高效液相色谱法测定粮食中玉米赤霉烯酮及其代谢物

采用甲醇-水提取,C18小柱净化,反相HPLC荧光检测器测定了玉米、面粉、小麦样品中的玉米赤霉烯酮及其代谢物和黄曲霉B1。对样品预处理和高效液相色谱测定条件进行了优化,ZON,α-ZOL,β-ZOL和AFTB1的线性范围分别为5.0μg/L~146g/L,25.0μg/L~200g/L,25.0μg/L~160g/L和0.4μg/L~2.0g/L,检出限分别为0.5、2.5、2.5和0.04μg/kg;加标回收率在80.0%~110.0%范围内;日间相对标准偏差为4·5%~9.2%,日内精密度2.7%~7.4%。本方法灵敏准确,易于推广,适用于粮食中玉米赤霉烯酮及其代谢物的检测。     

多功能柱净化-高效液相色谱法检测谷物中的玉米赤霉烯酮

建立了玉米和小麦中玉米赤霉烯酮（ZEN）的多功能柱净化-高效液相色谱检测方法。样品经乙腈-水混合溶剂（V（乙腈）：V（水）=84：16）提取,通过多功能净化柱（MFC）进行一次性净化,以Symmetry^R C18柱为分离柱,甲醇-水（V（甲醇）：V（水）=68：32）为流动相进行高效液相色谱分离和检测。玉米赤霉烯酮的质量浓度在0．01～4．0μg／mL范围内呈良好线性,相关系数为0．9996。检出限为0．04μg／g,在0、04—5．0mg／kg添加范围内的回收率为87．5％～98．6％,相对标准偏差为1．5％～8．3％。     

多功能针式过滤器净化超高效液相色谱法测定大米和花生中玉米赤霉烯酮

建立了一种基于多功能针式过滤器净化的超高效液相色谱测定大米和花生中玉米赤霉烯酮的方法。样品经乙腈提取,采用多功能针式过滤器通过式净化,以超高效液相色谱法测定其中的玉米赤霉烯酮,色谱柱为ACQUITY UPLC BEH C18柱（100 mm×2.1 mm,1.7μm）,以甲醇-乙腈-水（体积比为8∶46∶46）为流动相等度洗脱,流量为0.3 mL/min,用荧光检测器检测,色谱峰面积外标法定量。玉米赤霉烯酮的质量浓度在2.5～500 ng/mL范围内与色谱峰面积线性关系良好,相关系数不小于0.999 5。大米和花生样品的方法检出限分别为15.0、30.0μg/kg,定量限分别为50.0、100.0μg/kg。样品加标回收率为77.11%～93.65%,测定结果的相对标准偏差为0.49%～4.95%(n=6)。该方法简便快速,适用于大米和花生中玉米赤霉烯酮的日常检测。     

麦类中玉米赤霉烯酮的快速检测

提出了高效液相色谱荧光检测法测定麦类样品中玉米赤霉烯酮的方法.样品经乙腈-水(84 16,体积比)提取,多功能柱净化,C18色谱柱(4.6 mm×250 mm,5um)分离,水-乙腈-甲醇(46 46 8,体积比)混合溶液作流动相,流速1.0 mL·min-1.玉米赤霉烯酮的质量浓度在20.0～5 000 ug·L-1范围内,峰面积与玉米赤霉烯酮浓度之间呈线性关系,样品在50.0,100.0,1 000.0 ng·g-1添加水平的平均回收率分别为74.6%,75.5%,73.5%,相对标准偏差为9.7%～11.5%(n=8)之间,方法的测定限(10S/N)为50.0 ng·g-1.     

粮食中玉米赤霉烯酮和镰刀菌氧萘满酮高效液相色谱分析方法的研究

建立了粮食样品中镰刀菌氧萘满酮（ＴＤＰ－１）和玉米赤霉烯酮（ＺＥＡ）的高效液相色谱－荧光检测分析方法。粮食样品在碱性和中性条件下用乙腈－水（３：１,Ｖ／Ｖ）提取,经正己烷脱脂,过Ｆｌｏｒｉｓｉｌ柱净化,最后用反相高效液相色谱－荧光检测法测定。该法的最低检出灵敏度：玉米赤霉烯酮为５×１０＾－９ｇ,镰刀菌氧萘满酮为１×１０＾－９ｇ。玉米中镰刀菌氧萘满酮和玉米赤霉烯酮的回收率分别为１００．２％和９２．４     

