纳米金增强低温等离子体离子化质谱快速检测苯胺

低温等离子体(Low-temperature plasma,LTP)作为敞开式离子源满足环境检测要求的样品原位、快速检测,然而水体的基体效应使得检测灵敏度不足。该文将金纳米颗粒溅射于针灸针表面后,结合低温等离子体离子源进行检测,实现了质谱信号的增强效应,可以快速灵敏检测水体中的苯胺。结果表明,相比于无纳米金修饰的针尖,苯胺检测信号强度增强了43倍,且具有较低的背景噪音。苯胺的质量浓度与质谱信号强度在1~50μg/L范围内呈线性关系,检出限(LOD)为0.64μg/L,每个样品检测时间约为1 min。该方法简单易操作,结合便携式质谱有望用于环境水体及突发性水源污染事件中苯胺的检测。     

一测多评法在舒肝益脾颗粒中的应用

建立了舒肝益脾颗粒一测多评(QAMS)的高效液相色谱(HPLC)测定方法,并验证该方法在质量控制中的适用性与准确性.该方法能较好地分离舒肝益脾颗粒中3种主要成分(五味子醇甲、五味子甲素、五味子乙素),并以五味子醇甲为内参物,建立五味子甲素、五味子乙素与其之间的相对校正因子(fs/k),用fs/k计算各代表性成分含量,实...     

Quantitative analysis of multicomponents by a single marker and quality evaluation of Venenum Bufonis from different geographical origins

Bufadienolides are the main bioactive components of Venenum Bufonis (VB) and have been widely used to treat different types of human cancers for decades. The bufadienolide content in VB varies significantly in materials from different geographical origins. In this work, a new strategy for the quality assessment of VB was developed through quantitative analysis of multi‐components by single marker (QAMS). Cinobufagin was selected as the internal reference substance; seven bufadienolides were separated and simultaneously determined based on relative correction factors. The correlation coefficient value (r ≥ 0.9936) between QAMS and the normal external standard method proved the consistency of the two methods. According to the outcomes of 30 batches of VB samples, the contents of the seven bufadienolides were used for further chemometric analysis. All of the samples of VB from various geographical origins were divided into three categories based on hierarchical cluster analysis and radar plot, which indicated the crucial influence of geographical origins on VB. This study showed that QAMS combined with chemometristry could be used to comprehensively evaluate and effectively control the quality of VB from different geographical origins.     

Qualitative analysis of Psoraleae Fructus by HPLC‐DAD/TOF‐MS fingerprint and quantitative analysis of multiple components by single marker

A variety of bioactive substances may account for the recognized efficacy and wide clinical application of Psoraleae Fructus in China. A high‐performance liquid chromatography–diode array detector (HPLC‐DAD) fingerprint method was developed to present the comprehensive phytochemical profile of the crude drug. Thirteen major compounds were separated and identified by HPLC coupled with time‐of‐flight mass spectrometry (HPLC/TOF‐MS), namely psoralenoside (PO), isopsoralenoside (IPO), psoralen (PS), isopsoralen (IPS), neobavaisoflavone (NBF), bavachin (BC), corylin (CN), bavachromene (BCM), psoralidin (PD), isobavachalcone (IBC), bacachinin (BCN), corylifol A (CA) and bakuchiol (BK). Then quantitative analysis of multiple components by single marker (QAMS) was applied in content determination of PO, IPO, PS, IPS, BC, IBC, BCN, CA and BK, with NBF as the internal standard. The calculation results indicated no significant difference from the traditional external standard method (p > 0.05, RSD < 2.62%), suggesting that QAMS is a reliable and convenient method for content determination of multiple chemical compositions, especially when there is a shortage of reference substances. In conclusion, simultaneous qualitative and quantitative analysis of Psoraleae Fructus may be fulfilled through the newly proposed method of QAMS combined with HPLC‐DAD/TOF‐MS fingerprint.     

