超高效合相色谱-串联质谱法快速测定枸杞中甜菜碱含量（英文）

枸杞作为传统的中药材,具有滋补肝肾、益精明目的功效,亦可作为功能性食品被广泛食用。基于超高效合相色谱-质谱(UPC~2-MS),建立了一种快速、灵敏地定量测定枸杞中甜菜碱(指标化合物)含量的新方法。采用ACQUITY UPC2BEH 2-EP色谱柱(150 mm×2.1 mm,1.7μm),以0.7 mL/min超临界CO_2-甲醇(80∶20,v/v)等度洗脱,成功分离了枸杞中的甜菜碱。在该分离过程中,0.1%(v/v)甲酸作为改性剂;背压为1.31×10~7Pa;柱温为40℃;进样体积为1μL;保留时间为3 min。质谱检测工作于电喷雾(ESI)正离子模式和选择离子监测(SIR)模式。在上述条件下得到线性回归方程。其相关系数为0.999 2,检测范围为0.5～50.0μg/mL,检出限为0.013μg/mL。随后,通过对精度、重复性、稳定性和加标回收率(平均值96.3%)的分析,验证了该方法的有效性。最后,将所建立的方法应用于11批样品的分析。结果表明,该方法可较好地用于枸杞的质量控制与分析评价。     

Simultaneous determination of phytoestrogens in different medicinal parts of Sophora japonica L. by capillary electrophoresis with electrochemical detection

A high-performance capillary electrophoresis with electrochemical detection (CE-ED) method has been developed for the determination of phytoestrogens from the pericarps and seeds of Sophora japonica L. in this work. Genistin, genistein, rutin, kaempferol and quercetin are important bioactive constituents in these plants. The effects of several factors such as the acidity and concentration of running buffer, the separation voltage, the applied potential and the injection time on the CE-ED procedure were investigated. Under the optimum conditions, the five analytes could be well separated within 18 min in a 75 cm length capillary (i.d. 25 microm) at the separation voltage of 16 kV in a 50 mmol L(-1) borax running buffer (pH 9.0). A 300 microm diameter carbon disk electrode was used as the working electrode positioned carefully opposite the outlet of the capillary in a wall-jet configuration at the potential of +950 mV (vs SCE). Detection limits (S/N = 3) ranged from 1.1 x 10(-7) to 2.8 x 10(-7) g mL(-1) for all fi ve analytes. This method was successfully used to analyse dried Flos sophorae immaturus, pericarps and seeds of dried Fructus sophorae after a relatively simple extraction procedure, and the assay results were satisfactory.     

Electrochemical Determination of Synephrine by Hydrophilic Interaction Liquid Chromatography Using a Zwitterionic Monolith Column

A sensitive method for the electrochemical determination of synephrine (SYN) by hydrophilic interaction liquid chromatography (HILIC) has been developed. Optimal chromatographic separation and high sensitive determination by HILIC with electrochemical detection (HILIC‐ECD) was achieved using a sulfobetaine‐type zwitterionic monolith column (100×1.02 mm, i.d.), a mixture of 10 mM sodium phosphate (pH 4) and acetonitrile (20 : 80, v/v) as mobile phase, and a glassy carbon working electrode which was applied with a potential at +1.0 V vs. Ag/AgCl. The chromatographic peak height of SYN was proportional to the concentration from 5.0 µg/L to 1.0 mg/L (r=0.999). The detection limit of SYN (S/N=3) was 3.7 pg on the column. Moreover, the present HILIC‐ECD could be applied to the accurate and precise determination of SYN in Aurantii nobilis Pericarpium. In conclusion, we have demonstrated that an ECD is one of useful detection methods applicable to HILIC.     

Rapid extraction and determination of 25 bioactive constituents in Alpinia oxyphylla using microwave extraction with ultra high performance liquid chromatography with tandem mass spectrometry

A novel and rapid microwave extraction and ultra high performance liquid chromatography with tandem mass spectrometry method was developed and validated for the simultaneous determination of 25 bioactive constituents (including two new constituents) in Fructus Alpinia oxyphylla. The optimized conditions of the microwave extraction was a microwave power of 300 W, extraction temperature of 80°C, solvent‐to‐solid ratio of 30 mL/g and extraction time of 8 min. Separation was achieved on a Waters ACQUITY UPLC^® HSS C₁₈ column (2.1 mm× 50 mm, 1.8 μm) using gradient elution with a mobile phase consisting of acetonitrile and 1 mM ammonium acetate at a flow rate of 0.2 mL/min. This is the first report of the simultaneous determination of 25 bioactive constituents in Fructus Alpinia oxyphylla by ultra high performance liquid chromatography with tandem mass spectrometry. The method was validated with good linearity, acceptable precision and accuracy. The validated method was successfully applied to determine the contents of 25 bioactive constituents in Fructus Alpinia oxyphylla from different sources and the analysis results were classified by hierarchical cluster analysis, which indicated the effect of different cultivation regions on the contents of constituents. This study provides powerful and practical guidance in the quality control of Alpinia oxyphylla and lays the foundation for further research of Alpinia oxyphylla.     

