Research On The Material Basis And Quality Control Of Ciwujia Granules

Ciwujia granules are made from75%ethanol extract. It has been used for invigorating the spleen and kidney, replenishing the vital essence and calmming the nerves. The research focused on the material basis and quality control of Ciwujia granules. The chemical constituents from the EtOAc-soluble fraction of75%EtOH extract of Acanthopanax senticosus and Ciwujia granules were studied. And we examined the antioxidant activities of the compounds we got. Furthermore, we have established a method for the simultaneous determination of five components in Ciwujia granules by HPLC with different UV wavelengths. The main results are described as follows:1. The EtOAc-soluble fraction was isolated by column chromatography and RP-preparative HPLC. Their structures were identified by spectroscopic methods (MS,’H-NMR,13C-NMR). Twenty-one compounds were isolated from the EtOAc extract of Acanthopanax senticosus and identified as quercitin (1), quercetin (2), kaempferol (3), hyperoside (4), rutin (5), acacetin (6), daidzin (7),3’-methoxydaidzin (8), puerarin (9),3’-methoxypuerarin (10),4’-methoxypuerarin (11), syringaldehyde (12), syringic acid (13), glucosyringic acid (14), isofraxidin (15),2,3di(3’,4’-methyl-endioxy-benzyl)-2-buten-4-olide (16),l-sesamin (17), methylpluviatolide (18),4’-hydroxy-2’-methoxycinnamaldehyde (19), diisobutyl phthalate (20), and dibutyl phthalate (21). And compounds6-11,14,16,18-21are isolated from the plants in Acanthopanax (Deche et Planch.) Miq. for the first time. We also have identified8constituents in Ciwujia granules by using HPLC-PDA/ESI-MSn. These compounds were protocatechuic acid, eleutheroside B, chlorogenic acid, caffeic acid, eleutheroside E, hyperoside, isofraxidin and quercetin.2. We examined the antioxidant activities by DPPH radical-scavenging assay of eighteen compounds, eleutheroside B, eleutheroside E, chlorogenic acid and protocatechuic acid. The results showed that compounds1-5,10,13, chlorogenic acid and protocatechuic acid have significant DPPH radical-scavenging activities and compounds8and15have weaker activities.3. We have established a method for the simultaneous determination of five components (protocatechuic acid, eleutheroside B, chlorogenic acid, eleutheroside E and isofraxidin) in Ciwujia granules by HPLC with different UV wavelengths. The results showed that protocatechuic acid, eleutheroside B, chlorogenic acid, eleutheroside E and isofraxidin had good linearity in the ranges of0.935-187μg/mL (r=1.0000),1.52-303μg/mL (r=1.0000),2.74-547μg/mL (r=1.0000),3.62-723μg/mL (r=1.0000), and0.510-102μg/mL (r=1.0000), respectively. The average recoveries of high, median, and low concentrations were between97.8%-102.6%,98.0%-103.5%, and96.8%-104.8%and RSD were between0.8%-4.0%,1.3%-4.0%, and0.9%-4.0%, respectively. We also detected the five components in Ciwujia Granules from different manufacturers and batches.