# Commission Regulation (EC) No 152/2009 of 27 January 2009 laying down the methods of sampling and analysis for the official control of feed (Text with EEA relevance), Division 5.4: HPLC determination

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

The following conditions for isocratic elution are offered for guidance; other conditions may be used, provided they yield equivalent results (see also observations 9.1 and 9.2):
Liquid chromatographic column (4.2):
125 mm x 4 mm, C18, 3 μm packing or equivalent
Column temperature:
Room temperature
Mobile phase (3.22):
3,0 g acetic acid (3.18) + 900 ml water (3.1) +50,0 ml solution (3.21) of 1,1,1-trichloro-2- methyl-2-propanol (3.19) in methanol (3.8) (1 g/100 ml). Adjust pH to 5,0 using ethanolamine (3.20). Make up to 1 000 ml with water (3.1)
Flow rate:
1 ml/min.
Total run time:
approx. 34 min.
Detection wavelength:
excitation: 280 nm, emission: 356 nm.
Injection volume
20 μl

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Source: https://www.legislation.gov.uk/eur/2009/152/annex/III/division/G/division/5/division/5.4 (the official version)
Page: https://openlegislation.uk/eu-regulation-2009-152/annex-III/division-G/division-5/division-5.4
Contains public sector information licensed under the Open Government Licence v3.0. Source: legislation.gov.uk, The National Archives.