当归多糖ASP3的甲基化分析

通过部分酸水解和甲基化分析, 结合GC-MS测试手段, 对当归多糖ASP3的糖链结构进行了研究. 采用0.2 mol/L三氟乙酸(Trifluoroacetic acid, TFA)对ASP3进行了部分酸水解, 对水解前后的多糖组分进行甲基化分析. 结果显示: ASP3的糖残基主要由D-GalpA, D-Galp, L-Araf和L-Rhap组成, 主链由1,4-D-GalpA连接, 形成“光滑区”半乳糖醛酸聚糖, 由1,4-D-GalpA通过O-4位与1,2-; 1,2,4-L-Rhap的O-2位交替连接形成含有较多分支的“毛发区”鼠李半乳糖醛酸聚糖. 58.8%的Rhap残基发生O-4位取代(1,2,4-L-Rhap). 由T-, 1,5-, 1,3,5-Araf和T-, 1,3-, 1,3,6-, 1,4-, 1,4,6-D-Galp聚合形成的阿拉伯半乳聚糖、半乳聚糖以及阿拉伯聚糖是ASP3侧链的主要组成, 通过Rhap残基的O-4位与主链相连.     

Evaluation of antioxidants in Dong quai (Angelica sinensis) and its dietary supplements

The antioxidant activity (AA), total phenolic content (TPC) and total flavonoids content (TFC) in Dong quai (DQ, Angelica sinensis) raw materials and dietary supplements (DS) containing this plant were determined using the CUPRAC, FRAP and fluorescence methods. The antioxidant activity for DQ aqueous extracts revealed by CUPRAC was (1330.45 ± 1.30) μmol Trolox equivalent (TE) per 100 g of dry mass (DM), whereas the antioxidant activity as determined by FRAP was (1813.9 ± 2.0) μmol of TE per 100 g of DM. Lower values were noted for the fluorescence method than for CUPRAC and FRAP (ranging from (35.96 ± 0.3) to (304.6 ± 1.4) μmol of TE per 100 g of DM). The highest TPC values were determined for an aqueous extract of DQ ((3330.3 ± 2.3) μmol of TE per 100 g of DM), while TFC for ethanolic extracts of DQ was ((146.50 ± 0.5) mg of quercetin equivalent (QE) per 100 g of DM). Cinnamic acid, isomers of benzoic acid and derivatives of quercetin were analysed by HPLC-PDA. The ferulic acid concentration in an ethanolic extract of DQ was (21.83 ± 0.07) mg per 100 g of DM. Of the flavonols detected, rutin exhibited the highest concentration in ethanolic extract of DQ ((3.32 ± 0.13) mg of QE per 100 g of DM). Other phytochemicals (alkaloids, saponins, flavonoids, anthraquinones, tannins, steroids, etc.) were identified by phytoscreening colour reaction. The results were analysed by principal component analysis (PCA), cluster analysis and one-way ANOVA tests.     

当归多糖ASP3及其水解产物的NMR光谱分析

对当归多糖ASP3的糖链结构进行分析. 分别采用0.2 mol/L三氟乙酸(Trifluoroacetic acid, TFA)和内切-α-(1→4)-聚半乳糖醛酸酶(EndoPG)对ASP3进行部分酸水解和酶水解, 并对水解前后多糖组分的1D和2D NMR光谱特征进行分析. 实验结果表明, ASP3是一种果胶多糖. GalpA和Rhap位于多糖分子的主链, 由1→4-D-GalpA相连形成的“光滑区”(半乳糖醛酸聚糖)是其主要组成部分; 由α-(1→4)-GalpA通过O4位与α-(1→2)-和α-(1→2,4)-Rhap的O2位交替连接所形成的重复单元[→4)-α-GalpA-(1→2)-α-Rhap-(1→]构成具有较高分支的“毛发区”(富含中性糖侧链的鼠李半乳糖醛酸聚糖). Galp和Araf是中性糖侧链的主要组成, 通过Rhap残基的O4位与主链相连. 非还原性末端T-β-Galp, β-(1→3)-, β-(1→3,6)-, β-(1→4)-, β-(1→4,6)-Galp聚合形成以β-(1→3,6)-Galp为分支点的β-(1→6)-半乳聚糖和以β-(1→4,6)-Galp为分支点的β-(1→4)-半乳聚糖. T-α-Araf, α-(1→5)-Araf和α-(1→3,5)-Araf聚合形成以α-(1→3,5)-Araf为分支点的α-(1→5)-阿拉伯聚糖. 此外, 由α-(1→5)-阿拉伯聚糖通过α-(1→3)连接与β-(1→6)-半乳聚糖末端聚合形成阿拉伯半乳聚糖.     

