基于LC-MS/MS成分分析结合高效液相色谱法评价泻白散基准样品质量

目的：建立泻白散基准样品特征图谱方法,以LC-MS/MS指认各特征峰并对其进行归属,建立基准样品各组成药味质控指标的含量测定方法综合评价其质量。方法：以色谱柱Hypersil ODS²(250 mm×4.6 mm,5μm)、乙腈-0.1%磷酸/甲酸梯度洗脱、检测波长280nm、流速1.0 ml/min、柱温25℃、进样量10μl进行指纹图谱研究,以同一色谱条件使用LC-MS/MS联用进行峰归属并进行化学成分分析,以指认的各药味专属性峰建立基准样品质控指标含量测定方法。结果：泻白散基准样品特征图谱重复性良好,供试品溶液48 h内稳定,多批次检测显示泻白散指纹图谱共有峰14个,相似度均＞0.9;峰归属确认其中5个归属于炒桑白皮,5个归属于炙甘草,3个归属于地骨皮;并通过LC-MS/MS指认出其中11个色谱峰;选取桑皮苷A、甘草酸、枸杞素B分别建立含量测定方法,结果显示方法均具有专属性,重复性良好,加样回收率分别为94.9%、93.5%、95.2%。结论：该指纹图谱方法经验证可行,可有效鉴别泻白散三味饮片;结合LC-MS/MS成分分析可有效选取基准样品各药味的质控成分;建立的检测桑皮苷A、甘草酸、枸杞素B含量测定方法经方法学考察确认可行。     

舒心片的质量标准研究

目的：建立中药舒心片中丹酚酸B含量的测定方法。方法：采用高效液相色谱法（HPLC）测定舒心片中丹酚酸B的含量,色谱柱为Agilent TC-C₁₈（2）色谱柱（4.6 mm×250 mm,5 μm）,以乙腈-0.1%的磷酸溶液（25∶75）为流动相;检测波长为286 nm;柱温为25℃;流速为1.0 ml/min。结果：供试品和对照品溶液中丹酚酸B与他峰分离度良好;在0.050～1.642 μg之间丹酚酸B呈良好线性关系,R=0.9993,丹酚酸B平均加样回收率为97.58%,RSD=0.25%。结论：经方法学验证,本法可用于舒心片的质量控制。     

基于指纹图谱和含量测定评价脑得生丸的质量

目的：以高效液相色谱(HPLC)指纹图谱结合含量测定方法,为科学评价脑得生丸的质量提供理论依据。方法：采用HPLC建立脑得生丸的指纹图谱,色谱柱为 Inertsil ODS-SP（4.6 mm×250 mm,5μm）;流动相为乙腈-0.1%甲酸溶液,梯度洗脱;柱温 30℃;检测波长为254 nm;流速为1 ml/min。采用HPLC方法测定脑得生丸中人参皂苷 Rg1,色谱柱为 HypersilODS2（4.6 mm×250 mm,5μm）;流动相为乙腈-0.05%磷酸（19∶81）,等度洗脱;检测波长为 203 nm;流速为 1.0 ml/min;进样量为10 μl。基于指纹图谱和含量测定结果,对脑得生丸的质量进行评价。结果：建立了脑得生丸的指纹图谱,确定了11个共有峰,15批脑得生丸样品相似度均大于0.97。人参皂苷 Rg1的线性方程为 y=13600x-9.5581,线性范围是 19.2 μg/ml～1.920 mg/ml,相关系数是0.9992。15批脑得生丸中有5批样品为过期产品,仅批次为161003样品（过期5个月）含量检测不合格,其他均符合质量标准要求。结论：该方法简便、准确、重复性好,为脑得生丸质量的有效评价与整体控制提供了实验依据。     

海洋药用生物系列HPLC化学指纹图谱研究 Ⅱ.三斑海马HPLC化学指纹图谱研究

采用高效液相色谱(HPLC)法建立三斑海马的参考指纹图谱.运用DAD-Agilent 1100高效液相色谱仪,色谱条件:Agilent Zorbax XDB C18色谱柱(4.6 mm×250 mm;5.0 μm),乙腈-0.1%磷酸水流动相梯度洗脱,流速1.0 mL·min-1,检测波长200 nm,参比波长360 nm,柱温30 ℃.结果显示建立的指纹图谱包含11个共有峰,方法的精密度、重现性和稳定性试验RSD值均小于5.0%.研究表明所建立的三斑海马的参考指纹图谱可用于药材的真伪鉴别.     

