Accepted Manuscript

Extraction of edible avocado oil using supercritical CO2 and a CO2/ethanol mixture as
solvents

Sayuri C.S. Corzzini, Helena D.F.Q. Barros, Renato Grimaldi, Fernando A. Cabral

PII: S0260-8774(16)30309-0
DOI: 10.1016/j.jfoodeng.2016.09.004
Reference: JFOE 8649

To appear in: Journal of Food Engineering

Received Date: 27 February 2016

Revised Date: 29 August 2016
Accepted Date: 3 September 2016

Please cite this article as: Corzzini, S.C.S., Barros, H.D.F.Q., Grimaldi, R., Cabral, F.A., Extraction of
edible avocado oil using supercritical CO2 and a CO2/ethanol mixture as solvents, Journal of Food
Engineering (2016), doi: 10.1016/j.jfoodeng.2016.09.004.

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 ACCEPTED MANUSCRIPT
1 Extraction of edible avocado oil using supercritical CO2 and a CO2 / ethanol
2 mixture as solvents

4 Sayuri C.S. Corzzinia, Helena D.F.Q. Barrosa, Renato Grimaldib, Fernando A.

5 Cabrala*

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6

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7 Department of Food Engineering, University of Campinas UNICAMP, 13083-862

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8 Campinas SP, Brazil

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9 Department of Food Technology, University of Campinas UNICAMP, 13083-862
10 Campinas SP, Brazil

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12 * Corresponding author. Tel.: +55 19 3521 4030; fax: +55 19 3521 4027.

13 E-mail address: facabral@unicamp.br (F.A. Cabral)

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21 Abstract

22 Avocado oil is similar to olive oil, it can be used as an ingredient in functional

23 foods since it contains substantial amounts of vitamins and phytosterols and its

24 triacylglycerols contain high contents of unsaturated fatty acid. The present study

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25 sought to evaluate the technical viability of extraction of avocado oil by supercritical

26 technology using GRAS solvents (Generally Recognized as Safe). Freeze-dried

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27 avocado (Persea Americana) pulp with 65% lipids was subjected to supercritical

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28 extraction in a fixed bed at temperatures of 40, 60, and 80 C and pressures of 200,

29 300 and 400 bar, using supercritical carbon dioxide (scCO2) as a solvent in a first step,

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30 and a scCO2 / ethanol mixture (93/7 w/w) in a second step. Extraction curves (mass of
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31 oil extracted versus extraction time) were obtained from the overall extraction yields,

32 and from this curve the solubility of oil in scCO2 was calculated. The extraction
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33 appeared to be technically viable to obtain up to 98% oil recovery. The solubility values

34 correlated well with the Chrastil equation, and presented similar order of magnitude to
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35 the solubility of oils reported in literature.

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37 Keywords: Avocado; Supercritical extraction; Solubility

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40 1. Introduction

41 Avocado is one of the few fruits containing oil as a main component; it

42 possesses high levels of unsaturated fatty acids in its triacylglycerol

43 composition (about 70% of oleic acid) and substantial amounts of compounds

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44 with health benefits such as tocopherols, phytosterols, and lutein. The

45 unsaponifiable fraction of avocado oil has regenerative properties for the skin

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46 (Gunstone, 2006).

47 Avocado oil is thermally stable, with high glycerol decomposition

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48 temperature (smoke point of 255 C), which allows f or its use in frying and

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49 baking. Because it is extracted from the fruit pulp, it is similar to olive oil with
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50 respect to its physicochemical properties, particularly the fatty acids

51 composition of its triacylglyerols, which may serve as a condiment for fresh

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52 salads (Kurlaender et al., 2004; Berasategi et al., 2012; Canto et al, 1980). The

53 -9 fatty acids appear to be the substances responsible for the beneficial

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54 effects on human health in preventing cardiovascular diseases (Rebollo et al.,

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55 1998). Avocado oil is easily absorbed by the skin, being used as a vehicle for
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56 medicinal substances and perfume absorption. It is of great value to the

57 cosmetics industry due to the ease of emulsion formation and for soap
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58 manufacture (Gunstone, 2006).

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59 Although avocado oil extraction with organic solvents presents high

60 extraction yield, the extraction process can cause environmental impacts.

61 Mechanical extraction by cold pressing or centrifugation procedures are clean

62 technologies, however a low extraction yield is obtained (Kurlaender, 2004).

