Skip to main content
SpringerLink
Log in

Advances in various techniques for isolation and purification of sterols

  • Review Article
  • Published:
Journal of Food Science and Technology Aims and scope Submit manuscript

Abstract

Plants consist of triterpenoids such as phytosterols (PT) (C29H50O) with steroidal nuclei, including sitosterol, stigmasterol, brassicasterol and campesterol. They are hydrophobic but soluble in alcohol and other organic solvents and are isolated from industrial waste deodorizer distillates of various edible oil industries. They exist as free PT or their ester derivatives in soybean, rice, wheat, oat, cottonseed and corn fiber, and other cereals and grains. Conventional isolation techniques such as solvent extraction, distillation, evaporative fractionation, saponification and chemical esterification are employed for isolation and purification of PT. The present article reviews the various advanced separation techniques like solvent crystallization, supercritical fluid extraction, high speed counter-current chromatography and enzymatic process as strategic methods to isolate and purify sterols.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1

Similar content being viewed by others

References

  • Abbas C, Rammelsberg AM, Beery K (2008) Extraction of phytosterols from corn fiber using green solvents, Google Patents

  • Abbas K, Mohamed A, Abdulamir A, Abas H (2008b) A review on supercritical fluid extraction as new analytical method. Am J Biochem Biotechnol 4(4):345–353

    Article  CAS  Google Scholar 

  • Agostini F, Bertussi R, Agostini G, Atti dos Santos A, Rossato M, Vanderlinde R (2012) Supercritical extraction from vinification residues: fatty acids, α-tocopherol, and phenolic compounds in the oil seeds from different varieties of grape. Sci World J 2012:1–9

    Article  CAS  Google Scholar 

  • Ayre A, Ghude K, Mane P, Nemade M, Gosavi S, Pathare A, Lad A (2013) Supercritical fluid extraction-a green paradigm in the area of separation science. Asian J Biomed Pharm Sci 3(23):1–7

    CAS  Google Scholar 

  • Berry WW, Sutterlin WR, Tegen MG, Ryan L (2018) Methods for the separation or purification of vitamin E, tocopherols and tocotrienols from lipid mixtures. Google Patents

  • Böszörményi A, Szarka S, Hethelyi E, Gyurjan I, Laszlo M, Simándi B, Szőke É, Lemberkovics É (2009) Triterpenes in traditional and supercritical-fluid extracts of Morus alba leaf and stem bark. Acta Chromatogr 21(4):659–669

    Article  CAS  Google Scholar 

  • Bruni R, Medici A, Guerrini A, Scalia S, Poli F, Muzzoli M, Sacchetti G (2001) Wild Amaranthus caudatus seed oil, a nutraceutical resource from Ecuadorian flora. J Agric Food Chem 49(11):5455–5460

    Article  CAS  PubMed  Google Scholar 

  • Carmona MA, Jiménez C, Jiménez-Sanchidrián C, Peña F, Ruiz JR (2010) Isolation of sterols from sunflower oil deodorizer distillate. J Food Eng 101(2):210–213

    Article  CAS  Google Scholar 

  • Chitra J, Ghosh M, Mishra H (2017) Rapid quantification of cholesterol in dairy powders using Fourier transform near infrared spectroscopy and chemometrics. Food Control 78:342–349

    Article  CAS  Google Scholar 

  • Chuang YH, Ju YH, Widjaja A (2006) Separation of campesterol and β-sitosterol from a sterol mixture. Sep Sci Technol 41(13):3027–3038

    Article  CAS  Google Scholar 

  • Copeland D, Belcher M (2003) Methods for treating deodorizer distillate. WO03080778

  • Dunford N, King J (2000) Phytosterol enrichment of rice bran oil by a supercritical carbon dioxide fractionation technique. J Food Sci 65(8):1395–1399

    Article  CAS  Google Scholar 

  • Eller F, Moser J, Kenar J, Taylor S (2010) Extraction and analysis of tomato seed oil. J Am Oil Chem Soc 87(7):755–762