HPLC Fingerprint with Multi-components Analysis for Quality Consistency Evaluation of Traditional Chinese Medicine Si-Mo-Tang Oral Liquid Preparation

Si-Mo-Tang(SMT) oral liquid preparation, a traditional Chinese medicine, was prepared from four crude herbal drugs, Fructus Aurantii Submaturus, Radix Aucklandiae, Semen Arecae and Radix Linderae Aggregatae. A combinative method using HPLC fingerprint and quantitative analysis was developed and validated for quality consistency evaluation of SMT. Individual HPLC chromatograms were evaluated against the mean chromatogram generated via a similarity evaluation computer program. Data from chromatographic fingerprints were also processed with principal component analysis(PCA) and hierarchical cluster analysis(HCA). Additionally, six components (naringin, isonaringin, hesperidin, neohesperidin, norisoboldine and potassium sorbate) in SMT were simultaneously determined to interpret the quality consistency. For fingerprint analysis, 20 peaks were selected as the characteristic peaks to evaluate the similarities of 26 SMT collected from different manufacturers. Among the 20 characteristic peaks, 10 peaks were assigned to be naringin, hesperidin, neohesperidin, isonaringin, neoeriocitrin, tangeretin, nobiletin, norisoboldine, 5-(ethoxymethyl)furan-2-carbaldehyde and potassium sorbate, respectively. The results of similarity analysis, PCA and HCA, indicate that the samples from different manufacturers were consistent with each other in composition. The results from the quantitative data show that the contents of six compounds were significantly different in SMT oral liquid preparations from different manufacturers. The combinative method of chromatographic fingerprint with quantitative analysis developed here offered an efficient way for the quality consistency evaluation of the traditional Chinese medicine SMT.     

高效液相色谱法结合一测多评法对隔山消发酵前后3种苯乙酮类化合物含量变化的分析

采用高效液相色谱法结合一测多评法测定了隔山消发酵前后2,4-二羟基苯乙酮、2,5-二羟基苯乙酮和白首乌二苯酮等3种苯乙酮类化合物的含量。分别制备发酵前后的隔山消样品溶液,色谱分析中采用Thermo C18色谱柱作为固定相,用不同比例的水(A)、乙腈(B)和甲醇(C)的混合液作为流动相进行梯度洗脱,达到上述3种组分的良好分离。采用二极管阵列检测器在相同的检测波长(265nm处)分别测定3种组分的峰面积。分别用一测多评法(QAMS)和外标法计算所测定组分的含量,结果表明两种计算方法所得结果相似性极高,据此认为在隔山消发酵前后的样品溶液中各组分的质量评价中采用QAMS是可行的。按上述方法分析了同一批隔山消样品12份,取其中6份经发酵处理,对所得结果的平均值进行了比较,未经发酵的样品溶液中2,4-二羟基苯乙酮的质量分数为0.208mg·g^-1、2,5-二羟基苯乙酮的质量分数为0.152mg·g^-1,与经发酵的样品溶液中的质量分数(2,4-二羟基苯乙酮的质量分数为0.235mg·g^-1、2,5-二羟基苯乙酮的质量分数为0.180mg·g^-1)相比呈升高趋势,而白首乌二苯酮的质量分数由未经发酵样品溶液中的0.678mg·g^-1下降为发酵样品溶液中的0.543mg·g^-1。此外,研究结果还表明,不同含量的2,5-二羟基苯乙酮和白首乌二苯酮相对于2,4-二羟基苯乙酮(参照物)的相对校正因子和相对保留时间基本相同。     

Carotenoid Contents of Lycium barbarum: A Novel QAMS Analyses,Geographical Origins Discriminant Evaluation,and Storage Stability Assessment

Given the standard substances of zeaxanthin and its homologues obtained from Lycium barbarum L. (LB) are extremely scarce and unstable, a novel quantitative analysis of carotenoids by single marker method, named QAMS, was established. Four carotenoids including lutein, zeaxanthin, β-carotene, and zeaxanthin dipalmitate were determined simultaneously by employing trans-β-apo-8′-carotenal, a carotenoid component which did not exist in LB, as standard reference. Meanwhile, β-carotene, another carotenoid constituent which existed in LB, was determined as contrast. The QAMS methods were fully verified and exhibited low standard method difference with the external standard method (ESM), evidenced by the contents of four carotenoids in 34 batches of LB samples determined using ESM and QAMS methods, respectively. HCA, PCA, and OPLS-DA analysis disclosed that LB samples could be clearly differentiated into two groups: one contained LB samples collected from Ningxia and Gansu; the other was from Qinghai, which was directly related to the different geographical location. Once exposed under high humidity (RH 75 ± 5%) at a high temperature (45 ± 5 °C) as compared with ambient temperature (25 ± 5 °C), from day 0 to day 28, zeaxanthin dipalmitate content was significantly decreased, and ultimately, all the decrease rates reached about 80%, regardless of the storage condition. Our results provide a good basis for improving the quality control of LB.     

