Discovery of cancer cell proliferation inhibitors from Salviae miltiorrhizae radix et rhizoma by a trace peak enrichment approach

Salviae miltiorrhizae radix et rhizoma is a traditional herbal medicine with anti‐cancer activities. In this work, a trace peak enrichment approach combined with a cell proliferation assay was applied for screening cancer cell proliferation inhibitors from the extract of S. miltiorrhiza. A set of 123 peak fractions were prepared, and by comprehensive screening, 21 tanshinones were screened out as cancer cell proliferation inhibitors and their structures were tentatively identified by liquid chromatography coupled with quadrupole time‐of‐flight tandem mass spectrometry analysis. The inhibitory activities of nine available screened tanshinones were validated, with their IC₅₀ values ranging from 0.63 to 28.40 μM, indicating their activities strongly inhibit the proliferation of cancer cells. This study presents tanshinones that are potential cancer cell proliferation inhibitors and may explain the anti‐cancer activity of S. miltiorrhiza.     

Simultaneous qualitative and quantitative analysis of commercial bistorta rhizome and its differentiation from closely related herbs using TLC and HPLC-DAD fingerprinting

A qualitative and quantitative analysis method was established to improve quality assessment standards for Rhizoma Polygoni Bistortae (Polygonum bistorta L.) and differentiate commercial bistorta rhizome from closely related herbs by TLC and HPLC-DAD fingerprinting. Three compounds including phenolic acid and flavane were identified by comparison with standard compounds and quantified simultaneously by HPLC-DAD simultaneously. A comprehensive validation of the method that included sensitivity, linearity, repeatability and recovery was conducted. Paris polyphylla SM., a herb often mixed with Polygonum bistorta L. in China due to their same popular name "Caoheche" in history, was successfully distinguished by thin-layer chromatography (TLC) fingerprinting of the petroleum-soluble fraction. Polygonum paleaceum WALL., another herb often mixed with Polygonum bistorta L. due to their similar external appearances, was distinguished by HPLC fingerprinting.     

野生与栽培羌活药材挥发油含量及组分的比较分析

分析野生和栽培羌活药材挥发油含量及其组分的变化,为羌活药材的引种栽培及进一步的开发利用提供了依据。采用水蒸气蒸馏法提取羌活药材中的挥发油,通过GC-MS对挥发油成分进行分析鉴定;利用聚类分析对测定结果进行分类;通过相关分析研究海拔对羌活药材挥发油含量的影响。6份药材样品挥发油含量范围为1.60～7.98 mL/100g,6份羌活药材挥发油成分经GC-MS分析,共鉴定出57个化合物,共有成分12个;基于挥发油成分种类及相对含量的聚类分析显示了羌活药材挥发性成分种类及含量的种间差异;相关分析结果表明,海拔高度与挥发油含量存在显著正相关。野生和栽培羌活药材挥发油含量存在差异,总体上野生羌活药材的挥发油含量高于栽培种;羌活药材挥发油含量随海拔升高而增加。     

山豆根和北豆根挥发性成分的对比分析

采用GC-MS对山豆根和北豆根的挥发性化学成分进行了分析。通过谱图检索和解析,从山豆根中确定了35种挥发性成分,从北豆根中确定了31种。其中北豆根中的化合物类型较多,含氮化合物和含氧杂环化合物明显较北豆根多。两种中草药中量最多的都是脂肪酸,但山豆根为十八碳二烯酸、十六碳酸和六碳酸;北豆根为十三碳烯酸。     

液相色谱指纹图谱在泽泻动态研究中的应用

应用指纹图谱技术,结合指标成分测定,采用高效液相色谱法,乙腈-水梯度洗脱,流速1．0mL／min,柱温20℃,色谱图光谱采集范围200～800nm,探讨了泽泻动态积累的规律。以泽泻标准对照药材为参照,用指标成分24-乙酰泽泻醇A和23-乙酰泽泻醇B进行了定位和测定,找出了21个共有峰,其中12号和20号峰分别为24-乙酰泽泻醇A和23-乙酰泽泻醇B。结果表明：泽泻样品指纹图谱及指标成分测定可为泽泻的最佳采收期提供科学依据。     

