低场核磁共振技术在鱼用冻干疫苗保护剂筛选中的应用

为筛选适合鱼类冻干疫苗中使用的保护剂,采用低场核磁共振技术(LF-NMR)对冻干后不同保护剂中的水分分布和迁移规律进行了分析。根据水中氢质子的弛豫时间,可将保护剂中的残留水分为结合水、不易移动水和自由水3种形态。结果显示:冷冻干燥过程中二次干燥温度对甘露醇和蔗糖中残留水的分布有较大影响,随着温度从4℃升高到37℃,大部分自由水解吸而小部分转化为结合水。当冻干后的样品暴露在相对湿度43%和温度25℃的环境中时,大部分保护剂吸收水分,并将其固定为结合水,而甘露醇中的部分结合水则转变为自由水,并解吸到环境中。不同冻干保护剂中的水分分布和迁移并不相同,代表其具有不同的水分结合能力。该研究结果有助于鱼用冻干疫苗中保护剂的筛选和冻干工艺的优化,以便获得更加稳定的疫苗产品。厦门海洋高技术产业基地创业创新共享服务平台项目(16PFW008SF15);;福建省高职院校海洋生物应用技术协同创新中心(2017);;“十三五”北海市海洋经济创新发展示范项目(Bhsfs003,Bhsfs009);;2010海洋公益行业科研专项(201005020-1

Determination of Squalene by Ultra Performance Liquid Chromatography

应用超高效液相色谱法,建立了角鲨烯的测定方法。采用uPlC bEH C18(2.1MMx50MM,1.7μM)色谱柱考察了流动相有机相比例、流速及柱温对角鲨烯样品分离的影响,确定了最佳色谱条件:等度洗脱,流动相为甲醇-水(95∶5,体积比),流速0.4M l/MIn,柱温35℃,检测波长210nM。该方法下,角鲨烯在0.2-10μg/M l范围内浓度和面积呈现良好的线性关系,相关系数r为0.9998;检测限为0.1ng,定量限分别为0.25ng,精密度试验rSd(n=6)为0.36%,平均回收率为100.82%(rSd=0.99%)。本方法快速、简单、可靠、灵敏、重复性好,可用于角鲨烯有关样品的快速检测。The article is researching determination of squalene by Ultra performance liquid chromatography(UPLC).The analysis was performed on an Acquity UPLC BEH C18 column(2.1mm×50mm,1.7μm).The influence of flow rate,the proportion of organic phase in mobile phase,and column temperature on the separation of squalene was comprehensively studied.The optimal separation condition was as follows:a mobile phase consisting of methanol and water(95∶5,v/v) with a isocratic elution profile,UVdetection wavelength at 210 nm,0.4 m L/min of flow rate for mobile phase,35℃ of column temperature.On the above condition,good linear was observed in the linear range of 0.2-10 μg/m L,and the correlation coefficient r was 0.9998.The limits of detection(S/N=3) were 0.1ng and the limits of quantification(S/N=10) were 0.25 ng.The average recovery rates was 100.82%(RSD=0.99%).The RSD of repetition was 0.36%(n=6).This method was simple,accurate and sensitive with a good reproducibility.It is suitable for fast detection of squalene.厦门海洋研究开发院共建项目(2014); 海洋生物技术产业化中试技术研发公共服务平台(12PZP001SF10); 广东海洋经济发展区域示范项目(GD2012-D01-001

Studies on determination of glucosamine by HPLC-ELSD

目的:建立氨基葡萄糖含量测定的高效液相色谱分析法。方法:样品采用高效液相色谱进行测定,使用Waters Carbohydrate色谱柱,流动相为乙腈-水=70:30,用蒸发光散射检测器(ELSD)检测。结果:方法的进样量线性范围为2.0～12.0μg,相关系数为0.9997,回收率为96.0%～97.7%。结论:此法简便、快速、准确,无需衍生反应,是测定氨基葡萄糖含量的有效方法,并可用于硫酸氨基葡萄糖含量的测定。Objective:A HPLC method for the determination of glucosamine was developed.Methods:The content of glucosamine was determined by HPLC on a water carbohydrate column(4.6 mm×250 mm,5 μm) with acetonitrile-water(70:30) as mobile phase and ELSD detector.Results:The linear range of calibration curve was 2.0～12.0 μg(r=0.9997),the recovery was 96.0%～97.7%.Conclusion:The method is simple,rapid,accurate and without derivation.It is a effective method for determination of glucosamine and can be used for determination of glucosamine sulfate.厦门市重大科技项目(3502Z20051025

