Distribution Characteristics of Polycyclic Aromatic Hydrocarbons in Seawater from the South Yellow Sea

2007年9月进行了2007南黄海海洋环境调查,并采集各个调查站位的表层、中层、底层海水及表层沉积物。利用液-液萃取法对采集到的样品进行前处理,并用气相色谱-质谱法检测海水中的多环芳烃。结果显示,监测站位中表层海水中总多环芳烃的含量范围是15.76--233.39ng/l,南黄海中部海水中多环芳烃的组成以2环的萘为优势组分;各种多环芳烃的含量与国际生物学组织或国家制定的评价水生生物暴露于水体的安全食用标准相比虽未超标,但个别组分如荧蒽的含量已达到生态毒理评价标准。Marine surface sediments and sea water samples from the surface, middle and bottom layers were collected from the South Yellow Sea of China.The samples were pretreated by liquid-liquid extraction and the 16 polycyclic aromatic hydrocarbons (PAHs) in the samples were determined by gas chromatography-mass spectrometry (GC-MS).The results show that the range of total PAHs is 15.76～233.39 ng/L in surface water and PAHs in the water samples from central South Yellow Sea are dominated by naphthalene (2 rings).Content of PAHs compounds don′t exceed the safety guidelines proposed by International Biology Organization or governments, but the individual compound, such as fluoranthene content has reached eco-toxicological evaluation criteria.国家海洋局节能减排专项资助;中国海监技术支撑项目资助;海洋公益性项目资助(200705011

Application of Accelerated Solvent Extraction Technique for Analysis of Active Components in Traditional Chinese Medicinal Herbs

以两种药材为研究实例,对加速溶剂萃取法(ASE)在中药材有效成分提取研究中的应用进行了简要介绍。采用正交试验法考察了提取丹参中丹酚酸B的提取条件(萃取温度、静态萃取时间、萃取溶剂以及料液比),确定了较好的实验条件。比较了ASE、水蒸气蒸馏法、超声波提取法及索氏提取法对木香挥发油的提取效果,结果表明ASE对木香挥发油的提取效果最好。The application of accelerated solvent extraction(ASE) technique for the Analysis of active components in traditional Chinese medicinal herbs was introduced by using two kinds of herbs as examples.The conditions including extraction temperature,static extraction time,the ratio of material to solvent and solvent of ASE for extraction of salvianolic acid B in Salvia miltiorrhiza were optimized by orthogonal experiments,and the optimal conditions were obtained.Different extraction methods(ASE,steam distillation,ultrasonic wave and Soxhlet extraction) were used to extract volatile oil in Aucklandia lappa Decne.Results of the comparative experiments indicated that ASE was the most effective method in this case.All the results from these studies demonstrate that ASE is indeed a powerful tool in the preparation of herbal extracts for downstream chromatographic analysis.青岛“2004将才计划”(04-3-JJ-11);; 共建生物医药研发测试中心(LS-05-KJZX-76)资

Content of gentiopicroside and loganic acid in Radix gentianae and their fingerprints

To develop a HPLC-DAD-ESI-TOF/MS analysis method for the determination of gentiopicroside and loganic acid in Radix gentianae samples and for the research of their fingerprints.The samples were extracted using ASE for 10 min under 100 ℃ and 9.65 MPa,and divided into water phase and chloroform phase and analyzed them with HPLC-DAD-ESI-TOF/MS method respectively.Based on this method,the HPLC fingerprints of Radix gentianae were established.Comparing the spectrogram and mass spectrum of the chromatogram peak with the reference value,three compounds in water phase were identified as gentiopicroside,asafetida acid and loganic acid.There is no report of the compounds in chloroform phase.The content of gentiopicroside and loganic acid in samples of different groups were determined,separately.The fingerprints were compared by the software of the similarity evaluation system for chromatographic fingerprint.The water phase fingerprint congruence coefficients of samples from six different areas were above 0.90,however,the chloroform phase fingerprint congruence coefficients were within 0.62-0.99.This method can be used for determination of potent component in Radix gentianae and its quality control.Radix gentianae from different producing areas have the largest diversities,and the diversities embodied in the content of chloroform phase compounds.青岛“2004将才计划”(04-3-JJ-11);; 国家海洋局青年基金资助项目(2005602);; 崂山区政府区校共建生物医药研发测试中心资助(LS-05-KJZX-76)

Determination of Gentiopicroside and Loganic Acid in Radix Gentianae by MEKC and MEEKC Mode

