Set-up State Demonstration Center of Experimental Teaching for Life-science by Education Reforming and Construction

该文介绍厦门大学生命科学学院创建“国家级生命科学实验教学示范中心”的实践经验。2000年以来,厦门大学生命科学学院教学改革与建设并举,全面进行教学管理体制改革,建立了“实验教学中心”的实验教学管理新体制;同时集中投入教学经费,购置先进仪器,改善教学实验室条件。“实验教学中心”以提高学生实验技术和科研能力为目标,综合改革实验教学体系。经过5年实践,培养出实验技能扎实的生物学研究型基础人才,取得一批实验教学成果。The experience for constructing the state demonstration center for life-science experimental teaching in Xiamen University was introduced in this paper.Since 2000,the School of Life Sciences in Xiamen University had reformed the administration system for experimental teaching and founded the experimental teaching center.Meanwhile,The school concentrated the education funds to improve the laboratory conditions and gain many modern instruments.Furthermore,The experimental teaching center integrated experimental courses to reform the experimental teaching system for enhancing the experimental technique and scientific research ability of students.After five years of practice,the School of Life Sciences in Xiamen University had educated many foundation talents with proficient biological techniques and bear some fruits in experimental teaching.国家基础科学人才培养基金项目(批号:J0530124

荧光猝灭响应的溶解氧有机改性溶胶-凝胶传感膜的构筑

利用二甲基二甲氧基硅烷为有机改性剂,制备溶胶-凝胶,并通过对氧荧光猝灭指示剂4,7-二苯基-1,10-邻菲咯啉钌(Ru(dpp)3(ClO4)2)的包埋,构筑以Ru(dpp)3(ClO4)2为荧光指示剂的氧传感敏感膜.利用蓝光发光二极管(LED,λmax=475nm)为激发光源,考察了氧敏感膜的荧光行为,并以此进行了水体中溶解氧含量的检测.传感膜对水溶液溶解氧的最低检出限为0.2μg/mL,测定的相对标准偏差为2%,线性范围为0.5~16μg/mL,响应时间(t95)为60s,敏感膜使用寿命大于10个月

BOD Measurement Based on Oxygen Fluorescent Quenching Rate Method

利用氧荧光猝灭速率的方法,结合自行构建的BOD光纤传感装置进行海水中生化需氧量(BOD)含量检测。考察了四种筛选的海洋耗氧菌种在四甲基硅氧烷(TMOS)、二甲基二甲氧基硅烷(Di Me-DMOS)和聚乙烯醇(PVA)包埋固定情况下,对不同浓度的葡萄糖-谷氨酸(GGA)标准溶液的荧光响应情况。BOD敏感膜的荧光响应在0·2~30mg/L浓度范围内呈良好的线性关系,对2mg/L标准溶液测定的相对标准偏差为2·5%(n=6),响应时间(t95%)为4·0min,BOD敏感膜使用寿命大于12个月。实际海水样品检测表明,利用BOD敏感膜检测得到的结果与国标BOD5方法之间存在较好的一致性。Based on oxygen fluorescent quenching rate method,a BOD optical sensing film immobilized four kinds of microorganisms from seawater in an Ormosils-PVA matrix was applied for BOD measurement of seawater. The responding characteristic of fluorescent quenching is researched. The BOD sensing film contributed a good linear relationship between concentration and fluorescent response in the concentration range of 0.2 to 30mg/L. Measurements were taken for 4min followed by 10min recovery time in 2mg/L glucose/glutamate (GGA) BOD standard solution. Some other experimental conditions, such as pH, measurement temperature, the concentration of sodium chloride, were studied. BOD values estimated by this optical BOD sensing film correlate well with those determined by the conventional BOD_5 method for seawater samples.国家863计划(2001AA635100);; 福建省自然科学基金(D0410001)资助项

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

Fluorescent Characters of Optical BOD Sensing Film

利用自行构建的BOD光纤传感装置对海水中BOD含量进行了检测,考察了海洋耗氧菌种在四甲 基硅氧烷(TMOS)、二甲基二甲氧基硅烷(DiMe-DMOS)和聚乙烯醇(PVA)包埋情况下,对BOD的荧光响应 行为以及耗氧菌种的加入量对BOD浓度的响应情况。传感膜对5 mg·L-1 BOD溶液测定的相对标准偏差为 1.2％(n=6),响应时间为3．2 min,敏感膜使用寿命大于12个月。实际海水样品检测表明,利用BOD光纤 传感检测的结果与国标BOD5方法,存在较好的一致性。We characterized an optical BOD sensing film used to immobilize three kinds of microorganisms from seawater. The sensing film for BOD measurement consists of an optical oxygen sensor and the microorganisms immobilized on poly( vinyl alcohol) ormosils matrix. The fluorescent responses of the BOD sensing film were obviously affected by the thickness of the oxygen film and the concentration of microorganisms in the film. Some other experimental conditions, such as pH, measurement temperature, and the concentration of sodium chloride, were studied. BOD values estimated by this optical BOD sensing film correlate well with those determined by the conventional BOD5 method for seawater samples.国家自然科学基金委员会与香港研究资助局联合科研基金 国家“863”计划海洋领域课题(2003AA635100)资助项