Quantitative Determination of Click Reaction in the Presence of Glycine Derivatives and in Dilute Solution

The click reaction is one of the latest techniques for the functional analysis of bioactive compounds and the analysis makes novel concepts and strategies for medicinal chemistry. N-methylated glycine derivatives have inhibitory activity for the click reaction in direct Cu(I) system because of decrease of Cu(I) concentration. The Cu(I) concentration recovered effectively by sodium ascorbate. Quantitative determination of click reaction at various ligand concentrations revealed that the decrease in reaction yields was observed in a substrate concentration-dependent manner.

Quantitative Determination of Tetrachlorantraniliprole by(1)^H NMR Spectroscopy with Internal Standard Method

[Objective]This study was to establish a rapid,specific and simple method for quantitative determination of tetrachlorantraniliprole by 1H NMR.[Method]1H NMR spectroscopy was acquired with deuterium DMSO as the solvent and maleic acid as internal standard under the conditions of temperature 25℃,pulses width 8.0μs,delay time 5 s,and scanning times 8.[Result]The hydrogen proton peaks of tetrachlorantraniliprole(δ=10.55)and maleic acid(δ=6.27)were taken as quantitative peaks.The peak area ratio y(As/Ar)and mass ratio x(ms/mr)were linearly regressed,and the correlation coefficient was 0.9999.The RSD value of repeatability test was 0.38%,and the RSD value of stability test was 0.77%.The content of tetrachlorantraniliprole was determined as 99.6%.[Conclusion]1H NMR spectroscopy can be used for quantitative determination of tetrachlorantraniliprole without standard reference,which is rapid,accurate and simple.

A Mathematical Calculation Model Using Biomarkers to Quantitatively Determine the Relative Source Proportion of Mixed Oils

It is difficult to identify the source（s） of mixed oils from multiple source rocks, and in particular the relative contribution of each source rock. Artificial mixing experiments using typical crude oils and ratios of different biomarkers show that the relative contribution changes are non-linear when two oils with different concentrations of biomarkers mix with each other. This may result in an incorrect conclusion if ratios of biomarkers and a simple binary linear equation are used to calculate the contribution proportion of each end-member to the mixed oil. The changes of biomarker ratios with the mixing proportion of end-member oils in the trinal mixing model are more complex than in the binary mixing model. When four or more oils mix, the contribution proportion of each end-member oil to the mixed oil cannot be calculated using biomarker ratios and a simple formula. Artificial mixing experiments on typical oils reveal that the absolute concentrations of biomarkers in the mixed oil cause a linear change with mixing proportion of each end-member. Mathematical inferences verify such linear changes. Some of the mathematical calculation methods using the absolute concentrations or ratios of biomarkers to quantitatively determine the proportion of each end-member in the mixed oils are deduced from the results of artificial experiments and by theoretical inference. Ratio of two biomarker compounds changes as a hyperbola with the mixing proportion in the binary mixing model, as a hyperboloid in the trinal mixing model, and as a hypersurface when mixing more than three end- members. The mixing proportion of each end-member can be quantitatively determined with these mathematical models, using the absolute concentrations and the ratios of biomarkers. The mathematical calculation model is more economical, convenient, accurate and reliable than conventional artificial mixing methods.

Simultaneous Determination of Five Components in Operculina turpethum

[Objectives]This study was conducted to establish a method for the simultaneous determination of caffeic acid, rutin, ononin, luteolin, and apigenin in Operculina turpethum(L.) S. Manso. [Methods]With ononin from O. turpethum as the internal reference, the five components were separated by HPLC, and the contents of various components were calculated according to the relative correction factors of ononin with caffeic acid, rutin, luteolin, and apigenin. Meanwhile, the calculated results of quantitative analysis of multi-components by single marker(QAMS) were compared with the determined values of the external standard method. [Results] The linear relationship of the five components in their respective ranges was good(r=0.999 9). The average recovery was in the range of 97.48%-101.05%, and the RSD values were in the range of 1.04%-2.71%. The results obtained by QAMS were close to those obtained by the external standard method. [Conclusions] The method is accurate, stable and adaptable, and can be used for the determination of five flavonoids in O. turpethum.

