Isolation of α-Amylase Inhibitors from Kadsura longipedunculata Using a High-Speed Counter-Current Chromatography Target Guided by Centrifugal Ultrafiltration with LC-MS.
In this study, a high-speed counter-current chromatography (HSCCC) separation method target guided by centrifugal ultrafiltration with high-performance liquid chromatography-mass spectrometry (CU-LC-MS) was proposed. This method was used to analyze α-amylase inhibitors from Kadsura longipedunculata extract. According to previous screening with CU-LC-MS, two screened potential α-amylase inhibitors was successfully isolated from Kadsura longipedunculata extract using HSCCC under the optimized experimental conditions. The isolated two target compounds (with purities of 92.3% and 94.6%) were, respectively, identified as quercetin-3-O-rhamnoside (1) and protocatechuic acid (2) based on the MS, UV, and ¹H-NMR spectrometry data. To verify the inhibition of screened compounds, the inhibitory activities of quercetin-3-O-rhamnoside (1) and protocatechuic acid (2) on α-amylase were tested, and it demonstrated that the experimental IC50 values of quercetin-3-O-rhamnoside (1) and protocatechuic acid (2) were 28.8 and 12.5 μmol/L. These results proved that the hyphenated technique using CU-LC-MS and HSCCC was a rapid, competent, and reproductive method to screen and separate potential active compounds, like enzyme inhibitors from the extract of herbal medicines.
College of Chemistry and Chemical Engineering, Central South University, Changsha 410083, Hunan, China. hn_cenyin@csu.edu.cn.
Yin
Y
Cen
Institute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences, Changsha 410205, Hunan, China. aipingxiao@yahoo.com.
Aiping
A
Xiao
College of Chemistry and Chemical Engineering, Central South University, Changsha 410083, Hunan, China. xqchen@csu.edu.cn.
Xiaoqing
X
Chen
Institute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences, Changsha 410205, Hunan, China. liuliangliang@caas.cn.
Liangliang
L
Liu
D002498:Centrifugation; D002845:Chromatography; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000302:isolation & purification; D062385:Hydroxybenzoates / Q000737:chemistry / Q000302:isolation & purification; D032135:Kadsura / Q000737:chemistry*; D011794:Quercetin / Q000031:analogs & derivatives / Q000737:chemistry / Q000302:isolation & purification; D014462:Ultrafiltration; D000516:alpha-Amylases / Q000037:antagonists & inhibitors*
D004791:Enzyme Inhibitors; D062385:Hydroxybenzoates; C009091:protocatechuic acid; D011794:Quercetin; D000516:alpha-Amylases
HSCCC; Kadsura longipedunculata; ultrafiltration; α-amylase