HPLC determination of ferulic acid in rat plasma after oral administration of Rhizoma Chuanxiong and its compound preparation

An HPLC method is described for determination of ferulic acid in rat plasma. The concentration of ferulic acid in rat plasma was determined after deproteinization with acetonitrile using sulfamethoxazole as internal standard. Chromatographic separations were performed on a C(18) stationary phase with a mobile phase composed of acetonitrile-water (16:84, v/v) with 1% glacial acetic acid. The UV detection wavelength was set at 320 nm. The method was successfully applied to the determination of pharmacokinetic parameters in rat plasma after oral administration of Rhizoma Chuanxiong and and its compound preparation Suanzaoren decoctions. The calibration curve was linear over the range 0.0510-4.08 micro g/mL in rat plasma. Within-day and between-day precisions were less than 4.5% RSD. Mean recovery was determined as 96.9%. The limit of quantitation was 0.0510 micro g/mL. The pharmacokinetic parameters of the two preparations were different significantly (p < 0.05), which may attribute to the effects of other ingredients present in Suanzaoren decoction.     

Assay of free ferulic acid and total ferulic acid for quality assessment of Angelica sinensis

Activity of Chinese Danggui (DG), the processed root of Angelica sinensis (Oliv.) Diels, is linked to the ferulic acid content but the stability of ferulic acid during extraction for medicinal use is not known. The stabilities of ferulic acid and coniferyl ferulate were evaluated in the extracts of DG using a variety of extraction solvents. These included various combinations and proportions of methanol, water, formic acid, 1 M aqueous hydrochloric acid and 2% sodium hydrogen carbonate (NaHCO3) in water. Coniferyl ferulate was found liable to hydrolyze into ferulic acid in neutral, strongly acidic and basic solvents, where heat and water could facilitate this hydrolysis. However, the hydrolysis was relatively resisted in weakly organic acid. Based on the stability evaluation, two new terms, namely: free ferulic acid and total ferulic acid, were suggested and defined. Free ferulic acid refers to the natural content of ferulic acid in herbs. Total ferulic acid means the sum of free ferulic acid plus the amount of related hydrolyzed components. Meanwhile, the high-performance liquid chromatographic (HPLC) method was developed to assay free ferulic acid and total ferulic acid in DG using methanol-formic acid (95:5) and methanol-2% NaHCO3 in water (95:5) as extraction solvents, respectively. Ten DG samples were investigated on their contents of free and total ferulic acid. The results indicated that the amount variety of free ferulic acid was larger than that of their counterparts, and the ratio of total ferulic acid to free ferulic acid was 4.07 +/- 2.73 (mean +/- SD, n = 10). The chemical assay of DG using total ferulic acid content would be a better choice to assess the herbal quality and was recommended.     

孔雀石绿光度法测定阿魏酸钠

在pH=4.5的B-R缓冲介质中,阿魏酸可与孔雀石绿反应生成具有正吸收峰和负吸收峰的离子缔合物,其最大正吸收波长位于612nm,最大负吸收波长位于650nm,阿魏酸的质量浓度在0.2~3.9mg/L(正、负吸收)范围内与吸光度A呈线性关系,表观摩尔吸光系数ε分别为3.95×104 L/(mol·cm)(正吸收)和1.20×104 L/(mol·cm)(负吸收)。该方法用于市售阿魏酸钠药物中阿魏酸钠的测定,结果满意。     

Structural elucidation of new ferulic acid-containing phenolic dimers and trimers isolated from maize bran

Four new phenolic dimers and trimers that contain ferulic acid moieties were isolated from the alkaline hydrolyzate of insoluble maize bran fiber and their structures were established by 1D/2D NMR and mass spectrometry. The biological role of one dimer remains unclear whereas the dimeric vanillin-ferulic acid-cross-product probably represents an oxidative degradation product from the corresponding diferulate. Both ferulic acid dehydrotrimers are able to cross-link polysaccharide chains. However, the 5-5/8-O-4(H₂O)-triferulic acid may be a cross-link in its identified structure whereas we assume that the identified 8-O-4/8-5(non-cyclic)-triferulic acid arose from a natural 8-O-4/8-5(cyclic)-triferulate analog during the saponification process.     