63 Extraction with supercritical carbon dioxide may be a suitable alternative, since

64 scCO2 is a green solvent suitable for extracting low polarity compounds such as

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65 oils and fats, besides producing a product (oil) and byproduct (defatted cake)

66 free of any traces of organic solvents, with high extraction yield.

67 Mostert et al. (2007) extracted avocado oil from the Fuerte variety using

68 scCO2 as a solvent at 37 oC and 350 atm, and they found overall yields of

69 62.9% and 66.7% (dry basis). Botha and McCrindle (2003) extracted 94% of the

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70 oil within 2 h at 350 atm. In a previous work (Barros et al, 2016), 88% of the oil

was extracted at 50oC and 400 bar from avocado pulp of the Hass variety,

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72 which corresponds to an overall yield of 57% from a pulp containing 68% lipids.

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73 This oil was used as a co-solvent of the scCO2 for extraction of capsanthin from

74 red bell pepper, and this was performed by simultaneous extraction of oil from

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avocado and capsanthin from red bell pepper. This process could extract 88%
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76 of oil from avocado and up to 50% of the capsanthin from red bell pepper, and
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77 still produce an avocado cake of commercial value, containing about 25% lipids.

78 CO2 is a nonpolar molecule, thus the scCO2 can extract nonpolar and low
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79 polarity substances, such as lipids. Co-solvents such as ethanol can also be

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80 used in different proportions to modify the polarity of the scCO2 / ethanol

81 mixture, and therefore increase the solubility and improve extraction of other
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82 substances such as tocopherols. To evaluate the extraction efficiency in terms

of yield and extraction time, extraction curves (mass of solvent versus extraction
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time) were obtained for the different temperature and pressure conditions,
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85 which are directly related to the solute concentration in the raw material and the

86 solubility of the solute in the solvent. The extraction curves present three

87 stages: 1) constant extraction rate (CER), characterized by a graphically

88 straight line, which is most soluble fraction of the extract. In this stage, mass

89 transfer by convection is predominant; 2) falling extraction rate (FER), in which

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90 failure begins to occur in the solute layers on the surface surrounding the

91 particles or the number of broken cells is not uniform; both convection and

92 diffusion are important at this stage; 3) diffusion controlled stage (DC), in which

93 the surface adsorption sites are exhausted. The extraction process is controlled

94 by diffusion of the solvent inside the particles and diffusion of the dissolved

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95 solute from within the particle to the particle surface. The use of a co-solvent

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96 together with scCO2, for example a small amount of ethanol, may improve the

97 efficiency of lipids extraction because it increases the solubility and facilitates

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98 the extraction of the less nonpolar substances. Scientific literature does not

99 present extraction curves, solubility values and tocopherol contents for avocado

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oil in all ranges of temperature and pressure, nor extraction values using
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101 ethanol as a co-solvent.
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102 In this context, with the aim of obtaining complete extraction of oil from

103 avocado, a two-step process was proposed for extraction of the main
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104 compounds and the minor solubility lipidic compounds. This study seeks to
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105 evaluate the extraction of avocado oil using supercritical carbon dioxide as the

106 solvent in a first step, followed by a second step using a CO2 / ethanol mixture.
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107 Furthermore, technical feasibility was evaluated based on the results of overall

extraction yield, oil solubility and tocopherol levels.

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110 2. Materials and Methods

111 2.1. Raw material and its characterization

112 Ripe avocados from Persea americana species and commercial avocado oil

113 (Jaguacy Brasil, Bauru/SP, Brazil) were acquired from local market (Campinas,

114 SP, Brazil). The avocado pulp was separated and frozen at -80 C in an Ultra-

115 freezer (Glacier NU-9483 Ultra-Low Temperature Freezer, NuAire, Plymouth,

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116 MN USA) and subjected to freeze drying (Liobras L101, SP, Brazil). After

117 freeze-drying, the pulp was ground in a domestic blender (Arno, Brazil). The

118 particle size distribution was measured by the ASAE method (ASAE, 1997),

119 using a series of Tyler sieves (Model 1868, Bertel, SP, Brazil), with sizes

120 ranging from 8 to 48 mesh, which are too wide, but because the pulp has a high

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121 oil content agglomerate formation is possible, so these sieves were used to

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122 avoid this problem. Total volatiles were determined by the gravimetric method

