微波辅助萃取/高效液相色谱串联质谱法对山银花中活性成分含量的测定

建立了微波辅助萃取/高效液相色谱串联质谱法(MAE/HPLC-MS/MS)同时测定山银花中10种活性成分含量的方法。山银花药材采用MAE萃取,萃取溶剂为乙醇-水(7∶3),固液比1∶30,萃取温度70℃,萃取时间10 min。采用HPLC-MS/MS测定萃取液中活性成分的含量,色谱柱采用Agilent Poroshell120 SB-C18(100 mm×2.1 mm,2.7μm),以0.5%甲酸-乙腈为流动相进行梯度洗脱,负离子多重反应离子监测模式检测。在优化条件下,10种成分的定量分析在10 min内完成。结果表明,10种活性成分的线性范围为0.05～500 mg/L,相关系数(r)不低于0.996 9,检出限和定量下限分别在69～4 413μg/kg和231～14709μg/kg范围,回收率为94%～105%。采用该方法检测6个不同产地的山银花样品,10种活性成分的含量在3.98～14 356.31 mg/kg范围。该方法快速、准确,可有效地用于山银花药材的质量控制。     

高效液相色谱法测定野菊花中蒙花苷的含量

建立高效液相色谱法测定野菊花中蒙花苷含量的方法。采用KromosilTM C18十八烷基硅烷键合硅胶色谱柱,甲醇-水-乙酸(体积比为55∶44.5∶0.5)为流动相,流速为0.8mL/min,柱温为室温,检测波长为334nm。蒙花苷含量在11.2~89.6μg/mL范围内,线性关系良好(r=0.9998),方法的相对标准偏差为0.51%(n=5),平均加标回收率为98.92%。     

杭白菊与野菊花的微量元素比较研究

采用火焰原子吸收光谱法对同属不同种菊科植物杭白菊及野菊的Ni,Zn,Mn,Cu,Mg,Fe,Ca和Pb等无机元素进行了分析测定。该方法的加标回收率为94.20%～110.50%,RSD＜4.12%,具有良好的准确度和精密度。结果表明：杭白菊中的Ni,Zn,Fe,Pb的含量较高,而野菊花的Mn,Ca含量较高。此测定结果可为探讨杭白菊和野菊花中元素含量与其药效的相关性提供科学数据。     

不同产地金银花药材的HPCE指纹图谱分析

以40mmoL/L硼砂溶液为缓冲液,利用高效毛细管电泳(HPCE)法,建立不同产地金银花的HPCE指纹图谱并测定其中绿原酸的含量。工作电压为17kV,温度为25℃,压力进样时间为6s。不同产地金银花药材中绿原酸的含量为3.2%4.4%。通过对HPCE指纹图谱共有峰数据进行判别分析和聚类分析,可以将不同产地的金银花进行判别分类。HPCE指纹图谱可以用于较全面地考察金银花药材的质量。     

固相萃取/液相色谱-质谱/质谱法测定山银花中5种主要黄酮苷元的含量

该文通过含有盐酸的乙醇溶液回流水解并提取,HLB固相萃取柱净化,液相色谱-质谱/质谱法检测,建立了山银花中槲皮素、木犀草素、山萘酚、芹菜素和黄芩素5种黄酮苷元含量的测定方法。实验以芦丁、木犀草苷、紫云英苷、野漆树苷和黄芩苷5种黄酮苷为代表开展研究,山银花样品经50%的乙醇溶液(含10%浓盐酸)回流2 h水解黄酮苷,同时对黄酮苷元进行提取,HLB固相萃取柱净化,采用Mightysil RP-18色谱柱分离,液相色谱-质谱/质谱法检测(电喷雾离子源、多反应监测模式、负离子扫描),外标法定量测定水解后的5种黄酮苷元含量。方法的定量下限(S/N=10)为0.005 g/kg(槲皮素),0.01 g/kg(木犀草素和芹菜素)和0.05 g/kg(山萘酚和黄芩素)。在0~1.0 g/kg范围内,5种黄酮苷元的线性相关系数均大于0.995;在山银花样品中对待测物进行3种加标水平的回收实验(加标水平相当于水解后槲皮素和木犀草素含量为:0.10、0.20、0.40 g/kg,山萘酚、芹菜素和黄芩素含量为:0.05、0.10、0.20 g/kg),方法的平均回收率70.4%~104%;相对标准偏差为4.0%~12%。该方法实现了山银花中多种主要黄酮苷元含量的同时测定,且对研究山银花药效及与黄酮类化合物的关系具有重要意义。     

Binding study of Flos Lonicerae Japonicae with bovine serum albumin using centrifugal ultrafiltration and liquid chromatography

The screening and analysis of bioactive components in traditional Chinese medicines (TCMs) is very important not only for the quality control of Chinese herbs but also for elucidating the therapeutic principles. This study developed a new method for screening and analyzing bioactive compounds from TCMs using centrifugal ultrafiltration coupled with high-performance liquid chromatography. The method was successfully applied in the binding study of Flos Lonicerae Japonicae with bovine serum albumin (BSA), and 11 compounds were found to be bound with the BSA. Eight of them were positively identified as chlorogenic acid, caffeic acid, rutin, quercetin-3-O-glucoside, luteolin-7-O-glucoside, lonicerin, 3, 5-di-O-caffeoyl quinic acid and 3,4-di-O-caffeoyl quinic acid. Another three compounds were tentatively identified as two isomers of chlorogenic acid and one isomer of di-O-caffeoyl quinic acid by comparing the UV data and MS data with the previous reports. Based on modern pharmacological study, these compounds are the major bioactive components in Lonicera japonica. Therefore, the proposed method could be a good approach to predicting the potential bioactivities of multiple compounds in TCMs simultaneously.     

