Study on HPLC Fingerprint of Chinese Marine Medicine Dried Scallop

目的建立海洋中药干贝高效液相色谱(HPlC)指纹图谱分析方法 ,为其质量控制和药材鉴别提供依据。方法采用反相AllTIMA-C18柱,水(A)和乙腈(b)为流动相,梯度洗脱,流速为0.8Ml/MIn,检测波长210nM,柱温20℃,进样体积30μl;采用高效液相色谱二极管阵列检测器-质谱联用技术(HPlC-dAd-MS)对其特征峰进行快速分析;采用相似度计算软件用于10批干贝药材的相似度评价。结果初步建立了干贝药材的HPlC指纹图谱,10批干贝药材指纹图谱有9个共有峰,相似度均在0.85以上;HPlC-dAd-MS分析获得的4个特征峰的紫外光谱和质谱信息,可用于干贝药材指纹图谱特征峰以及共有峰的准确确定。结论该方法准确、简单、稳定性好,为干贝药材的真伪鉴别及质量控制提供了科学依据。Objective To establish a chromatographic fingerprint of Dried Scallop,a kind of Chinese marine medicine,by high performance liquid chromatography (HPLC),and apply for quality assessment and control of medicine materials of Dried Scallop.Methods Separation was performed on an Alltima-C18 (250 mm × 4.6 mm,5μm),with mobile phase consisting of water (A) and acetonitrile (B) with gradient elution at the flow rate of 0.8 ml/min.The UV detection wavelength was 210 nm,column temperature was 20℃.Characteristic peaks of the fingerprint could be rapidly analyzed by high performance liquid chromatography-mass spectrometry (HPLC-MS),and the software of similarity analysis was used to calculate the similarity of all the Dried Scallop samples.Results All samples tested contained the same nine peaks,and the similarities of 10 batches of Dried Scallop were more than 0.85.MS and UV spectra of the 4 characteristic peaks obtained using HPLC-MS could be used to accurately confirm the characteristic and common peaks in fingerprint of Dried Scallop.Conclusion This method is simple,accurate and stable,and can provide scientific reference for the identification and quality control of Dried Scallop.国家自然科学基金(No.20602009;20905017);海洋公益性行业科研专项(No.200705011;200805039

Evaluation of DPPH Free Radical Scavenging Capability of Hippocampus japonicus Kaup. and Their HPLC Fingerprint

目的采用体外1,1-二苯基苦基苯肼(dPPH)抗氧化模型对海马提取物的抗氧化性质进行评价,并建立小海马HPlC特征指纹图谱,用于小海马药材的鉴别及质量评价。方法利用离线dPPH抗氧化评价体系对海马不同提取物进行评价,结果表明,海马水提物抗氧化能力最强,在此基础上又探明了海马水提物抗氧化能力随时间和浓度的变化规律,为海马抗氧化活性提供了科学依据。依据抗氧化活性实验结果,建立了海马水提物HPlC特征指纹图谱分析方法。结果海马水提物大部分化合物达到基线分离,方法的精密度、重现性、稳定性良好;建立小海马药材HPlC指纹图谱,采用中药指纹图谱相似度计算软件,对小海马进行真伪辨别和质量评价。结论表明该方法简捷、有效,是小海马药材鉴别及质量控制的有效方法。OBJECTIVE To evaluate the antioxidant activity of the extracts of Haima,and to establish the HPLC fingerprint of Haima for the discrimination and the quality evaluation of Haima.METHODS Antioxidant properties of Haima extracts with different solvent were assayed in terms of antioxidant activity by scavenging activities on 1,1-diphenyl-2-picrylhdrazyl(DPPH).The effect of time and concentration of Haima extract on the antioxidant activity was also studied.Secondly,a chromatographic fingerprint method was developed.RESULTS The antioxidant activity of water extract of Haima was higher than all the other extracts of Haima.The developed HPLC method was simple,accurate and reliable for the development of Haima fingerprint.Ten Haima samples collected from different medicine store were analyzed and the Haima HPLC fingerprint was established.The similarity of the HPLC chromatogram was performed for authentication and quality control of Haima.CONCLUSION The HPLC fingerprinting techniques have high potential in authentication or source-tracing types of applications.国家自然科学基金重点项目(20235020);青岛市共建生物医药研发测试中心资助项目(LS-05-KJZX-76

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

Simultaneous determination of 7 nucleosides in Asterias rollestoni using reversed-phase high performance liquid chromatography