Research progress of integrated membrane process on high-value development of fish processing by-products

集成膜工艺不仅可以发挥各种膜过程的优势,而且利用集成的协同作用,能够更高效地解决水产加工副产物高值化开发中的分离纯化难题,同时具有节能、低耗、清洁等优点。文章分析了集成膜工艺在水产加工副产物高值化开发中的典型应用,并探讨了实现集成膜工艺产业化应用的关键环节。Integrated membrane process,making use of both individual and synergic advantages of membrane operations,provided more effective separation solution for high-value development of fish processing by-products and had merits of energy saving,low consumption,clean production and so on.Reviewed the typical applications of integrated membrane process on high-value development of fish processing by-products,further discussed the key issues of achieving the industrial application of integrated membrane process.福建省海洋与渔业厅重点项目(闽海渔合同[2010]2-21号);厦门海洋研究开发院项目(K11102

基于二维石墨烯纳米材料优化高分子分离膜的研究进展

高分子分离膜在渗透性与选择性之间一直存在此长彼消的trade-off效应,且抗污染性能、化学稳定性能也亟待提升。有机-无机杂化膜能综合有机高分子材料和无机材料的优点,近年来,纳米材料的快速发展对于传统高分子膜材料具有重要的推动作用,尤其是基于二维石墨烯纳米材料开发新型复合分离膜成为备受关注的研究热点。然而,石墨烯化学稳定性高,其表面呈惰性状态,与其它介质的相互作用较弱,且石墨烯片层之间存在较强的范德华力,易聚集而难溶于水及膜溶剂,严重阻碍了其在高分子分离膜材料中的应用。氧化石墨烯(GO)在其表面和边缘引入了大量含氧极性基团,有助于摆脱了片层间强大的π-π堆积相互作用力,因此,GO在水及膜溶剂中具有良好分散性能;同时,大量的含氧基团也为设计与制备改性石墨烯提供了丰富的反应位点;此外,GO还具有可规模化制备、成本较低的优点,使得GO在高分子分离膜材料中的应用备受青睐。针对聚偏氟乙烯、聚砜与聚醚砜等常用膜材料疏水性强、易污染的缺点,将它们分别与GO、改性GO、或复合纳米材料等共混,通过浸没沉淀相转化法制备混合基质膜,可以有效改善混合基质膜的亲水性、膜孔结构、膜表面粗糙度、荷电性能等,从而提升混合基质膜的渗透性能与抗污染性能,甚至赋予抑菌等新功能。在聚酰胺复合膜的超薄分离层或多孔支撑层中引入适量的GO或改性GO,通过增强超薄分离层的亲水性能、荷电性能以及优化超薄分离层的结构,从而提升纳米复合膜的选择渗透性能、抗污染性能以及耐氯性能。此外,利用GO的静电、氢键、范德华力、π-π等非共价键相互作用力,或者利用GO活性位点与交联剂反应实现共价键连接,可以通过层层组装法制备水平取向、高效堆叠的高通量GO层状膜。本文归纳了基于物理共混、界面聚合、层层组装等常用制膜方法,将二维石墨烯纳米材料填充、交联、吸附、沉积而负载或包裹在高分子膜基质中,实现改善与优化高分子分离膜结构与性能的应用研究进展,并对二维石墨烯纳米材料在高分子分离膜的工业化应用所面临的挑战和应用前景进行了展望。厦门市产学研协同创新及科技合作项目(3502Z20172008);;海洋三所基本科研业务费专项资金资助项目(2016036);;厦门海洋高技术产业基地创业创新共享服务平台项目(16PFW008SF15);;海洋中试技术研发与检测公共服务平台建设(Bhsfs009);;厦门大学校长基金(No.20720170027

