姜黄素新型纳米乳药动学与生物等效性研究

建立高效液相色谱法(HPLC)测姜黄素新型纳米乳(Curcumin novel nano emulsion,CNNE)中姜黄素(Curcumin,Cur)在大鼠体内的血药浓度,通过比较相同给药计量下Cur和CNNE在大鼠体内的血药浓度考察CNNE的药代动力学行为,并比较二者生物等效性。采用水滴法制备CNNE,灌胃给予CNNE与Cur后,采用HPLC法测定Cur的血药浓度,绘制药物浓度物-时间曲线(AUC),DAS2.1.1软件计算药代参数与生物等效性。CNNE与Cur的房室模型AUC(0-∞)分别为(3 439.27±180.63)(μg·h)/L、(482.07±42.14)(μg·h)/L,CNNE与Cur的非房室模型AUC(0-∞)分别为(3 410.20±154.09)(μg·h)/L、(446.66±44.02)(μg·h)/L,CNNE的相对生物利用度为713.44%、763.49%。结果表明,将Cur制备成CNNE后促进了Cur的吸收,CNNE的生物利用度提高了约7倍。CNNE与Cur具有生物不等效性。     

双去甲氧基姜黄素与姜黄素的药代动力学比较

考察了SD大鼠口服灌喂给予双去甲氧基姜黄素和姜黄素药代动力学特征。将雄性SD大鼠12只随机分为2组,分别灌服双去甲氧基姜黄素和姜黄素混悬液后,于不同时间点大鼠眼底静脉丛取血,HPLC法测定血浆中双去甲氧基姜黄素和单体姜黄素的浓度,DAS 2.1.1药动学软件计算药动学参数。结果显示,双去甲氧基姜黄素和单体姜黄素的主要药动学参数Tmax为0.75 h和0.25 h;Cmax为(62.90±2.64)μg/L和(61.64±4.30)μg/L;AUC0~72 h为(281.75±3.61)μg/(L·h)和(171.79±32.18)μg/(L·h);AUC0~∞为(291.31±15.15)μg/(L·h)和(190.46±43.81)μg/(L·h)。表明双去甲氧基姜黄素和单体姜黄素相比有更好的生物利用度。     

格列吡嗪控释片在Beagle犬体内的药动学和相对生物利用度研究

目的建立高效液相色谱-串联质谱(HPLC-MS/MS)测定血浆中格列吡嗪的方法,研究格列吡嗪控释片在Beagle犬体内药动学和相对生物利用度。方法 8只Beagle犬双周期随机交叉单剂量口服10 mg受试格列吡嗪控释片和市售格列吡嗪控释片,HPLC-MS/MS测定血药浓度,计算两者的药动学参数及相对生物利用度,并进行生物等效性评价。结果单次口服10 mg格列吡嗪控释片受试制剂和参比制剂的主要药动学参数AUC0-32h分别为2940.3±947.1,3195.5±1294.3μg/L·h;Cmax分别为298.85±153.24,282.51±122.37μg/L;Tmax分别为6.8±1.8,8.0±4.0 h;平均滞留时间(MRT0-32h)分别为12.5±2.9,11.9±2.5 h;终末半衰期(t1/2z)分别为12.4±7.0,7.2±5.7 h。结论两种制剂具有生物等效性。     

HPLC-MS/MS法测定大鼠血浆中酸枣仁皂苷A含量及其药动学研究

建立高效液相色谱-质谱串联(HPLC-MS/MS)法检测大鼠血浆中酸枣仁皂苷A的含量,并应用于药动学研究。大鼠静注(4 mg/kg)或灌胃(20 mg/kg)给药,不同时间取血,血浆样品经处理后,采用HPLC-MS/MS法测定酸枣仁皂苷A含量,色谱柱为Waters YMCTM ODS-AQ S-5 120A(2.0×100 mm)分析柱,以电喷雾离子化串联质谱(ESI)及多反应监测扫描模式(MRM)进行检测,流动相为甲醇-水(0.1%甲酸)=50∶50,流速为0.3 m L/min,柱温为30℃,根据测定结果求算药物的药动学参数。酸枣仁皂苷A在40 ng/m L~4 000 ng/m L的浓度范围内线性关系良好(r2=0.999 1),各浓度水平的精密度及稳定性的RSD均小于15%,提取回收率均大于85%。大鼠注射给药后主要药动学参数分别为Ke0.28/h,t1/22.55 h,AUC0→t2 839.89 h·ng/m L,AUC0→∞3 201.51 h·ng/m L,灌胃给药后主要药动学参数分别为Ke0.51/h,t1/21.35 h,AUC0→t206.02 h·ng/m L,AUC0→∞211.13 h·ng/m L。经计算,酸枣仁皂苷A在大鼠体内的生物利用度为1.32%。本试验所建立的检测方法快速简便、精密度好、灵敏度高及稳定性好,适用于酸枣仁皂苷A在大鼠体内血药浓度测定及药代动力学的研究。     

