亲水作用色谱柱-高效液相色谱法测定鸡蛋中氨丙啉的残留量

目的建立亲水作用色谱柱-高效液相色谱法检测鸡蛋中氨丙啉残留的分析方法。方法样品加入10 mL5%三氯乙酸水溶液进行提取,离心后取一半上清液过HLB柱净化。色谱柱为Poroshell 120 HILIC-Z (100 mm×3.0 mm, 2.7μm),流动相为5 mmol/L乙酸铵-0.1%甲酸水溶液和乙腈,等度洗脱, 5 mmol/L乙酸铵-0.1%甲酸水溶液:乙腈=20:80(V:V),流速为0.4 mL/min,进样量为20μL,检测波长267 nm。结果氨丙啉在0.35~10.0μg/mL呈良好的线性关系, r大于0.999。方法的最低定量限为250μg/kg。氨丙啉在250~2000μg/kg的浓度添加水平上,其回收率在85.0%~96.7%,批内、批间的相对标准偏差小于6%。结论该方法具有简便快捷、灵敏度高等特点,适用于鸡蛋中氨丙啉的残留检测。

新型弱阴离子交换色谱柱-高效液相色谱法同时测定苯代谢产物的方法

目的:建立了苯及其同系物体内代谢产物粘糠酸、苯乙醛酸、马尿酸的弱阴离子交换-HPLC测定方法,并进行了方法学研究。方法:采用Acclaim Mixed-Mode WAX-1(150 mm×2.1 mm,5μm)弱阴离子交换色谱柱,以甲醇-50 mmol/L磷酸氢二钾(18∶82)为基础流动相,以波长时间程序分别在254 nm下检测粘糠酸、苯乙醛酸,224nm下检测马尿酸,探讨了弱阴离子交换色谱柱分离粘糠酸、苯乙醛酸及马尿酸的影响因素。结果:流动相的pH值、离子强度、有机相比例对上述三种代谢物产生的分离产生明显影响。结论:在确定的色谱条件下,从分离度、对称性、重复性、线性4个方面来评价,弱阴离子交换色谱柱分析酸性可离子化化合物要优于常规C18色谱柱。

高效液相色谱-库仑电化学检测法定量测定小鼠血浆中CTN986

目的 建立血浆中CTN986的定量测定方法，为药物代谢研究提供技术手段。方法 采用HPLC-库仑电化学色谱方法，Diamonsil^TM（钻石）C18（4．6mm×200mm，5μm）色谱柱，0．2mol·L^-1磷酸二氢钠-甲醇（60：40）为流动相，Coulochem Ⅲ电化学检测器，Model 5010 Analytical cell为检测手段，以峰面积定量。结果检测下限为150μg·L^-1，定量下限为390μg·L^-1，方法线性范围为0．390—12．5，12．5—50mg·L^-1；在1．563，6．25，12．5，25mg·L^-1 4种质量浓度平均回收率为53．43％；日内、日间平均RSD为1．03％，2．28％。结论 该方法灵敏度高，方法操作简便，可满足药动学研究的要求。

亲水色谱-高效液相色谱法测定保健品中维生素C的含量

目的建立一种快速准确测定保健品中维生素C含量的高效液相色谱方法。方法以片剂、胶囊剂和颗粒剂多种基质的保健品为研究试样,以50%乙腈(含2%乙酸)为溶剂,制备每毫升含10μg维生素C的溶液作为标准品溶液。试样经粉碎研细混匀后,用50%乙腈(含2%乙酸)超声提取保健品中的维生素C,进样待测。采用亲水色谱Hilic柱(250 mm×4.6 mm,5μm),以乙腈-100 mmol/L乙酸铵水溶液(75∶25,V/V)为流动相,流速为0.8 ml/min,检测波长为268 nm,以外标法定量。结果维生素C的进样量在10 ng^1 500 ng时,与峰面积呈良好的线性关系,线性方程为y=6.824 82x-57.578 17,r=0.999 8。多种保健品的回收率为98.75%~103.09%,平均回收率为100.90%,检出限为0.28μg/ml。结论本法前处理简单、分析速度快、灵敏、准确、专属性强,可用于保健品中维生素C含量的测定。

