槟榔壳多酚组分及抗氧化活性的测定

建立 测定槟榔壳中多种酚类物质的高效毛细管电泳方法,分析不同浓度和不同pH值硼酸缓冲液对10种标准品的分离效果,最后确定最佳缓冲液为0.1 mol/L,pH 9.0的硼酸缓冲液,紫外检测波长为280 nm,分离电压为20 kV。方法简便快速,能在20 min之内将10种酚类物质完全分离开,检测限为0.5～4.5 mg/L。此外,进一步测定了槟榔壳多酚的抗氧化活性,选用3个评价抗氧化能力的指标,即：对DPPH自由基的清除能力、对还原能力以及对ABTS自由基的清除能力。     

茶树酯型儿茶素水解酶鉴定及其检测体系的建立

本试验利用体外酶学方法,结合薄层层析（TLC）、高效液相色谱（HPLC）和液相色谱-串联质谱（LC-MS/MS）分析,首次从茶树中检测到活性较高的酯型儿茶素水解酶（Galloylated Catechins Hydrolase,GCH）的存在。在酯型儿茶素水解酶催化下,酯型儿茶素发生水解反应,生成没食子酸（GA）和非酯型儿茶素。试验确立了酯型儿茶素水解酶的最适检测体系,在2.5mL反应体系中包含0.2mmol/L酯型儿茶素、2.4mmol/L抗坏血酸、粗酶提取液若干（含0.1mg酶蛋白）和0.1mol/L磷酸缓冲液（pH6.5）,在30℃下,反应30min。此外,试验利用硫酸铵分级沉淀、阴离子交换层析和凝胶过滤层析对该酶进行了初步纯化。     

茶氨酸的衍生化及毛细管电泳定量技术

采用胶束电动力学毛细管电泳技术定量茶氨酸,以2,4-二硝基氟苯为衍生化试剂,建立茶氨酸定量的新方法。运行缓冲液为pH9.8的0.03βmol/L四硼酸钠缓冲液体系,分离电压为28kV,分离温度为17℃,在360βnm下进行检测。研究了不同pH、反应时间、反应温度、缓冲液浓度对衍生化反应的影响,确立了衍生化反应的最佳条件。证实了4℃和25℃环境下衍生化产物可以稳定保存5天。测定了茶氨酸衍生化方法的性能指标：线性范围为0.2~5βmmol/L（r²=0.993）,最低检测限为0.05βmmol/L。     

茶多酚对茶食品中还原糖检测方法的影响

为寻找准确测定茶面制品消化产物中还原糖含量的方法，选取了表没食子儿茶素没食子酸酯（EGCG）、没食子酸、原花青素和阿魏酸4种典型酚类物质，分别以单独的酚类、酚类与葡萄糖共混、酚类与淀粉酶解物共混体系为样品，研究酚类物质对3,5-二硝基水杨酸（DNS）法、葡萄糖氧化酶/过氧化物酶（GOPOD）法和荧光辅助糖电泳（FACE）法定量测定还原糖的影响。结果发现，阿魏酸对DNS法无影响，EGCG、没食子酸和原花青素可与DNS反应显色，表明其会影响DNS法的准确性；4种酚类物质均显著降低了GOPOD法测定的葡萄糖结果，而FACE法不受酚类影响且能直观表征淀粉酶解物中低聚还原糖分布。因此，FACE法在测定茶面制品及其酶解消化物中还原糖含量方面有较好的应用价值。     

儿茶素在酸性环境中的稳定性研究

本文研究了儿茶素在pH2.6-6.5各柠檬酸缓冲液中的稳定性。结果表明，儿茶素在pH3.6-6.5之间的缓冲溶液中比较稳定，放置18h后，儿茶素最低回收率为81．9％。将pH2.6、pH3.6、pH4.6、pH5．6的儿茶素溶液的pH值分别调到pH6.5，再放置18h后，儿茶素回收率最高为93．0％，最低可达84．7％，说明大部分的儿茶素在强酸性环境中仅发生非离子键结合的反应。     