Untargeted metabolomics analysis to unveil the chemical markers for the differentiation among three Gleditsia sinensis-derived herbal medicines by ultra-high performance liquid chromatography/quadrupole time-of-flight mass spectrometry

Precise identification and differentiation among those congeneric Traditional Chinese Medicines (TCMs) or derived from the same plant trend to be more challenging, particularly in the absence of appearance characteristics. Three TCMs, involving Gleditsiae Sinensis Fructus (GSF), Gleditsiae Fructus Abnormalis (GFA), and Gleditsiae Spina (GS), recorded in Chinese Pharmacopoeia (2020 edition) are derived from Gleditsia sinensis, but prescribed for different clinical uses. The documents aimed to compare their chemical differences are rare, to date. An untargeted metabolomics approach, based on ultra-high performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UHPLC/QTOF-MS), was elaborated to unveil the potential chemical markers to differentiate among GSF, GFA, and GS. Good chromatographic separation of all the GSF/GFA/GS components was achieved within 33 min by utilizing a BEH C18 column, while data-independent MS^E in the positive mode was selected for profiling the metabolic features. Notably, the high-mass saponins (1300–2500 Da) gave unique protonated precursors ([M + H]⁺) in the positive ESI mode, compared with those complicated ion species occurring in the negative mode. Pattern recognition chemometrics analysis of 45 batches of G. sinensis samples could unveil 70 significantly altered ions assigned as 46 potentially differential components. The positive/negative high-accuracy MS² data analysis, phytochemical isolation/NMR analysis, and searching of an in-house library of G. sinensis, were utilized for structural elucidation. Three compounds (saikachinoside A, locustoside A, and locustoside B) rich in GSF could be the markers to differentiate from GFA/GS, while four components were characteristic for GS. These results obtained can greatly benefit the quality control of TCMs derived from G. sinensis.     

Quality control of medicinal herbs Fructus gardeniae, Common Andrographis Herb and their preparations for their active constituents by high-performance liquid chromatography-photodiode array detection-electrospray mass spectrometry

Dehydroandrographolide, andrographolide and geniposide are the main active constituents of many herbal medicines, e.g., Fructus gardeniae, Common Andrographis Herb. They are used as the markers to control the quality of such herbal medicines and their herbal preparations. In this paper, a simple and sensitive high-performance liquid chromatographic (HPLC) method coupled with photodiode array detection (DAD) and electrospray mass spectrometry (ESI/MS) were developed to determine the three compounds simultaneously in extracts of medicinal herbs and herbal preparations produced by different companies. The extracts were separated on a C₁₈ reversed phase HPLC column, with a gradient solvent system, the time for the separation of the three target analytes was 10 min. The abundance ions were recorded using selected ion monitoring (SIM) mode with m/z 297.3, 297.3 and 411.1 for dehydroandrographolide, andrographolide and geniposide, respectively. The limit of detection for dehydroandrographolide, andrographolide and geniposide were 20, 30 and 150 ng mL⁻¹, respectively. The proposed method was successfully applied to the determination of the contents of the compounds in related to medicinal herbs and preparations.     

巴马火麻仁木脂素酰胺类提取物的鉴定及清除自由基活性的研究

将巴马火麻仁木脂素酰胺类的粗提物采用硅胶柱层析法以V(氯仿)∶V(甲醇)＝85∶15洗脱分离, 分离产物经快原子轰击质谱(FAB-MS)、核磁共振(NMR)鉴定为大麻酰胺A (cannabisin A). 以1,1-二苯基-2-苦基肼自由基(DPPH•)、超氧阴离子自由基 、羟自由基(OH•)三种不同自由基体系研究火麻仁木脂素酰胺类的粗提物、大孔树脂纯化后的精提物及cannabisin A的活性清除作用. 结果表明在一定的剂量范围内, 三者均有显著的活性清除作用, 且与剂量呈明显量效关系. 精提物的清除作用最强, 尤其是在OH•体系中, 其清除活性为粗提物的13倍.     

连翘的毛细管电泳指纹图谱研究

采用毛细管区带电泳法,以75 mmol/L硼砂溶液(用0.1 mmol/L氢氧化钠溶液调pH 9.7)为背景电解质,运行电压14 kV,检测波长228 nm,重力进样15 s(高度9 cm),建立了连翘药材的毛细管电泳指纹图谱(CEFP)。将10个不同产地的连翘药材进行比较,按各电泳峰共有率fi≥70%作为选择电泳指纹峰的依据,确定连翘的毛细管电泳指纹峰为29个。在方法的精密度和重复性试验中,各指纹峰的相对迁移时间的相对标准偏差(RSD)不大于5%,相对峰面积的RSD约为2%~15%。10个产地的连翘药材的CEFP与其对照CEFP的相似度均不低于0.94。同时,采用指纹图谱信息量指数I和相对信息量指数Ir评价了10个产地连翘药材的质量差异。     

固相萃取-高效液相色谱法测定枸杞中的类胡萝卜素

研究了固相萃取富集和预分离,高效液相色谱测定枸杞中的类胡萝卜素的方法;枸杞中的类胡萝卜素用WatersXterraTMRP18固相萃取小柱预分离,以WatersXterraTMRP18(3.9×150mm,5μm)液相色谱柱为固定相,甲醇 四氢呋喃(4 1)为流动相分离,用二极管矩阵检测器检测,检测波长为450nm。方法标准回收率为95%～103%。用该方法测定了几种枸杞样品中的类胡萝卜素。     

污泥上培植的报春花、冬葵中重金属的原子吸收测定

本文研究报春花、冬葵两种植物对污泥中重金属的吸收情况,样品用(1 4)HClO4 HNO3进行了消化处理,然后在实验选定的最佳的工作条件下,用火焰原子吸收光谱法对铅、汞、铬、镉重金属元素进行了测定。结果表明:种植冬葵和报春花前后,污泥样品重金属含量明显下降。