Simultaneous determination of seven flavonoids in Epimedium by liquid chromatography-tandem mass spectrometry method

In this paper, a sensitive and specific liquid chromatography-electrospray ionisation-mass spectrometry （LC-ESI-MS） method has been developed and validated for the identification and determination of seven flavonoids, namely epimedin A, epimedin B, epimedin C, icariin, sagittatoside B, 2＂-O-rhamnosyl icariside II, and baohuoside I in Epimedium from different sources.

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.

Simultaneous determination of ten components in Dianxiankang Capsules by HPLC QAMS

Objective:To establish an high performance liquid chromatography-quantitative analysis of multi components by single marker(HPLC-QAMS)method for simultaneous determination of gastrodin,parishin E,parishin B,parishin,tenuifolin,onjisaponin B,methylophiopogonanone A,methylophiopogonanone B,β-asarone andαasarone in Dianxiankang Capsules.Methods:Waters Symmetry C_(18)column was used with acetonitrile-0.05%phosphoric acid solution as mobile phase for gradient elution.Multiwavelength switching detection.The contents of gastrodin,parishin E,parishin B,parishin,onjisaponin B,methylophiopogonanone A,methylophiopogonanone B,β-asarone andα-asarone were calculated by relative correction factor.At the same time,the contents of 10 components in 12 batches of Dianxiankang Capsules were determined by external standard method(ESM).Results:An HPLC-QAMS method was established tenuifolin as the internal reference substance was established.The relative correction factors of gastrodin,parishin E,parishin B,parishin,onjisaponin B,methylophiopogonanone A,methylophiopogonanone B,β-asarone andα-asarone were 0.8238,0.7239,1.0229,1.1881,0.7272,1.3108,0.9314,0.6549 and 1.0572,respectively.The relative correction factors had good repeatability and no significant difference with ESM(P>0.05).Conclusion:HPLC-QAMS can be used for simultaneous determination of multi-index components in Dianxiankang Capsules.

乙型肝炎病毒基因B型及基因C型两种国家标准品研制

目的研制乙型肝炎病毒(HBV)基因B型及基因C型两种国家标准品。方法收集我国不同地区的HBV感染者血浆样品,经HBV五项、DNA病毒检测,从中筛选出11份基因分型标准品候选品样品。经HBV序列测定,进化树分析,初步确定2份样品分别为HBV B型及C型标准品候选品。分别将2份候选品离心、分装后,进行HBV基因分型验证测定,并进行定量的协作标定测定。结果确认筛选出HBV基因B型及基因C型标准品,其HBV DNA浓度分别为4.67×10~7IU/mL(CV为3.6%)及3.66×10^(8)IU/mL(CV为2.9%)。装量检查,大于0.5 mL/支,符合规定。均匀性检查,两者P值分别为0.428(F值为1.140)及0.420(F值为1.173),分装的标准品支与支之间浓度均匀无差异。将两种标准品在不同方式保存(反复冻融、4℃保存、室温及37℃放置)后,测定结果与-80℃保存样品测定结果相比较,绝对偏差均在±0.2的范围内,方差分析P值均大于0.1;另外,-20℃保存12个月后与-80℃保存样品两者测定数据分析,B型P值为0.237(F值为1.934)、C型P值为0.173(F值为2.737),P值均大于0.1,稳定性验证符合规定。结论制备了HBV基因B型及基因C型两种国家标准品,为我国HBV基因分型试剂的质量控制和标准化提供了依据。

不同月龄双线紫蛤数量性状的相关性和通径分析

为研究双线紫蛤形态性状与体质量之间的关系,随机选取6、12和18月龄双线紫蛤(Soletellina diphos)各50个个体,分别测量壳长(SL)、壳高(SH)、壳宽(SW)、体质量(BW)共4个数量性状,采用相关分析和通径分析的方法分别计算不同月龄群体对体质量的通径系数和决定系数。相关分析结果显示,6、18月龄群体均是壳长与体质量的相关系数最大(P<0.01)。根据多元回归分析结果,建立了3个月龄群体形态性状对体质量性状的多元回归方程:BW_(6)=-1.336+0.020SL+0.146SW+0.081SH,R^(2)=0.976;BW_(12)=-29.701+0.584SL+0.695SW,R^(2)=0.972;BW_(18)=-88.160+1.140SL+1.660SW,R^(2)=0.911。通径分析结果表明,6月龄群体中影响体质量的主要因素为壳宽(0.466),12和18月龄群体则为壳长(0.753,0.705),决定系数分析结果与通径分析结果相一致。本研究结果为双线紫蛤种质资源保护与利用提供了基础资料。