123 (AOAC, 1997), in which a 2.0 g sample was dried in an oven (Marconi, MA-

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124 030/12, SP, Brazil) at 105 C under 600 mm Hg vacuu m (Marconi pump, MA-

125 057-13, SP, Brazil) until reaching constant weight. Moisture content was

126
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determined by the Karl-Fisher method (KF Metrohm 701 Titrino) (AOCS, 1998),
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127 consisting of an oven (832 KF Thermoprep) using a sample of approximately
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128 0.4 g at 105 C for 30 min, and nitrogen flow rate of 50.0 mL / min (1.01 bar and

129 0 C).
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130 Total lipids were determined by the Soxhlet method (AOCS, 1997) in
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131 triplicate, using hexane as the solvent (150 mL) (Synth, Brazil, lot 116071) and
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132 about 5.0 g of the crushed freeze-dried sample. The solvent was removed from

133 the extracts by evaporation in a rotaevaporator (Marconi, MA-058, Brazil) at 42

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134 C under vacuum of 600 mmHg in vacuum oven (oven MA 030-12, and pump
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135 MA-057-13, Brazil) until reaching constant weight (variation less than or equal

136 to 3.0 mg).

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138 2.2. Reagents

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140 Carbon dioxide 99.5 % w/w (White Martins Gases Industriais, Brazil, lot

141 062 C/13), hexane 98.5 % w/w (Synth, Brazil, lot 116071), and ethanol 99.8 %

142 w/w (xodo, Brazil, lot AE10296RA) were used as solvents in the different

143 extraction steps.

144

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145 2.3. Extraction procedures

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146 2.3.1. Extraction curves

147 Avocado samples were subjected to two-step extraction in a fixed bed

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148 extractor, the first step using scCO and the second step using a mixture of

149 ethanol and scCO2, whose methodology has been discussed in detail elsewhere

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(Bitencourt et al., 2014, Paula et al., 2013, Garmus et al., 2015).
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151 The bed was packaged with 5.0 g of the sample spread horizontally,
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152 where the solvent flowed through bed and the extract was collected at

153 predetermined intervals. The first step lasted about 3 hours (9 samples), in
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154 which the scCO flow rate was 2.5 g / min, while the second step lasted 1
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155 hours (6 samples), in which the first solvent (at the same flow rate) was mixed
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156 with ethanol at a flow rate of approximately 0.20 g / min, resulting in a

157 CO/ethanol ratio of 93/7 w/w. The pressure and temperature conditions were
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158 200, 300, and 400 bar, and 40, 60, and 80 C. All e xtractions were performed in
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159 triplicate.

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161 2.3.2. Determination of the global extraction yield

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163 The global extraction yield was used to compare the different extraction

164 steps and different temperature and pressure conditions, obtained by the ratio

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165 between the extract mass at the end of the process and the mass of the matrix

166 subjected to extraction. Considering that the experiments were carried out in

167 triplicate, the overall yield was a result of the arithmetic mean of the

168 experimental values.

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mass of oil
169 Overall Yield (%) = 100% = f (T , P) (1)
mass of raw material

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170

171 2.3.3 Calculation of solubility

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172 After obtaining the extraction curves and considering the non-extractable

173 solid portion of the matrix as inert, the solubility of avocado oil in scCO2 as a

174
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function of the temperature and pressure was obtained from the slope of the
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175 extraction curves for the constant extraction rate, as illustrated in Fig. 1. It

considered the (mass of oil) versus (mass of solvent), calculated from the
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177 quotient between the mass of oil sampled and the total mass of carbon dioxide
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178 used, according to Eq. (2).

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mass of oil
179 Y= = f (T , P) (2)
mass of CO 2
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181 Fig. 1.
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183 2.4. Correlation of oil solubility in scCO2

184 The values for solubility as a function of temperature and pressure were

185 correlated by the Chrastil equation (Chrastil, 1982), which correlates the solute

186 solubility with the CO2 density (kg/m3) and temperature T (K) of the system,

187 where k, a, and b are adjustable parameters. The original Chrastil equation can

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188 ()
also be rewritten in terms of the solubility in the mass fraction Y (kg solute / kg

189 solvent), according to Eq. (3):

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190 Y = k 1 exp + b (3)
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191 where density of the pure CO2 under the different temperature and pressure

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192 conditions was calculated using a computer program, based on an accurate

193 equation of state for carbon dioxide, the Huang equation (Huang et al., 1985)

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194 which reproduces the experimental values of IUPAC (Angus et al., 1976).