Development of Anhydrotetrodotoxin Certifi ed Reference Material

研制4,9-脱水河豚毒素国家标准样品。以河豚鱼卵巢为原料,提取制备4,9-脱水河豚毒素,采用红外光谱(Ir)、高分辨质谱和核磁共振谱(nMr)进行结构确证。样品分装成140瓶后,采用柱后衍生–高效液相荧光法进行均匀性、稳定性检验和定值分析。从样品中随机抽取15瓶进行均匀性检验,经f检验表明在95%的置信区间范围内样品均匀性良好;稳定性考察按照40℃加速试验稳定性(6个月)进行,结果表明在考察期间内样品稳定性良好;标准样品经国内8家具有分析资质的实验室进行协同定值,并评定了定值结果的不确定度,4,9-脱水河豚毒素标准样品定值结果为97.77%,相对扩展不确定度为0.4%(k=1.96)。该标准样品达到国家标准样品的技术要求,可用于有关脱水河豚毒素的方法校正和质量控制。4,9-anhydrotetrodotoxin certified reference material was developed.4,9-anhydrotetrodotoxin was made from the ovarium of pufferfish.The structure of anhydrotetrodotoxin certified reference material was comfirmed by mass spectrometry and NRM,infrared spectroscopy(IR).4,9-anhydrotetrodotoxin was divided into 140 bottles.The homogeneity and stability testing and quantitative analysis were evaluated by post column derivation high performance liquid chromatography–fl uorescent detection(HPLC–FLD) method.According to analysis procedure of homogeneity,15 bottles of sample were randomly taken from 140 bottles,and the results were validated by F-test statistical method.The stability inspection was carried on the short-term(6 months),and the results indicated that the period for anhydrotetrodotoxin of storage was 6 months at 40℃.A cooperative certifi cation was conducted with 8 qualifi ed laboratories.The certifi ed purity value of the reference material of 4,9-anhydrotetrodotoxin was 97.77% with relative expaned uncertainty of 0.40%(k=1.96).The reference material can conform to the technical requirement of the certified reference material.The material was intended for use in the method validation and quality control regarding anhydrontetrodotoxin.广东海洋经济发展区域示范项目(GD2012-D01-001); 海洋生物产业化中试技术研发公共服务平台(12PZP001SF10); 厦门市重大科技创新平台项目(3502Z20111001); 国家科技支撑计划(NO.2011BAK04B09

基于二维石墨烯纳米材料优化高分子分离膜的研究进展

高分子分离膜的渗透性与选择性之间存在此长彼消的Trade-off效应,且其抗污染性能、化学稳定性能也亟待提升。有机-无机杂化膜综合了有机高分子材料和无机材料的优点。近年来,纳米材料的快速发展对传统高分子膜材料具有重要的推动作用,尤其是基于二维石墨烯纳米材料开发的新型复合分离膜成为备受关注的研究热点。然而,石墨烯化学稳定性高,其表面呈惰性状态,与其他介质的相互作用较弱,且石墨烯片层之间存在较强的范德华力,易聚集而难溶于水及膜溶剂,严重阻碍了其在高分子分离膜材料中的应用。氧化石墨烯(GO)在石墨烯的表面和边缘引入了大量的含氧极性基团,有助于摆脱片层间强大的π-π堆积相互作用力,因此,GO在水及膜溶剂中具有良好的分散性能;同时,大量的含氧基团也为设计与制备改性石墨烯提供了丰富的反应位点;此外,GO还具有可规模化制备、成本较低的优点,使得GO在高分子分离膜材料中的应用备受青睐。针对聚偏氟乙烯、聚砜与聚醚砜等常用膜材料疏水性强、易污染的缺点,将它们分别与GO、改性GO或复合纳米材料等共混,通过浸没沉淀相转化法制备混合基质膜,可以有效改善混合基质膜的亲水性、膜孔结构、膜表面粗糙度、荷电性能等,从而提升混合基质膜的渗透性能与抗污染性能,甚至赋予其抑菌等新功能。在聚酰胺复合膜的超薄分离层或多孔支撑层中引入适量的GO或改性GO,通过增强超薄分离层的亲水性能、荷电性能以及优化超薄分离层的结构,从而提升纳米复合膜的选择渗透性能、抗污染性能以及耐氯性能。此外,利用GO的静电、氢键、范德华力、π-π等非共价键相互作用力,或者利用GO活性位点与交联剂反应实现共价键连接,可以通过层层组装法制备水平取向、高效堆叠的高通量GO层状膜。本文归纳了基于物理共混、界面聚合、层层组装等常用制膜方法,将二维石墨烯纳米材料填充、交联、吸附、沉积,从而负载或包裹在高分子膜基质中,实现改善与优化高分子分离膜结构与性能的应用研究进展,并对二维石墨烯纳米材料在高分子分离膜的工业化应用中所面临的挑战和前景进行了展望。厦门市产学研协同创新及科技合作项目(3502Z20172008);;\n海洋三所基本科研业务费专项资金(2016036);;\n厦门海洋高技术产业基地创业创新共享服务平台项目(16PFW008SF15);;\n海洋中试技术研发与检测公共服务平台建设(Bhsfs009);;\n厦门大学校长基金(20720170027)~