Chemical composition analysis of Angelica sinensis (Oliv.) Diels and its four processed products by ultra-high-performance liquid chromatography coupled with quadrupole-orbitrap mass spectrometry combining with nontargeted metabolomics

Angelica sinensis (Oliv.) Diels. has been used for women to enrich the blood, prevent and treat blood deficiency syndrome in Traditional Chinese Medicine for thousands of years. Wine-processed Angelica sinensis, soil-processed Angelica sinensis, oil-processed Angelica sinensis, and charred-processed Angelica sinensis are the most significant four processed products used in Chinese clinic. However, there have been few studies aimed at comparing their chemical differences. Ultra-high-performance liquid chromatography coupled with quadrupole-orbitrap mass spectrometry combining with nontargeted metabolomics was applied to investigate the diversity of processed products of Angelica sinensis. A total of 74 compounds with the variable importance in the projection value more than 1.5 and P less than 0.05 in ANOVA were highlighted as the compounds that contribute most to the discrimination of Angelica sinensis and four processed products. The results showed the metabolic changes between Angelica sinensis and its four processed products, there were 19 metabolites, 3 metabolites, 6 metabolites, and 45 metabolites were tentatively assigned in soil-processed Angelica sinensis, wine-processed Angelica sinensis, oil-processed Angelica sinensis, and charred-processed Angelica sinensis, respectively. These results suggested that the proposed metabolomics approach was useful for the quality evaluation and control of processed products of Angelica sinensis.     

基于质量标志物的当归抗炎功效近红外快速评价

该文通过筛选当归中对NF-κB具有抑制活性的质量标志物(Q-marker),建立了一种利用近红外光谱技术(NIRS)快速评价当归抗炎功效的方法。采用UPLC/Q-TOF技术结合NF-κB双荧光素酶报告基因对当归中具有NF-κB抑制活性的成分进行筛选,并通过单体、整体及等效性验证确立了关键Q-marker。以UPLC分别对多批当归药材中的Q-marker进行含量测定,建立其NIRS定量模型,并进一步将Q-marker的含量与当归的NF-κB抑制活性相关联,构建"量-效"拟合函数。结果表明,绿原酸(X₁)、洋川芎内酯I(X₃)和Z-藁本内酯(X₄)为NF-κB抑制活性的关键Q-marker,其含量波动与整体抗炎活性(Y)的变化相吻合,符合拟合方程:Y=16.13-14.84X₁+7.981X₃+0.112 6X₄,且基于NIRS的预测效果良好。该方法通过整合分析实现了基于关键Q-marker的当归药材抗炎功效的快速评价,为中药材品质的快速分析提供了新的研究范式...     

A flow-injection mass spectrometry fingerprinting method for authentication and quality assessment of Scutellaria lateriflora-based dietary supplements

Scutellaria lateriflora, commonly known as skullcap, is used as an ingredient in numerous herbal products. However, it has been occasionally adulterated/contaminated with Teucrium canadense and/or Teucrium chamaedrys, commonly known as germander, due to the morphological similarities between the two genera. The latter contains hepatotoxic diterpenes. Despite the potential hepatotoxicity introduced by germander contamination, analytical methodologies for the authentication and quality assessment of S. lateriflora-based dietary supplements have not been reported. In this study, a flow-injection/mass spectrometry fingerprinting method in combination with principal component analysis was used to survey S. lateriflora-based dietary supplements sold in the USA.     