目的建立胶束电动毛细管色谱(MEKC)和微乳液毛细管电动色谱(MEEKC)分析龙胆药材中龙胆苦苷和马钱子苷酸含量的方法。方法采用加速溶剂萃取法(ASE)对龙胆药材进行提取,萃取温度:100℃,压力:9.65MPa,萃取时间:10min。采用未涂层熔融石英毛细管(内径75μm,有效长度50cm)。分别考察了两种分离模式下电泳介质的构成和电泳过程中的各操作参数对样品分离过程的影响,优化了MEKC和MEEKC的分析条件,在各自对应的缓冲液体系下,MEKC和MEEKC分离电压分别为30和22kV,柱温均为25℃,检测波长均为238nm。结果在选定的工作条件下,龙胆苦苷和马钱子苷酸与其他组分达到了基线分离,两种成分的浓度与其响应信号值之间具有较好的线性相关性,加标回收率在96.3%～105.1%之间,检测限均低于10mg·L-1,对6处不同产地的龙胆药材进行了分析,并对测定结果进行了t检验,结果表明,两种模式下,测定结果之间不存在显著性差异,而不同产地的龙胆药材的龙胆苦苷和马钱子苷酸含量之间存在较大差异。结论本方法简便,准确,快速,重现性较好,可用于龙胆药材有效成分的含量测定和质量控制。OBJECTIVE To develop MEKC and MEEKC modes for the determination of gentiopicroside and loganic acid in extracts of Radix Gentianae. METHODS The analyte was extracted from Radix Gentianae samples by accelerated solvent extraction, and the extraction conditions were optimized. Separation and determination were carried out on a bared fused silica capillary(50 cm×75 μm) with corresponding buffer. The run voltage of MEKC and MEEKC were 30kV and 22 kV respectively. Detection wavelength of DAD was at 238 nm and column temperature was 25 ℃. RESULTS The developed ASE-HPCE method was simple and reliable for the determination of gentiopicroside and loganic acid in Radix Gentianae samples with a broad linear dynamic range, a recovery range of 96.3%～105.16%,and the detection limit was below 10 mg·L-1. The contents of gentiopicroside and loganic acid in six samples from different regions were determined by the developed method in two modes. T-test value indicated that the determined contents of gentiopicroside and loganic acid by MEKC and MEEKC were consistent. CONCLUSION The method is simple,accurate,rapid and with good reproducibility.It can be used to determine active components in Radix Gentianae.国家自然科学基金重点项目(20235020);; 青岛“2004将才计划”(04-3-JJ-11);; 崂山区校区共建生物医药研发测试中心资助(LS-05-KJZX-76

Comparison of Recoveries of PAHs Obtained by Ultrasound-assisted Extraction from Wet and Freeze-dried Marine Sediment

以美国环保局(EPA)优先控制的16种多环芳烃(polycyclic aromatic hydrocarbons,PAHs)为研究对象,建立了湿法超声波辅助萃取技术提取沉积物中痕量PAHs的可靠方法,对萃取剂、萃取功率、萃取次数和萃取时间进行了优化。并与传统干法超声辅助萃取进行了比较,湿法超声波辅助萃取的PAHs回收率为57%～125%,而干法超声波辅助萃取PAHs的回收率为48%～113%,对相对分子质量小的PAHs如萘～芴的回收率,湿法明显高于干法。该文建立的湿法超声辅助萃取适用于沉积物和泥土中痕量PAHs的快速测定。An ultrasound-assisted extraction method for the extraction of 16 trace PAHs(USEPA priority pollutant) from wet(i.e.untreated) marine sediment was developed.The effects of solvent type,power of ultrasound,and extraction time were studied and optimized.The developed method was compared with the traditional ultrasound-assisted extraction for freeze-dried sediment.The results showed that recoveries of sixteen PAHs extracted from the wet marine sediment ranged from 57% to 125%,whereas those extracted from the freeze-dried marine sediment ranged from 48% to 113%.The results also indicated that the recoveries of PAHs with smaller relative molecular mass(e.g.naphthalene and fluorine) extracted from the wet sediment were higher than those from the freeze-dried sediment.Thus the developed method was appropriate for rapid determination of trace PAHs in marine sediment and soil.海洋公益性行业科研专项(200705011);; 中国海监技术支撑体系项目;; 2008年海洋环境保护及节能减排专

Concentration and distribution of polycyclic aromatic hydrocarbons in surface sediments collected in the Southern Yellow Sea

