesn’t require any human or animal trials. Edition 2012;51:5308–32. 6. Gupta A K, Mishra R. Lal R K, Characterization and REFERENCES Utilization of Agronomical Traits in Turmeric (Curcuma 1. Selvam R, Lalitha S, Gayathri R, Angayarkanni N. The Longa L.). Industrial Crops and Products 2015;77:708– Anti-Oxidant Activity of Turmeric (Curcuma Longa). J of 12. Ethnopharmacology 1995;47 :59–67. 7. Yang, Fusheng, Giselle P,Lim, Begum A N, Oliver J, 2. Akbarian A, Abolghasem G, Hassan K, Ali G, Sajad M, Ubeda,Mychica R, Simmons, Surendra S, Curcumin

Journal of critical reviews 724

ISOLATION OF CURCUMINOIDS FROM TURMERIC USING NON CHLORINATED SOLVENTS

Inhibits Formation of Amyloid β Oligomers and Fibrils, Controlled Trial.Phytotherapy Research 2014; 28(10): Binds Plaques, and Reduces Amyloid in Vivo.J of 1461–67. Biological Chemistry 2005;280(7): 5892–5901. 11. Mandal,Vivekananda, Mohan Y, Hemalatha S, Microwave 8. Dane, S., Welcome, M.O.A case study: Effects of wet Assisted Extraction of Curcumin by Sample-Solvent Dual cupping therapy in a male with primary infertility (2019) Heating Mechanism Using Taguchi L9 Orthogonal Design Journal of Complementary Medicine Research, 10, pp. Journal of Pharmaceutical and Biomedical Analysis 2008; 155-161. 46(2): 322-327. 9. Xu Y, Ku B,Cui L,Li X, Barish P A, Foster C T, Ogle W O, 12. Zhan,Yin P, Hua Zeng X, Ming Zhang H, Hang Li, High- .Curcumin Reverses Impaired Hippocampal Neurogenesis Efficient Column Chromatographic Extraction of and Increases Serotonin Receptor 1A MRNA and Brain- Curcumin from Curcuma Longa. Food Derived Neurotrophic Factor Expression in Chronically Chemistry,2011;129(2): 700–703. Stressed Rats Brain Research 2007;1162(1): 9–18. 13. Sahne, Foozie, Mohammadi M, Najafpour D, 10. Panahi,Yunes,Alireza,Saadat,Fatemeh,Beiraghdar,Amirho Moghadamnia A A , Enzyme-Assisted Ionic Liquid ssei, Sahebkar, Adjuvant Therapy with Bioavailability- Extraction of Bioactive Compound from Turmeric Boosted Curcuminoids Suppresses Systemic (Curcuma Longa L.): Isolation, Purification and Analysis Inflammation and Improves Quality of Life in Patients of Curcumin,Crops Industrial and Products 2017;95: with Solid Tumors: A Randomized Double-Blind Placebo- 686–94.

Journal of critical reviews 725