高效液相色谱法测定重组人纽兰格林的电荷变异体

目的:重组蛋白在表达和纯化过程中,甚至在存放过程中,都有可能产生降解、聚集,错误折叠、翻译后修饰(如:糖基化、氧化、脱酰胺),从而形成不同的表面电荷,影响产品的稳定性、活性和安全性。该研究旨在确定重组人纽兰格林原液中是否存在电荷变异体并对其定量。方法:采用极端的理化变性条件,制备重组人纽兰格林的电荷变异体;用HPLC(阳离子交换色谱柱,Tosoh Bioscience LLC,TOSOH TSK gelSP-STAT(4.6mm×10cm),7μm)分析重组人纽兰格林中的电荷变异体。结果:纯度99%以上的重组人纽兰格林原液,经酸破坏、高温条件下,产生了相对保留时间为1.1,质谱分子量为5786.28,N末端序列未发生变化的电荷变异体。重组人纽兰格林浓度在0.0156 mg/mL^1.25 mg/mL范围内,浓度与峰面积呈现良好的线性关系,y=17187x-309.09 R2=0.9998。专属性好:在流动相A、流动相B、水、重组人纽兰格林缓冲液中无重组人纽兰格林及电荷变异体峰。回收率:重组人纽兰格林在低(0.5mg/mL)、中(1.0mg/mL)、高(1.25mg/mL)3个浓度的回收率分别为99.0%、99.9%、100.7%。重复性:以重组人纽兰格林峰面积计算RSD为0.57%。最低检测限:电荷变异体的最低检测限为:0.16μg。最低定量限:电荷变异体的最低定量限为0.2μg。结论:该方法用于测定重组人纽兰格林电荷变异体,无杂质干扰,方法准确、可靠。

超高效液相色谱串联质谱法监测血清苯妥英浓度及室间质量评价

目的建立一种灵敏、快速、稳定的超高效液相色谱串联质谱(UPLC-MS/MS)法测定人血清中苯妥英的浓度,并探讨其在苯妥英治疗药物监测(TDM)中的应用。方法在血清添加同位素内标苯妥英-D10,经乙腈沉淀蛋白后,以Shim-pack XR-ODSⅢ(50 mm×2.0 mm,1.6μm)为分析柱进行反相色谱分离,以0.1%乙酸水溶液、乙腈为流动相进行梯度洗脱,流速为0.3 mL·min-1;以电喷雾离子化串联四级杆质谱、正离子多反应监测进行定量检测,监测离子反应分别为m/z 253.3→182.2(苯妥英),m/z 263.3→192.3(内标苯妥英-D10)。用建立的方法参加2018年上半年卫生部临床检验中心茶碱TDM室间质量评价。结果 UPLC-MS/MS法检测苯妥英的线性范围为0.016~2.051 mg·L-1,批内和批间精密度分别为1.68%~4.98%和1.49%~3.82%,准确度均为85%~115%。样本的提取回收率≥93%,基质效应≥97%。室温放置6 h、自动进样器放置18 h的稳定性均良好。UPLC-MS/MS法测定结果参与室间质评样本均合格。结论该UPLC-MS/MS法采用同位素内标法,具有前处理简单、检测快速、特异性好和灵敏度高的特点,测定结果符合全国室间质量评价要求,可用于苯妥英的临床TDM。

生长抑素及制剂中多聚体的定量研究

目的:建立高效液相色谱-分子排阻色谱法(HPLC-SEC)测定生长抑素及制剂中的多聚体,并进行了方法学验证。方法:采用TSKgel G2000SWXL色谱柱(7.8 mm×300 mm,5μm),流动相为乙腈-水-三氟乙酸(20∶80∶0.1,体积比),线性等度洗脱,流速0.5 mL·min^(-1),柱温25℃,进样量为100μL;紫外检测器,检测波长为215 nm;标准曲线法和面积归一化法计算多聚体含量。结果:生长抑素二聚体的检测限为5.2 ng,定量限为15.6 ng,线性范围为0.1~5μg·mL^(-1)(r=0.999 6),平均回收率(n=3)为100.3%(RSD=0.52%)。结论:该方法快速、灵敏、准确,能够对生长抑素及制剂中的多聚体进行质控。

HPLC法测定地龙注射液中苯酚的含量

目的 建立高效液相色谱法测定地龙注射液中苯酚的含量。方法 采用高效液相色谱法,色谱柱为Agilent C_(18)色谱柱,流动相为甲醇-水(60∶40),检测波长270 nm,流速1.0 mL·min^(-1),柱温30℃,进样量10μL。结果 苯酚浓度在26.59~425.44μg·mL^(-1)范围呈良好的线性(r=1);平均回收率(n=9)为98.52%;样品中苯酚含量为标示量的54%~60%。结论 此方法简便、准确,可用于地龙注射液中苯酚的含量测定。

