Abstract
A reproducible analytical method using reverse-phase high liquid performance chromatography combined with UV detecting was developed for the quantitative determination of four compounds isolated from the ethanol extract of Phaseolus angularis seeds (PASE): oleanolic acid (1), oleanolic acid acetate (2), stigmasterol (3) and β-sitosterol (4). This method was fully validated in terms of linearity (r2 > 0.999), accuracy (98.5%–100.8%), precision (<0.92%), LOD (<0.0035 mg/mL), and LOQ (<0.0115 mg/mL). The effects of the PASE and isolated compounds 1–4 on TLR4 activation were tested in THP1-Blue cells. Among the tested substances, compound 2 showed potent inhibitory activity with an IC50 value of 3.89 ± 0.17 µM.
Highlights
Phaseolus angularis seeds (PAS) as a very important crop in East Asia that has mainly been cultivated in Korea, Japan, northern and central China for many centuries
We found that the ethanol extract of PAS showed strong TLR4 inhibitory activity in human monocyte THP1-Blue cells (IC50 < 30 μg/mL)
An HPLC-UV method was developed for the quantitation of four compounds isolated from the ethanol extract of PAS: oleanolic acid (1), oleanolic acid acetate (2), stigmasterol (3) and β-sitosterol (4)
Summary
Phaseolus angularis seeds (PAS) as a very important crop in East Asia that has mainly been cultivated in Korea, Japan, northern and central China for many centuries. They are traditionally used as diuretic, antidote and a remedy for dropsy and beriberi [1]. The inhibitory effect of PASE on the TLR4 activation and TLR4-mediated inflammatory response was reported [10]. We isolated four constituents from PASE to identify the active compounds that are responsible for the inhibitory effect on TLR4 activity. This paper describes the quantification and validation of isolated compounds with TLR4 inhibitory effects from PASE
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