紫杉醇长循环脂质体的制备及其药动学研究

目的制备紫杉醇长循环脂质体并观察其在大鼠体内的药动学.方法用薄膜分散法制备紫杉醇长循环脂质体;HPLC法测定血浆中紫杉醇的药物浓度;用3P97程序计算药动学参数.结果脂质体粒径为(106.5±1.7)nm,包封率为(87.5士2.5)%,紫杉醇长循环脂质体和紫杉醇注射液的血药浓度-时间曲线均符合二室模型,药动参数t1/2β分别为(7.32±1.24)h和(0.32±0.06)h,AUC分别为(23.85±1.20)μg·h·mL-1和(5.08±0.45)μg·h·mL-1.结论与紫杉醇注射液相比,紫杉醇长循环脂质体显著延长了紫杉醇在血液循环系统中的驻留时间,具有长效作用.     

复方甲巯咪唑软膏在大鼠体内的药动学研究

目的建立大鼠血清及甲状腺组织中甲巯咪唑(methimazole)的高效液相色谱(HPLC)测试方法及液相色谱-串联质谱(LC-MS/MS)测试方法,并用于研究复方甲巯咪唑软膏在大鼠体内的药动学性质。方法将240只Wistar大鼠按体重随机分成4组:复方甲巯咪唑软膏高、中、低剂量(分别为25,9,1.5 mg/kg,按甲巯咪唑含量计)组及单方甲巯咪唑软膏(9 mg/kg)组。分别于颈部皮肤涂抹给药后不同时间点取血清与甲状腺组织(n=6)。采用HPLC与LC-MS/MS两种方法测定大鼠血清与甲状腺组织中的甲巯咪唑浓度,DAS2.1.1软件统计药动学参数。结果大鼠单次颈部涂抹复方甲巯咪唑软膏高、中、低剂量及单方甲巯咪唑软膏后甲巯咪唑血清AUC_(0-12h)分别为38.24±7.12,10.37±3.10,0.25±0.07,15.32±4.96 mg/(L·h);C_(max)分别为9.50±1.43,2.88±0.43,0.07±0.01,3.78±0.36mg/L。大鼠单次颈部涂抹复方甲巯咪唑软膏高、中、低剂量及单方甲巯咪唑软膏后甲巯咪唑在甲状腺组织中的AUC_(0-12h)分别为221.91±42.74,117.17±25.22,75.23±12.75,139.28±35.31 ng/(mg·h);C_(max)分别为63.29±9.60,32.34±10.20,20.71±6.09,42.33±10.71 ng/mg。结论甲巯咪唑在大鼠体内呈现线性动力学特征。相同剂量下,复方制剂与单方制剂靶部位暴露水平接近,复方制剂中氢化可的松对甲巯咪唑的药动学特征影响小,两者可配伍协同发挥药效。     

莫达非尼片在健康人体的药代动力学研究

目的在中国健康男性志愿者中进行莫达非尼片的药动学研究。方法 30名男性健康志愿者随机分为3个剂量组:单次口服给药低、中、高剂量(100,200,400mg)组;多次口服给药组,每天给药1次,每次200 mg,连续给药7 d。采用高效液相色谱法(HPLC)测定血浆中莫达非尼浓度。应用DAS2.0软件进行数据处理,求算药动学参数。结果莫达非尼片单次口服给药低、中、高剂量组主要药动学参数AUC0-t分别为(33.49±9.31),(82.52±20.88),(173.72±38.60)mg·h/L;AUC0-∞分别为(36.05±10.51),(87.70±24.42),(189.33±53.81)mg·h/L;Cmax分别为(2.44±0.35),(5.45±0.93),(9.90±1.13)mg/L;tmax分别为(2.35±0.85),(2.35±1.00),(2.70±0.89)h;t1/2分别为(14.08±3.31),(14.52±2.48),(15.99±3.59)h;多次口服给药组主要药动学参数分别为:AUCss为(88.60±28.84)mg·h/L;AUC0-t为(124.80±48.71)mg·h/L;AUC0-∞为(136.53±60.80)mg·h/L;Css_max为(6.99±1.68) mg/L;Css_min为(1.98±1.02)mg/L;tmax为(1.95±0.96)h;t1/2为(16.26±3.03)h。单次口服给药3个剂量药动学参数AUC0-∞,Cmax,t1/2z,CLz,Vz经方差分析后组间差异无统计意义(P0.05);多次口服给药达稳态时的AUC0-t,AUC0-∞与单次给药相比有所升高,差异具有统计学意义(P0.05)。结论单次口服莫达非尼片在中国健康男性受试者体内的代谢规律符合线性药动学特征。     

