separation techniques in oils & fats science
TRANSCRIPT
Novel Separation Techniques for Isolation of Fatty Acids and Oil By-Products
National Conference on
“Newer Oleo-chemicals: Production & Industrial Application”
Organised By
Centre of Excellence on
‘Applied Research, Training & Education in Lipid Science’Department of Oil & Paint Technology,
Harcourt Butler Technological Institute, KanpurJanuary, 10-11, 2015
Sadanand Patel & Sanjay Kr Singh
Department of Oil & Paint Technology
HBTI, Kanpur
Contents
Conclusion
By products
Enzymatic & Urea Complexion
Counter Current Chromatography
Need & Fundamentals
Need???
Potential health benefits of bioactive compounds
Challenge of refineries
Profitability of processing operations
.
.
Separation
Density
Boiling Point
Melting Point
Centrifugal
Separation
Distillation
Crystallization
Principle of Separations
Molecular Properties
Bulk Properties
Melting Point Boiling Point Density
Position & Nature of Bonds Packing Structure Molecular Dimensions
Liquid Chromatography
Packing Materials Type
Adsorption
Ion Exchange
Gel Filtration
Non Packing Materials Type
Centrifugal Field
Gravitational
Preparative Liquid Chromatography
Counter Current Chromatography
This technique includes:
Liquid-liquid partition,
Countercurrent distribution of solute mixture
between two immiscible liquid phases,
In the most distinct variants of CPC, one liquid phase remains
stationary while the second solvent phase passes through the
stationary phase solvent.
Cost Effective: 10-20%
Efficient: 99.5 – 99.8%
Faster: 2-3 times
Flexible
Easy Scale Up
One single separation /purification techniqueadapted to samples rangingfrommg to several kg.
Bousquet et al*.EPA and DHA from micro algal oil used heptane as the stationary phase and acetonitrile/ water (3% v/v) as the mobile phase.
*O. Bousquet, N. Sellier, and F. L. Goffic, Chrotographia, 39, 40–44 (1994).
Advantages of CPC/CCC & Application
Enzymatic Separations
Long and highly unsaturated
fatty acids show high
resistance for lipases at attack
on ester bonds due to large
stearic hindrance and thus
method is widely used in
separation of 3-PUFA from
marine oils.
Bottino et al.* have illustrated themechanism of resistance of lipasestoward long-chain o3-PUFA inmarine oils.
*N. R. Bottino, G. A. Vandenberg, and R. Reiser, Lipids, 2, 489–493 (1967)
Pure Urea crystallizes intetragonal structure withchannels of 5.67 A° diameter
Crystal channel diameter: 5.6 A0
Fatty acid equivalent diameter: 6.5 – 13.5 A0
Urea complexation
In presence of long
straight-chain molecules,
it crystallizes in hexagonal
structure with inner
channels of 8–12 A°
diameter
Source: A. E. Smith, Acta Cryst., 5, 224–235
TAGs + Alc. KOH
Saponifiable
Unsaponifiable
Almost pure FFAs and Subjected to
Urea Complexation
FFAs
+
Alcohol
Urea
UIC
Two fraction are developed and then UIC fraction is evaporated to separate urea and fatty acids
Step: 1
Step: 2
Urea fractionation has been most frequently used for thepurification of fatty acids from fish oil [1] and from several vegetaloils like, blackcurrant oil [2] borage oil [3]; rapeseed oil [4].
Sources:
1. Ratnayake, WMN et al., Fat Sci Technol, 90, 381, 1988.
2. Traitler H et al., J. Am Oil Chem Soc, 65, 755, 1988.
3. Shimada Y et al., J Am Oil Chem Soc, 75, 1539, 1988.
4. Hayes DG et al., J Am Oil Chem Soc, 75, 1403, 1998.
Advantages & Applications
Crystallization is technically not feasible to separate high unsaturatedfatty acids from moderately high unsaturated fatty acids. e.g.: C18:3 &C18:5 can't easily separated as they have negative melting point, insuch cases UIC is robust tool.