Gas Chromatographic/mass spectrometric analysis of azines

Chromatographia - We have studied the gas chromatographic behaviour of several 3-methyl-2-benzothiazolone-azines. In HPLC analysis all the unsymmetrical azines show double peaks, whereas in HRGC...     

Chromatographic separation of strontium from barites for mass spectrometric analysis

Summary A simple Chromatographic method for the separation of strontium in barites containing 500 times excess barium is described. The strontium fraction is sufficiently pure for mass spectrometer studies of strontiumisotopes. The resin used is Dowex-50 and the column dimensions are 1.1 by 8 cm. The column is eluted at room temperature with 1.5M hydrochloric acid at a flow rate of 1 ml/min. Effect of column temperature on the elution peaks is pointed out.

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Zusammenfassung Eine einfache chromatographische Methode zur Abtrennung von Strontium, aus Baryten mit 500fachem Bariumüberschuß wurde angegeben. Die Strontiumfraktion ist genügend rein für massenspektrometrische Untersuchungen der Sr-Isotopen. Dowex-50 dient als Austauscher in 1,1×8-cm-Säulen. Die Elution erfolgt bei Zimmertemperatur mit 1,5-m Salzsäure bei einer Durchflußgeschwindigkeit von etwa 1 ml/min. Der Einfluß der Säulentemperatur auf die Elutionspeaks wird hervorgehoben.

     

当归挥发性有机物的SPME/GC-MS分析

采用不同极性的SPME萃取吸附了当归的挥发性有机物,并用GC-MS定性和相对定量,比较了不同萃取时间和不同萃取温度下所获得的挥发性有机物的差异。结果表明:藁本内酯和α-蒎烯是其主要成分。随着萃取时间的延长,α-蒎烯含量逐渐减少,藁本内酯含量逐渐升高;随着萃取温度的升高,低沸点化合物含量逐渐减少,高沸点化合物含量逐渐增加。     

Selective Acid Hydrolysis Condition for the Composition and Linkage with a Fructofuranosyl Backbone of a Polysaccharide from Angelica sinensis （Oliv） Diels

Angelica sinensis is a well-known Chinese traditional herbal medicine. Its roots have been widely used over two thousand years for tonifying blood, promoting blood circulation, regulating menorrhea and subduing inflammation1. The polysaccharide of the ASD…     

Gas chromatographic/mass spectrometric and microbiological analyses on irradiated chicken

Ionizing radiation is widely used as treatment technique for food preservation. It involves among others reduction of microbial contamination, disinfestations, sprout inhibition and extension of shelf life of food. However, the commercialization of irradiated food requires the availability of reliable methods to identify irradiated foodstuffs. In this paper, we present results on the application to irradiated chicken of this method, based on the detection, in muscle and skin samples, of the peaks of ions 98 Da and 112 Da, in a ratio approximately 4:1, typical of radiation induced 2-dodecylcyclobutanones (2-DCB). Aim of the work was also to study the time stability of the measured parameters in samples irradiated at 3 and 5 kGy, and to verify the efficacy of the treatment from a microbiological point of view. Our results show that, one month after irradiation at 3 kGy, the method is suitable using the skin but not the muscle, while the measured parameters are detectable in both samples irradiated at 5 kGy. The microbial population was substantially reduced even at 3 kGy.     

