ABSTRACT
The aim of this study was to develop a simple, sensitive ultra performance liquid chromatography mass spectrometry (UPLC-MS/MS) method for the determination of syringaresinol, N-trans-feruloyltyramine, chelerythrine chloride, sinomenine, coptisine chloride, sanguinarine, chelidonine, magnolflorine, allocryptopine, protopine, farrerol, stylopine and dihydrosanguin-arine in Tong'an injection (TAI), which could be used for the quality control of TAI. The UPLC analysis was performed on Agilent Zorbax SB-Aq column (2.1 mm×150 mm,3.5 μm), with 0.1% formic acid solution (A) -acetonitrile (B) as the mobile phase for gradient elution (0.01-2 min, 5%B; 2-8 min, 5%-30%B; 8-11 min, 30%-95%B; 11-13 min, 95%B; 13-13.1 min, 95%-5%B; 13.1-14 min, 5%B). The flow rate was 0.5 mL•min⁻¹, and the column temperature was 25 ℃; multiple reaction monitoring (MRM) was performed in electrospray ion source positive ion mode for quantitative determination. The calibration curves for the above thirteen compounds showed good linear relationship in corresponding mass concentration range (r>0.999 0). The average recovery rate of the compounds ranged from 95.70% to 104.8%, with RSD of less than 1.9%. The contents of thirteen active components in 10 batches of TAI were 0.021 2-0.029 0, 0.001 7-0.002 3, 0.000 9-0.001 3, 5.952-6.205 2, 0.195 4-0.240 5, 0.002 0-0.002 9, 0.693-0.798 2, 0.069 3-0.078 2, 0.089 29-0.102 9, 0.386 5-0.420 1, 0.014 3-0.015 9, 0.755 3-0.842 1, and 0.008 2-0.011 2 g•L⁻¹ respectively. Methodology validation proved that this method was simple, rapid, sensitive and accurate, which can be used to provide reference for the comprehensive evaluation of TAI quality. The determination results of 10 batches of TAI showed the content of each batch had no significant difference. The results may provide a basis for the quality control of TAI.
ABSTRACT
This study aimed at investigating the antiviral constituents from the active fractions of Tong-An (TA) injection.In this study,the active constituents of TA injection were screened by LPS-induced PGE2 production mode to detect the contents of PGE2.The chemical constituents were isolated by HP-20 macroporous resin,silica gel column chromatography,ODS column chromatography,Sephadex LH-20 column chromatography and preparative and semi-preparative HPLC.The structures were identified by spectral data and physicochemical property.As a result,the 95％ ethanol eluate of TA injection on the macroporous adsorption resin column was proved to be the active fraction of TA injection.Seventeen compounds were isolated from TA injection and identified as syringaresinol (1),N-Trans-Feruloyltyramine (2),chelerythrine (3),sinomenine (4),coptisine (5),sanguinarine (6),chelidoniny (7),magnoflorine (8),allocryptopine (9),protopine (10),farrerol (11),dihydrosanguinarine (12),heptadec-(9Z)-enoic acid (13),chlorogenic acid (14),cryptochlorogenin acid (15),3,5-di-O-caffeoylquinic acid (16) and 4,5-di-O-caffeoylquinic acid (17).PGE2 inhibitory activities of these compounds were determined,among which six compounds presented inhibitory activities against PGE2.It was concluded that all the isolated compounds from TA injection were firstly reported with the favorable inhibitory activities of compounds 2,5,9,10,11,12 against PGE2.