Off-line coupling of new generation centrifugal partition chromatography device with preparative high pressure liquid chromatography-mass spectrometry triggering fraction collection applied to the recovery of secoiridoid glycosides from Centaurium erythra

- New CPC device used for pre-purification of Centaurium erythraea methanolic extract.
- Final step of isolation by PrepHPLC hyphenated with mass triggering detector.
- Quick and efficient recovery sequence of valuable pure secoiridoid glycosides.

A purification sequence including a Gilson CPC 250 PRO device coupled to PrepHPLC hyphenated with a MS triggering fraction collector was applied to isolate secoiridoid glycosides from a complex methanolic extract of Centaurium erythraea. This species is widely used for ethnomedicinal purposes around the Mediterranean Sea.The solvent system ethyle acetate/ethanol/water 7.5/3/5 was determined using shake-flask method targeting swertiamarin, the major secoiridoid of the extract. Optimization of CPC experimental parameters enabled the injection of 4 g of extract with a flow rate of 40 mL/min at 3000 rpm to provide a secoiridoid glycosides enriched fraction. 130 mg of this latter was submitted to a second step of purification by preparative HPLC (gradient water/formic acid (19:1) (A) and methanol (B) as follows: 0 min, 85% A; 8 min, 60% A; 12 min, 55% A; 35 min, 55% A; 40 min, 10% A; 50 min, 10% A; 52 min, 85% A; 55 min, 85% A) to give swertiamarin (36 mg, yield 27.7%, purity 98.2%). Other secoiridoid glycosides (sweroside, gentiopicroside, secologanol, secoxyloganin) were also isolated in minor amounts. As these monoterpene derivatives are responsible for several biological activities, their quick recovery with high yield and purity may serve as a model for further scale-up and industrial development.