Differentiation of strains from the Bacillus cereus group by RFLP‐PFGE genomic fingerprinting

Bacillus mycoides, Bacillus pseudomycoides, Bacillus weihenstephanensis, Bacillus anthracis, Bacillus thuringiensis, and Bacillus cereus belong to the B. cereus group. The last three species are characterized by different phenotype features and pathogenicity spectrum, but it has been shown that these species are genetically closely related. The macrorestriction analysis of the genomic DNA with the NotI enzyme was used to generate polymorphism of restriction profiles for 39 food‐borne isolates (B. cereus, B. mycoides) and seven reference strains (B. mycoides, B. thuringiensis, B. weihenstephanensis, and B. cereus). The PFGE method was applied to differentiate the examined strains of the B. cereus group. On the basis of the unweighted pair group method with the arithmetic mean method and Dice coefficient, the strains were divided into five clusters (types A–E), and the most numerous group was group A (25 strains). A total of 21 distinct pulsotypes were observed. The RFLP‐PFGE analysis was successfully used for the differentiation and characterization of B. cereus and B. mycoides strains isolated from different food products.     

Determination of phenolic compounds in medicinal herbs by reversed-phase HPLC

An HPLC procedure is proposed for the determination of 12 phenolic compounds in plant tissues by reversed-phase HPLC with gradient elution and UV detection. The influence of pH, composition of the mobile phase, concentration of the organic modifier, and temperature on the separation of gallic, protocatechuic, trans-ferulic, trans-caffeic acid, rutin, quercetin, dihydroquercetin, and (_)-epicatechin for 30 min is studied. The lower limit of quantification of phenolic compounds is 1–2.5 μg/L. The procedure was applied to the determination of phenolic compounds in aqueous extracts of Hypericum perforatum; its sample was found to contain protocatechuic acid, (_)-epicatechin, and also rutin.     

Evaluation of Chinese medicinal herbs fingerprinting by HPLC-DAD for the detection of toxic aristolochic acids

Aristolochic acids are known to contribute to various renal disorders; therefore, expanding the availability of analytical methodology to detect these compounds is important in order to assess the quality of Chinese herbal medicines in which they can be found. Twelve medicinal herbal samples were procured from various sources and extracted in duplicate prior to a "fingerprint" analysis using conventional HPLC-DAD. Multivariate analysis was performed on the entire chromatographed fingerprints. The resulting output was a partial least-square discriminant analysis model, which was able to evaluate the potential presence of aristolochic acids I and II as well as providing an individual herbal "fingerprint". The results of this study provide evidence that the presence of aristolochic acids contained within certain herbal extractions could be detected using a simple method, although some limitations apply to this method for quality control, since newly detected samples for aristolochic acid (positives) will need further confirmation with purity checks or MS hyphenation.     

Botanical Source Differentiation of Podophyllum Resin by HPLC

Abstract

A high-performance liquid chromatographic method to determine botanical sources of Podophyllum resin via approximate podophyllotoxin content is described. North American resin contains about 10% podophyllotoxin, the Indian variety about 40%. Samples are dissolved in mobile phase solution and analyzed by HPLC using normal-phase chromatography with detection at 280 nm. Estimates of podophyllotoxin content were made using commercially available references.     

Separation and detection of closely related endorphins by liquid chromatography-electrochemistry

The specific aim of this study was to investigate the selectivity of liquid chromatography--electrochemistry for minor variations in the structure of small endorphins. Using isocratic mobile phases, chromatographic conditions were established for the separation of a series of closely related endorphins. Hydrodynamic voltammetry showed that each peptide exhibited a characteristic oxidative behavior that was also reflected in peak current ratios. Changes in a small moiety altered both the chromatographic behavior and electroactivity of these neuropeptides.     