利巴韦林含片的人体药动学及生物等效性研究

目的:建立人血浆中利巴韦林的HPLC-MS分析方法,用以测定18名健康男性受试者舌下含服不同厂家的利巴韦林含片后的血药浓度,估算受试制剂和参比制剂的药动学参数,评价两种制剂是否生物等效。方法:采用双周期随机交叉试验设计。分别给予18名男性健康受试者试验制剂或参比制剂80mg,采集静脉血样,血浆样品去蛋白后用HPLC/MS/MS法检测药物浓度。计算药动学参数,判定两制剂是否生物等效。结果:测定利巴韦林的线性范围为2～500ng/mL(r2为0.9944),平均回收率>90%,日内RSD和日间RSD均<10%。测得血浆中两种制剂的利巴韦林的主要药代动力学参数tmax、Cmax、t1/2、AUC0-72和AUC0→∞分别为:(1.1±0.5)、(1.1±0.4)h,(249±89)、(232±65)ng/mL,(34±11)、(34±11)h,(2828±1215)、(2685±1096)ng.h.mL-1,(3600±1568)、(3416±1379)ng.h.mL-1。以AUC0-72计算,利巴韦林含片的相对生物利用度平均为(106±16)%。结论:本方法更简便、准确,灵敏度得到很大提高。两种制剂的利巴韦林药代动力学参数无统计学差异,具有生物等效性。

Polyphenols from Vitis thunbergii Sieb. et Zucc.

Aim Isolation and structural elucidation of the constituents from the aerial part of Vitis thunbergii . Methods To isolate chemical constituents, column chromatography and HPLC were used. Physico chemical characterization and spectroscopic analysis were employed for structural identification. Results Eleven polyphenols were isolated and identified. Conclusion Compound 1 is a new compound and its structure was characterized to be 3,5 dimethoxyl 4 hydroxyl phenylpropanol 9 O β D glycopyranoside.

Optimization of Chromatographic Conditions for Detecting Ellagic Acid in Pomegranate Peels Using HPLC Method

[Objective] This study aimed to optimize the chromatographic conditions for detecting ellagic acid in pomegranate peels using HPLC method. [Method] By using 0.2 mg/ml ellagic acid standard solution, on the basis of single-factor experiment and orthogonal experiment, chromatographic conditions （mobile phase ratio, flow rate, col- umn temperature） for detecting ellagic acid using HPLC were optimized. Based on the optimal chromatographic conditions, the ellagic acid content in experimental pomegranate peels was determined. [Resull] The optimal chromatographic conditions for detecting ellagic acid in pomegranate peels using HPLC method are： 1.2% phos- phoric acid：acetonitrile=85：15, column temperature of 35 ℃, and flow rate of 1.0 ml/min. The linear regression equation of ellagic acid is： y=2.9e＋0.6x＋4.4e＋5 （FF=9 999）. Ac- cording to the standard addition recovery test, the average recovery rate of ellagic acid is 98.20%, and RSD is 0.60%. Under above optimized chromatographic condi- tions, ellagic acid can be well separated from other interfering components in pomegranate peels, with shorter peak time and ideal effect, which is convenient for the detection in production practices. [Conclusion] This study laid the foundation for detecting ellagic acid in pomegranate peels using HPLC method.

HILIC-HPLC法测定山药中尿囊素的含量

目的 建立HILIC-HPLC法测定山药中尿囊素的含量,用于中药材及中成药中尿囊素的含量测定。方法 色谱柱为Thermo Syncronis HILIC（250 mm×4.6 mm,5μm）,流动相为乙腈-水（90：10）,流速为1.0 mL min-1,柱温30℃,检测波长为191 nm。结果 尿囊素在16 min左右出峰。在0.100 4～1.004μg与峰面积线性关系良好（r=0.999 6）,其平均回收率（n=6）为101.7%（RSD为1.9%）。结论 本法灵敏、准确、专属性强,能准确有效的控制山药中尿囊素的含量,并将出峰时间推迟至16 min,不易与其他峰重叠而满足中药材及中成药中尿囊素含量测定的需要。