基于XRD内标法测定LiF-BeF_2熔盐体系中UF_3含量

在氟化物熔盐相直接将UF6中转化为UF_4是熔盐堆燃料盐的合成及重构的候选工艺之一。其中,UF_(3)是该工艺过程的重要中间产物,其含量是开展工艺研究的关键参数之一。本文采用X射线衍射(X-ray Diffraction,XRD)内标法测量氟化物体系中的UF_(3)含量,分析了不同组分的固态熔盐样品,建立了UF_(3)含量的测量分析方法。首先以刚玉(α-Al_(2)O_(3))为内标,获得了UF_(3)在1.00~10.00 wt%含量范围时,LiF-BeF_(2)-UF_(3)熔盐的XRD峰高度内标曲线(R=0.986)和峰面积内标曲线(R=0.995)。然后应用这两条内标曲线测量已知含量的LiUF_(5)和UF_(3)固体混合样品,结果表明峰面积内标曲线具有更好的准确度,测量相对误差不大于8.7%。最后分别对快速冷却的LiF-BeF_(2)-UF_(3)固态熔盐样品和自然冷却的LiF-BeF_(2)-UF_(3)-LiUF_(5)固态熔盐样品进行测量,测量结果的相对误差不大于5.4%。以上结果表明:本文建立的XRD内标法可以用于混合氟盐样品的UF_(3)分析,并具有较好的测量精度和重复性。

Determination of 14 Chemical Constituents in the Traditional Chinese Medicinal Preparation Huangqin-Tnag by Hight Performance Liquid chromatography

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超高效液相色谱-三重四级杆质谱联用法同时测定荔枝中4种细胞分裂素

【目的】应用超高效液相色谱-三重四级杆质谱联用仪(UPLC-MS/MS),建立一种同时测定荔枝反式玉米素核苷(tZR)、异戊烯基腺嘌呤核苷(IPR)、二氢玉米素(DHZ)、异戊烯基腺嘌呤(IP)4种细胞分裂素含量的方法。【方法】0.25 g荔枝样品用乙腈-水(80∶20,体积比)溶液浸提8 h,经Bond Elut Plexa PCX固相萃取柱纯化,2.5%氨水甲醇洗脱后过0.22μm有机滤膜检测。选取XSelect HSS T3色谱柱,以甲醇和5 mmol·L^(-1)甲酸铵水溶液为流动相进行7 min梯度洗脱,采用电喷雾正离子(ESI+)模式电离,选择反应监测(MRM)模式对细胞分裂素进行定量。【结果】4种细胞分裂素的检出限和定量限分别低于18.12和60.39 pg·g^(-1),在0.05~50 ng·mL^(-1)质量浓度范围内呈良好的线性关系,线性相关系数(r2)大于0.999。在高、中、低浓度三种加标水平下,4种细胞分裂素的平均回收率为80.0%~108.2%,标准偏差为0.8%~15.5%。采用建立的方法可以同时检测出荔枝果皮、果肉、种子、幼果、叶片和果柄离区中4种细胞分裂素含量。【结论】该方法简便、快速、灵敏、准确,适用于荔枝不同组织中细胞分裂素含量的测定。