建立了罗氏海盘车中7种核苷化合物的反相高效液相色谱分析测定方法。采用超声波辅助提取,选用两根不同的C18色谱柱串联,以甲醇和0.2%(体积分数)乙酸/水溶液为流动相梯度洗脱分离。优化的色谱条件为:柱温为室温,检测波长为260nM,流速为0.8Ml/MIn,进样量为20μl。结果表明,7种核苷化合物在一定的浓度范围内线性关系良好,次黄嘌呤和胸苷的线性范围为0.65~40Mg/l,尿苷、黄嘌呤和肌苷的线性范围为0.80~40Mg/l,胸腺嘧啶的线性范围为1.15~40Mg/l,鸟苷的线性范围为0.50~40Mg/l。样品中7种核苷化合物的加标回收率为90.00%~105.00%,相对标准偏差为0.72%~3.23%。该方法操作简便、灵敏度高、重复性好,回收率高,适用于罗氏海盘车中7种核苷类成分的同时分析,也可用于罗氏海盘车的质量控制和综合评价。A method for the simultaneous determination of 7 nucleosides in Asterias rollestoni was devel-oped using reversed-phase high performance liquid chromatography ( RP-HPLC) .Analytes were extracted by ultrasonic-assisted extraction and separated on two different C18 columns,which were connected in se-ries,under the gradient elution with the mobile phases of methanol and 0.2% ( v/v) acetic acid/water at room temperature.The chromatographic conditions were as follows: flow rate,0.8 mL/min; detection wavelength,260 nm; injection volume,20 μL.Under the optimized conditions,good linear relationships between the values of mass concentrations and the peak areas of hypoxanthine,uridine,xanthine,thy-mine,inosine,guanosine and thymidine were observed in the ranges of 0.65-40,0.80-40,0.80-40,1.15-40,0.80-40,0.50-40,and 0.65-40 mg/L,respectively.The relative standard devia-tions were around 0.72%-3.23% and the recoveries were around 90.00%-105.00%.The results showed that the developed method is sensitive,accurate and reproducible.It is suitable for the analysis of nucleosides in Asterias rollestoni with high recoveries and it is expected to be used for the quality control and evaluation of Asterias rollestoni.国家自然科学基金项目(20905017);海洋公益性行业科研专项(200705011;200805039);海洋局青年基金项目(2010140);海洋一所基本科研业务专项(GY-022008T32;2010G25);中国科学院实验海洋生物学重点实验室开放基金课

Primary Exploration on Quality Evaluation of Holothurian Sold on the Market by HPLC Fingerprint

以10批不同产地刺参HPlC指纹图谱中的6个共有峰为评价指标,结合相似度分析,对不同刺参质量进行评价和检验。结果表明:不同批次刺参样品的6个色谱峰在指纹图谱分析过程中色谱行为相同,峰面积大,特征性强,能反映刺参的固有化学特征。结合相似度分析,可用于刺参质量评价。An optimized high performance liquid chromatography (HPLC) method was developed for the analysis of holothurian and the HPLC fingerprint was established from 10 batches of the holothurians from different habitats.The HPLC fingerprint showing 6 common characteristic peaks was used to explore the quality evaluation of holothurian and distinguish from the fakes with the similarity analysis.This method is accurate and reliable,providing a scientific basis for the quality control of holothurian and can be used to evaluate the quality of holothurian sold on the market.我国近海海洋综合调查与评价908专项(908-02-05-04):海洋药用生物资源评价和《中华海洋本草》编纂;海洋一所基本科研业务专项(GY-022008T32):海洋绿藻活性成分分析、鉴

Determination of five poisonous elements in holothurian sold on the market by Microwave Digestion with ICP—MS

建立了微波消解—电感耦合等离子体质谱法测定海参中Cu,AS,Cd,Hg,Pb 5种有毒元素含量的方法,采用In元素为内标,利用其回收率对以上5种元素的测定结果进行校正,该方法变异系数(CV%)为0.2%--4.0%,In元素加标回收率为96.7%--104.5%,说明该方法灵敏度高,结果准确可信。利用该方法对10批不同市售海参中5种有毒元素含量进行了测定,结果表明10批市售海参中的AS含量均超标,部分批次海参中的Cd、Pb超标,说明市售海参确实存在有毒元素超标的问题。A method based on microwave sample digestion and inductively coupled plasma-mass spectrometry(ICP—MS) detection was established for determination of Cu,As,Cd,Hg and Pb in holothurian sold on the market.Indium was used as internal standard element to calibrate the results of determination of these five elements.Determination limits of these five elements were in the range of 0.006～0.063 μg/g,coefficient of variation(CV%)were in the range of 0.2%～4.0%,and the recovery of In was in the range of 96.7%～104.5%.The method has been applied to determine the five poisonous elements in ten holothurian samples sold on the market.The results indicated that the over-standard problem of poisonous elements in holothurian sold on the market existed.The content of total arsenic was over standard in all the ten samples,but the over standard problem of lead and cadmium was only found in part of the samples.国家自然科学基金(20675021);中国近海海洋综合调查与评价908专项(908-02-05-04)资

塔城南湖草场自然资源考察

有关单位通过野外考察和室内分析研究，基本查清了新疆塔城南湖地区的水、土、气、草、林等主要自然资源的数量、质量、分布规律、成因类型和畜牧业现状；并对这些自然资源进行了类型比较，绘制了十一幅相应的类型图、资源图和分区图，撰写了十三篇考察报告；此外，对水、土、草、林等资源的合理利用和改良建设提出了初步的规划设想意见。它对指导当前以种草植树为中心的改良建设工作，尽快改变生产面貌具有重要价值

Applications of HPLC-MS in the Rapid Identification of Components in Marine Medicinal Organisms