双贝胶囊的质量标准

目的建立双贝胶囊的质量标准。方法采用薄层色谱法对双贝胶囊中土贝母、浙贝母、连翘进行定性鉴别;采用高效液相色谱法对双贝胶囊中土贝母苷甲进行定量分析。结果薄层色谱鉴别分离效果好,专属性强;土贝母苷甲含量在0.182~0.912 mg·m L-1范围内线性关系良好,r=0.9997;平均加样回收率99.31%,RSD 0.49%。结论该方法简单、准确、专属性强、重复性好,可有效控制双贝胶囊的质量。

Fucoxanthin Content of Five Species Brown Seaweed from Talango District, Madura Island

Fucoxanthin content of five brown seaweed species of East Java collected from Talango district （Madura sea-Jawa sea） namely Padina australis, Turbinaria conoides, Sargassum cinereum, Sargassum filipendula and Sargassum echinocarpum were determined and samples were collected using Global Positioning System （GPS）. Fucoxanthin from those samples was determined using Thin Layer Chromatography （TLC）, High Pressure Liquid Chromatography （HPLC） and Spectrophotometer UV-Vis and the results were then calculated using Seely equation. The pigments separation using HPLC gave a better result compared to TLC, and beside fucoxanthin otlher pigments namely chlorophyll and β carotene were also identified in these samples. Four points of sampling location were chosen using GPS and spot with coordinate positions observed were 113.94444° EL-7.08795° SA; 113.94231° EL-7.08913° SA. The results showed that fucoxanthin content were as follow： Padina australis 0.2674± 0.0046 mg/g; Turbinaria conoides 0.2134± 0.0100 mg/g; Sargassum filipendula 0.1957 ± 0.0173 mg/g; Sargassum cinereum 0.1640 ± 0.0092 mg/g and Sargassum echinocarpum 0.1576 ± 0.0001 mg/g. It can be concluded that Padina australis and Turbinaria conoides contain the most amount of fucoxanthin and it is possible to be cultivated as edible brown seaweed fucoxanthin resources.

Economic and Qualitative Traits of Italian Alps Saffron

Saffron, obtained from the flower stigmas of Crocus sativus L., is one of the most expensive food spices. The introduction of saffron in alpine areas could help to broaden and diversify the activities of mountain multifunctional farms, with a positive impact on economy and land management. According to ISO 3632(2010/2011), saffron can be classified into three categories of quality(I, II, III) depending on the concentration of the three main metabolites responsible for its characteristic colour, flavor and aroma: Crocin, Picrocrocin and Safranal. This study represents the first investigation of the quality of saffron produced in the Italian Alps evaluated with spectrophotometry, HPLC, solid-phase microextraction(SPME), and gas chromatographic analysis combined with mass spectrometry(GC/MS). The experiments used Crocus sativus stigmas produced in 2012-2013 in different areas of the Central Italian Alps were located at an altitude between 720 and 1200 m a.s.l.. Results obtained were compared to commercial saffron. The analyses confirmed that all samples can be classified in the first quality category according to the ISO classification. This high quality is also confirmed by HPLC analysis. Moreover, the SPME-GC/MS analysis identified some differences in the aromatic profile of saffron samples, in particular regarding safranal concentration. A preliminary assessment of the economic viability of high quality saffron production for local markets was also performed. Our study provides valid information regarding the quality and economic sustainability of saffron production in the alpine area confirming this crop as a good candidate for a new source of income for multifunctional farms in mountain areas.

HPLC法测定五指毛桃中补骨脂素含量及质量评估

目的探究采用色谱柱高效液相色谱(HPLC)法测定五指毛桃中包含的补骨脂素含量程序优化的方法,并对其质量实施评估。方法获取来自不同地区的10批五指毛桃作为研究样本,并将其进行编号为1#~10#,采用HPLC法对1#~10#五指毛桃中的补骨脂素含量加以检测,并对其质量进行评估。结果补骨脂素的对照品在0.5~290.0 mg/L(r=0.9997)这一范围内同峰面积之间的线性关系为良好,经平均加样回收试验得出平均加样回收率能够达到99.56%,相对标准偏差(RSD)为1.926%。本次对10个样本进行补骨脂素含量检测得出,含量最高为0.069%,最低含量为0.043%。结论经过程序优化后的HPLC法对补骨脂素的提取更加简单快捷,可行性表现突出,并且能够保证结果的准确性和稳定性,能够对五指毛桃的质量评估提供有效依据,应予以推广。