分子印迹固相萃取和电喷雾质谱法联用测定茶叶中四种儿茶素

为了对茶叶样品中微量儿茶素进行快速准确分析,利用分子印迹固相萃取(MIPs-SPE)与电喷雾质谱(EIMS)联用技术进行了茶叶中四种儿茶素—表儿茶素(EC)、表没食子儿茶素(EGC)、表儿茶素没食子酸酯(ECG)、表没食子儿茶素没食子酸酯(EGCG)的分离测定。首先以表儿茶素(EC)为模板分子,采用紫外聚合方法合成分子印迹聚合物(EC-MIPs),功能单体为甲基丙烯酸(MAA),交联剂为乙烯二醇二甲基丙烯酸酯(EDMA),引发剂为偶氮二异丁腈(AIBN)。利用EC-MIPs作为固定相制备EC-MIPs-SPE柱,对茶叶样品进行分离萃取后采用EIMS对洗脱液中儿茶素单体进行检测,分别与常规C18和非分子印迹聚合物(NIP)固相萃取方法进行比较。结果表明EC-MIPs-SPE法对儿茶素类单体有特异性识别能力,对茶中主要干扰物质咖啡因、茶碱的结合能力远小于儿茶素分子。最佳萃取条件为:水相上样,用50%(v/v)甲醇/水溶液淋洗除去非特异性结合干扰物,1%(v/v)醋酸的甲醇洗脱特异性结合的四种儿茶素单体。对真实茶叶样品测试发现:经过EC-MIPs-SPE法预富集,可以去除94.2%的咖啡因和全部茶碱干扰,同时可得四个主要儿茶素信号,相对强度分别为:EC(12.1%),EGC(8.2%),ECG(35.4%),EGCG(45.7%)。本法可特异性地识别和分离儿茶素单体,有效消除咖啡因和茶碱干扰,可用于茶叶中微量儿茶素分离鉴定。     

番石榴多酚成分分析及抑菌活性研究

采用UPLC-MS/MS法对番石榴多酚中部分酚类物质进行了定性分析。以抑菌圈直径为评价指标,测定了番石榴多酚的抑菌活性,并以金黄色葡萄球菌为指示菌,对其抑菌稳定性进行了研究。结果表明：番石榴多酚中含有没食子酸、儿茶素、槲皮素、原儿茶酸、山奈素和绿原酸等酚类物质。番石榴多酚对金黄色葡萄球菌、大肠埃希氏杆菌、单增李斯特菌、铜绿假单胞菌、腐败希瓦氏菌和枯草芽孢杆菌具有较好的抑制效果,除了单增李斯特菌和腐败希瓦氏菌的MIC低于5 mg/mL外,其余均低于2.5 mg/mL。番石榴多酚在pH4~5范围内处理后抑菌活性最强,其抑菌活性对温度、紫外照射具有稳定性,低盐度会促进其抑菌活性。     

茶多酚在化妆品中的研究进展

茶多酚又名茶单宁、茶鞣质,是茶叶中所含的一类多羟基酚类化合物的总称,简称为TP(Teaupolyphe- nol)。其主要成分为儿茶素类(黄烷醇类)、黄酮、黄酮醇类、花青素类、酚酸、缩酚酸类及聚合酚类等。其中儿茶素类化合物为茶多酚的主体成分,约占茶多酚总量的65%～80%。儿茶素类化合物主要包括表儿茶素、表没食子儿茶素、表儿茶素没食子酸醋和没食子儿茶素没食子酸醋四种物质。其结构为:     

茶儿茶素的高效液相色谱测定方法

应用高效液相色谱法;研究分离测定茶儿茶素的结果表明,采用茶样沸水浴振荡浸提20分钟,SEP-PAK C_(18)小柱净化试样的前处理方法和μBondapak C_(18)不锈钢柱,二甲基甲酰胺、甲醇、醋酸和水4成分系统流动相,流量、浓度线性梯度洗脱,柱温30℃等色谱条件。能使儿茶素组分在25分钟内得到理想的分离。应用紫外检测器(280nm)检测各儿茶素,最低检出限在5.7×10~(-8)g以下,添加回收试验,回收率在85.13%—97.56%之间。5种儿茶素组分回归方程的相关系数为0.9995—0.999     

不同品种荔枝果皮酚类物质组成及其抗氧化活性比较

为比较不同品种荔枝果皮酚类物质含量、组成及其抗氧化活性差异,用80%甲醇分别提取6个品种荔枝果皮中的酚类物质,测定其总酚和总黄酮含量。采用铁离子还原能力（FRAP）和自由基离子清除能力（ABTS）等方法评价其抗氧化活性。通过高效液相色谱（HPLC）鉴定不同的单体酚并分析其在不同品种荔枝果皮之间的差异。结果表明,不同品种荔枝果皮的酚类物质含量为28.69~68.48 mg/g,不同品种荔枝果皮的酚类物质含量和抗氧化能力存在显著的差异（P<0.05）。酚类物质的含量、FRAP和ABTS抗氧化活性能力均以‘荔枝王’为最高,分别为68.48 mg/g_、546.31 μmol/g和511.25 μmol/g。液相色谱结果鉴定出‘糯米糍’中的8种单体酚,而含量相对较高的是A型原花青素三聚体和原花青素A2。研究结果可为荔枝加工废弃物荔枝果皮的开发利用提供理论依据。     