一测多评法同时测定参芪二皮冬茶颗粒中6种成分的含量

目的 建立一测多评法同时测定参芪二皮冬茶颗粒中毛蕊异黄酮葡萄糖苷、阿魏酸、迷迭香酸、橙皮苷、橘皮素、川陈皮素的含量。方法 分析采用Agilent Eclipse Plus-C_(18)色谱柱(4.6 mm×250 mm, 5μm);流动相乙腈-0.1%磷酸,梯度洗脱;体积流量1.0 mL/min;柱温25℃;检测波长260、330 nm。以橙皮苷为内标,计算其他5种成分的相对校正因子,测定其含量。结果 6种成分在各自范围内线性关系良好(r≥0.999 0),平均加样回收率99.82%~101.18%,RSD 1.26%~2.25%。一测多评法所得结果与外标法接近。结论 该方法简便可靠,重复性好,可用于参芪二皮冬茶颗粒的质量控制。

一测多评法同时测定天麻头风灵胶囊中10种成分的含量

目的建立一测多评法同时测定天麻头风灵胶囊中天麻素、巴利森苷E、紫丁香苷、巴利森苷B、巴利森苷C、阿魏酸、巴利森苷A、蒙花苷、哈巴俄苷、肉桂酸的含量。方法分析采用GL Science Inertsil^(TM)ODS-3色谱柱(150 mm×4.6 mm,5μm);流动相乙腈-0.1%磷酸,梯度洗脱;体积流量1.0 mL/min;柱温30℃;检测波长220、280 nm。以紫丁香苷为内标,计算其他9种成分的相对校正因子,测定其含量。结果10种成分在各自范围内线性关系良好(r≥0.9997),平均加样回收率98.53%~102.22%,RSD 1.26%~2.68%。一测多评法所得结果与外标法接近。结论该方法准确、专属性强,可用于天麻头风灵胶囊的质量控制。

基于β-环糊精的高矿化度水中曲拉通X-405的检测新方法

油田压裂用水矿化度高,为了降低压裂用水对紫外光谱法检测聚乙二醇叔辛基苯基醚(曲拉通X-405)产生的影响,采用添加适量β-环糊精(β-CD)作为屏蔽剂来消除压裂用水的干扰作用。研究结果表明,压裂用水能明显降低曲拉通X-405的吸光度和临界胶束浓度(cmc),在压裂用水中,100.0 mg/L曲拉通X-405在最大吸收波长224 nm处的吸光度降幅达6.67%,曲拉通X-405的cmc也由蒸馏水中的103.978 mg/L降低至97.056 mg/L。加入与曲拉通X-405相同物质的量的β-环糊精后,压裂用水对曲拉通X-405吸光度的干扰作用显著降低,100.0 mg/L曲拉通X-405在224 nm处的吸光度降幅仅为1.99%,在20.0~160.0 mg/L范围内,曲拉通X-405在蒸馏水中及在压裂用水中均没有形成胶束,曲拉通X-405的检测回收率也由88.3%~97.4%改善至97.5%~99.6%。等摩尔连续变化法、FT IR及^(1)H NMR结果表明,在水溶液中曲拉通X-405与β-CD按物质的量比1∶1形成包结物,曲拉通X-405分子可以分别从β-CD分子空腔的宽、窄口径两端进入β-CD内腔,包结物的屏蔽作用是降低压裂用水对曲拉通X-405的干扰作用的根本原因。该方法为准确检测高矿化度压裂用水中的曲拉通X-405提供了新参考。

一测多评法同时测定心脑康胶囊中9种成分

目的 建立一测多评法同时测定心脑康胶囊中丹参素钠、丹酚酸B、芍药苷、阿魏酸、羟基红花黄色素A、2,3,5,4′-四羟基二苯乙烯-2-O-β-D-葡萄糖苷、斯皮诺素、甘草苷、葛根素的含量。方法 分析采用SVEA C_(18)色谱柱(250 mm×4.6 mm, 5μm);流动相乙腈-0.2%甲酸,梯度洗脱;体积流量0.5 mL/min;柱温30℃;检测波长280、403 nm。以葛根素为内标,计算其他8种成分的相对校正因子,计算其含量。结果 9种成分在各自范围内线性关系良好(r≥0.999 4),平均加样回收率99.83%~101.18%,RSD 1.04%~1.91%。一测多评法所得结果与外标法接近。结论 该方法准确稳定,重复性好,可用于心脑康胶囊的质量控制。