Screening and analysis of an antineoplastic compound in Rhizoma Chuanxiong by means of in vitro metabolism and HPLC-MS

A new screening and analysis method that combines in vitro metabolism with high-performance liquid chromatography-mass spectrometry (HPLC-MS) was developed for the screening and analysis of an antineoplastic compound, coniferyl ferulate, which is present in the rhizome of Rhizoma Chuanxiong. Infrared (IR), ultraviolet visible spectroscopy (UV-Vis), nuclear magnetic resonance (NMR) and element analysis were used to identify the molecular structure of coniferyl ferulate. The quantitative analysis of coniferyl ferulate in different extracts of Rhizoma Chuanxiong was carried out, and the metabolism of coniferyl ferulate was investigated by in vitro incubation with rat liver homogenate. The metabolite of coniferyl ferulate, ferulic acid ethyl ester, was identified by HPLC-MS, UV-Vis and IR. In addition, antineoplastic activities of coniferyl ferulate and ferulic acid ethyl ester were detected by the MTT assay. The observed inhibition rate of coniferyl ferulate on the activity of HeLa cells was over 80% at 5.4 ng μl⁻¹. However, its metabolite, ferulic acid ethyl ester, showed no antineoplastic activity in vitro.      

正交试验法优选都梁凝胶贴剂的提取工艺

采用超声法优选都梁凝胶贴剂中白芷、川芎药对的最佳提取工艺.按照L9（34）正交试验设计,以欧前胡素、阿魏酸的转移率及干浸膏得率为综合评价指标,用高效液相色谱法进行含量测定.结果表明,都梁凝胶贴剂中白芷、川芎药对的最佳超声提取工艺为：8倍量的80％乙醇提取2次,每次超声40min;此时浸膏中欧前胡素的平均含量为0.1379％,阿魏酸的平均含量为0.1410％.本实验所建立的高效液相色谱法的准确度、精密度与重现性良好;提取工艺简便,合理.     

毛细管电泳同时分离测定阿魏酸、异阿魏酸的研究

建立了同时分离测定阿魏酸、异阿魏酸的毛细管电泳(CZE)新方法。以20 mmol/L硼砂为背景电解质,体积分数15%异丙醇为有机改性剂,分离电压为20 kV,在219 nm波长下紫外检测。对硼砂浓度、有机溶剂体积分数、分离电压等因素对分离的影响做了系统的研究,最后确立了阿魏酸、异阿魏酸的最佳分离条件。阿魏酸、异阿魏酸分别在2.40~24.0μg/mL、1.80~18.0μg/mL范围内线性关系良好(r=0.9995和r=0.9991),回收率分别为96.61%~101.9%,98.80%~101.8%。方法已用于升麻中阿魏酸、异阿魏酸的测定。     

脱除阿魏酸后车前子多糖的溶液性质、构象特征及固体形貌

分别采用Na OH和阿魏酸酯酶对车前子多糖(PLCP)进行脱除阿魏酸处理,得到的多糖分别记为PLCP-FAS和PLCP-FAE.通过基本理化性质测定和单糖组成分析比较了处理前后多糖的基本结构特征,并采用乌式黏度计、高效体积排阻色谱(HPSEC)和扫描电子显微镜(SEM)等方法表征了多糖的溶液性质、构象特征及固体形貌.结果表明,经Na OH或阿魏酸酯酶处理后多糖的单糖组成基本不变,但糖含量和糖醛酸含量增加,表观黏度显著下降;PLCP-FAS和PLCP-FAE在0.1 mol/L Na Cl和0.5 mol/L Na OH中的特性黏度基本相同;HPSEC分析结果表明,PLCP-FAE的重均分子量(Mw)为3.17×106,PLCP-FAS的Mw为2.83×106,PLCP-FAS和PLCP-FAE的持续围长值一致,但相同分子量下PLCP-FAS的特性黏度低于PLCP-FAE;SEM测定结果显示,PLCP-FAS和PLCP-FAE仍保留一定线性链特征.上述结果表明,脱除阿魏酸的大粒车前子多糖呈现半柔顺链的构象特征,样品固体形貌仍保留一定的线性链特征.     

高效液相色谱法同时测定参茸补酒中芍药苷、阿魏酸的含量

建立一种高效液相色谱双波长法同时测定参茸补酒中芍药苷、阿魏酸的含量。该方法采用Symmetry C18色谱柱(150×3.9mm,5μm),流动相为乙腈 水 冰乙酸(12∶88∶1,V/V),检测波长分别为323nm(通道1)、230nm(通道2)。结果显示:芍药苷、阿魏酸分别在21.4～214.0和1.65～16.50μg·mL-1范围内本法有良好的线性关系;平均回收率分别为98.1%(RSD=1.28%)和97.5%(RSD=1.73%)。本法操作简便,快速,重现性好,结果准确,可作为参茸补酒的质量控制方法。