液相色谱法测定鱼肌肉中孔雀石绿、结晶紫及其代谢物

孔雀石绿(MG)和结晶紫(GV)曾作为渔药使用,在鱼体内可分别代谢为无色孔雀石绿(LMG)及无色结晶紫(LGV),具有潜在的致癌性,已被许多国家禁用。本文研究了同时检测孔雀石绿、结晶紫及其代谢物无色孔雀石绿和无色结晶紫的液相色谱方法,建立了检测前处理方法及色谱条件。经过液液萃取和丙基磺酸阳离子树脂纯化,采用C18分离柱后串联二氧化铅柱氧化衍生的方法,同时检测母体药物及其代谢物,回收率>70%,重复性的相对标准偏差<15%,定量限分别为2μg/kg及1μg/kg。经样品实测,在一份鲈鱼样品中检测到无色孔雀石绿残留97μg/kg。     

小儿扶脾颗粒指纹图谱研究

目的：建立湖南时代阳光药业股份有限公司小儿扶脾颗粒的高效液相色谱法（HPLC）指纹图谱分析方法。方法：采用反相高效液相色谱法,Ulimate-XB C_{18色谱柱（4.6mm×250mm,5μm）,乙腈-0.1%磷酸溶液梯度洗脱,柱温30℃,检测波长330nm。结果：以橙皮苷为参照峰,建立了小儿扶脾颗粒HPLC指纹图谱,确立了12个共有峰,10批样品之间的共有峰相似度>0.9。结论：该方法准确,精密度、重复性较好,可作为湖南时代阳光药业股份有限公司小儿扶脾颗粒质量控制的依据。}     

护心康片及其组方药物多波长HPLC指纹图谱色谱峰相关性研究

目的：建立护心康片多波长HPLC指纹图谱,并研究复方与组方药物指纹图谱峰的相关性。方法：采用C_{18色谱柱（4.6mm×200mm,5μm）;柱温35℃;0.1%磷酸溶液（A）-乙腈（B）为流动相梯度洗脱;流速1.0ml/min;检测波长分别为203nm（检测三七和黄芪）、270nm（检测丹参酮类）、286nm（检测丹参酚酸类和黄芪）。结果：建立了23个特征峰的护心康片HPLC指纹图谱,并对其特征峰进行了组方药物归属分析。结论：该方法能揭示复方主要色谱峰在组方药物中的归属,有效控制护心康片的质量。}     

山银花配方颗粒高效液相色谱特征指纹图谱研究

目的：建立山银花配方颗粒高效液相色谱（HPLC）特征指纹图谱。方法：采用Scienhome Column Kromasil C_{18色谱柱（4.6 mm×250mm,5μm）,以乙腈-0.4%磷酸水溶液为流动相进行梯度程序洗脱（0～20min,3%→15%乙腈;20～45min,15%→24%乙腈;45～55min,24%→40%乙腈;55～65min,40%→3%乙腈;65～75min,3%乙腈）,检测波长238nm,流速1.0ml/min,柱温30℃,进样量5μl,检测时间75min。结果：由10批实验室自制样品建立山银花配方颗粒HPLC特征指纹图谱,确定了8个共有峰,已解析出5个共有峰成分。结论：本方法灵敏度高、稳定性和重复性好,所建立的特征指纹图谱可用于山银花配方颗粒的鉴别和质量评价。}     

四甲丸质量标准定量方法研究

目的：建立四甲丸中2种活性成分哈巴俄苷、肉桂酸的含量测定方法。方法：色谱条件为C18色谱柱（4.6mm×250mm,5μm）;流动相为乙腈-1%醋酸水溶液,体积比为25∶75,等度洗脱;流速：1mL/min,柱温：25℃;检测波长：278nm。结果：在该色谱条件下,哈巴俄苷、肉桂酸分离良好,质量浓度分别在5.36～134.06μg/mL（r=0.9999）、5.28～131.95μg/mL（r=0.9997）时,与其峰面积线性关系良好。加样回收率分别为103.6%（1.20%）、100.4%（1.30%）（n=6）。结论：本试验所建立的方法简便、准确、重复性好,可用于四甲丸的质量控制。     

滨海湿地典型耐盐植物盐地碱蓬的HPLC化学指纹图谱

建立药用耐盐植物盐地碱蓬(Suaeda salsa)的HPLC指纹图谱。运用DAD-Agilent 1100高效液相色谱仪,色谱条件:Agilent Zorbax XDB C18柱(4.6 mm×250 mm;5.0μm)色谱柱,1.0 mL.min-1流速,320 nm检测波长,360 nm参比波长,10μL进样量,30℃柱温,乙腈(A)-0.1%磷酸水(B)梯度洗脱。建立了盐地碱蓬的HPLC指纹图谱,包含16个共有峰,指纹图谱特征明显,化学信息完整。方法的系统适应性测定结果符合指纹图谱的技术规范。结论:首次建立的盐地碱蓬HPLC参考指纹图谱可用于盐地碱蓬的快速真伪鉴别。     