Antibacterial and antifungal properties of crude extracts and isolated compounds from Lychnophora markgravii

Abstract

Antimicrobial activity of dichloromethane and ethanol extracts and five compounds: pinostrobin (I), pinocembrin (II), tectochrysin (III), galangin 3-methyl ether (IV) and tiliroside (V) isolated from Lychnophora markgravii aerial parts against fifteen microorganisms was determined. The structures of these compounds were elucidated based on ESI-MS and NMR spectroscopic data. Both extracts showed antimicrobial activity against several tested microorganisms. Pinostrobin, tectochrysin and galangin 3-methyl ether showed the strongest antibacterial and antifungal effects.     

Application of high-performance liquid chromatography-electrospray ionization time-of-flight mass spectrometry for analysis and quality control of Radix Astragali and its preparations

A high-performance liquid chromatography coupled with electrospray ionization time-of-flight mass spectrometry (HPLC-ESI-TOF-MS) method has been developed for qualitative and quantitative analysis of isoflavonoids and saponins, as well as for the quality control of Radix Astragali and its preparations. The selectivity, reproducibility and sensitivity are compared with HPLC with diode array detection (DAD) and evaporative light scattering detection (ELSD). Limits of detection and quantification fell in ranges of 9-40 and 23-103 ng/mL for 13 analytes with a injection of 10 microL samples, and all calibration curves showed good linear regression (r(2)>0.9938) within the test range. The assay was successfully utilized to analyze the 13 marker components in 20 samples of Radix Astragali products. The quantitative results demonstrated that samples from different localities and manufacturers showed different quality. Advantages, in comparison with conventional HPLC-DAD-ELSD, are that reliable identification of target compounds could be achieved by accurate mass measurements (<3 ppm) along with characteristic retention time, extracted ions chromatograms using a narrow mass window for quantification ensure that the chromatographic peaks are free from background or co-elutes interference, and the great enhancement in selectivity and sensitivity allows identification and quantification of low levels of constituents in complex Radix Astragali matrixes.     

Fingerprint Studies of Radix Scutellariae by Capillary Electrophoresis and High Performance Liquid Chromatography

Two methods based on capillary electrophoresis (CE) and high performance liquid chromatography (HPLC) are described to establish fingerprints of Radix Scutellariae simultaneously. In order to choose an appropriate extraction method, Radix Scutellariae samples extracted by different methods were determined by HPLC. The contents of baicalin, the quality marker of Radix Scutellariae, as well as the number of peaks in the chromatograms were determined to evaluate the extraction methods. 10 batches of Radix Scutellariae collected from different regions in China were applied to establish the fingerprints. Eleven common peaks were isolated within 12 min by CE. The fingerprints obtained with HPLC consisted of 14 common peaks within 40 min. The two proposed methods demonstrated good stability and reproducibility with RSD less than 4% for relative migration time in CE and retention time in HPLC. Finally, the data from the 10 batches of Radix Scutellariae by CE and HPLC were all processed with two kinds of mathematical methods including correlation coefficient and the included angle cosine. The fingerprints of Radix Scutellariae established with CE and HPLC are suitable to identify and differentiate samples by geographical origin and can used for quality control.     

GC-MS identification of the flavonoid aglycones isolated from propolis

Summary Between six and nine flavonoids have been identified by GC-MS in five samples of propolis and in one sample of bud exudate from poplar (Populus nigra), all collected in Poland. On the basis of experimental data and data obtained by use of Drylab software, the optimum temperature programme was found for separation of reference samples of pinostrobin chalcone, pinocembrin, tectochrysin, genkwanin, chrysin, galangin, 5-hydroxy-4′,7-dimethoxyflavone, and pilloin.     

