Bioassay‐guided isolation of an active compound with protein tyrosine phosphatase 1B inhibitory activity from Sargassum fusiforme by high‐speed counter‐current chromatography

Abstract

A rapid and efficient method using high-speed counter-current chromatography was established for the bioassay-guided separation of an active compound with protein tyrosine phosphatase 1B inhibitory activity from Sargassum fusiforme. Under the bioassay guidance, the ethyl acetate extract with the best IC value of 0.37 ± 0.07 μg/mL exhibited a potential protein tyrosine phosphatase 1B inhibitory activity, which was further separated by high-speed counter-current chromatography. The separation was performed with a two-phase solvent system composed of n-hexane/methanol/water (5:4:1, v/v). As a result, dibutyl phthalate (19.7 mg) with the purity of 95.3% was obtained from 200 mg of the ethyl acetate extract. Its IC was 14.05 ± 0.06 μM, which was further explained by molecular docking. The result of molecular docking showed that dibutyl phthalate enfolded in the catalytic site of protein tyrosine phosphatase 1B. The main force between dibutyl phthalate and protein tyrosine phosphatase 1B was the hydrogen bond interaction with Gln266. In addition, hydrogen bond, van der Waals force and hydrophobic interaction with the amino acids (Ala217, Ile219, and Gly220) were also responsible for the stable protein-ligand complex.

Bioassay‐guided separation and identification of the anticancer composition from Curcuma longa L. by the combination strategy of methanol gradient countercurrent chromatography and ultra‐high‐performance liquid chromatography coupled with high‐resolution mass spectrometry

“…At this point, methanol gradient CCC based on HepEMWat solvent systems was proven to be an efficient method for the fractionation and separation of active components from C. longa L. Except for the attractive curcuminoids, one labdancetype diterpene dialdehyde galanal A was screened out. In the previous studies, CCC was also widely used for the bioassay-guided separation of components with diverse activities, but the polarity of ingredients was usually in a much smaller range [41][42][43]. In addition, though gradient CCC based on an ethyl acetate-methanol-water solvent system could elute compounds with large range of Kvalues [44], the scope of application was much smaller than gradient CCC based on HepEMWat solvent systems.…”

Bioassay-Guided Separation of Centipeda minima Using Comprehensive Linear Gradient Centrifugal Partition Chromatography

“…In order to solve these problems, countercurrent separation could provide an excellent alternative, such as centrifugal partition chromatography (CPC) or high-speed counter-current chromatography (HSCCC). CPC is a type of support-free liquid–liquid partition chromatography, with a low risk of sample denaturation, as well as total sample recovery, and large load capacity that has been widely used for the separation of constituents with bioactivity [ 6 , 7 , 8 , 9 , 10 ]. Despite the above-mentioned advantages, the CPC-mediated bioactivity-guided separation has several complementary points: (i) most solvent systems are limited to separating constituents with a narrow polarity range, (ii) optimizing a biphasic solvent system is a time-consuming process, as the partition coefficient of each component of the two layers should be compared in different ratios of the various solvents using HPLC or TLC.…”

A multilayer coil in type-I counter-current chromatography

“…The multilayer coil separation column was widely used in the system of type-J CCC with high performance [16, 17]. In order to simplify the column design and increase the sample loading capacity the multilayer coil was applied to type-I counter-current chromatography in the present study.…”