Microbial Biomass and Organic Nutrients in the Deep-Sea Sediments of the Central Indian Ocean Basin

In order to assess the impact of deep-sea mining on the in situ benthic life, we measured the microbial standing stock and concentration of organic nutrients in the deep-sea sediments of the Central Indian Ocean Basin in the Indian pioneer area. Sediments were collected using box core and grab samples during September 1996. The total bacterial numbers ranged from 10 10 -10 11 cells per g -1 dry weight sediment. There was a marginal decrease in the number of bacteria from surface to 30 cm depth, though the subsurface section registered a higher number than did the surface. The highest numbers were encountered at depths of 4-8 cm. The retrievable number of bacteria were two orders less in comparison with the direct total counts of bacteria. An almost homogeneous distribution of bacteria, total organic carbon, living biomass, and lipids throughout the depth of cores indicates active microbial and benthic processes in the deep sea sediments. On the other hand, a uniform distribution of total counts of bacteria, carbohydrates, and total organic carbon in all the cores indicates their stable nature and suggests that they can serve as useful parameters for long-term monitoring of the area after the benthic disturbance. Further studies on temporal variability in this region would not only verify the observed norms of distribution of these variables but would also help to understand restabilization processes after the simulated benthic disturbance.     

Assessing the Validity of Seismo-Acoustic Predictor Equations for Obtaining Seabed and Sub-Surface Sediment Physical Properties

The interdependence between the seismo-acoustic properties of a marine sediment and its geotechnical/physical parameters has been known for many years, and it has been postulated that this should allow the extraction of geotechnical information from seismic data. Though in the literature many correlations have been published for the surficial layer, there is a lack of information for greater sediment depths. In this article, a desktop study on a synthetic seafloor model illustrates how the application of published near-surface prediction equations to subsurface sediments (up to several tens of meters burial depth) can lead to spurious predictions. To test this further, acoustic and geotechnical properties were measured on a number of sediment core samples, some of which were subjected to loading in acoustically-equipped consolidation cells (oedometers) to simulate greater burial depth conditions. For low effective pressures (representing small burial depths extending to around 10 meters subsurface), the general applicability of established relationships was confirmed: the prediction of porosity, bulk density, and mean grain size from acoustic velocity and impedance appears generally possible for the investigated sedimentary environments. As effective pressure increases through, the observed relationships deviate more and more from the established ones for the near-surface area. For the samples tested in this study, in some instances increasing pressure even resulted in decreasing velocities. There are several possible explanations for this abnormal behavior, including the presence of gas, overconsolidation, or bimodal grain size distribution. The results indicate that an appropriate depth correction must be introduced into the published prediction equations in order to obtain reliable estimates of physical sediment properties for greater subsurface depths.     

Geoacoustic Inversion via Genetic Algorithm and Its Application to Manganese Sediment Identification

An acoustic inversion method using a wide-band signal and two near field receivers is proposed and applied to multiple layered seabed models including a manganese sediment. The inversion problem can be formulated into a probabilistic model comprised of signals, a forward model, and additive noise. The forward model simulates wide-band signals, such as chirp signals, and is chosen to be the source-waveletconvolution plane wave modeling method. The wavelet matching technique, using weighted least-squares fitting, estimates the sediment sound-speed and thickness on which determination of the possible numerical ranges for a priori uniform distribution is based. The genetic algorithm is applied to a global optimization problem to find a maximum a posteriori solution for determined a priori search space. Here the object function is defined by an L 2 norm of the difference between measured and modeled signals. Not only the marginal pdf but also its statistics are calculated by numerical evaluation of integrals using the samples selected during importance sampling process of the genetic algorithm.     

海上大直径钢管桩打桩过程中桩周土强度弱化研究

海上大直径钢管桩打桩过程中,桩周土体受到强烈扰动而发生强度弱化,掌握桩周土体强度弱化规律对于准确预测打桩过程、保证工程安全具有重要意义。为研究土体强度弱化规律,开展了环剪试验模拟打桩对桩周土体的扰动,测试土体强度随剪切速率的变化规律,建立了描述土体强度弱化规律的拟合公式,引入到打桩分析软件中。研究结果表明：土体的强度折减程度不仅与土体本身的性质有关还受到土体的埋深和剪切速率的影响,埋深越深土体强度折减程度越低,剪切速率越高土体强度折减越高,在打桩分析中可采用这里推荐的线性折减方法来模拟不同深度处土体强度的折减规律。     

Interannual Sea Level Variations and Annual Tides in the Northwestern Pacific

Specific properties of the interannual sea level variations and annual tides in the Northwestern Pacific were studied. Several tide stations were monitored. The monthly mean sea level for the year of 1995 was analyzed at each tide station. A seismic event in 1995, some tectonic activity around the subject area, and the Kuroshio (the oceanic western boundary current) may possibly contaminate results which would have occurred from the astronomical annual tide alone.     