凝胶渗透色谱净化超高效液相色谱-串联质谱法检测甘草及其提取物中的11种氨基甲酸酯类农药残留

建立了甘草及其提取物中11种氨基甲酸酯类农药多残留的超高效液相色谱-串联质谱(UPLC-MS/MS)分析方法。以11种氨基甲酸酯类农药为目标分析物,样品经乙腈超声提取、凝胶渗透色谱(GPC)净化后,用UPLC-MS/MS检测。以甘草及其提取物为例,分别进行了0.02、0.04和0.1 mg/kg 3个添加浓度的11种目标分析物的加标回收率实验,甘草中11种目标分析物的回收率为72.2%～94.0%,相对标准偏差为0.7%～7.8%;甘草提取物中11种目标分析物的回收率为73.8%～94.7%,相对标准偏差为1.5%～12.7%。该方法灵敏度高、准确度好,符合农药多残留检测的技术要求,适用于甘草、黄芪等中药材及其提取物中氨基甲酸酯类农药残留的检测。     

Separation and determination of closely related lantibiotics by micellar electrokinetic chromatography

A sensitive micellar electrokinetic chromatography (MEKC) method was developed for the separation and determination of four closely related lantibiotics: gallidermin, cinnamycin, duramycin and nisin. Factors affecting the separation of the lantibiotics such as pH, phosphate buffer concentration, SDS concentration and wavelength for UV detection were investigated. By optimizing these experimental conditions, successful separation was achieved between class 1A lantibiotics (nisin and gallidermin) and class 1B lantibiotics (duramycin and cinnamycin). The four lantibiotics were separated within 12 min in 50 mM phosphate buffer at pH 3.95 ± 0.1 containing 80 mM SDS with UV detection of 214 nm. The LOD (S/N = 3) were 61 ng/mL for gallidermin, 57 ng/mL for cinnamycin, 55 ng/mL for duramycin and 58 ng/mL for nisin. The method was successfully applied to real samples such as fermentation broth, bovine colostrum and predrop beer. This method yielded satisfactory results, with quantitative recoveries of spiked lantibiotics in the three samples ranging from 86.1 to 99.6%.     

Quantification of eight active ingredients in crude and processed radix polygoni multiflori applying miniaturized matrix solid‐phase dispersion microextraction followed by UHPLC

A rapid, efficient, and green sample preparation method has been developed to extract eight active ingredients (gallic acid, catechins, epicatechin, polydatin, 2,3,5,4′‐tetrahydroxystilbene‐2‐O‐β‐d ‐glucoside, resveratrol, emodin, and physcion) in radix polygoni multiflori by miniaturized matrix solid‐phase dispersion microextraction. Simple and sensitive ultra high performance liquid chromatography combined with ultraviolet detection has been applied to analyze the multiple compounds. The best results were obtained by adding 25 mg sample into 25 mg adsorbent and grinding for 2 min with disorganized silica as adsorbent and 1 mL 150 mM 1‐dodecyl‐3‐methylimidazolium bromide as a green eluting solvent. Good linearity (r² > 0.998) for each analyte was obtained by this method. The intra‐day and inter‐day precision (RSD) were both below 5.31%, and the recoveries of the analytes ranged from 93.3 to 100.0%. This simple miniaturized matrix solid‐phase dispersion microextraction method for analyzing the compounds in radix polygoni multiflori needs a short time and requires little sample and reagent. Thus, this method is far more eco‐friendly and efficient than traditional extraction methods (reflux and ultrasound‐assisted extraction). The present investigation provided a promising method for the fast preparation and discrimination of chemical differences in crude and processed radix polygoni multiflori.     

中药大黄蒽醌类物质电喷雾质谱裂解规律研究

大黄是一味临床常用药材,化学成分复杂,主要药效成分为蒽醌类衍生物.其中1,8-二羟基蒽醌类衍生物如大黄素(Emodin)、大黄酚(Chrysophanol)、大黄酸(Rheni)、大黄素甲醚(Physcion)、芦荟大黄素(Aloe-emodin)等在中药中分布较广泛.这类物质及其甙类具有泻下、抗菌、抗癌等多种生理活性.对于大黄蒽醌类物质的分离、含量测定、药理研究等一直是一个非常活跃的领域,但对大黄蒽醌类衍生物的电喷雾质谱裂解规律的研究还未见报道.本文我们应用电喷雾离子阱质谱(ESI-MS)研究了5种大黄素型蒽醌衍生物的裂解规律.为进一步开展大黄蒽醌类物质在生物体内的代谢规律等做了有益的探讨.     