Production of Aflatoxins from Aspergillus flavus in Liquid Medium

Aflatoxins are produced mainly by Aspergillus flavus and Aspergillus parasiticus, and can be found in many grains such as peanuts, soybeans and com. This study aimed to qualitatively and quantitatively evaluate the production of aflatoxin in liquid media using strains of Aspergillus flavus obtained from peanuts marketed in the city of Fortaleza, CEo Strains of Aspergillus flavus were inoculated into a liquid medium malt extract and after 2 days inoculated into a second medium containing sucrose 5%, MgSO4·7H20 0.1%, KH2PO4 1%, ZnSO4·7H2O 0.0176 g, and cultured for 3 more days. The media were kept at room temperature ranging from 24°C to 32 °C with agitation of 130 rpm and aeration of 4.17 Llmin. Qualitative analysis was performed by thin layer chromatography and quantitatively by high performance liquid chromatography with fluorescence detection, demonstrating the production of aflatoxin B I （588 mg/L） and B2 （929 mg/L）.

Ultrafiltration of Shiitake Polyphenol with Ultrasound Enhancement

Ultrafiltration （UF） experiments were subjected to the concentration of Shi- itake polyphenol（SP） with ultrasound （US） enhancement, by which the effects of the main parameters such as ultrasonic power, transmembrane pressure, temperature and axial flow-rate on the membrane fluxes were analyzed, and then the SP of the ultrafiltrate after follow-up resin adsorption by HPLC was investigated. The results in- dicated that US could significantly enhance the membrane fluxes. The optimal con- ditions obtained was as follows： 0.60 W/cm2 US power, axial flow-rate of 80 L/h, operation temperature 30 ~C, transmembrane pressure 0.10 MPa. US increased the absorption capacity of the absorbent resin during the following stage. The HPLC analysis also showed that catechins were purified to some extent as the ultrafiltrate was treated by US and macroporous resin, indicating the technology of US en- hancement coupled with UF showed the potential for concentration and purification of SP by absorbent resins.

Changes of Metabolites and Macro- and Micro-elements in Hungarian Potatoes under Organic and Conventional Farming

To study the effect of cultivation system （conventional and organic） on potato tuber components, 42 parameters （anti-nutritives, micro- and macro-elements and 23 metabolites） were studied in three multi-resistant Hungarian potato varieties for three years in 2007-2009. Discriminant analysis of data proved that all investigated factors （farming technology, genotype and season/year） had significant effect on metabolites. Reversed phase high performance liquid chromatographic （HPLC-DAD, HPLC-MS） and elemental analysis methods have been adapted to quantify the major components, i.e., steroidal alkaloids, nitrite, nitrate, sugars, amino acids and micro-elements, in potato tubers. The absolute amount and changes of tuber components were influenced differentially by the technology, genotype and season in a complex manner. Any examined component, except nitrate content, was found to be significantly higher or lower consistently in relation to the production practice （organic or conventional） during the three years trials. Under the examined circumstances, no consequent positive effect of organic fanning on the total amount of anti-nutritive components, vitamins or micro- and macro-elements of potato tubers could be proven.

Removal of Pharmaceutically Active Compounds in Sequencing Batch Reactor

Biological treatment efficiency of six pharmaceutical compounds （acetazolamide, metronidazole, opipramol, piracetam, salicylamide and tinidazole） was evaluated using lab-scale Sequencing Batch Reactor （SBR）. Comparative biological degradation processes of two types of activated sludge from municipal and pharmaceutical industry sewage treatment plants were examined. Three different organic loadings （0.05 g COD/g MLSS.d, 0.1 g COD/g MLSS.d and 0.2 g COD/g MLSS-d） and reaction time on the efficiency of Active Pharmaceutical Ingredient （API） decomposition were examined. Chemical oxygen demand, non-purgeable organic carbon as well as ammonium nitrogen contents were monitored by standard methods. Percentage of API decomposition was analysed by High Performance Liquid Chromatography （HPLC）. The overall API removal efficiency was strictly dependent on the type of activated sludge origin. The main biodegradation products were identified using HPLC-MS,1H NMR and 13C NMR methods as e.g. （{4-[3-（5H-dibenzo[b,f]azepin-5-yl]piperazin-l-yl}methanamine） and （2-amino-1,3,4-thiadiazol-5-sulfonamide） for opipramol and acetazolamide respectively.