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3 protocols using n heptane

1

Biofuel Production from Waste Cooking Oil

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The waste oil for this work was a blend of 95 vol% of soy-bean and 5 vol% of corn oils, supplied by the restaurant “La poissonerie du port” in Oran (Algeria) and it was heavily used for frying fish. The temperature observed during the fish frying was in the range of 100 to 150 ºC. Five liters of oil sample were collected from a collecting drum in which the waste frying oil was dumped in once every day and kept at a room temperature for one day to enable settling. Two liters of this waste frying oil were heated up to 50 °C for 10 min to rule out the presence of water. Thereafter, the oil was filtered through a filter composed of cotton cloth, filter paper and around 1 cm of fine quartz sand to remove slag and impurities in a plastic container (see Figure 1S in Supplementary Material).
Sodium metal (Rhone-Poulenc, Paris, France), absolute ethanol, citric acid, n-heptane, medium boiling point petroleum ether and sodium chloride (Panreac, Barcelona, Spain) were used without further purification. The feedstock oil was tested for humidity by FT-IR spectroscopy in a Nicolet 6700 instrument (Fischer Scientific, USA) placing an oil drop between two sodium chloride discs (Aldrich, USA, 5.40 mm thick, 24.97 mm o.d.). The oil acidity index was determined after the standard method EN 14104.
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2

Comprehensive Chemical Reagents Sourcing

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Dichloromethane 99.9 % (ref.: 32,222), Toluene 99.7 % (ref.: 32,249) and 2-Propanol 99.9 % (ref.: 59,300) were purchased from Honeywell, Germany. Methanol (ref.: 412,722), HPLC-GOLD-Ultragradient grade, was brought from Carlo Erba reagents, Italy. n-Hexane 95 % (ref.: 363,242), high performance chromatography grade, and phenol crystalline (ref: 144852.1211) and n-heptane (ref.: 162062.1611) were provided by PanReacAppliChem, Spain. sulfuric acid reagent (ref: 34,632), orange reagent (ref: 131130.1612), sulfuric acid 95.0–97.0 % (ref: 30,743), bovine serum albumin (BSA) (ref: A9647), sodium hydroxide 98 % (ref: 30,620), sodium carbonate (ref: 222,321), potassium sodium tartrate tetrahydrate (ref: 217,255), copper (II) sulphate pentahydrate (ref: 209,198), Folin&Ciocalteu's phenol reagent (ref: F9252), magnesium sulphate monohydrate (ref: 434,183), anhydrous sodium sulphate (ref: 239,313) and fuming hydrochloric acid (ref: 84,418), high analytical reagent grade, were supplied by Sigma – Aldrich, USA. ASTM® D2887 Reference Gas Oil (ref: 48,873) and chloroform-d (ref: 225,789) provided also by Sigma Aldrich, USA.
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3

Histological Analysis Materials and Reagents

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All materials and reagents employed for the histological analyses were supplied by Bio-Optica Milano S.p.a. (Milan, Italy). The following reagents were all provided by Sigma-Aldrich (Milan, Italy): sulfuric acid (95–98% purity), sodium hydroxide, boric acid, hydrogen chloride, benzoic acid, and phosphate-buffered saline (PBS). The Kjeldahl catalyst was supplied by Carlo Erba (Milan, Italy). n-Heptane was purchased from PanReac AppliChem (Barcelona, Spain). Potassium hydroxide, chloroform, methanol, sodium carbonate anhydrous, Folin Ciocalteau’s reagent, stock standard solution of Re (1000 mg L−1 in 2% HNO3), gallic acid, and stock standard solutions of Sc, Ge, In, and Bi (1000 mg L−1 in 2% HNO3) were from Fluka (St. Gallen, Switzerland). Nitric acid (65%, v/v) was of Suprapur grade (Mallinckrodt Baker, Milan, Italy). Ultrapure water (<5 mg L−1 TOC) was obtained from a Barnstead Smart2Pure 12 water purification system (Thermo Scientific, Milan, Italy).
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