SSR荧光标记毛细管电泳检测法在水稻DNA指纹鉴定中的应用

 以16个杂交水稻不育系、恢复系及组合为材料,初步建立了水稻品种SSR荧光标记毛细管电泳检测方法。与常规凝胶电泳检测方法相比,荧光标记毛细管电泳检测方法可以读出目标DNA片段的准确大小,检测数据更为精确,检测效率更高。常规凝胶电泳检测方法验证表明,利用SSR荧光标记毛细管电泳检测方法进行水稻品种DNA指纹鉴定,方法可行、结果可靠。     

毛细管柱气相色谱法测定胡椒中有机磷农药残留量

应用毛细管柱气相色谱技术分析胡椒中有机磷农药敌敌畏、乐果和马拉硫磷的残留量。采用弹性石英毛细管柱AC10 （30m×0.32mmx0.25μm）,火焰光度检测器（FPD）。结果表明:样品的色谱图未出现与标准溶液色谱图中敌敌畏、乐果、马拉硫磷相一致的色谱峰,未检出相应农药残留量,3种农药的平均回收率分别为74.6％（RSD=7.9％）、 76.4％（RSD=7.3％）、81.0％（RSD=8.3％）;检出限分别为4.0,5.0,8.0μ·kg-1。分析工作质量控制均符合要求。     

An HPLC Method for Microanalysis and Pharmacokinetics of Marine Sulfated Polysaccharide PSS-Loaded Poly Lactic-co-Glycolic Acid (PLGA) Nanoparticles in Rat Plasma

This study was aimed at developing a sensitive and selective HPLC method with postcolumn fluorescence derivatization for the detection of propylene glycol alginate sodium sulfate (PSS) in rat plasma. Plasma samples were prepared by a simple and fast ultrafiltration method. PSS was extracted from rat plasma with d-glucuronic acid as internal standard. Isocratic chromatographic separation was performed on a TSKgel G2500 PWxL column with the mobile phase of 0.1 M sodium sulfate at a flow rate of 0.5 mL/min. Analyte detection was achieved by fluorescence detection (FLD) at 250 nm (excitation) and 435 nm (emission) using guanidine hydrochloride as postcolumn derivatizing reagent in an alkaline medium at 120 °C. The calibration curve was linear over a concentration range of 1–500 μg/mL, and the lower limit of detection (LLOD) was found to be 250 ng/mL. This validated method was applied successfully to the pharmacokinetic study of PSS and PSS-loaded poly lactic-co-glycolic acid (PLGA) nanoparticles (PSS-NP) in rat plasma after a single intravenous (PSS only) and oral administration (PSS and PSS-NP). Significant differences in the main pharmacokinetic parameters of PSS and PSS-NP were observed. The relative bioavailability of PSS-NP was 190.10% compared with PSS which shows that PSS-NP can improve oral bioavailability.     

离子对色谱法测定茶叶中的茶氨酸

通过添加十二烷基磺酸钠(SDS)为离子对试剂,以乙腈-水(磷酸)为流动相,在200nm的检测波长条件下,建立了一种快速测定茶叶中茶氨酸的分析方法。茶氨酸的保留时间为6.27min;在0.04~20μg范围内,茶氨酸峰面积与进样量之间有良好的线性关系,回归方程为:Y=1.51466×106X+3.28706×105(r2=0.99901);当S/N=3时,最低检测浓度为1.15ng。所建方法快速,简便,准确,可用于茶叶中茶氨酸含量测定。     

Rapid determination of glucose,fructose and sucrose in potato tubers by capillary gas chromatography

Summary A method has been developed for the rapid quantification of glucose, fructose and sucrose in potato tubers by capillary gas chromatography. Baseline separation of trimethylsilyl derivatives of β-D(−) fructose, α-D(+) glucose β-D(+) glucose, myoinositol (internal standard) and sucrose was achieved in under 10 min with a detection range of 10 ng to 3 μg per component. Use of an on-column injector resulted in the retention of a proportion of injected sucrose by the column, despite an oven starting temperature of 140°C. This did not occur with a splitless injection system (inlet temperature 220°C).     

