中药材中5种拟除虫菊酯农药残留量的测定

研究中药材中5种拟除虫菊酯农药残留量的测定方法。用石油醚-丙酮混合溶剂超声提取,固相萃取法净化,用气相色谱-电子捕获检测器（ECD）测定。5种拟除虫菊酯农药在3种代表性中药材的加标回收率为72．6％-135．0％,相对标准偏差为3．6％。27％;方法的检出限为0．001—0．005μg／g。方法灵敏度高,选择性好,操作较为简便,适用于植物性（根、茎类）药材中5种拟除虫菊酯农药残留量的测定。     

毛细管气相色谱法测定中药材中多种拟除虫菊酯农药残留量

研究了气相色谱法测定中药材中多种拟除虫菊酯类残留量的分析方法。样品经正己烷．丙酮混合液[V（正己烷）：V（丙酮）=1：1]提取,氟罗里硅土层析柱净化,采用毛细管色谱柱分离,GC-ECD可同时测定8种拟除虫菊酯类农药残留量,回收率为76．6％～104．2％。     

Determination of five toxic alkaloids in two common herbal medicines with capillary electrophoresis

Calycosin was purified from an ethyl acetate extract of the root of Astragalus membranaceus Bge. var. mongholicus (Bge.) Hsiao by high-speed counter-current chromatography. The separation was performed in two steps with a two-phase solvent system composed of n-hexane-chloroform-methanol-water (1:3:3:2, v/v). From 200 mg of the crude extract, 14.8 mg of calycosin was obtained at over 99% purity as determined by HPLC analysis, and its chemical structure was confirmed by MS, 1H and 13C nuclear magnetic resonance.     

氢化物发生-原子荧光光谱法测定中草药中的微量铅

采用水煎煮法提取中草药,HNO3-H2O2密闭消解体系消解样品,应用氢化物发生-原子荧光光谱法测定中草药及其提取液中的微量铅;优化了仪器工作及实验条件,荧光强度与Pb质量浓度在0.49～400 μg/L范围内呈线性关系,相关系数:0.9996,检出限:0.49 μg/L;发现部分提取液中Pb量比原中草药样品中Pb量有明显降低.     

Chip-based high-throughput screening of herbal medicines

At present, high-throughput screening (HTS) programs in drug discovery rely mainly on compound libraries from combinational chemistry. Similarly, natural flora has been used as a prominent origin for new and potent herbal drugs. Herbal medicines have been used worldwide for thousands of years to cure many diseases. As such, herbal secondary metabolites show a remarkable structural diversity that supplements chemically synthesized compound analogs in drug discovery screening. Unfortunately, there is often a considerable deterioration in the quality of herbal drugs in such screening programs as there are time-consuming manual processes involved in the isolation of active ingredients from the highly complex mixtures of herbal plant products. The quality and quantity of herbal samples are critical for the success of HTS programs. In the recent past, there have been substantial improvements in HTS due to the miniaturization and integration of microchip (e.g., Herbochip(?), DNA chip, protein chip, cell chip, etc.)-based technologies so as to design herbal drugs that compete with synthetic drug analogs. Here we will review various technologies used for HTS of herbal medicines. Finally, we will summarize our efforts to develop a novel chip-based HTS assay to explore the antioxidant and radioprotective properties of herbal plants.     

Determination of polyphenol components in herbal medicines by micellar electrokinetic capillary chromatography with Tween 20

A simple and convenient method of micellar electrokinetic capillary chromatography (MEKC) using polyoxyethylene sorbitan monolaurate (Tween 20) to form single micelle and methanol as a buffer additive was introduced for the simultaneous determination of five polyphenols, including scopoletin, rutin, esculetin, chlorogenic acid and caffeic acid. A running buffer solution of pH 9.3, 20 mmol/L sodium tetraborate containing 64 mmol/L Tween 20 and 9% (v/v) methanol was adopted in the separation. Because rutin and esculetin were difficult to be separated by capillary zone electrophoresis (CZE) and SDS-based MEKC, Tween 20-based MEKC was adopted and the polyphenols were separated satisfactorily. The proposed method was used to determine the polyphenol components in the herbal medicine of Cortex fraxini. The separation mechanism of Tween 20-based MEKC for the polyphenols was discussed preliminarily.     