高效液相色谱法测定硝酸甘油气雾剂定量阀门中5种抗氧剂含量与迁移量研究

目的 建立同时测定硝酸甘油气雾剂定量阀门中5种常见抗氧剂含量的高效液相色谱(HPLC)法,并考察抗氧剂在其中的迁移情况。方法 色谱柱为Inertsil?ODS-SP柱(150 mm×4.6 mm,5μm),流动相为水-四氢呋喃-乙腈(10∶30∶60,V/V/V),流速为1.5 mL/min,柱温为30℃,检测波长为280 nm,进样量为20μL。结果 抗氧剂168、抗氧剂330、抗氧剂1010、抗氧剂1076、抗氧剂2,6-二叔丁基对甲酚(BHT)的质量浓度均在0.2~250μg/mL范围内与峰面积线性关系良好(r≥0.999 8,n=6);检测限分别为1.58,0.27,0.38,0.94,0.24μg/mL;定量限分别为3.84,0.91,1.44,3.02,0.60μg/mL;精密度、稳定性、重复性试验结果的RSD均小于2.0%(n=6);回收率分别为100.67%,99.97%,99.93%,99.60%,99.63%,RSD分别为0.47%,0.21%,0.93%,0.44%,0.60%(n=9)。硝酸甘油气雾剂定量阀门组件材料(内垫圈、阀杆、阀室、吸管)中抗氧剂168、抗氧剂330、抗氧剂1010、抗氧剂1076的检出量均低于其评价阈值(AET),均未检出抗氧剂BHT。加速试验0个月、3个月、6个月,定量阀门组件材料中抗氧剂168、抗氧剂330、抗氧剂1010、抗氧剂1076、抗氧剂BHT的迁移量均低于其AET。结论 该方法操作简单、灵敏度高、结果准确、重复性和稳定性均较好,可用于硝酸甘油气雾剂定量阀门中5种抗氧剂的含量测定及迁移量研究。

橄榄油中生物酚准确定量的瓶颈及对策

橄榄油中的生物酚是具有广泛生物活性的极其复杂的极性酚类物质,不仅化学结构异质、性质多不稳定、异构化程度高,而且橄榄多酚等歧义概念的长期共存,使得生物酚始终没有理想的测定方法,一定程度上影响了橄榄油的品质控制及贸易。为了寻求突破橄榄油中生物酚测定瓶颈的对策,为橄榄油中生物酚的准确定量提供理论参考,综述了多酚、酚类化合物、橄榄油多酚与生物酚等类似术语使用范围的异同,分析了欧洲食品安全局(EFSA)关于橄榄油多酚健康声称中术语模糊、缺失适用分析方法的争议,从裂环烯醚萜类化合物的复杂异构体与衍生物、色谱基线分离不可行、标准品缺失、单体酚法与总酚法的不兼容性4个方面阐述了橄榄油中生物酚准确定量的瓶颈,并对突破瓶颈的4个具有潜力的研究方向进行展望,即实现全回收率与零人工产物的直接进样法,提升分辨力与准确性的离子淌度质谱(IM-MS),降低基质效应与提升灵敏度的二维液相色谱(LC×LC),加速裂环烯醚萜类标准品研制。橄榄油生物酚准确定量的瓶颈将长期存在,方法突破有赖于更深入的基础研究与创新技术的应用。

定量核磁共振技术在中药质量控制中的应用进展

中药质量控制是保证中药安全有效、质量稳定的基础,是中药标准化、现代化、国际化的前提,选用合适的分析方法在中药质量控制中具有重要意义。与薄层色谱、气相色谱、高效液相色谱等常规分析方法相比,定量核磁共振技术(qNMR)样品前处理简单、分析检测速度快、无需自身对照品,集定性鉴别和定量分析于一体,适用于中药中各类成分的检测分析,或将作为一种有力的分析技术,应用于中药质量控制。对qNMR的基本原理、定量方法和药典应用进行总结,重点介绍了qNMR在中药材、中药制剂、中药对照品及中药生产过程质量控制中的应用进展,提出了qNMR存在的若干问题并展望了其发展前景,旨在为采用qNMR控制中药质量提供参考。