高效液相色谱测定石榴皮水提取物中4种多酚化合物的含量

建立了石榴皮水提取物中没食子酸、安石榴甙A、安石榴甙B和鞣花酸4种多酚化合物的高效液相色谱(HPLC)分析方法.样品经前处理并过滤后,采用HPLC检测,外标法定量.在最佳分析条件下,上述4种多酚化合物的线性范围分别为7.48 ～ 149.50、15.36～153.55、27.65 ～276.45、7.13 ～ 114....     

一种测定中药复方制剂中活性成分的计算分析方法

提出了一种测定中药复方制剂中活性成分含量的计算分析新方法。该方法将主 成分分析和广义秩消因子分析集成用于处理HPLC／DAD数据，并改进了传统的IND因 子指示函数，适用于定量计算分析复杂混和物体系中的待测成分。将其用于测定复 方丹参制剂中三七皂苷R1的含量，计算分析结果表明，本方法明显优于广义秩消因 子法，同时证明IND函数的改进提高了计算分析准确度。     

Quality evaluation of Radix Stemonae through simultaneous quantification of bioactive alkaloids by high-performance liquid chromatography coupled with diode array and evaporative light scattering detectors

A high-performance liquid chromatography coupled with diode array detection and evaporative light scattering detection (HPLC-DAD-ELSD) method was developed to simultaneously quantify six major bioactive alkaloids belonging to different structure types in Radix Stemonae, Bai-Bu in Chinese, a traditionally used antitussive and insecticidal medicinal material in China and other countries of Southeast Asia. Diode array detector (DAD) with the wavelengths at 307 and 260 nm was used to monitor the conjugated system of protostemonine (2) and maistemonine (4), respectively, whereas evaporative light scattering detector (ELSD) was employed to detect croomine (1), stemoninine (3), neotuberostemonine (5) and tuberostemonine (6), the other four analytes with no or poor chromophores. The assay was validated to be sensitive, precise and accurate, with a detection limit of 3.64-0.04 microg/mL depending on the individual analytes. The overall intra- and inter-day variations were less than 9.3%, and the overall recoveries higher than 91.2%, respectively. The correlation coefficients of the calibration curves were better than 0.996 for all analytes. The newly established method was successfully utilized to determine six major components in the genuine sources of Radix Stemonae: Stemona japonica, S. sessilifolia and S. tuberosa. Significant variations of contents of these components were demonstrated in samples of these three species. This simple, rapid, low-cost and reliable method is suitable for the routine quality control of herbal medicines containing bioactive components with different structure types such as Radix Stemonae.     

Determination of five sesquiterpenoids in Xingnaojing injection by quantitative analysis of multiple components with a single marker

A quantitative analysis of multiple components with a single‐marker method was established for the simultaneous determination of five sesqutiterpenoids in Xingnaojing injection. This method was established with Xingnaojing injection determined by high‐performance liquid chromatography coupled with diode array detection. The durability and system suitability of the established method were evaluated, and the reliable relative correction factors were obtained with curdione selected as an internal reference. The contents of the five components in all Xingnaojing injections were determined by external standard method and the contents of curcumenone, curcumenol, curzerenone, and germacrone were also calculated with the obtained relative correction factors. Then, relative error was investigated to estimate the difference of the two methods. As a result, the established new method possesses good adaptability, and there is no significant difference between the two methods, except for the content of curzerenone in eight samples. To put the established method into practice, the limits of quantitation of the established method of the five components were proposed and defined. Thus, the developed methodology can also be utilized to the quality evaluation of Xingnaojing injection, in spite of the difference found in the content of curzerenone between the external standard method and the newly established method.     