量子点探针应用于粮油中黄曲霉毒素免疫检测的研究

为改良检测技术,拓宽量子点探针在农产品安全领域的应用范围,制备了用于粮油中黄曲霉毒素检测的2种量子点探针,即碳量子点(CQD)探针和石墨烯量子点(GQD)荧光免疫探针,研究2种探针应用于黄曲霉毒素检测中的可行性。采用绿色合成方法,发现以0.375g/m L柠檬酸水溶液中获得的石墨烯量子点荧光强度最强。利用1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(EDC·HCL)和N-羟基琥珀酰亚胺(NHS),将碳量子点和GQD与抗黄曲霉毒素的单克隆抗体1C11进行共价偶联,发现偶联前后量子点的荧光光谱变化趋势一致,而且偶联后均保持了较好的荧光特性,因此均可作为黄曲霉毒素免疫检测的探针,且与抗体偶联后的碳量子点的荧光特性优于石墨烯量子点。     

Transparent Polycaprolactam Electrospun Nanofibers Doped with 1,10-phenanthroline Optical Sensor for Colorimetric Determination of Iron (II) and Vitamin C

A novel optical chemical sensor based on a transparent electrospun nanofibrous scaffold, composed of polycaprolactam (PA6) and 1,10-phenanthroline (Phen), deposited on a glass slide and impregnated with polyvinyl alcohol (PVA) was coupled with UV-vis spectrophotometry and used for colorimetric determination of ferrous ion (Fe²⁺) and ascorbic acid (AA). The electrospun nanofibers were characterized using scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FT-IR) techniques. The main factors affecting performance of the optical sensor (PA6-Phen/PVA@glass) including Phen dosage, pH of sample solution, electrospinning time, polymer solution flowrate, electrospinning voltage, and PVA amount were investigated and the related optimum values were obtained. The analytical merits of the sensor for quantitative determination of Fe²⁺ and ascorbic acid was evaluated. The limit of detection (LOD) and limit of quantification (LOQ) for Fe²⁺ were 1 and 3 μg mL^-1, respectively. The linear dynamic range (LDR) was 3–150 μg mL^-1 with the determination coefficient (R²) of 0.991. The relative standard deviation (RSD %, n=3) for a solution of 60 μg mL^-1 was 5.4 %. For determination of AA, LOD and LOQ were obtained equal to 0.5 and 2 μg mL^-1, respectively. The linear dynamic range was in the 2–200 μg mL^-1 range with a R2 of 0.994. The RSD % at 100 μg mL^-1, n=3) was equal to 7.0 %. The sensor was applied successfully to the detection of Fe(II) and AA in real water samples and aspirin tablets.     

Metabolite profiling of colored rice (Oryza sativa L.) grains

HPLC analysis of dehulled red, black and non-colored indica and japonica rice subspecies revealed significant differences in the contents of the anthocyanins cyanidin-3-glucoside and peonidin-3-glucoside. The rice materials were subjected to a comparative capillary gas chromatography-based metabolite profiling approach. The employed extraction and fractionation protocol enables the analysis of a broad spectrum of lipophilic and hydrophilic low molecular weight constituents from different chemical classes. The method covers not only primary metabolites (e.g. sugars, fatty acids) but also nutritionally relevant constituents (e.g. α-tocopherol, γ-aminobutyric acid). Statistical assessment of the data via principal component analysis and agglomerative hierarchical clustering revealed a distinct grouping of the different colored rice subspecies on the basis of their metabolite profiles. Compared to non-colored and red rice, black rice exhibited, in particular, higher levels of fatty acid methyl esters, free fatty acids, organic acids and amino acids.     

SSR荧光标记毛细管电泳法与国家冬油菜区试指纹鉴定平台的构建

为了提高油菜品种指纹检测的精确性及未来构建大规模油菜新品种指纹数据库的需求,应用SSR荧光标记毛细管电泳检测法构建国家冬油菜区试指纹鉴定平台。以40对荧光引物对2012-2013年度163份国家冬油菜区试参试品种（系）进行分析。结果共检测到42个等位位点和131个等位变异。其中A基因组（n1～n10）检测位点25个,C基因组(n11～n19)检测位点17个。每个位点等位变异数从2到6不等,平均为3.02。42个检测位点的PIC值变化范围在0.10～0.69之间,平均值为0.36。其中引物BRGMS171的杂合度、PIC值分别高达0.67、0.70,可考虑作为以后区试杂交种纯度鉴定的核心标记。SSR位点的纯合度分析得出本年度18份常规种平均纯合度为81.9%,145份杂交种为57.9%。以131个等位变异计算品种（系）间DICE相似系数,163份品种（系）间平均遗传相似系数变幅为0.607～0.765,变幅最大的为品种（系）FC03（0.438～0.879）,变幅最小为品种（系）宜油21（0.611～0.806）。