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195

196 2.5. Acidity (Free Fatty Acid)

197
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The free fatty acid (FFA) levels of all oil samples were determined by
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198 titration with sodium hydroxide in an automatic titrator (Metrohm 808, Herisan,

199 Switzerland), according to the IUPAC methodology 2.201 (IUPAC, 1987).

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200 Approximately 0.1 g of oil was diluted in 40 mL of a 1:1 ether:ethanol solution

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201 which was then titrated with a standard NaOH solution, under agitation; the
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202 titration was automatically finalized when reaching the potenciometric end point.

203 All analyses were performed in triplicate and the acidity calculated by Eq. (4).
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204 acidity =
(Vtitration Vbrank ) N NaOH .M oleic (4)
10 Wsample
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205 where Vtritation, Vbrank, NNaOH, Moleic and Wsample are: NaOH volume consumed in
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206 titration of 40 ML; NaOH volume consumed in titration of 40 ML of ether/ethanol

207 without oil; normality of the NaOH titrant; molecular mass of oleic acid and the

208 mass of oil. The results were expressed as percentage of oleic acid.

209 2.6. Tocopherols levels

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210 To determine the tocopherol levels of all oil samples, 0.5 g of the sample

211 was diluted in 10 mL of hexane. Quantification was performed using an external

212 standardization curve with the 4 tocopherol isomers (alpha, beta, gamma, and

213 delta) and 5 different sample concentrations within the range of interest, based

214 on the tocopherol standard set cat 613424. Liquid chromatography (HPLC) was

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215 performed under the following operation conditions: Isocratic pump Perkin

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216 Elmer Series 200; detector: Fluorescence detector 200a Perkin Elmer Series;

217 excitation wavelength of 290 nm, emission wavelength of 330 nm; column:

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218 Hibar RT 250-4 Li Chrosorb Si 605 mm; Mobile Phase - Hexane / Isopropanol

219 (99/1), 1.0 mL / min.

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220 3. Results and discussion
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222 3.1. Characterization of the freeze-dried avocado pulp
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223 Table 1 shows the characterization of the freeze-dried avocado pulp.

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224 After grinding, the freeze-dried pulp presented an average diameter of 0.95
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225 0.13 mm, corresponding to the following fractions: 0.3% (2.569 mm), 9.3%

226 (1.817 mm), 27.2% (1.177 mm), 52.3% (0.833 mm), 10.1 % (0.589 mm) and
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227 0.7% (0.382 mm). The reduction in diameter results from the milling process

228 that breaks the cell walls, ensuring that more solute comes into direct contact
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229 with the solvent. The smaller particle diameter leads to a shorter diffusion path.
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230 Therefore, there is less resistance to diffusion of the solvent into the particle

231 surface. Thus, for both convection and diffusion, a smaller diameter results in

232 less resistance to mass transfer (BIRD et al., 2004), but in this study due to the

233 high lipid content of the pulp, the mean diameter used was larger than normal

234 so as to avoid agglomeration.

235 Table 1

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236

237 The low moisture content of 2.62% determined by the Karl Fischer

238 method was suitable, because although water may be a co-solvent in some

239 supercritical extraction processes, it adversely affects the extraction of various

240 substances including lipid compounds. Therefore, reduced moisture content in

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241 the sample to be extracted is an important factor for a good extraction yield. The

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242 difference between total volatiles (3.6%) and moisture (2.62%) obtained by the

243 Karl Fischer method indicates approximately 1% volatiles in addition to water.

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244 This difference is expected, since the oven drying method accounts for other

245 volatiles besides water. Avocado oil, if extracted carefully without damage,

246
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contains a very low percentage of free fatty acids (%FFA) (<0.5% as oleic
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247 acid) (Wong et al., 2010). In this work, the avocado oil acidity was 0.16%

oleic acid, less than the recommended maximum value of 0.5% and also
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248

249 less than the value of 0.19% citric acid (equivalent to 0.83% oleic acid)
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250 obtained by Buelvas, Salgado (2012), and of 1.9 mg KOH/g (equivalent to

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251 0.95% oleic acid) obtained by Botha (2004).