Structural characterization of a glucan from Angelica sinesis (Oliv) Diels

Angelica sinesis (Oliv) Diels is a traditional Chinese drug used clinically in the treatment of gynecological diseases. Its polysaccharide fraction been reported to have several immunomodulating, antitumor, antiradiation activities^[1] and a recent paper showed it could obviously act on the hemopoietic system^[2]. Haruki Y. et al. have elucidated the chemical structure of a α(1→4) linked D-glucan from Angelica acutiloba^[3]. We now describe the structure of a water soluble glucan, AS-1, which was isolated and purified from the water extract of Angelica sinesis (Oliv) Diels. It is only composed of D-glucose and its mean molecular weight is estimated to be 5.3×10⁵. The absorption peak of 846cm^-1 on IR, the chemical shift of δ 99.98 ppm for C-1,and the 168 Hz of ¹J_C-H suggested that the D-glucose residues were α-linked. Methylation analysis (Table 1),periodate oxidation, Smith degradation, ¹H NMR and ¹³C NMR (Table 2) showed that it is α(l→6) linked D-glucan to which are attached two glucosyl side chains, Glc-(l→3)-Glc→1 and Glc-(1→4)-Glc→1, both at 3-O of the glucosyl residues of every 14th glucose units of the main chain. The structure was further proved by using HMBC,HMQC and DQF-COSY techniques. The details of its pharmacological results will be published elsewhere.     

Headspace Solid-phase Microextraction–Gas Chromatographic–Mass Spectrometric Analysis of the Essential Oils of Two Traditional Chinese Medicines, Angelica pubescens and Angelica sinensis

Angelica pubescens and Angelica sinensis belong to the Umbelliferae family and both are used as traditional Chinese medicines. In the present study, headspace solid-phase microextraction (HS-SPME) with gas chromatography-mass spectrometry (GC-MS) was used for the analysis of the volatile constituents present in their roots. Eighty-seven compounds in Angelica pubescens and thirty-six compounds in Angelica sinensis were identified by GC-MS. Their relative contents were calculated by the peak area ratio. HS-SPME was compared to steam distillation (SD) by analyzing the volatile constituents of Angelica sinensis root. A good agreement between results obtained with both techniques was found. As a conclusion, HS-SPME is a powerful tool for determining the volatile constituents present in the TCMs.     

Investigation of the elemental composition and chemical association of several elements in fulvic acids dietary supplements by size-exclusion chromatography UV inductively coupled plasma mass spectrometric

Four fulvic acid dietary supplement samples were obtained for this study with the intention of investigating the elemental composition and association of fulvic acids found in fulvic acid supplements. This was achieved by coupling size-exclusion chromatography (SEC) sequentially with UV-vis and inductively coupled plasma mass spectrometric (ICP-MS) detectors. The combination of UV and ICP-MS offered highly sensitive and selective detection. This technique was used in the present study to initially investigate the chemical association of several different elements including, Cr, Co, Ca, Fe, I, Mg, Zn, Se, Cu, Mn, Mo, As, Hg, Pb, and Ag, by observing the elution profile of the fulvic acids obtained with UV detection and matching their retention times with the peaks measured with ICP-MS. The results found based on this type of analysis suggest that there was some association of the elements to the fulvic acids. It was also of interest to observe the stability of these complexes upon human digestion; therefore a gastric digestion was mimicked. In the fulvic acid dietary supplement samples studied, fulvic acids were present in the samples and there was elemental association based on the retention time overlap in the UV as well as the ICP-MS. The fulvic complexes found in the samples were of a low molecular weight As a result of the digestion the SEC-ICP-MS chromatographic profile in some of the samples changed, which may infer that the elemental association had changed.     

Evaluation of major active components in St. John's Wort dietary supplements by high-performance liquid chromatography with photodiode array detection and electrospray mass spectrometric confirmation

A RP-HPLC method with photodiode array detection and LC-electrospray ionization (ESI) MS confirmation was established for the determination of major active components in St. John's Wort dietary supplement capsules. The samples alternatively were extracted with ethanol-acetone (2:3) using a 55 degrees C water-bath shaker or an ambient temperature ultrasonic bath. Extracts were separated by RP-C18 chromatography using a 95-min water-methanol-acetonitrile-trifluoroacetic acid gradient. The major components were identified by photodiode array detection and then confirmed by LC-ESI-MS. The quantification of components was performed using an internal standard (luteolin). This method may serve as a valuable tool for the quality evaluation of St. John's Wort dietary supplement products.     