    Article  CAS  Google Scholar 

  • Fabian C, Gunawan S, Kasim NS, Chiang C-L, Ju Y-H (2009) Separation of nonpolar lipid from soybean oil deodorizer distillate by stirred batch-wise silica gel adsorption-desorption. Sep Sci Technol 44(7):1621–1637

    Article  CAS  Google Scholar 

  • Fang L, Liu Y, Yang B, Wang X, Huang L (2011) Separation of alkaloids from herbs using high-speed counter-current chromatography. J Sep Sci 34(19):2545–2558

    Article  CAS  PubMed  Google Scholar 

  • Ferdosh S, Haque Akanda MJ, Ghafoor K, Ali ME, Kamaruzzaman B, Shaarani S, Islam Sarker MZ (2018) Techniques for the extraction of phytosterols and their benefits in human health: a review. Sep Sci Technol 53(14):2206–2223

    Article  CAS  Google Scholar 

  • Fernandes P, Cabral J (2007) Phytosterols: applications and recovery methods. Biores Technol 98(12):2335–2350

    Article  CAS  Google Scholar 

  • Fornari T, Torres CF, Señoráns FJ, Reglero G (2009) Simulation and optimization of supercritical fluid purification of phytosterol esters. AIChE J 55(4):1023–1029

    Article  CAS  Google Scholar 

  • Ghosh R (2006) Principles of bioseparations engineering. World Scientific Publishing Company, Singapore

    Book  Google Scholar 

  • Gossé B, Amissa AA, Anoh Adjé F, Bobélé Niamké F, Ollivier D, Ito Y (2005) Analysis of components of Neem (Azadirachta indica) oil by diverse chromatographic techniques. J Liq Chromatogr Relat Technol 28(14):2225–2233

    Article  CAS  Google Scholar 

  • Gunawan S, Ismadji S, Ju Y-H (2008a) Design and operation of a modified silica gel column chromatography. J Chin Inst Chem Eng, 39(6):625–633

    Article  CAS  Google Scholar 

  • Gunawan S, Kasim NS, Ju Y-H (2008b) Separation and purification of squalene from soybean oil deodorizer distillate. Sep Purif Technol 60(2):128–135

    Article  CAS  Google Scholar 

  • Holen B (1985) Rapid separation of free sterols by reversed-phase high performance liquid chromatography. J Am Oil Chem Soc 62(9):1344–1346

    Article  CAS  Google Scholar 

  • Hu L, Llibin S, Li J, Qi L, Zhang X, Yu D, Walid E, Jiang L (2015) Lipase-catalyzed transesterification of soybean oil and phytosterol in supercritical CO2. Bioprocess Biosyst Eng 38(12):2343–2347

    Article  CAS  PubMed  Google Scholar 

  • Huang L, Zhong T, Chen T, Ye Z, Chen G (2007) Identification of β-sitosterol, stigmasterol and ergosterin in A. roxburghii using supercritical fluid extraction followed by liquid chromatography/atmospheric pressure chemical ionization ion trap mass spectrometry. Rapid Commun Mass Spectrom 21(18):3024–3032

    Article  CAS  PubMed  Google Scholar 

  • Huang L, Cao Y, Xu H, Chen G (2011) Separation and purification of ergosterol and stigmasterol in Anoectochilus roxburghii (wall) Lindl by high-speed counter-current chromatography. J Sep Sci 34(4):385–392

    Article  CAS  PubMed  Google Scholar 

  • Hwang SH, Jang JM, Lim SS (2012) Isolation of fucosterol from Pelvetia siliquosa by high-speed countercurrent chromatography. Fish Aquat Sci 15(3):191–195

    CAS  Google Scholar 

  • Irmak S, Solakyildirim K, Hesenov A, Erbatur O (2010) Study on the stability of supercritical fluid extracted rosemary (Rosmarinus offcinalis L.) essential oil. J Anal Chem 65(9):899–906

    Article  CAS  Google Scholar 

  • Ito V, Martins P, Batistella C, Maciel MW (2005) Tocopherols and phytosterols concentration from soybean oil deodorizer distillate. Presentado en 4th Mercosur Congress on process systems Engineering and 2nd Mercosur Congress on Chemical Engineering, Rio de Janeiro