Deep-Sea Sediment Compression Curves: Some Controlling Factors, Spurious Overconsolidation, Predictions, and Geophysical Reproduction

Oedometer tests have been carried out on 70 undisturbed surficial clays (at approximately 250 mm below the mudline), mostly collected by free-fall corers from sites widely scattered throughout the deep-sea North Atlantic. Acoustic measurements were also made, initially on contiguous samples and ultimately on the same sample using a geophysically instrumented oedometer which also collected electrical resistivity data. Apart from those quiescent areas below the carbonate compensation depth, such as north of the West Indies where very fine clays exist, most of the samples are silty clays whose geotechnical-geophysical properties are dependent on the type of clay minerals present (and their ability to take in moisture), the sand-size fraction, and the quantity of carbonate present. Thus the pure clays have high compressibilities which decrease on the addition of coarse particles, while the converse is true for the acoustic parameters, these increasing with the sand fraction. Using the notion of the intrinsic compression line for all samples, and comparison to it of the measured compression curves, it is clear that, contrary to some previously held ideas, most deep-sea clays are normally consolidated; the addition of carbonate has the effect of creating an open, stronger sediment skeleton. Interestingly, where information is available, the variation with depth of a sample's acoustic velocity follows the void ratio pressure relationship of the compression curve. This allows the construction of an in-situ sediment compression curve using the in-situ geophysical observations.     

Flow Cytometric Analysis of Coastal Lagoon Bacterioplankton and Picophytoplankton: Fixation and Storage Effects

The effect of different fixation and storage protocols on the flow cytometric (FCM) simultaneous analysis of bacterioplankton and phytoplankton in coastal seawater samples (Mediterranean coastal lagoons) was investigated. FCM measurements (cell number, fluorescence and scatter characteristics) were obtained through DAPI staining. Three fixatives [glutaraldehyde (GA), formaldehyde (FA) and paraformaldehyde (PFA)] and two storage (3 months duration) methods (5 °C and −196 °C) were tested. Two dominant populations were detected in studied samples: bacteria and eukaryotic picophytoplankton. Adding fixatives (2% final concentration) appears necessary to obtain FCM exhaustive counts of all the bacteria and phytoplanktonic cells. This was related to the permeation effect of fixatives which allowed a better DAPI staining of the cells. Maximum fluorescence, i.e. optimal staining of the cells was obtained with FA or PFA, and significant lower fluorescences with GA. Fixed samples stored at 5 °C induced rapid cell loss. Only storage in liquid nitrogen of samples fixed with FA or PFA, allows mid-term (≥4 months) preservation of bacteria or picophytoplankton cell numbers, and limited evolution of DAPI-induced fluorescence and scatter characteristics.     

Phylogenetic diversity and phenotypic characterization of cultivable bacterioplankton isolated from polar oceans

A set of 27 marine planktonic bacteria isolated from the polar regions was characterized by 16S rDNA sequencing and physiological and biochemical testing. More than half of these bacteria were positive for caseinase, gelatinase and 13-glucosidase, and could utilize glucose, maltose or malic acid as carbon source for cell growth. Twelve isolates expressed nitrate reduction activities. Except for one antarctic isolate BSwlO175 belonging to Actinobacteria phylum, these isolates were classified as γ-Proteobacteria, suggesting that γ-Proteobacteria dominated in cultivable marine bacterioplankton at both poles. Genus Pseudoalteromonas was the predominant group in the Chukchi Sea and the Bering Sea, and genus ShewaneUa dominated in cultivable bacterioplankton in the Prydz Bay. With sequence similarities above 97%, genus Psychrobacter was found at both poles. These 27 isolates were psychrotolerant, and significant 16S rDNA sequence similarities were found not only between arctic and antarctic marine bacteria （ 〉 99% ）, but also between polar marine bacteria and bacteria from other aquatic environments （ ≥ 98.8% ）, including temperate ocean, deep sea, pond and lake, suggesting that in the polar oceans less temperature-sensitive bacteria may be cosmopolitan and have a bipolar, even global, distribution at the species level.