Determination of anabolic steroids in dietary supplements by liquid chromatography-tandem mass spectrometry

Nineteen different dietary supplements, ordered through the internet and intercepted by the Belgian pharmaceutical inspection at the post office, were analyzed by means of liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the presence of anabolic steroids. After a methanolic extraction the samples were screened for the presence of 49 compounds. This resulted in almost 60% of the samples being suspected of containing one of these 49 anabolic compounds and being subjected to a confirmatory product ion scan. In all of these suspected samples we were able to confirm at least one anabolic steroid with concentrations between 0.01 and 2.5 mg unit(-1) (unit: one capsule or tablet or for liquids: the prescribed dose). The anabolic steroids that was mostly encountered was testosterone (50%) followed by beta-boldenone (25%). These results once more confirm the dubious reputation of over-the-counter dietary supplements.     

Chromatographic and mass spectrometric methods for determination of lysergic acid diethylamide (LSD) and metabolites in body fluids.

Continued illicit use of the potent psychedelic drug lysergic acid diethylamide (LSD) has stimulated efforts to develop effective analytical methods for detection of the drug and its metabolites in body fluids from suspected LSD users. Recently reported methods based on gas and liquid chromatography, combined with single- and multiple-stage mass spectral analysis, now permit accurate detection and quantitation of LSD at sub-nanogram/milliliter concentrations.     

A multiple electrospray interface for parallel mass spectrometric analyses of compound libraries

A parallel spray interface for mass spectrometry is described. This new electrospray interface enables effluent flow streams from an array of HPLC columns to be sampled independently and sequentially on a chromatographic time-scale. Unlike our previously reported parallel LC-MS interface, which incorporated a dual-sheath spray interface accommodating up to four flow streams that are sampled simultaneously, this new interface permits up to four columns to be sampled sequentially by means of a stepping motor and rotating plate assembly. Effluent flow streams from an array of four HPLC columns are connected to a parallel arrangement of electrospray needles co-axial to the mass spectrometer entrance aperture. Within the needle assembly, the individual spray tips are oriented in a circular array, where each needle is situated 90 degrees relative to one another for four-column operation. An eight-column system is described with needles spaced at 45 degree intervals. In between the needle assembly and the mass spectrometer entrance aperture is a Teflon disk with a through-hole that is mounted to a stepping motor assembly. By precisely controlling the stepping of the motor assembly, it is possible to sample each of the spray positions multiple times per second. By operating the quadrupole mass spectrometer in the single ion monitoring (SIM) mode, it was possible to acquire data at each of the spray positions during the course of the elution of compounds from the HPLC column array while maintaining both good sensitivity and peak shape. Preliminary results suggest this technique will be useful for high throughput combinatorial library analysis and profiling.     

Chromatographic separation and mass spectrometric identification of positional isomers of polyethylene glycol-modified growth hormone-releasing factor (1-29)

A one-step chromatographic method capable of separating all isomers of polyethylene glycol (PEG)-growth hormone-releasing factor (GRF) (1-29) conjugates was developed. The unmodified GRF (1-29) and seven different isomers of PEG-GRF (1-29) conjugates were separated by using a simple reversed-phase HPLC method depending on the differences of hydrophobicity due to the number and site of PEG attachment. The PEGylation sites of all isomers of PEG-GRF (1-29) conjugates were identified by determining the molecular masses of the Lys-C digested fragments with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. This study is a first report for the separation of all PEG-conjugate isomers and would be useful for further studies to find the promising conjugate by evaluating biological activity and stability of each isomer.