Single standard substance for the determination of nine volatile components in the distillate of Fructus Gardeniae and Radix Curcumae (an intermediate of Xingnaojing Injection)

Xingnaojing Injection is a traditional Chinese medicine extensively used for stroke and cerebral ischemia. For better in‐process quality control of Xingnaojing Injection, a method for the analysis of its intermediate (i.e., the distillate of Fructus Gardeniae and Radix Curcumae ) is needed to monitor and optimize the hydrodistillation extraction process. In this work, nine major volatile components in the intermediate were identified: isophorone, 4‐methylene‐isophorone, curcumenone, curcumenol, curdione, curzerenone, furanodienone, curcumol, and germacrone. A quantitative analysis of multi‐component with a single‐marker method based on high‐performance liquid chromatography with diode array detection was developed for the simultaneous determination of the nine components. In this method, only curdione was needed as the reference substance, and the other eight components were determined using their relative correction factors to curdione. In the method validation, good linearity (r  > 0.9999), sensitivity, repeatability, and accuracy (recoveries within 95.3–105.4%) were shown. The repeatability and robustness of the relative correction factors were studied with different column temperatures, flow rates, detection wavelengths, columns, and instruments. In sample analyses, consistent results between the proposed method and the external standard method were shown. The proposed method provides a comprehensive and low‐cost tool for the quality assessment of the intermediate of Xingnaojing Injection.     

LC Determination of Five Flavonoid Aglycones in the Tibetan Medicinal Plant Oxytropis falcata Bunge

A simple, efficient and accurate liquid chromatographic method was established to determine five flavonoid aglycones, 7-hydroxy flavonone, pinocembrin, 2′,4′-dihydroxy chalcone, 2′-hydroxy-4′-methoxy chalcone and pinostrobin in the whole plant powder of Oxytropis falcata Bunge. These five compounds were separated on an Agilent Zorbax Eclipse XDB-C₈ column (150 × 4.6 mm, 5 μm). Mobile phases were composed of water containing 0.1% v/v formic acid and acetonitrile using gradient elution. The established method was validated for linearity, accuracy, precision, limit of detection and quantitation, repeatability and stability.

     

Determination of major components from Radix Achyranthes bidentate using ultra high performance liquid chromatography with triple quadrupole tandem mass spectrometry and an evaluation of their anti‐osteoporosis effect in vitro

Ecdysterone and saponins are the most characteristic components of Radix Achyranthes bidentate, which acts on the human body to promote collagen synthesis and stimulates cell growth. However, the relationship between these components and the differentiation of MC3T3‐E1 osteoblastic cells is unknown. We developed a rapid ultra high performance liquid chromatography with triple quadrupole tandem mass spectrometry method for direct determination of one ecdysterone and four saponins in crude and salt‐processed Radix Achyranthes bidentate. The method was interrogated in terms of linearity, intra‐ and inter‐day precision, repeatability, stability and recovery. The method was linear within the concentration ranges of 0.003–336 μg/mL for β‐ecdysterone, 0.0035–130 μg/mL for 25S‐inokosterone, 0.004–423 μg/mL for ginsenoside Ro, 0.0036–66 μg/mL for chikusetsusaponin IV and 0.0044–111 μg/mL for chikusetsusaponin IVa. The intra‐ and inter‐day precisions were all within 2.7%. The standard addition method determined recovery rates for each component (98.7–102.5%). The method was successfully applied to simultaneously quantify five components in ten batches of crude and salt‐processed Radix Achyranthes bidentate. Subsequently, the examination of these extracts on the differentiation of MC3T3‐E1 osteoblastic cells were carried out. Finally, the relationships between the contents of five components and their anti‐osteoporosis effect were investigated by using canonical correlation analysis.     

Quality Assessment of Cardiotonic Pills by HPLC Fingerprinting

An HPLC method has been developed for the fingerprinting and quantitative analysis of cardiotonic pills. A standard fingerprint containing 11 common peaks at three wavelengths was constructed from ten batches of pills to evaluate batch-to-batch consistency. In addition, the amounts of three marker compounds were also determined to evaluate the quality of the quantitative analysis. Chromatographic fingerprints at three wavelengths, along with the content of three marker compounds were found to be suitable for quality assessment. Fufang Danshen Pian (cardiotonic tablets), a traditional dosage form is produced from three kinds of Chinese medicinal herbs for the prevention and treatment of angina pectoris and coronary heart disease. Fufang Danshen Pian mainly contained an additional group of liposoluble components besides salvianolic acid B from Salvia miltiorrhiza Bunge while cardiotonic pills only contained water-soluble components. Therefore, the fingerprint accompanied with marker compounds can be used to assess the quality of the cardiotonic pills.