# Commission Regulation (EEC) No 2568/91 of 11 July 1991 on the characteristics of olive oil and olive-residue oil and on the relevant methods of analysis, Division 6: SEPARATION OF THE ALCOHOLIC FRACTION BY HPLC

Extent: E+W+S+N.I.

The unsaponifiable obtained from Part 1 is dissolved in 3 mL of the mobile phase (4.14), filter the solution with a syringe filter (3.10) and reserve.
Inject 200 μL of the filtered unsaponifiable solution in the HPLC (3.8).
Run the HPLC separation at 0,8 mL/min, discard the first 5 min. and collect in 25 mL conical flasks (3.11) between the 5 and 10 min. for aliphatic and triterpenic alcohols and between 11 and 25 min for sterols and erythrodiol and uvaol (Note 5).
The separation can be monitored with an UV detector at 210 nm wavelengths or a refractive index detector (see Figure 6).
The fractions are evaporated until dryness and prepared for chromatographic analysis.

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Source: https://www.legislation.gov.uk/eur/1991/2568/annex/XIX/part/2/division/6 (the official version)
Page: https://openlegislation.uk/eu-regulation-1991-2568/annex-XIX/part-2/division-6
Contains public sector information licensed under the Open Government Licence v3.0. Source: legislation.gov.uk, The National Archives.