高效液相色谱快速测定草本植物饮料中28种外源性药物和内源性成分

建立了高效液相色谱快速测定草本植物饮料中28种外源性药物和内源性成分的分析方法。样品经甲醇-水（70：30,v/v）超声提取后,使用Thermo Accucore C18色谱柱（100 mm×4.6 mm,2.6 μm）,以甲醇、乙腈和20 mmol/L乙酸铵溶液（pH 4.2）为流动相进行梯度洗脱,流速为1.2 mL/min,柱温为35℃,采用二极管阵列检测器,检测波长为254和220 nm,外标法定量。结果表明,28种成分在1~100 mg/L范围内线性关系良好,相关系数（r）均不小于0.999。液体样品和固体样品的检出限（LODs）分别为1~10和20~200 mg/kg。28种成分在液体和固体样品中的加标回收率分别为88.8%~118.6%和92.7%~112.3%,相对标准偏差（RSDs）分别为0.1%~6.7%和0.1%~6.4%。按上述方法检测草本植物饮料456批,检出阳性样品55批次,检出率为12.1%。该法简便快速、准确可靠,适用于草本植物饮料中28种成分的测定。     

CE and CEC analysis of phytochemicals in herbal medicines

CE and CEC, due to their versatility and high efficiency, have attracted great interest in the analysis of phytochemicals in herbs and their preparations. Previously, we reviewed the analysis of phytochemical bioactive compounds by CE in 2006 (Electrophoresis 2006, 27, 4808-4819) or CEC in 2010 (Electrophoresis 2010, 31, 260-277). This review followed the previous studies and covered the literature published since 2006 for CE and 2009 for CEC (excluding those mentioned in the two previous reviews), which emphasized the development of CE and CEC techniques in phytochemical analysis. In addition, sample preparation and detection were also discussed.     

Analytical supercritical fluid extraction of Chinese herbal medicines

Summary An analytical supercritical fluid extraction (SFE) technique, followed by GC/MS, was developed to separate and determine the volatile components in Chinese herbal medicine. Three kinds of herbs, frankincense, myrrh, andEvodia rutaecarpa were extracted and analyzed. The extraction was carried out using supercritical fluid CO₂ at 20 MPa and 50°C. The main factors affecting the efficiency and selectivity of the extraction are discussed. The results revealed the potential of supercritical fluid extraction as an analytical procedure for the study of medicinal plants.     

Antioxidants in Chinese herbal medicines: a biochemical perspective

Recently, intense interest has focused on the antioxidant properties of natural products. In particular, Chinese herbal medicines (CHM) have become hot topics for life science researchers since many are reported to possess cardioprotective compounds, many of which remain to be identified. Indeed, the exact mechanisms by which CHM work remain unknown. Although many of these herbal remedies are undoubtedly efficacious, few have been scientifically investigated for their active chemical constituents and biological activities. We have previously reported higher activities of antioxidant defence enzymes such as superoxide dismutase, catalase, glutathione peroxidase and glutathione S-transferases in the liver of rats treated with the herb Salvia miltiorrhiza in a model of acute myocardial infarction. Using well established in vitro antioxidant assays employing 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) and diphenyl-l-picrylhydrazyl (DPPH) we have shown that in addition to elevating endogenous antioxidant enzyme activity, Salvia miltiorrhiza and other CHM traditionally used for cardiovascular disorders (such as Rhizoma ligustici, Herba leonuri, Radix achyranthis bidentatae, and Camellia sinensis) contain potent antioxidant moieties in addition to their phenolic constituents. Furthermore, these novel non-phenolic components are effective inhibitors of oxidative reactions mediated by the inflammatory oxidants, peroxynitrite,hypochlorous acid and hydroxyl radical as well as iron-dependent lipid peroxidation. In this review, we discuss the various antioxidant properties of CHM in the context of their biochemical mechanisms.     