  • Kagliwal LD, Pol AS, Patil SC, Singhal RS, Patravale VB (2012) Antioxidant-rich extract from dehydrated seabuckthorn berries by supercritical carbon dioxide extraction. Food Bioprocess Technol 5(7):2768–2776

    Article  CAS  Google Scholar 

  • Kasim NS, Gunawan S, Yuliana M, Ju Y-H (2010) A simple two-step method for simultaneous isolation of tocopherols and free phytosterols from soybean oil deodorizer distillate with high purity and recovery. Sep Sci Technol 45(16):2437–2446

    Article  CAS  Google Scholar 

  • Ketenoglu O, Tekin A (2015) Applications of molecular distillation technique in food products. Ital J Food Sci 27(3):277–281

    CAS  Google Scholar 

  • Khatoon S, Rajan RR, Krishna AG (2010) Physicochemical characteristics and composition of Indian soybean oil deodorizer distillate and the recovery of phytosterols. J Am Oil Chem Soc 87(3):321–326

    Article  CAS  Google Scholar 

  • Kirwan D, Orella C (2002) Crystallization in the pharmaceutical and bioprocessing industries. In: Myerson AS (ed) Handbook of industrial crystallization, 2nd edn. Elsevier, Amsterdam, pp 249–266

    Chapter  Google Scholar 

  • Kong Z, Rinehart KL, Milberg RM, Conway W (1998) Application of high-speed countercurrent chromatography/electrospray ionization mass spectrometry (HSCCC/ESIMS) in natural products chemistry. J Liq Chromatogr Relat Technol 21(1–2):65–82

    Article  CAS  Google Scholar 

  • Kotnik P, Škerget M, Knez Ž (2006) Kinetics of supercritical carbon dioxide extraction of borage and evening primrose seed oil. Eur J Lipid Sci Technol 108(7):569–576

    Article  CAS  Google Scholar 

  • Kumar P, Murali-Krishna C (2019) Optical techniques: investigations in oral cancers. In: Panta P (ed) Oral cancer detection: novel strategies and clinical impact. Springer, Cham, pp 167–187

    Chapter  Google Scholar 

  • Leo L, Rescio L, Ciurlia L, Zacheo G (2005) Supercritical carbon dioxide extraction of oil and α-tocopherol from almond seeds. J Sci Food Agric 85(13):2167–2174

    Article  CAS  Google Scholar 

  • Liu D, Shi J, Posada LR, Kakuda Y, Xue SJ (2008) Separating tocotrienols from palm oil by molecular distillation. Food Rev Int 24(4):376–391

    Article  CAS  Google Scholar 

  • Liu G, Xu X, Hao Q, Gao Y (2009) Supercritical CO2 extraction optimization of pomegranate (Punica granatum L.) seed oil using response surface methodology. LWT Food Sci Technol 42(9):1491–1495

    Article  CAS  Google Scholar 

  • Mingyai S, Srikaeo K, Kettawan A, Singanusong R, Nakagawa K, Kimura F, Ito J (2018) Effects of extraction methods on phytochemicals of rice bran oils produced from colored rice. J Oleo Sci 67(2):135–142. https://doi.org/10.5650/jos.ess17122

    Article  CAS  PubMed  Google Scholar 

  • Moreau RA, Whitaker BD, Hicks KB (2002) Phytosterols, phytostanols, and their conjugates in foods: structural diversity, quantitative analysis, and health-promoting uses. Prog Lipid Res 41(6):457–500

    Article  CAS  PubMed  Google Scholar 

  • Myerson A (2002) Handbook of industrial crystallization. Butterworth-Heinemann, Oxford

    Google Scholar 

  • Nagao T, Kobayashi T, Hirota Y, Kitano M, Kishimoto N, Fujita T, Watanabe Y, Shimada Y (2005) Improvement of a process for purification of tocopherols and sterols from soybean oil deodorizer distillate. J Mol Catal B Enzym 37(1–6):56–62