采用气相色谱与质谱联用(gC/MS)技术,在一个航次内对南黄海表层沉积物中16种优先监控的PAHS的污染状况进行了调查,采用菲/蒽、荧蒽/芘、荧蒽/(荧蒽+芘)、吲哚芘/(吲哚芘+苯并(g,H,I)苝)等特定比值对PAHS来源进行了分析.结果表明,南黄海表层沉积物中检出PAHS的总含量为90.4~732.65ng·g-1,各站点均以4~6环为主;与其它站位相比,倾废区的H01站位受到PAHS污染较为明显,无论是16种PAHS总量还是高分子量组分最高值都出现在该站点,虽然该海区沉积物中PAHS的含量没有超出生物影响低值,但苯并(b)荧蒽、吲哚芘和苯并(g,H,I)芘等一些没有最低安全标准的PAHS也有不同程度的检出,对海洋生物具有潜在的毒副作用.PAHS可能来源于原油、生物和煤燃烧造成的污染.The concentrations of sixteen representative Polycyclic Aromatic Hydrocarbons(PAHs) in surface sediment samples collected from the Southern Yellow Sea were determined by GC-MS.The distribution and potential sources of PAH contamination in the region were investigated.The types of PAHs found in the surface sediments were mainly 4～6 ring PAHs,and they are unevenly distributed among the different sampling sites covering an area of 139 km2.The area is known for continuous waste dumping activities in recent years,and indeed sites adjacent to such activities were found to have high levels of PAHs.Some high molecular weight PAH compounds such as benzo$$b$$fluoranthene,dibenzo$$a,h$$anthracene and benzo$$g,h,i$$perylene were detected in some samples.The health implication of these contaminants needs careful assessment since toxicity guidelines are not available for these species.The potential sources of PAHs are discussed using various source-specific PAH indexes such as Phenathrene/Anthracene、Fluoranthene/Pyrene、Fluoranthene/(Fluoranthene+Pyrene) and Indeno [1,2,3-cd] pyrene/(Indeno[1,2,3-cd] pyrene+Benzo [g,h,i] perylene).The composition of the observed PAHs reflects that they originate from both petroleum utilization and incomplete combustion.海洋公益性行业科研专项(No.200705011);中国海监技术支撑体系项目;2008年海洋环境保护及节能减排专项---

Application of oil spill identification by GCMS——A case study

通过对中国近海海域一个具体海面溢油事例的研究,提出了通过气相色谱及气质联用技术进行海面溢油鉴别的方法。采用GC-FID对油样中的正构烷烃的主要组分进行定性分析,利用GC-MS对油中难降解的生物标记化合物:霍烷及其他五环三萜类化合物、规则甾烷及重排甾烷类化合物、三芳甾类化合物、多环芳烃进行分析,计算相应的比值并采用"t-分布"方法进行溢油样品和可疑溢油源样的相关分析。结果如下:(1)从GC-FID色谱图可以看出,YY2679与KY0631、KY0632的色谱图极其相似,与其余四个可疑油源的色谱图差异较大。六个可疑油源的R值在0.04~1.95之间,只有KY0631和KY0632的R值小于0.1,KY0633~KY0636与YY2679的差异不是由风化引起的,可以排除,由此可以推断出YY2679溢油样品可能来源于KY0631或KY0632两个可疑溢油源。(2)采用21个生物标志物指标进行相关性判别,KY0631可疑溢油源在比值点的区间范围为95%和98%时,有多个点x或y误差棒均未跨过直线y=x。KY0632可疑溢油源在比值点的区间范围为95%时,各点的x或y误差棒均跨过直线y=x,由此推断YY2679溢油样品和可疑油源KY0632是同一油源。On the basis of a case study,a method to identify the spilled oil at sea was introduced.N-alkane distribution patterns of unknown oil were recognized by gas chromatography-mass spectrometry determined the distribution of a suit of petroleum biomarkers hopanes,triterpanes,regular steranes diasteranes,triaromatic steroids and polycyclic aromatic hydrocarbons.The calculated diagnostic ratios for selected biomarker in the source of the spilled oil were done.The correlation studies compared the spill sample and suspected source were made.The results showed three samples nearly identical GC chromatographic profiles,n-alkane distribution patterns,as well as nearly identical diagnostic ratios.This implies that they were most likely from the same source.In 95%confidence limit,all diagnostic ratios of sample KY0632 is exactly the same as for YY2679,so conclusion that the sample of KY2679 and YY0632 are come from the same source

大孔吸附树脂分离纯化龙胆药材中龙胆苦苷和马钱子苷酸的研究

目的:建立利用大孔吸附树脂对龙胆药材中龙胆苦苷和马钱子苷酸进行富集和分离纯化的方法。方法:采用加速溶剂萃取法对龙胆药材中两种有效成分进行高效提取,比较了D301,AB-8,D101,XDA-1四种大孔树脂对龙胆苦苷和马钱子苷酸的吸附性能,最终确定采用D301型大孔对脂对二者进行富集吸附,对其工艺参数进行优化,全程采用高效液相色谱进行目标化合物浓度检测。结果:化优后的工艺参数为:上样浓度:0.2 g/mL,最大上样量:0.25 g龙胆药材/g树脂,最佳静态吸附时间:8 h,采用8%和55%的乙醇溶液对龙胆苦苷和马钱子苷酸分别进行洗脱;龙胆苦苷和马钱子苷酸分别富集在8%和55%的乙醇洗脱液中,洗脱液浓缩后冷冻干燥,可得到纯度分别为74.3%和80.9%的粗产物,龙胆苦苷和马钱子苷酸的回收率分别为70.11%和67.82%。结论:此法效率较高,操作简便,即可用于实验室制备少量的难以购置的标准品,也可进行放大研究,用于工业生产。国家自然科学基金重点项目(20235020);; 青岛“2004将才计划”(04-3-JJ-11);; 共建生物医药研发测试中心(LS-05-KJZX-76)资

