ISO 7358-2021 pdf download.Essential oils of bergamot, lemon, bitter orange and lime, fully or partially reduced in bergapten- Determination of bergapten content by high-performance liquid chromatography (HPLC). 62 HPLC system 6.2.1 LIquid chromatograph. 622 Column for HPLC 1 normal phase, made of stainless steel, of length between 150 mm .md 250 mm. having an internal diameter between 4 mm and 5 mm arid packed wIth .m stationary phase consisting aigranuLited silica tiC IIPL.C qualily. with a grain iize of approximately 5 pm. 6.2.3 Column for HPLC in reversed phase, made of stainless steel, of length 150 mm (or 250 mm), having an internal diameter 4,6 mm and packed with a stationary phase C18 type with a grain size of approximately 3.5 pm (or 5 pm). (or example Zorbax Eclipse Ptusi C: 18: 3.5 pm (4,6 a 150) mm ilent. 6.2.4 A ternary pump system enabling programmed solvent gradients. 62.5 Solvent degasslng system (npional), (or example ultrasonic tank. 6.2.6 t)elecllon system. .iditistablc to wavelengths o1254 rim or 312 nrn. diode-array IN spectromrtrbc detection system. 62.7 Recorder and (optional) Integrator, suitable for this HPLC system, 7 Sample preparation Prepare the test sample as specified In 150 356. Dissolve any solid deposit by moderate heating A Procedure 9.1 OperatIng conditions Adlust the flow rate of the mobile phase (52.L4) so as to obtain good separation of the peaks corresponding to berg.ipten and coumarin from other essential oil omponrllts detectable by the LIV detector (626). The flow rate is typically between I mi/mm and 1.5 mI/mm. Follow the procedure specified in ISO 8432. 8.2 Detenninatlon 8.2.1 IIPLC In normal phase 8.2.1.1 Internal standard method 9.2.1.1.1 Optimization of HPL( chroniatographic conditions in normal phase 8.2.1.1.1.1 SeparatIon Verify that the bergapten is well separated From the other constituents of the essential oil in the chromatograms obt.iinvd, Next. verily thit the Inis’rnal standard. coum.irin (5.2) dsw not mask ‘r coincide with any constituent of the essential oil. Determine the retention times of the bergapten and coumarin.ISO 7358 pdf download.
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