Diagnosis of public programs focused on herbal medicines in Brazil

The present study is aimed to diagnose the current public programs focused on herbal medicines in Brazil by means of in loco visits to 10 programs selected by means of questionnaires sent to 124 municipalities that count on herbal medicine services. The main purpose of the implementation of program programs is related to the development of medicinal herbs. 70% of them are intended for the production of herbal medicines and 50% are aimed to ensure the access of the population to medicinal plants and or herbal medicines. The initiative of the implementation of these programs was related to the managers (60%). The difficulties in this implementation were due to the lack of funding (100%) of the programs. In 60% of the programs, the physicians did not adhere to herbal medicine services due to the lack of knowledge of the subject. Training courses were proposed (80%) to increase the adhesion of prescribers to the system. Some municipalities use information obtained from patients to assess the therapeutic efficiency of medicinal plants and herbal medicines. Of the programs underway, cultivation of medicinal plants was observed in 90% and 78% of them adopt quality control. In most programs, this control is not performed in accordance with the legal requirements. The programs focused on medicinal plants and herbal medicines implemented in Brazil face some chronic problems of infrastructure, management, operational capacity and self-sustainability, which can be directly related to the absence of a national policy on medicinal plants and herbal medicines.     

微波消解-FAAS法测定大别山区三种中药中的微量元素

采用微波消解-火焰原子吸收光谱(FAAS)法对大别山区3种中药(何首乌、黄精和拐枣)中的微量元素进行测定。用微波消解仪处理样品,对消解条件及消解程序进行优化,在最佳微波消解条件下,结合FAAS法测定上述3种中药中Ca、Mg、Cu、Zn、Fe、Mn 6种元素的含量。结果表明,微波消解的最佳条件为:何首乌和拐枣采用HNO3-H2O2混酸体系,配比分别为5∶2和5∶3,固液比1∶20,最高功率(700W)时的保温时间分别为8min和6 min;黄精采用HNO3-HCl,混酸体系,配比5∶2,固液比1∶28,保温时间为8 min。所建立的定量分析方法回收率为95.63%～101.36%,相对标准偏差小于3.0%,线性相关系数大于0.999,方法检出限为0.18～2.20 ng/mL。3种中药中均含有丰富的微量元素,尤其是黄精中Fe、Ca和Mn的含量较高;何首乌中Mg、Zn和Fe含量相对较高;拐枣中Fe和Ca含量相对较高。研究结果可为上述3种中药中微量元素的含量与其药效的相关性提供科学依据。     

色谱指纹图谱在中药质量控制中的应用

介绍了色谱指纹图谱在中药质量控制体系,即中药真实性鉴定(化学指纹图谱)、有效性评价(谱效学、生物指纹图谱、代谢指纹图谱)以及安全性评价方面的应用。目前化学指纹图谱在中药的真实性鉴定方面已比较成熟;谱效学、生物指纹图谱和代谢指纹图谱由于与药效活性相关,在中药的有效性评价中已体现出优势;色谱指纹图谱在中药的安全性评价方面的应用也愈来愈受到重视。     

Role of counter-current chromatography in the modernisation of Chinese herbal medicines

This review focuses on the growing popularity of using counter-current chromatography (CCC), with its liquid stationary phase, as one of the prime methods for isolating compounds from Chinese herbal medicines (CHMs). 198 publications are reviewed covering 108 different plant species from 56 plant families. These describe the isolation of 354 different molecules across a wide range of polarities, chemical classes and molecular weights (in the range 100–1000 Da). The suitability of CCC for the separation of active compounds from CHM, the phase systems used, how CCC has developed in China, compounds isolated, CCC instrumentation, performance, operational issues and innovations, all supported by detailed cross-referencing, are described. It is concluded that CCC is making an increasingly important contribution to the modernisation of Chinese herbal medicines.     

食品中亚硫酸盐的离子色谱法测定

建立了食品中亚硫酸盐的离子色谱检测方法. 样品采用40 mmol/L NaOH溶液提取, 甲醛作稳定剂, 经ENVI-Carb活性碳小柱除去提取液中的色素, 石油醚除去提取液中的油脂, 用配有电导检测器的离子色谱仪测定. 以AS9-HC为色谱柱, 流动相为8 mmol/L Na2CO3-2.5 mmol/L NaOH, 亚硫酸盐的残留量在0～6.0 mg/L的范围内线性关系良好, 相关系数为0.9989, 相对标准偏差为1.3%～9.1%, 回收率在88.4%～98.1%之间.     