By - products
Component Sunflower Cottonseed Soybean
Unsaponifiables, % 39 42 33
Tocopherols, % 9.3 11.4 11.1
a-tocopherol, % 5.7 6.3 0.9
Sterols, % 18 20 18
Stigmasterol, % 2.9 0.3 4.4
Gums Deodorizer distillate (DOD) Waxes Soap Stock
Deodorizer distillate (DOD)
Tocols from Deodorizer distillate (DOD)
Tocols = Tocopherols + Totrienols
A combination of molecular distillation (MD), ethanol
fractionation, chemical alcoholysis, and ion-exchange
chromatography
MD may not produce a high purity tocopherol because very
similar molecular weights of Tocols and Sterols
By Ethanol fractionation purity can be enhanced as sterols are
insoluble, whereas Tocopherols are soluble in ethanol.
Esterification of alcohols
Series of Distillation
FAME
Tocols
Esters Residues
Barnicki et al. *
Patented a solvent less process, comprised of an esterificationreaction to reduce the volatility of alcohols followed by a series ofdistillation steps, where components boiling higher and lower thantocopherols are separated from tocopherols.
*S. D. Barnicki, C. E. Sumner, Jr., and H. C. Williams, U.S. Patent 5,512,691, 1996.
Sterols are polycyclic alcohols,
Unsaponifiable fraction & quality
depends on temperature, duration,
quantity of stripping steam, and
the extent of vacuum
Phytosterols from DOD
•Either by hydrolysis or trans-esterification
Breaking of ester bonds
•Re-esterification of Phytosterols occurs during methyl ester
Trans-esterification •Phytosterols
content up to 50% by removing FAMEs
Distillation
Instead of Distillation we can go for Crystallization
(physical), solvent extraction (chemical), or crystallization
with additives via adduct formation and separation
(physicochemical)
Organic solvents or solvent mixtures composed of low- and
high-polarity solvents and water are used for crystallization
of Phytosterols
Quality & Quantity of feed and final product are the majorparameters for selection of isolation techniques
Present day demand for a variety of fatty acids, including long-chain polyunsaturated fatty acids (PUFA), may require acombination of different processes and further steps ofseparation
Bulk properties as well as molecular properties both should becombined to isolate the product within the specifications
Good amount of experimental work & processes feasibilityanalysis is required in order implement novel techniques onindustrial scale
CCC/CPC techniques can be used for isolation of oleo-chemicalsat commercial scale
Conclusion
Carbon
No.Chloroform Benzene Cyclohexane Acetone
Ethanol
95%
Acetic
acidMethanol
10 3260 3980 3420 4070 4400 5670 5100
12 830 936 680 605 912 818 1200
14 325 292 215 159 189 102 173
16 151 73 65 53.8 49.3 21.4 37
18 60 24.6 24 15.4 11.3 1.2 1
Solubility
Volatility
Component Mol. Wt. Rel. Volatility
Fatty Acids 280 2.5
Squalene 411 5
Tocopherols 410 1
Sterols 415 0.6
Sterols esters 675 0.04
TAGs 885 small
Systematic Name Trivial NameShorthand
designation
Molecular
wt.
Melting point
(0C)
Dodecanoic Lauric 12:00 200.3 44.2
Tetradecanoic Myristic 14:00 228.4 53.9
Hexadecanoic Palmitic 16:00 256.4 63.1
Heptadecanoic Margaric
(Daturic)
17:00 270.4 61.3
Octadecanoic Stearic 18:00 284.4 69.6
Eicosanoic Arachidic 20:00 312.5 75.3
Docosanoic Behenic 22:00 340.5 79.9
Tetracosanoic Lignoceric 24:00 368.6 84.2
Hexacosanoic Cerotic 26:00 396.7 88
Melting Point (M.Wt.)
Systematic Name Trivial NameShorthand
designation
Molecular
wt.
Melting point
(°C)
Cis-6-
octadecenoicPetroselinic 18:1 (n-12) 282.4 30
Cis-9-
octadecenoicOleic 18:1 (n-9) 282.4 16.2
Tr-9-octadecenoic Elaidic tr18:1 (n-9) 282.4 43.7
Cis-11-
octadecenoic
Vaccenic
(Asclepic)18:1 (n-7) 282.4 39
9,12-
octadecadienoic
Linoleic Acid18:2(n-6) 280.4 -5
6,9,12-
octadecatrienoic
G-linolenic
Acid18:3(n-6) 278.4
Melting Point (Unsaturation)