However, the previous reported yield of DPHC from the ethyl acetate fraction of ) chromatogram of IOEA.

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The MS was fitted with a Finnigan electrospray source and can analyze ions up to m/z 2000.

Xcalibur software (Finnigan MAT) was used for MS operation.

Thereafter, the frozen sample was freeze-dried before extraction.

All solvents used for the preparation of the crude sample for CPC separation were of analytical grade (Daejung Chemicals & Metals Co., Seoul, South Korea), and HPLC grade solvents were purchased from Burdick & Jackson (MI, USA).

The bottom aqueous phase was used as the stationary phase, whereas the top organic phase was employed as the mobile phase.

When the mobile phase appears from the column, it reaches a hydrostatic equilibrium (back pressure, 3.1 MPa), and then the emission from the CPC process is monitored in UV at 254 nm.

The powdered extract (68 g) was then mixed in water (1.0 L) and in a row separated with n-hexane (n-Hex), chloroform (CHCl) with some modifications.

For the HPLC, an Atlantis T3 column (3 μL, 3.0 × 150 mm column) (Waters, USA) was used, and the mobile phase was acetonitrile-water in gradient mode as follows: (0 → 40 min, → HPLC–DAD–ESI/MS analyses were carried out using a Hewlett–Packard 1100 series HPLC system equipped with a binary pump, a degasser, an autosampler, a DAD detector, and a column oven (Hewlett–Packard, Waldbronn, Germany) coadunate to a Finnigan MAT LCQ ion-trap mass spectrometer (Finnigan MAT, San Jose, CA, USA).

Preparative CPC is liquid–liquid separation without the use of a support, based on the dispersion of each component in two non-mixed liquid phases (Lee et al. In addition, the CPC system provides various technological merits, such as a short operation time, inexpensive product, higher yields and throughput, and reduced operating costs (Berthod and Armstrong , was collected from Seongsan located in the eastern part of Jeju Island, South Korea.

The sample was rinsed more than three times with tap water to remove the surface matters such as epiphytes, salt, and sand and then stored in a refrigerator at −20 °C after cautiously washing with tap water.

(500 g) was extracted three times with 70 % ethanol (Et OH) under stirring for 24 h at room temperature, then it was filtered.