252 An extraction yield of 65.2% was obtained by the Soxhlet oil extraction
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253 method, which is of the same order of magnitude as 72 5%, 61%, and 68%

254 obtained by Mostert et al., (2007), Meyer et al. (2008) and Barros et al. (2016),
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255 respectively. Avocados have different lipid levels depending on variety, climate,
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256 soil, and cultivation of the orchards. Moreover, because the externally-visible

257 signs of ripening of avocado are very difficult to be detected, in addition to the

258 long time in the tree before being harvested even in the state of full maturity, its

259 fat level may be greatly affected even in the earliest harvest due to deterioration

260 of lipids. In general, the more mature the fruit, the greater the lipid levels (Tango

261 et al., 2008).

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262

263 3.2. Kinetics and extraction yield

264 Table 2 shows the extraction yields in the two steps, and Figs 2 to 4

265 represent the average and standard deviations of all extraction curves for each

266 temperature and pressure condition, represented by the overall extraction yield

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267 versus S/F (solvent mass per mass of raw material). The theoretical maximum

268 overall extraction yield was adopted as 65.2%, obtained by the Soxhlet method.

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269 Table 2
270

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271 Fig. 2.
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273 Fig. 3
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275 Fig. 4.
276 As shown in Table 2 and Figures 2-4, in the first extraction step with
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277 scCO the yield increased with the increase in pressure at constant

278 temperature, but decreased with higher temperatures at pressures of 200 and
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279 300 bar. This behavior can be observed in other works from literature, for
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280 example in the work of Machmudah el al (2008) for extraction of rosehip seed

oil, where they observed that for the behavior of extraction yield with
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282 temperature there is an inversion at 345 bar. Below this pressure, the yield
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283 decreases with temperature and at higher pressures increases.

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284 At 40 C, practically all steps were equalized at the end of the extraction,

285 reaching a yield of approximately 64%. In the second extraction step, the slope

286 of the extraction curve increased, indicating greater solubility in the scCO2 /

287 ethanol mixture. On average, 94% and 97% recovery were obtained at 200 bar

288 and 300 bar, respectively.

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289 As can be seen in Fig. 3, the extraction yield increased with increasing

290 pressure at 60 C. However, it is clear that the ex tractions which presented

291 lower yields in first step, and had higher yields in second step. In general, the

292 two-step extractions at 300 and 400 bar recovered between 97 and 98% of the

293 oil in relation to the Soxhlet extraction.

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294 Fig. 4 shows the yield versus solvent mass curve after complete

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295 extraction (CO and CO + ethanol) at the three pressures, at 80 C. At th is

296 temperature, only two pressure conditions approached the yield obtained by the

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297 extraction method using organic solvent, with a great difference in yield

298 between the pressures. On average, at 400 and 300 bar, 98% and 97%

299
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recoveries were obtained in relation to the Soxhlet extraction. In contrast, at
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300 80C and 200 bar, the recovery was 26%, representing only 40% of the result
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301 obtained by Soxhlet extraction. In the first step with scCO, although the yield at

302 400 bar was much greater than that at 300 bar, similar yields were observed for
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303 both conditions in the second step with ethanol.

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304

305 3.3. Solubility of avocado oil in scCO2

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307 The solubility of avocado oil in scCO2 was calculated from the slope of
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308 the extraction curves in the constant extraction step, as shown in Table 3.
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310 Table 3
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312 The solubility of avocado oil in scCO2 as a function of temperature and

313 pressure was correlated by the Chrastil equation (1982), as written by equation

314 (3). The parameters k, a, and b were obtained by fitting the experimental data to

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315 the model, according to Eq. (4), using a computer program which minimizes an

316 objective function, using the modified Nelder and Mead simplex method (Nelder

317 and Mead, 1965).

3246.0967
318 ln(Y ) = 7.4198 ln( ) 38.0089 (4)
T

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319 The comparison between experimental values and those calculated by the

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320 model is shown in Table 3 and Fig. 5. In addition, a comparison between the

321 experimental values is shown in Fig 6 to illustrate the solubility behavior as a

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322 function of temperature and pressure.