    Article  CAS  Google Scholar 

  • Özdemir İS, Dağ Ç, Özinanç G, Suçsoran Ö, Ertaş E, Bekiroğlu S (2018) Quantification of sterols and fatty acids of extra virgin olive oils by FT-NIR spectroscopy and multivariate statistical analyses. LWT 91:125–132

    Article  CAS  Google Scholar 

  • Panpipat W, Xu X, Guo Z (2012) Towards a commercially potential process: enzymatic recovery of phytosterols from plant oil deodoriser distillates mixture. Process Biochem 47(8):1256–1262

    Article  CAS  Google Scholar 

  • Pei H, Ma X, Pan Y, Han T, Lu Z, Wu R, Cao X, Zheng J (2019) Separation and purification of lanosterol, dihydrolanosterol, and cholesterol from lanolin by high-performance counter-current chromatography dual-mode elution method. J Sep Sci 42(12):2171–2178. https://doi.org/10.1002/jssc.201900063

    Article  CAS  PubMed  Google Scholar 

  • Robinson PL, Cuff TJ, Parker JE III (2000) Isolation and purification of sterols from neutrals fraction of tall oil pitch by single decantation crystallization. Google Patents

  • Romo-Hualde A, Yetano-Cunchillos A, González-Ferrero C, Sáiz-Abajo M, González-Navarro C (2012) Supercritical fluid extraction and microencapsulation of bioactive compounds from red pepper (Capsicum annum L.) by-products. Food Chem 133(3):1045–1049

    Article  CAS  Google Scholar 

  • Rousková M, Heyberger A, Tříska J, Krtička M (2011) Extraction of phytosterols from tall oil soap using selected organic solvents. Chem Pap 65(6):805

    Article  CAS  Google Scholar 

  • Sajfrtová M, Ličková I, Wimmerová M, Sovová H, Wimmer Z (2010) β-sitosterol: supercritical carbon dioxide extraction from sea buckthorn (Hippophae rhamnoides L.) seeds. Int J Mol Sci 11(4):1842–1850

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Salas S, Chakraborty K (2018) First report of bioactive sterols from the muricid gastropod Chicoreus ramosus. Steroids 137:57–63

    Article  CAS  PubMed  Google Scholar 

  • Sánchez-Camargo ADP, Parada-Alonso F, Ibáñez E, Cifuentes A (2019) Recent applications of on-line supercritical fluid extraction coupled to advanced analytical techniques for compounds extraction and identification. J Sep Sci 42(1):243–257

    Article  PubMed  CAS  Google Scholar 

  • Savinova T, Diep NT, Voishvillo N, Andryushina V, Karpova N, Beletskaya I, Huy LD (2012) Extraction of a mixture of phytosterols from soybean processing by-product and its use in the manufacture of 9α-hydroxyandrost-4-en-3, 17-dione. Pharm Chem J 46(3):183–186

    Article  CAS  Google Scholar 

  • Schröder M, Vetter W (2012) Investigation of unsaponifiable matter of plant oils and isolation of eight phytosterols by means of high-speed counter-current chromatography. J Chromatogr A 1237:96–105

    Article  PubMed  CAS  Google Scholar 

  • Sethi N, Anand A, Sharma A, Chandrul KK, Jain G, Srinivasa KS (2009) High speed counter current chromatography: a support-free LC technique. J Pharm Bioallied Sci 1(1):8

    Article  Google Scholar 

  • Srividya N, Heidorn DB, Lange BM (2014) Rapid purification of gram quantities of β-sitosterol from a commercial phytosterol mixture. BMC Res Notes 7(1):182

    Article  PubMed  PubMed Central  Google Scholar 

  • Sujith Kumar M, Mawlong I, Singh D (2017) Phytosterol recovery from oilseeds: recent advances. J Food Process Eng 40(3):e12466

    Article  CAS  Google Scholar 

  • Teixeira A, Santos J, Crespo J (2011) Production of steryl esters from vegetable oil deodorizer distillates by enzymatic esterification. Ind Eng Chem Res 50(5):2865–2875