双去甲氧基姜黄素与姜黄素纳米乳药代动力学的比较研究

制备并考察双去甲氧基姜黄素纳米乳和单体姜黄素纳米乳的药代动力学特征。雄性SD大鼠口服灌胃分别给予双去甲氧基姜黄素纳米乳和单体姜黄素纳米乳后,于大鼠眼底静脉丛取血,采用HPLC法测定血浆中单体双去甲氧基姜黄素和姜黄素的浓度,DAS 2.1.1药动学软件计算药动学参数。得到双去甲氧基姜黄素纳米乳和单体姜黄素纳米乳主要药动学参数Tmax为1.50h和1.00 h;C_(max)为(85.87±2.53)和(85.60±2.30)μg/L;AUC0~72 h为(788.23±52.04)和(1 345.50±64.88)μg/L·h。表明姜黄素纳米乳的口服生物利用度比双去甲氧基姜黄素纳米乳更好。     

优化鸡粪微生态分解及添加降解液促进微藻过量合成生物油脂

以小球藻(Chlorella protothecoides)为出发菌株,优化鸡粪微生态分解及添加降解液促进微藻细胞生长和胞内油脂合成。通过控制溶解氧(DO)优化鸡粪微生态分解,获得高氮和低氮两种降解液。微藻培养过程分为细胞生长(0~120 h)和油脂合成(120~200 h)两个阶段,前期添加高氮降解液促进细胞生长,后期添加低氮降解液和葡萄糖补充氮源及能量过量合成油脂,实现细胞浓度和生物油脂产率最大化。C.protothecoides培养期间,0~120 h恒速流加(15 m L/(L·D))高氮降解液,120~200 h流加(15 m L/(L·D))低氮降解液和葡萄糖(1.5 g/(L·D)),培养结束(200h)时,油脂质量浓度高达5.28 g/L,分别比对照及不添加葡萄糖时油脂质量浓度增加74.8%和21.1%。结果表明,两阶段鸡粪降解液添加与葡萄糖再生ATP结合策略促进油脂合成是有效的。     

液相色谱-串联质谱法测定大鼠体内凤仙萜四醇苷B血药浓度及其药动学研究

目的建立液相色谱-串联质谱法(LC-MS/MS)测定大鼠血浆样品中凤仙萜四醇苷B浓度的方法,并进行药动学研究。方法取6只大鼠,尾静脉单剂量注射凤仙萜四醇苷B 1.0 mg/kg,分别于给药前和给药后2,5,20,40,60,90,120,180 min眼眶隐静脉丛采血,采用LC-MS/MS测定其血药浓度,并采用DAS计算药动学参数。结果凤仙萜四醇苷B在大鼠体内的主要药动学参数t_(1/2z)为0.518±0.114 h,C_(2min)为159.11±9.17 ng/m L,AUC0-t为142.86±23.53 ng·h/m L。结论本方法选择性好、灵敏度好、准确度高,可为下一步该化合物临床前药动学研究提供方法学依据。     

天门冬酰胺酶基因在黑曲霉中的同源表达

根据NCBI中的黑曲霉天门冬酰胺酶基因asp序列(GI:145231287)设计特异性引物,从黑曲霉CICC2462基因组中扩增asp基因编码区,构建其黑曲霉表达载体p SZHG-Asp。通过农杆菌介导法转化黑曲霉,筛选以asp基因置换糖化酶基因gla A的同源重组转化子。对转化子的表达产物进行SDS-PAGE分析和酶活检测。获得8株在gla A位点发生基因置换的同源重组转化子,并对其中4株的上清液进行检测。SDS-PAGE结果显示,在42 000处有目的蛋白质条带,重组菌株的目的蛋白质表达量为185~417μg/m L,发酵液最高酶活为21 111 U/m L。     