高效液相色谱-质谱联用(HPlC-MS)技术,将HPlC的高分离效能与MS的强大结构鉴定功能结合起来,已逐渐成为天然产物化学成分快速分离、鉴别的强有力手段。本研究采用高效液相色谱-电喷雾质谱联用(HPlC-ESI-MS)技术,成功建立了刺参皂苷快速鉴别方法,并对市售干刺参中的4种皂苷进行了鉴别。此外,应用该技术对罗氏海盘车中的13种化学成分进行了快速鉴别。为海洋药用生物化学成分快速鉴别提供了1种值得推广的新方法,为更好的开发利用海洋天然药物奠定了基础。In recent years,high performance liquid chromatography-mass spectrometry combined the high separation efficiency of HPLC with rapid identification function of MS had become a powerful tool for the rapid separation and identification of chemical components in natural products.In the present study,a high performance liquid chromatography-electrospray ionization mass spectrometry(HPLC-ESI-MS) method was developed for rapid identification of 4 saponin components in dry Holothurian sold on the market.On the other hand,13 compounds in Asterias rollestoni were also identified by HPLC-ESI-MS.The HPLC-MS method presented in this paper for the rapid identification of marine compounds was worth popularizing and to lay a foundation for the further exploration of marine natural products.国家自然科学基金项目(20675021;20905017);中国近海海洋综合调查与评价908专项项目(908-02-05-04);海洋局青年基金资

Study on the antioxidant activity of extracts from Enteromorpha prolifera

目的通过探讨浒苔提取物体外的抗氧化活性,为浒苔综合开发利用提供科学依据。方法采用AbTS自由基清除法对浒苔不同溶剂提取物的抗氧化活性进行评价,对不同浒苔样品水提物的抗氧化活性进行比较,并以咖啡酸和抗坏血酸作为阳性对照品对浒苔水提物的抗氧化能力进行量化。结果浒苔水提物的抗氧化活性最高,1Ml浒苔水提物溶液(浓度相当于5.0g新鲜浒苔提取后定容至100Ml)的抗氧化能力与1Ml2.55x10--(-3)Mg·Ml--(-1)的咖啡酸或1Ml2.89x10--(-3)Mg·Ml--(-1)的抗坏血酸相当,而提取方法、样品干燥与否、浒苔样品的差异性均对浒苔水提物的抗氧化活性有一定的影响。结论绿潮藻浒苔具有较强的抗氧化活性,可作为潜在的抗氧化剂进行开发利用。Objective To investigate the antioxidant activity of Enteromorpha prolifera by ABTS radical scavenging assay in vitro and provide scientific basis for exploitation of Enteromorpha prolifera.Methods The antioxidant activities of extracts by different solvents from Enteromorpha prolifera and water extracts from different Enteromorpha prolifera samples were investigated by ABTS radical scavenging assay.Moreover,the antioxidant ability of the water extract from Enteromorpha prolifera was evaluated by using caffeic acid and ascorbic acid as positive control standards.Results The water extract from Enteromorpha prolifera provided the highest radical scavenging activity,and the antioxidant capability of 1mL of water extract from Enteromorpha prolifera was equivalent to 1mL of 2.55 10--(-3) mg·mL--(-1) caffeic acid or 1mL of 2.89×10--(-3)mg·mL--(-1) ascorbic acid.However,the extraction methods,dry sample or not and different kinds of Enteromorpha prolifera samples had certain influence on the antioxidant activity of water extract from Enteromorpha prolifera.Conclusion There are abundant antioxidant compounds present in the aqueous extract of Enteromorpha prolifera,and it can be exploited as latent oxidation inhibitor.海洋公益性行业科研专项(200705011;200805039);国家自然科学基金(20602009;20905017);海洋一所基本科研业务专项(GY-022008T32;GY-022008G47)资

DEVELOPMENT of A DISCRIMINATION METHOD of BOHAI SEA SIMULATED OIL SPILLS BASED ON STABLE CHARACTERISTIC PEAK RATIO of HPLC

考察室外自然风化对原油液相色谱稳定特征峰分布的影响,提出基于溢油液相色谱稳定特征峰比值结合聚类分析和“T-检验“分析进行模拟溢油鉴别.结果表明,液相色谱稳定特征峰比值结合聚类分析可以初步推测模拟溢油样品来源,进一步采用特征峰比值和“T-检验“分析可以实现模拟溢油样与可疑原油样品的成功判别.The effect of weathering outdoors on the distribution of HPLC characteristic peaks is examined,and a methodology using HPLC stable characteristic peak ratios combined with several statistical data correlation analysis methods is developed for oil differentiation,including clustering analysis and Student′s t-test method.The origins of simulated oil spills can be speculated preliminarily through the method using HPLC stable characteristic peaks ratios combined with clustering analysis.Further,the oil spills and suspected sources can be successfully identified via using HPLC stable characteristic peaks ratios combined with Student′s t-test method.The methodology using reverse phase HPLC stable characteristic peaks ratios combined with multi-statistical analysis is expected to be a powerful tool for the discrimination of oil spill discrimination.海洋公益性行业科研专项(200705011;200805039);国家自然科学基金(20905017);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