The characterization of closely related polymeric materials by thermogravimetry—mass spectrometry

The potential of a new technique in forensic science, thermogravimetry—mass spectrometry, is illustrated by its application to the examination of acrylic fibres and white alkyd gloss paints, small samples of which are difficult to differentiate even by a combination of current techniques. A simple interface allowing the coupling of a magnetic sector mass spectrometer to a thermobalance is described. This combination allows up to five experimental parameters to be determined simultaneously either for classification of the material under investigation or for the detailed direct comparison of samples. The thermobalance allows the temperatures at which weight losses occur and their relative proportions to be determined; the basic parameter obtained from the mass spectrometer is the total ion current trace. From this, single qualitative mass spectra may be chosen for examination and ions, whose presence or absence may be diagnostic or which vary characteristically throughout the analysis, can be selected. The variations of these ions may then be examined by computer-generated mass thermograms (analagous to mass chromatograms in g.c.—m.s.) or, if computer facilities are not available, by multiple ion detection. The residue from the analysis can be used for trace element analysis, e.g. by energy-dispersive x-ray fluorescence spectrometry.     

Simultaneous chemical fingerprinting and quantitative analysis of crude and processed Radix Scrophulariae from different locations in China by HPLC

A validated liquid chromatography method was first developed to evaluate the quality of crude and processed Radix Scrophulariae extracts through establishing chromatographic fingerprint and simultaneous determination of five bioactive compounds, namely 5-hydroxymethylfurfural (5-HMF), acteoside, angroside C, harpagoside and cinnamic acid. The chromatographic were separated on an Agilent Zorbax Extend C(18) column (250 mm × 4.6 mm, 5 μm) and detected by diode array detector (DAD). Mobile phase was composed of (A) aqueous phosphoric acid (0.03%, v/v) and (B) acetonitrile using a gradient elution. Analytes were performed at 30 °C with a flow rate of 1.0 mL/min and UV detection at 280 nm. All calibration curves showed good linear regression (r(2) ≥0.9996) within the tested ranges, and the recovery of the method was in the range of 98.12-103.38%, with RSD values ranging from 0.6 to 2.8%. In addition, the contents of those five bioactive compounds in crude and processed Radix Scrophulariae prepared by different locations of China were determined to establish the effectiveness of the method. The results demonstrate that the developed method is accurate and reproducible and could be readily utilized as a suitable quality control method for the quantification of Radix Scrophulariae.     

Differentiation of wool and spruce waxes affected by environmental influences using fingerprinting by soft ionization mass spectrometry

Summary Wool wax from two sheep which showed different susceptibility towards fleece rot was analysed by field desorption mass spectrometry. In the mass range from m/z 360 to m/z 410 clear differences of ion abundances were found indicating lower proportions of oxidized cholesterone in the wax of the resastent sheep. This suggests that the weathering of the fleece wax could be a factor in relation to the cause of fleece rot.Epicuticular wax isolated from green and yellowish Norway spruce needles was analysed by field ionization mass spectrometry. The comparison of the both mass spectra in the range from m/z 70 to m/z 1000 demonstrates that the wax layer of conifer needles is altered by air pollution resulting in higher proportions of free n-fatty acids and estolides. This corresponds to a premature ageing of the epicuticular wax.Both examples demonstrate that soft ionization mass spectral fingerprints of raw waxes of animals and plants can be used for a rapid characterization and differentiation of untreated and complex, biological samples.

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Unterscheidung von Woll- und Fichten-Wachsen nach Umweltschädigung mit Fingerabdruck-Massenspektren nach schonender Ionisierung

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Field desorption mass spectrometry of lipids IV, for Part I–III, see refs. [5–7]