天冬酰胺酶纳米微球体外活性和稳定性

制备自组装天冬酰胺酶(Asparaginase,AAS)透明质酸-聚乙二醇(Hyaluronic acid-graftpoly(ethylene glycol),HA-g-PEG)/羟丙基-β-环糊精(Hydroxypropyl-beta-cyclodextrin,HPCD)纳米微球(self-assembly HA-g-PEG/HPCD hollow nanospheres loaded with AAS,AHHPs),并对AHHPs的体外活性及稳定性进行初步研究。制备并测定AHHPs的透射电镜、粒径、Zeta电位和包封率。再分别从最适温度、最适pH、热稳定性、贮存稳定性、酸碱稳定性、抗胰蛋白酶水解能力和血浆稳定性初步考察AHHPs的体外活性和稳定性。测得AHHPs的平均粒径为(367.43±2.72)nm,Zeta电位为(-15.70±1.25)mV,平均包封率为(66.03±3.81)%。AHHPs的最适温度为50℃,最适pH值为7.0;游离AAS的最适温度为60℃,最适pH值为7.5。体外稳定性的结果显示,同样条件下,AHHPs的体外稳定性明显比游离AAS的好。因此AHHPs能有效提高AAS的体外活性和体外稳定性。     

丙烯酰胺抑制剂对曲奇面团流变学和烘焙特性的影响

通过引入天冬酰胺酶和/或甜菊苷到曲奇配方中替代部分糖以抑制其生产过程中丙烯酰胺的生成,分析单独或同时添加这两种配料时曲奇面团的动态流变学特性、硬度和曲奇饼干的烘焙感官特性。结果表明：当单独添加天冬酰胺酶(1000 ASNU)时可降低曲奇样品中天冬酰胺含量(0.045mg/g)的67%,从而抑制95%丙烯酰胺的生成,且不会影响曲奇产品的烘焙特性。而天冬酰胺酶和甜菊苷同时添加时可抑制样品中96%丙烯酰胺的生成。动态流变学结果表明：引入天冬酰胺酶不会影响曲奇面团的流变学性质,而甜菊苷的加入会增加面团的弹性模量G＇和黏性模量G〃,这是因为甜菊苷替代面团中糖成分后促进曲奇面团面筋网络的形成。但是,部分糖被取代后会改变曲奇的一些烘焙特性,如：水分含量增加,曲奇饼干颜色变淡,延展率和破碎力降低。感官分析结果表明：感官评定者不能接受45%和60%糖取代量的曲奇,但可以接受15%或30%糖取代量的曲奇。     

Reliable identification of grapevine rootstock varieties using RAPD PCR on woody samples

Since grapevine ( Vitis spp .) rootstock material is being traded increasingly as disbudded woody material a lack of distinctive morphological features on such material necessitates an alternative and reliable means of identification. Methods described here were developed for rapid and efficient extraction of DNA from woody samples rich in phenolic compounds and polysaccharides, and for subsequent identification of varieties by RAPD PCR. Using these methods, and with the application of only one selected RAPD primer, we were able to differentiate sixteen rootstock varieties, including the seven varieties most commonly used in Germany. Problems commonly encountered with reproducibility of RAPD patterns were avoided by choosing primers with a dinucleotide sequence and a high G/C content that allowed a rather high annealing temperature of 45°C. Methods described here should also be useful for other horticultural crops, especially those with woody tissues rich in phenolic compounds and polysaccharides.     

An internet compendium of analytical methods and spectroscopic information for monomers and additives used in food packaging plastics

An internet website (http://cpf.jrc.it/smt/) has been produced as a means of dissemination of methods of analysis and supporting spectroscopic information on monomers and additives used for food contact materials (principally packaging). The site which is aimed primarily at assisting food control laboratories in the European Union contains analytical information on monomers, starting substances and additives used in the manufacture of plastics materials. A searchable index is provided giving PM and CAS numbers for each of 255 substances. For each substance a data sheet gives regulatory information, chemical structures, physico-chemical information and background information on the use of the substance in particular plastics, and the food packaging applications. For monomers and starting substances (155 compounds) the infra-red and mass spectra are provided, and for additives (100 compounds); additionally proton NMR are available for about 50% of the entries. Where analytical methods have been developed for determining these substances as residual amounts in plastics or as trace amounts in food simulants these methods are also on the website. All information is provided in portable document file (PDF) format which means that high quality copies can be readily printed, using freely available Adobe Acrobat Reader software. The website will in future be maintained and up-dated by the European Commission's Joint Research Centre (JRC) as new substances are authorized for use by the European Commission (DG-ENTR formerly DGIII). Where analytical laboratories (food control or other) require reference substances these can be obtained free-ofcharge from a reference collection housed at the JRC and maintained in conjunction with this website compendium.     