Identification of Bohai Sea crude oil based on characteristic fingerprinting by MSPD-RP-HPLC

建立渤海原油的高效液相色谱(HPlC)特征指纹图谱,为渤海原油的鉴别提供一种新方法。以氧化铝作为基质固相分散剂进行样品处理,采用HPlC法进行分析,双反相C18色谱柱串联,以乙腈-水为流动相,二元线性梯度洗脱,流速为1.0 Ml/MIn,柱温为20℃,双波长检测226 nM、254 nM,进样量10μl;采用所发展的方法对10批渤海原油样品进行分析,建立渤海原油的HPlC特征指纹图谱,结合相似度分析用于渤海原油的鉴别。该方法具有很好的精密度、重复性和稳定性,10批渤海原油特征指纹图谱有13个共有峰,结合相似度分析可以用于渤海原油样品的鉴别,说明HPlC特征指纹图谱是渤海原油鉴别的有力工具。Based on the high performance liquid chromatography(HPLC) characteristic fingerprinting,a new method for crude oil identification of Bohai Sea was developed.The sample pretreatment was done using matrix solid-phase dispersion,and Al2O3 was used as the dispersant.The chromatographic fingerprints were determined by injecting 10 L sample solution each time on two reverse phase C18 columns in series with the gradient elution solvent system composed of water and acetonitrile.The flow rate was 1.0 mL/min,the column temperature was maintained at 20℃ and the signals were acquired at 226 nm and 254 nm.The characteristic fingerprints of crude oil were established by analysis of ten representative samples derived from Bohai Sea,and identification was carried out by the similarity analysis of the characteristic fingerprints.The method was ideally suited for the fingerprinting of Bohai Sea crude oil with good precision,repeatability and stability.13 peaks were selected as the common peaks of the characteristic fingerprint and used for the similarity analysis,which could be applied to distinguish crude oil of Bohai Sea from that of other sources.The characteristic HPLC fingerprinting technique thus was a powerful tool for the identification of crude oil originated from Bohai Sea.海洋公益性行业科研专项(200705011;201005034-3);中国海监技术支撑体系项目;2008年海洋环境保护及节能减排专

Analysis of Triterpene Saponins in Semen Aesculi by High Performance Liquid Chromatography-Electrospray-Time of Flight-Mass Spectrometry

采用高效液相色谱/电喷雾飞行时间质谱联用技术(HPLC/ESI-TOF/MS),研究4种七叶皂苷的分子结构与裂解规律间的关系,并对娑罗子中的七叶皂苷类化合物进行鉴定。实验采用反相C18色谱柱,以乙腈-0.2%乙酸溶液为流动相,二元线性梯度洗脱,通过与电喷雾飞行时间质谱联用获得娑罗子中各皂苷成分的精确分子量和分子式;采用质谱碰撞诱导解离技术获得各化合物碎片裂解信息,结合文献对娑罗子中的14种皂苷类化合物进行了初步鉴定。研究表明,高效液相色谱-电喷雾飞行时间质谱联用技术是娑罗子中皂苷类化合物鉴别的有效工具。A convenient and user-friendly method based on high performance liquid chromatography(HPLC)and positive ion electrospray-time of flight-mass spectrometry(ESI-TOF/MS)has been developed for analysis and characterization of triterpene saponins in extract of Semen Aesculi.Samples were analyzed by a reversed-phase C18 column using a binary eluent of acetonitrile and water(with 0.2% acetic acid)under gradient conditions.The[M+H]+ or [M+Na]+ ions of saponins in Semen Aesculi extract were observed by ESI-TOF/MS on-line detection,and used to obtain accurate molecular weight and molecular formula information of saponins.Fourteen saponins in Semen Aesculi extract could be primary identified by accurate molecule weight information and fragmentation behavior obtained by collision induced dissociation experiment combined with literature review.This method has the advantages of simple operation,rapid measurement and accurate determination and it is a powerful tool for identification of saponins in Semen Aesculi.国家自然科学基金重点项目(No.20235020);; 青岛“2004”将才计划(No.04-3-JJ-11);; 共建生物医药研发测试中心(No.LS-05-KJZX-76)资