Variable selection for discriminating herbal medicines with chromatographic fingerprints

When discriminating herbal medicines with pattern recognition based on chromatographic fingerprints, typically, the majority of variables/data points contain no discrimination information. In this paper, chemometric approaches concerning forward selection and key set factor analysis using principal component analysis (PCA), unweighted and weighted methods based on the inner- and outer-variances, Fisher coefficient from the between- and within-class variations were investigated to extract representative variables. The number of variables retained was determined based on the cumulative variance percent of principal components, the ratio of observations to variables and the factor indicative function (IND). In order to assess the methods for variable selection and criteria levels to determine the number of variables retained, the original and reduced datasets were compared with Procrustes analysis and a weighted measure of similarity. Moreover, the tri-variate plots of the first three PCA scores were used to visually examine the reduced datasets in low dimensional space. Herbal samples were finally discriminated by use of Bayes discrimination analysis with the reduced subsets. The case study for 79 herbal samples showed that, the methods of forward selection associating the variables with the loadings closest to 0 and key set factor analysis were preferable to determine the representative variables. Procrustes analysis and the weighted measure were not indicative to extract representative variables. High matching between the original and reduced datasets did not suggest high prediction accuracy. Visually examining the PC1-PC2-PC3 scores projection plots with the reduced subsets, not all the herb samples could be separated due to the complexity of chromatographic fingerprints.     

阴离子交换色谱法测定硫磺熏制姜中亚硫酸盐

建立了阴离子交换色谱分离、抑制型电导检测分析硫磺熏制生姜(简称硫磺姜)中亚硫酸盐的方法。使用NJ-SA-4A阴离子交换分析柱,淋洗液为1.92mmol/L Na2CO3-1.80mmol/L NaHCO3-2%(体积分数)丙酮。通过在样品提取液中加入三乙醇胺,有效地维持了亚硫酸盐的稳定性、并能提高方法的选择性。SO23-在0.02～2.00mg/L范围内线性良好,相关系数为0.9998,峰面积的相对标准偏差为1.9%。方法的检出限为0.015mg/L,加标回收率为84.0%～93.5%。该方法为生姜的市场监测提供了有效的方法。     

Correction of retention time shifts for chromatographic fingerprints of herbal medicines

In this study, the combination of chemometric resolution and cubic spline data interpolation was investigated as a method to correct the retention time shifts for chromatographic fingerprints of herbal medicines obtained by high-performance liquid chromatography-diode array detection (HPLC-DAD). With the help of the resolution approaches in chemometrics, it was easy to identify the purity of chromatographic peak clusters and then resolve the two-dimensional response matrix into chromatograms and spectra of pure chemical components so as to select multiple mark compounds involved in chromatographic fingerprints. With these mark components determined, the retention time shifts of chromatographic fingerprints might be then corrected effectively. After this correction, the cubic spline interpolation technique was then used to reconstruct new chromatographic fingerprints. The results in this work showed that, the purity identification of the chromatographic peak clusters together with the resolution of overlapping peaks into pure chromatograms and spectra by means of chemometric approaches could provide the sufficient chromatographic and spectral information for selecting multiple mark compounds to correct the retention time shifts. The cubic spline data interpolation technique was user-friendly to the reconstruction of new chromatographic fingerprints with correction. The successful application to the simulated and real chromatographic fingerprints of two Cortex cinnamomi, fifty Rhizoma chuanxiong, ten Radix angelicae and seventeen Herba menthae samples from different sources demonstrated the reliability and applicability of the approach investigated in this work. Pattern recognition based on principal component analysis for identifying inhomogenity in chromatographic fingerprints from real herbal medicines could further interpret it.     

Discrimination of herbal medicines by molecular spectroscopy and chemical pattern recognition

The molecular spectroscopy (including near infrared diffuse reflection spectroscopy, Raman spectroscopy and infrared spectroscopy) with OPUS/Ident software was applied to clustering ginsengs according to species and processing methods. The results demonstrate that molecular spectroscopic analysis could provide a rapid, nondestructive and reliable method for identification of Chinese traditional medicine. It's found that the result of Raman spectroscopic analysis was the best one among these three methods. Comparing with traditional methods, which are laborious and time consuming, the molecular spectroscopic analysis is more effective.     

Liquid chromatographic separation of alkaloids in herbal medicines: Current status and perspectives

Alkaloids are a widespread group of basic compounds in herbal medicines and have attracted great interest due to various pharmaceutical activities and desirable druggability. Their distinctive structures make chromatographic separation fairly difficult. Peak tailing, poor resolution, and inferior column‐to‐column reproducibility are common obstacles to overcome. In order to provide a valuable reference, the methodologies and/or strategies on liquid chromatographic separation of alkaloids in herbal medicines proposed from 2012 to 2019 are thoroughly summarized.