Aroma characterization of various apricot varieties using headspace–solid phase microextraction combined with gas chromatography–mass spectrometry and gas chromatography–olfactometry

The characterization of the aromatic profile of several apricot cultivars with molecular tracers in order to obtain objective data concerning the aromatic quality of this fruit was undertaken using headspace–solid phase microextraction (HS–SPME). Six apricot cultivars were selected according to their organoleptic characteristics: Iranien, Orangered, Goldrich, Hargrand, Rouge du Roussillon and A4025. The aromatic intensity of these varieties measured by HS–SPME–Olfactometry were defined and classified according to the presence and the intensity of grassy, fruity and apricot like notes. In the six varieties, 23 common volatile compounds were identified by HS–SPME–GC–MS. Finally, 10 compounds, ethyl acetate, hexyl acetate, limonene, β-cyclocitral, γ-decalactone, 6-methyl-5-hepten-2-one, linalool, β-ionone, menthone and (E)-hexen-2-al were recognized by HS–SPME–GC–O as responsible of the aromatic notes involved in apricot aroma and considered as molecular tracers of apricot aromatic quality which could be utilized to discriminate apricot varieties.     

Tests of potential functional barriers for laminated multilayer food packages. Part I: Low molecular weight permeants

The advent of the functional barrier concept in food packaging has brought with it a requirement for fast tests of permeation through potential barrier materials. In such tests it would be convenient for both foodstuffs and materials below the functional barrier (sub-barrier materials) to be represented by standard simulants. By means of inverse gas chromatography, liquid paraffin spiked with appropriate permeants was considered as a potential simulant of sub-barrier materials based on polypropylene (PP) or similar polyolefins. Experiments were performed to characterize the kinetics of the permeation of low molecular weight model permeants (octene, toluene and isopropanol) from liquid paraffin, through a surrogate potential functional barrier (25 μm-thick oriented PP) into the food simulants olive oil and 3% (w/v) acetic acid. These permeation results were interpreted in terms of three permeation kinetic models regarding the solubility of a particular model permeant in the post-barrier medium (i.e. the food simulant). The results obtained justify the development and evaluation of liquid sub-barrier simulants that would allow flexible yet rigorous testing of new laminated multilayer packaging materials.     

Chlorohydrins of bisphenol A diglycidyl ether (BADGE) and of bisphenol F diglycidyl ether (BFDGE) in canned foods and ready-to-drink coffees from the Japanese market

BADGE.2HCl and BFDGE.2HCl were determined in 28 samples of ready-to-drink canned coffee and 18 samples of canned vegetables (10 corn, 5 tomatoes and 3 others), all from the Japanese market. HPLC was used as the principal analytical method and GCMS for confirmation of relevant LC fractions. BADGE.2HCl was found to be present in one canned coffee and five samples of corn, BFDGE.2HCl in four samples of canned tomatoes and in one canned corn. No sample was found which exceeded the 1mg/kg limit of the EU for the BADGE chlorohydrins. However the highest concentration was found for the sum of BFDGE.2HCl and BFDGE.HCl.H₂O at a level of 1.5mg/kg. A Beilstein test confirmed that all cans containing foods contaminated with BADGE.2HCl or BFDGE.2HCl had at lest one part coated with a PVC organosol.     

European priorities for research to support legislation in the area of food contact materials and articles

A strong science base is required to underpin the planning and decision-making process involved in determining future European community legislation on materials and articles in contact with food. Significant progress has been made in the past 5 years in European funded work in this area, with many developments contributing to a much better understanding of the migration process, and better and simpler approaches to food control. In this paper this progress is reviewed against previously identified work-areas (identified in 1994) and conclusions are reached about future requirements for R&D to support legislation on food contact materials and articles over the next 5 or so years.