Foreword

1Scope

2Normative references

3Terms and definitions

4Principle

5Reagents

6Apparatus

6.1Common laboratory equipment

6.2HPLC system

7Sample preparation

8Procedure

8.1Operating conditions

8.2Determination

8.2.1HPLC in normal phase

8.2.1.1Internal standard method

8.2.1.1.1Optimization of HPLC chromatographic conditions in normal phase

8.2.1.1.1.1Separation

8.2.1.1.1.2Quantity of internal standard

8.2.1.1.2Response factor K

8.2.1.1.3Determination of bergapten

8.2.1.2External standard method

8.2.2HPLC in reversed phase

8.2.2.1Preparation of the reference solutions for calibration

8.2.2.1.1General

8.2.2.1.2Stock solutions

8.2.2.1.3Working solutions

8.2.2.2Preparation of citrus essential oils samples

8.2.2.3Determination by HPLC and internal standardization of bergapten content in essential oils of bergamot

9Calculation — Normal phase or reversed phase HPLC

9.1Internal standard method

9.2External standard method

10Precision — Repeatability

11Test report

Annex ATypical chromatogram of the reference substances using high-performance liquid chromatography (HPLC) of reference compounds in reversed phase (informative)

Annex BBergapten UV-Vis spectrum (informative)

Bibliography