    Article  CAS  Google Scholar 

  • Uddin MS, Sarker MZI, Ferdosh S, Akanda MJH, Easmin MS, Bt Shamsudin SH, Yunus KB (2015) Phytosterols and their extraction from various plant matrices using supercritical carbon dioxide: a review. J Sci Food Agric 95(7):1385–1394

    Article  CAS  PubMed  Google Scholar 

  • Venturelli A, Brighenti V, Mascolo D, Pellati F (2019) A new strategy based on microwave-assisted technology for the extraction and purification of beeswax policosanols for pharmaceutical purposes and beyond. J Pharm Biomed Anal 172:200–205

    Article  CAS  PubMed  Google Scholar 

  • Watanabe Y, Nagao T, Hirota Y, Kitano M, Shimada Y (2004) Purification of tocopherols and phytosterols by a two-step in situ enzymatic reaction. J Am Oil Chem Soc 81(4):339–345

    Article  CAS  Google Scholar 

  • Xiao X-H, Yuan Z-Q, Li G-K (2013) Preparation of phytosterols and phytol from edible marine algae by microwave-assisted extraction and high-speed counter-current chromatography. Sep Purif Technol 104:284–289

    Article  CAS  Google Scholar 

  • Yan F, Yang H, Li J, Wang H (2012) Optimization of phytosterols recovery from soybean oil deodorizer distillate. J Am Oil Chem Soc 89(7):1363–1370

    CAS  Google Scholar 

  • Yan F, Huang Y, Wang Y, Li Q, He X (2019) Bioactive sterols and triterpenoids from the fruits of giant crepe-myrtle. Ind Crops Prod 130:363–370

    Article  CAS  Google Scholar 

  • Yang H, Yan F, Wu D, Huo M, Li J, Cao Y, Jiang Y (2010) Recovery of phytosterols from waste residue of soybean oil deodorizer distillate. Biores Technol 101(5):1471–1476

    Article  CAS  Google Scholar 

  • Zhang X, Cambrai A, Miesch M, Roussi S, Raul F, Aoude-Werner D, Marchioni E (2006) Separation of Δ5-and Δ7-phytosterols by adsorption chromatography and semipreparative reversed phase high-performance liquid chromatography for quantitative analysis of phytosterols in foods. J Agric Food Chem 54(4):1196–1202

    Article  CAS  PubMed  Google Scholar 

  • Zhang H, Cao X, Liu Y, Shang F (2017) Rapid recovery of high content phytosterols from corn silk. Chem Cent J 11(1):108

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhou Y, Chen F, Li Z (2002) Preparative separation of β-sitosterol by high speed countercurrent chromatography. J Liq Chromatogr Relat Technol 25(10–11):1693–1701

    Article  CAS  Google Scholar 

  • Zougagh M, Valcárcel M, Rıos A (2004) Supercritical fluid extraction: a critical review of its analytical usefulness. TrAC Trends Anal Chem 23(5):399–405

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors did not receive any funds for this work.

Author information

Author notes

  1. Piyush Kumar

    Present address: Department of Environmental Medicine and Public Health, Icahn School of Medicine at Mount Sinai, New York City, 10029, USA

  2. Sneha Dikshit and Sakshi Bubna have contributed equally to this work.

Authors and Affiliations

  1. Amity Institute of Biotechnology, Amity University Mumbai, Bhatan, Post-Somatane, Panvel, Navi Mumbai, Maharashtra, 410206, India

    Sneha Dikshit, Sakshi Bubna, Anand Gupta & Piyush Kumar

Authors
  1. Sneha Dikshit
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sakshi Bubna
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Anand Gupta
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Piyush Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Anand Gupta.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Dikshit, S., Bubna, S., Gupta, A. et al. Advances in various techniques for isolation and purification of sterols. J Food Sci Technol 57, 2393–2403 (2020). https://doi.org/10.1007/s13197-019-04209-3

Download citation

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13197-019-04209-3

Keywords

Search

Navigation