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324

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326 Fig. 6.
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327
328 The solubility of a solute depends on the solvent density and solute vapor
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329 pressure, as indicated by the Chrastil equation. The increase in temperature

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330 decreases the density of the solvent, but increases its vapor pressure, resulting

331 in a pressure condition where the effects are offset and the solubility behavior is
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332 reversed with temperature. Below the crossover pressure, solubility decreases

333 with the increase in temperature, and above this point the higher temperature
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334 increases the solubility. The crossover pressures for solubility of various
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335 substances in scCO2 have been reported by several authors, including 250 bar

336 for squalene (Martinez-Correa et a., 2010), 170 bar for coumarin (Rodrigues et

337 al, 2008), 300 bar for -oryzanol (Bitencourt et al., 2016), 140 bar for diuron

338 (Ciou and Su, 2015) and about 150 bar for phenolic compounds (Murga et al.,

339 2003). For mixtures containing similar compounds, which is the case of oils (a

340 mixture of triacylglycerols), the crossover pressure is approached (not found at

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341 a one specific point). Fig. 6 shows a crossover pressure of avocado oil in the

342 range from 350 to 375 bar. A crossover pressure of approximately 350 bar was

343 obtained for oils and fats, 300 bar for rice bran oil (Tomita et al., 2014), roughly

344 350 bar for Sacha inchi oil (Follegatti-Romero, 2009), 295 bar for coconut oil

345 (Zuknik et al., 2016), 280 bar for annatto seeds oil (Silva et al., 2008), and from

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346 250 to 300 bar for fish oil (Lopes et al., 2012).

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348 3.4. Tocopherol levels in the extracts

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349 Table 4 shows the tocopherol levels in extracts obtained in the first step

350 with pure scCO2, in the second step with the scCO2 / ethanol mixture and in a

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351 sample of commercial avocado oil. The main tocopherol present in avocado oil
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352 is the -tocopherol, consisting of approximately 20 mg per 100 gram of oil. The

353 tocopherol content was analyzed only in extracts obtained at 200 bar, in order
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354 to evaluate the fractionation of tocopherols and posterior extraction with ethanol

as the co-solvent. It is possible that at 400 bar tocopherols can be extracted.

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355

Pure scCO2 at 200 bar is not effective for extraction of tocopherols, but the
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357 addition of ethanol in a second step shows that even at the pressure of 200 bar,
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358 tocopherols are extracted. The extracts obtained at 400 bar were not analyzed.

359 In general, it is observed that extracts from the second extraction step with
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360 ethanol presented higher tocopherol concentrations, especially those obtained

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361 at 40 and 60 C.

362 Table 4
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364 4. Conclusion
365 Supercritical CO2 extraction has proven to be technically feasibility for

366 avocado oil extraction, especially at higher pressure conditions (400 bar) with

367 97-98% oil recovery. Except at 80 C and 200 bar, t he use of ethanol as a co-

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368 solvent in the second extraction step favored extraction of the remaining oil,

369 obtaining a differentiated fraction, enriched in tocopherols. This resulted in a

370 new opportunity in addition to producing a single special product, since a clean

371 byproduct was also produced. The Chrastil equation correlated well the

372 solubility values of avocado oil in scCO2, which corroborated the solubility

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373 values reported in literature.

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374

375 Acknowledgments

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376 The authors wish to thank the So Paulo Research Foundation FAPESP

377 for the financial support and the scientific initiation scholarship granted to Sayuri

378
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Clarice Shintani Corzzini (Process 2013/10932-8).
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379
380 5. References
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381 Angus, S., Armstrong, B., Ruck, k.M., 1976. Carbon Dioxide: International
382 Thermodynamic Tables of the Fluid State, vol 3. Pergamon Press, New
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383 York, p.266.

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384 ASAE. Standards, 1997. Method of Determining and Expressing Fineness of

385 Feed Materials by Sieving, ASAE S319.3, 547.
386 AOAC, 1997. Official Methods of Analysis, vol 16, Association of Official
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387 AnalyticalChemists. Washington, Dc.

388 AOCS, 1998. American Oil Chemists Society. Official Methods and
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389 Recommended Practices of the American Oil Chemist's Society, vol 1. 6th
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390 ed. AOCS Press Champaign, pp 1. (Ca 23-55).

391 AOCS, 1997. American Oil Chemists Society. Official methods and
392 recommended practices of the American Oil Chemists Society. Section I:
393 Physical and Chemical Characteristic of oil, fats and waxes, fourth Edition,
394 Champaign IL, USA, AOCS Press. v. 2. pp 54-83.
395 Berasategi, I., Barriuso, B., Ansonera, D., Astiasarm I., 2012. Stability of
396 avocado oil during heating: Comparative study to olive oil. Food Chem. 132,
397 439446.