Determination of Astaxanthin by High Performance Liquid Chromatography with Photodiode Array Detector

建立虾青素含量测定的高效液相色谱–光电二极管阵列法。采用PurOSPHEr STAr rP 18(4.6 MMx250MM,5μM)色谱柱,以甲醇–水(体积比为95∶5)为流动相,流速1.0 Ml/MIn,检测波长为482 nM,柱温为30℃,进样量为20μl。在所选定的液相色谱条件下,虾青素主峰与其它杂质峰分离良好,虾青素在0.2~16μg/Ml范围内线性良好,线性相关系数r=0.999 9,检出限为0.01μg/Ml,测定结果的相对标准偏差为0.42%(n=6),平均回收率为100.4%。该法分析快速准确、灵敏度高、重现性好。A method using high-performance liquid chromatography(HPLC) coupled with photodiode array detector(PDA) was established for determination of astaxanthin.The separation was performed on Purospher STAR RP 18 column(4.6 mm×250 mm,5μm) with the mobile phase of methanol–water(Volume ratio was 95∶5).The detector was photodiode array detector and the variable wavelength detector(VWD) was set at 482 nm.The column temperature was 30℃ and the injection volume was 20 μL.The calibration curve was linear in the range of 0.2–16 μg/mL,and the correlation coefficient was 0.999 9.The relative standard deviation of determination results was 0.42%(n=6).The average recovery was 100.4%, and the detection limit was 0.01 μg/mL.This method is characteristic of rapidity,accuracy,high sensitivity and good reproducibility.海洋生物产业化中试技术研发公共服务平台项目(厦海渔合[2013]19号

Development of Trehalose Certified Reference Material

研制了海藻糖国家标准样品。以食品级海藻糖粗品为原料,纯化制备海藻糖,采用红外光谱(Ir)、质谱和核磁共振谱(nMr)以及单晶衍射等方法进行结构确证。样品分装成400瓶后,采用离子色谱法进行均匀性、稳定性检验和定值分析。从样品中随机抽取15瓶进行均匀性检验,经f检验表明,在95%的置信区间范围内,样品均匀性良好。在40℃下,经过24个月稳定性考察,结果表明样品稳定性良好。标准样品经国内8家具有分析资质的实验室进行协同定值,并评定了定值结果的不确定度,海藻糖标准样品定值结果为99.72%,扩展不确定度为0.26%(k=1.96)。该标准样品达到国家标准样品的技术要求,可用于有关海藻糖的方法校正和质量控制。Trehalose certified reference material was developed.Trehalose was purified from the crude product.The structure of trehalose certified reference material was comfirmed by infrared spectroscopy(IR),mass spectrometry,NRM and XRD.Trehalose was divided into 400 bottles.The homogeneity and stability testing and quantitative analysis were evaluated by ion chromatography method.According to analysis procedure of homogeneity,15 bottles of sample were randomly taken from 400 bottles,and the results were validated by F-test statistical methods.The stability inspection was carried on the long-term(24 months),and the results indicated that the period for trehalose storage was 24 months at 40 ℃.A cooperative certification was conducted by 8 qualified laboratories.The certified purity value of the reference material of trehalose was 99.72%with relative expaned uncertainty of 0.26%(k=1.96).The reference material can conform to the technical requirement of the certified reference material,and it can be used for validation and quality control regarding trehalose.福建省科技计划项目(2013N01010205); 厦门市海洋经济创新发展区域示范项目(12PZP001SF10); 海洋公益行业科研专项(201005020–1); 厦门市科技计划项目(3502Z20122009