Construction of universal quantitative models for the determination of cefoperazone sodium/sulbactam sodium for injection from different manufacturers using near-infrared reflectance spectroscopy

To develop near-infrared （NIR） reflectance spectroscopic methods for the quantitative analysis of cefoperazone sodium/ sulbactam sodium from different manufacturers for injection powder medicaments. Various powders of cefoperazone sodium/ sulbactam sodium were directly analyzed by non-destructive NIR reflectance spectroscopy using the spectrometer EQUINOX55. Two quantitative methods via integrating sphere （IS） and fiberoptic probe （FOP） models were explored from 6 batches of commercial samples and 42 batches of laboratory samples at a content ranging from 30% to 70% for cefoperazone and 60% to 20% for sulbactam. The root mean square errors of cross validation （RMSECV） and the root mean square errors of prediction （RMSEP） of IS were 1.79% and 2.85%, respectively, for cefoperazone sodium, and were 1.86% and 3.08%, respectively, for sulbactam sodium; and those of FOP were 2.93% and 2.92%, respectively, for cefoperazone sodium, and were 2.23% and 3.01%, respectively, for sulbactam sodium. Based on the ICH guidelines and Ref. 12, the quantitative models were then evaluated in terms of specificity, linearity, accuracy, precision, robustness and model transferability. The non-destructive quantitative NIR methods used in this study are applicable for rapid analysis of injectable powdered drugs from different manufacturers.

Quantitative determination of polysaccharide in Curcuma wenyujin, a traditional Chinese medicine, by a phenol-sulfuric acid method

Curcuma wenyujin has been widely used as a traditional medicine in China. In this paper a strategy for the quantitative determination of the polysaccharide by a phenol-sulfuric acid method was described. Involved in three factors, 5% phenol volume, H2SO4 volume, and temperature of water bath, we adopted the L9（3）3 orthogonal array design to gain the optimal colorimetric method. 3.0 ml of polysaccharide solution, 1.0 ml, 5% phenol and 7.0 ml H2SO4 were mixed with constant stirring in a glass vessel, and then kept in a water bath at 40 ℃. After cooling to room temperature for 20 min, the absorbance values were recorded by the UV-2501 PC spectrometer at the wavelength range of 485 nm. The polysaccharide content in Curcuma wenyujin were 3.21%, 3.23%, 3.20%, 3.18~/0, 3.22% and 2.38%, respectively. All results showed that this method was adequate, valid and applicable, may be applied to the determination of other bacterial polysaccharide as well.

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 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.

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.

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

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Application of a quantitative ^1H NMR method for rapid extraction and determination of the content of paeonol in Cynanchum paniculatum

Cynanchum paniculatum(Bunge) Kitagawa is usually used as an herbal medicine for treating many diseases. Paeonol is the main active component, and its content is the key indicator for quality control of C. paniculatum. In the present study, we developed a rapid, accurate and precise method for quantitation of paeonol in C. paniculatum using 1 H NMR spectra. The deuterated solvent of methanol-d4 enabled satisfactory separation of the signals to be integrated in 1 H NMR spectrum. H-6(δ 7.78) of 1 H NMR spectrum of C. paniculatum was selected as the feature signal for quantitation, and trimesic acid(TMA) was selected as an internal standard. Validation of the quantitative method was performed in terms of linearity, specificity, repeatability and stability. This is the first time to report quantitative 1 H NMR(qHNMR) applied to determine the content of paeonol in C. paniculatum and showed a wider linearity range than the reported quantitation of paeonol in others. The simple extraction of paeonol from C. paniculatum was rapid and will prompt the application of the developed method. This work implied that qHNMR represented a feasible alternative to HPLC-based methods for quantitation of paeonol in C. paniculatum, and it was suitable for the quality control of C. paniculatum.

Simultaneous quantitative determination of eight index constituents and compatibility changes in Longchai Decoction by UPLC–Q-TOF-MS

The goal of this research was to develop a simple,rapid and sensitive method for simultaneous quantitative determination of salidroside,gardenoside,liquiritin,baicalin,wogonoside,wogonin,saikosaponin A and saikosaponin D in Longchai Decoction by ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry(UPLC–Q-TOF-MS),in order to control the quality of Longchai Decoction and to analyze the changes of chemical components before and after the compatibility of the component herb drugs.The chromatographic separation was performed on the Waters ACQUITY BEH C18 column(2.1 mm
100 mm,1.8μm)using the mixture of acetonitrile and 0.1%(v/v)methanoic acid as mobile phase with a gradient elution program at the flow rate of 0.3 mL/min and the column temperature of 301C.The eight components of the standards achieved baseline separation.Regression analysis revealed a linear relationship(r 240.9998)between the contents and the peak areas of the mixed standard substances.The average recovery rates were between 99.72%and 102.13%with RSD values were less than 2.82%(n¼5).The obtained results indicated that the content of index components were higher in co-decoction compared to mixed decoction.This method with a good resolution and high precision can be used for the quality control of Longchai Decoction.

Origin of potential errors in the quantitative determination of terahertz optical properties in time-domain terahertz spectroscopy

We demonstrate theoretically and experimentally how changes of a terahertz （THz） beam induced by the sample affect the accuracy of the determination of THz dielectric properties in THz time-domain transmission spectros- copy （TDTS）. We apply a Gaussian beam and the ABCD matrix formalism to describe the propagation of the THz beam in a focused beam setup. The insertion of the sample induces a focus displacement which is absent in the reference t without a sample. We show how the focus displacement can be corrected. The THz optical properties after focus displacement correction reported in this Letter are in quantitative agreement with those obtained using collimated beam THz-TDTSinpreviouswork.

Quantitative analysis of FRET assay in biology New developments in protein interaction affinity and protease kinetics determinations in the SUMOylation cascade

Forster resonance energy transfer （FRET） techniques have been widely used in biological studies in vitro andin vivo and are powerful tools for elucidating protein interactions in many regulatory cascades. FRET occurs between oscillating dipoles of two fluorophores with overlapping emission and excitation wavelengths and is dependent on the spectroscopic and geometric properties of the donor-acceptor pair. Various efforts have been made to develop quantitative FRET methods to accurately determine the interaction affinity and kinetics parameters. SUMOylation is an important post-translational protein modification with key roles in multiple biological processes. Conjugating SUMO to substrates requires an enzymatic cascade. Sentrin/SUMO-specific proteases （SENP） act as endopeptidases to process the pre-SUMO or an isopeptidase to deconjugate SUMO from its substrate. Here we also summarize recent developments of theoretical and experimental procedures for determining the protein interaction dissociation constant, Kd, and protease kinetics parameters, kcat and Kin, in the SUMOylation pathway. The general principles of these quantitative FRET-based measurements can be applied to other protein interactions and proteases.

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。

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。

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。

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。

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。