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398 Bird, R B., Stewart, W.E., Lightfoot, E.N., 2004. Fenmenos de transporte. Rio
399 de Janeiro: Livros Tcnicos e Cientficos.
400 Barros, H.D.F.Q., Coutinho, J.P., Grimaldi, R., Godoy, H.T., Cabral, F.A., 2016.
401 Simultaneous extraction of edible oil from avocado and capsanthin from red
402 bell pepper using supercritical carbono dioxide as solvente. J. Supercrit.
403 Fluids 107, 315-320.

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404 Bitencourt, R.B., Queiroga, C.L., Montanari Junior, I., Cabral, F.A., 2014.
405 Fractionated extraction of saponins from Brazilian ginseng by sequential

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406 process using supercritical CO2, ethanol and water. J. Supercrit. Fluids 92,
407 272281.

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408 Bitencout, R.G., Rammazzina Filho, W.A., Paula, J.T., Garmus, T.T., Cabral,
409 F.A., 2016. Solubility of -orizanol in supercritical carbono dioxide and
410 extraction from rice bran. J. Supercrit. Fluids 107, 196-200.
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491 240-244.
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494

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Table 1
Characterization of the freeze-dried raw material
Characteristics Results
Moisture (%) 2.62 0.01
Mean particle diameter (mm) 0.95 0.13
Volatile + moisture (%) 3.6 0.7
Soxhlet
Total lipids (%) 65.2 0.4
Acidity (oleic acid %) 0.16 0.01

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Table 2
Overall extraction yield in the first and second extraction steps
Overall Yield (%)
T (C) P (bar)
scCO2 scCO2+ethanol Total
200 39 8 23 2 62 8
40 300 61 2 2.9 0.6 63 2

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400 60 5 43 63 2
200 27 2 31 1 59 3
60 300 49 8 14 5 63 3

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400 62 1 1.9 0.6 64 1

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200 12 1 14 2 26 2
80 300 55 2 81 63 3
400 61 3 32 64 1
Soxhlet - -
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Table 3
Solubility values of avocado oil in scCO2, and comparison with the values
adjusted by the Chrastil equation.
(kg/m )
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T (C) P (bar) Experimental Chrastil Eq. (5)

(%)
exp calc *
Y (g/kg) Y (g/kg)
200 840.8 4.8 0.4 4.9 2.1
40 300 910.3 9.4 1.7 8.8 6.3

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400 956.2 12.9 4.1 12.7 1.6
200 724.5 3.9 1.6 3.0 23

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60 300 830.0 7.7 2.9 8.2 6.5
400 890.2 13.4 0.2 13.8 3.0

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200 595.4 1.2 0.1 1.2 0.0
80 300 745.5 6.2 0.2 6.4 3.2
400 822.8 14.7 0.6 13.3 9.5
* deviation calculated by the equation = Y( exp

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Table 4
Tocopherol levels in the mixture of extracts obtained at 200 bar. Extracts from
the first step with pure CO2 and extracts from the second step using ethanol as
the co-solvent.
Type of extract Concentration (mg tocopherol /100g oil)
mix of extracts Alfa Beta Gamma Delta Total
scCO2 200bar 0.0 0.0 0.910.03 2.110.07 3.00.1
mix (40 + 60 + 80oC)
scCO2 + ethanol 200 bar 20.21.1 0.140.02 0.330.08 7.490.48 28.21.7
mix (40 and 60oC)

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scCO2 + ethanol 13.40.9 0.0 1.840.42 0.190.03 15.41.4
80oC and 200 bar
Commercial avocado oil 21.01.5 0.130.09 0.640.18 0.0100.003 222

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Fig. 1. Illustrative calculation of avocado oil solubility in scCO2

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Fig. 2. Extraction curves at 40 C
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Fig. 3. Extraction curves at 60 C.

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Fig. 4. Extraction curves at 80 C.
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Fig. 5. Comparison between the experimental solubility values and the values
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Fig. 6. Solubility of avocado oil as a function of temperature and pressure

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The avocado oil extraction was technically feasible in one and in two steps on higher
pressures.

It was recovered until 98% of the oil when was used two steps extraction.

The crossover pressure from the oil solubility data was around 350 to 375 bar.

The extracts of second step were more concentrated in tocopherols.

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