Carbon nanotube/polystyrene composite electrode for microchip electrophoretic determination of rutin and quercetin in Flos Sophorae Immaturus

In this report, carbon nanotube/polystyrene (CNT/PS) composite electrodes have been fabricated as sensitive amperometric detectors of microchip capillary electrophoresis (CE) for the determination of rutin and quercetin in Flos Sophorae Immaturus. The composite electrode was fabricated on the basis of the in situ polymerization of a mixture of CNT and styrene in the microchannel of a piece of fused silica capillary under heat. The surface morphologies of the composite in the electrodes were observed by using a scanning electron microscope. The performance of this unique system has been demonstrated by separating and detecting rutin and quercetin. The new CNT-based CE detector offered significantly lower detection potentials, yielded substantially enhanced signal-to-noise characteristics, and exhibited resistance to surface fouling and hence enhanced stability. It demonstrated long-term stability and reproducibility with a relative standard deviation of less than 5% for the peak current (n = 20) and should also find a wide range of applications in conventional CE, flowing injection analysis, and other microfluidic analysis systems.     

Chemical Fingerprint for Identification and Quality Control of Saccharides in Danhong Injection Based on HPLC-ELSD with Chemometrics

Chemical fingerprinting has been widely used for the quality control of complex natural products including traditional Chinese medicines and botanical drugs. However, there is still lack of appropriate method to monitor the batch-to-batch consistency of hydrophilic constituents, such as saccharides and oligosaccharides. In the present study, we developed a novel approach based on high performance liquid chromatography with evaporative light scattering detector(HPLC-ELSD) for establishing saccharide fingerprinting of Danhong Injection(DHI), which is a widely used botanical drug for treating cardiovascular diseases. Major saccharides in DHI were isolated and four compounds including fructose, glucose and two oligosaccharides were identified. The structures of two novel saccharides named glycerol-1-O-galactfpyranosyl-(1→4)-O-arabmofuranoside, and glycerol-1-O-galactpyranosyl-(1→4)[O-arabmfuranosyl-(1→3)]-O-arabinofuranoside were confirmed by NMR, HR-ESI-MS and GC-MS for the first time. The establishment approach was successfully applied to distinguishing 12 batches of DHI from other botanical drugs with the aid of principle component analysis as well as to evaluating batch-to-batch consistency with the help of calculating similarity of tmgerprints. Our findings indicate that the chemical fingerprint of saccharides can be a useful tool for the quality control of complex natural products.     

金银花道地品与非道地品的FTIR光谱研究

利用FTIR光谱法分析了四个不同产地金银花的红外光谱图,对道地品(河南)和非道地品金银花进行了对比研究。结果表明,由于产地不同,其所含主要有效成分间的相对含量有所改变,反映在FTIR谱图上的吸收峰频率和形状都有所差异。四川产金银花与道地品的谱图从波数和吸收峰形状上都有较明显的差异,虽然山东产金银花与道地金银花齐名,但红外谱图也稍有差异。河北与山东产金银花谱图基本一致。道地品在1 734 cm-1处有一弱吸收,山东、河北两地在此处几乎无吸收;在1 522 cm-1处,道地品是一组弱小的吸收组峰,其他产地的吸收峰为强度略强的独立峰或分裂峰;在1 155～1 045 cm-1范围内,道地品的峰形较尖窄,山东、河北的吸收峰是较宽平的分裂峰。因此,利用FTIR光谱法不仅能快速、准确地鉴别道地药材,而且还可以用来监控药材的引种栽培,优良品种的提纯复壮。     

Development of microwave-assisted extraction followed by headspace solid-phase microextraction and gas chromatography-mass spectrometry for quantification of camphor and borneol in Flos Chrysanthemi Indici

In the work, microwave-assisted extraction (MAE) followed by headspace solid-phase microextraction (HS-SPME) and gas chromatography-mass spectrometry (GC-MS) was developed for quantitative analysis of the bioactive components of camphor and borneol in a traditional Chinese medicines (TCM) of Flos Chrysanthemi Indici. After systematical investigation, the optimal experimental parameters microwave power (400 W), irradiation time (4 min), fiber coating (PDMS/DVB fiber), extraction temperature (40 °C), extraction time (20 min), stirring rate (1100 rpm), and salt effect (no salt added) were investigated. The optimized method provided satisfactory precision (RSD values less than 12%), good recovery (from 86% to 94%), and good linearity (R² > 0.999). The proposed method was applied to quantitative analysis of camphor and borneol in Flos Chrysanthemi Indici samples from 11 different growing areas. To demonstrate the method feasibility, steam distillation was also used to analyze camphor and borneol in Flos Chrysanthemi Indici samples from these different growing areas. The very close results were obtained by the two methods. It has been shown that the proposed ME-HS-SPME-GC-MS is a simple, rapid, solvent-free and reliable method for quantitative analysis of camphor and borneol in TCM, and a potential tool for quality assessment of Flos Chrysanthemi Indici.