免疫亲和柱净化-高效液相色谱法同时检测鸡蛋中6种玉米赤霉醇类化合物残留量

建立了鸡蛋中6种玉米赤霉醇类化合物(α-玉米赤霉醇、β-玉米赤霉醇、α-玉米赤霉烯醇、β-玉米赤霉烯醇、玉米赤霉酮和玉米赤霉烯酮)残留量的免疫亲和柱净化-高效液相色谱检测方法。样品酶解后用叔丁基甲醚提取、氢氧化钠反萃取,经免疫亲和柱富集和净化后,采用高效液相色谱-紫外检测器进行测定。色谱柱: Agilent Eclipse XDB-C18(150 mm×4.6 mm, 3.5 μm);流动相: 甲醇-乙腈-水(50:15:35, v/v/v);流速: 1.0 mL/min;检测波长: 270 nm。结果表明,6种目标物在0.01～0.2 mg/L范围内线性关系良好,相关系数(r)≥0.9998,检出限(LOD,S/N≥3)为1.0 μg/kg,平均回收率为73.2%～95.7%,相对标准偏差小于8%。该方法灵敏度高、重现性好,适用于鸡蛋样品中痕量玉米赤霉醇类药物残留的测定。     

冬小麦越冬茎尖中的玉米赤霉烯酮

本文报道冬小麦在越冬期间,茎尖内出现与春化作用密切相关的物质,经硅胶薄层层析分离、高效液相色谱仪纯化、紫外吸收光谱分析、电子电离(E1)质谱以及质谱/质谱联机鉴定,确证这种物质是玉米赤霉烯酮。     

高效液相色谱法测定鸡肝中玉米赤霉醇的残留量

建立了高效液相色谱测定鸡肝中玉米赤霉醇的方法。样品经β-葡糖苷酸酶水解、乙醚提取、氢氧化钠抽提、C18小柱净化后,用Zorbax SB-Phenyl柱(250 mmx4.6 mm i.d., 5 μm)分离,以乙腈-0.3%三氟乙酸水溶液(体积比为40∶60)作流动相,流速为0.80 mL/min,于262 nm波长处检测。结果表明,样品的加标平均回收率为72.0%-80.4%,相对标准偏差为8.3%-13.9%,定量测定低限(LOQ)为5 μg/kg。方法已应用于实际样品的测定。另外,用高效液相色谱-质     

高效液相色谱–串联质谱内标法测定动物源性食品中玉米赤霉烯酮及其代谢产物

建立高效液相色谱–串联质谱法(HPLC–MS/MS)测定动物源性食品中玉米赤霉烯酮及其5种代谢产物(α-玉米赤霉烯醇、β-玉米赤霉烯醇、α-玉米赤霉醇、β-玉米赤霉醇、玉米赤霉酮)残留量。在样品中加入4种同位素内标(13C18–玉米赤霉烯酮,D7–α-玉米赤霉烯醇,D7–β-玉米赤霉烯醇,D5–α-玉米赤霉醇)后,经β-葡萄糖苷酶/硫酸酯酶酶解,用叔丁基甲基醚萃取,取上清液氮吹至近干后用三氯甲烷复溶,再用氢氧化钠溶液反向萃取,以HLB固相萃取柱净化后,用HPLC–MS/MS检测。结果表明,玉米赤霉烯酮及其代谢产物在1.0~100.0μg/L范围内线性关系良好,相关系数均大于0.996,方法的检出限为0.04~0.13μg/kg,定量限为0.11~0.43μg/kg。在1.0、4.0、10.0μg/kg三种加标浓度水平下,回收率为77.7%~105.5%,测定结果的相对标准偏差为4.8%~9.8%(n=6)。该方法准确、可靠,灵敏度高,适用于动物源性食品中玉米赤霉烯酮及其代谢产物的定量分析。     

高效液相色谱和高效液相色谱－质谱法测定粮食中互隔交链孢霉醇、互隔交链孢霉醇单甲醚及玉米赤霉烯酮

建立了粮食中互隔交链孢霉醇（ＡＯＨ）、互隔交链孢霉醇单甲醚（ＡＭＥ）和玉米赤霉烯酮（ＺＥＡ）三个毒素的ＨＰＬＣ定性定量分析法和粒子束ＬＣ－ＭＳ／ＥＩ￣＋鉴定方法。采用反相色谱,以８０％的甲醇水溶液作流动相,这三个毒素在Ｃ＿（１８）色谱柱上彼此间均获得较好的分离。方法对粮食中ＡＯＨ,ＡＭＥ两个毒素的回收率均在７９％以上。ＡＯＨ,ＡＭＥ两个毒素的最低检出限为１×１０￣（－９）ｇ。用所建方法对１００多个大骨节病病区、非病区的粮食样品进行了检测,结果为阴性。     

高效液相色谱荧光检测法测定蔬菜中残留的甲氨基阿维菌素苯甲酸盐

建立了甘蓝和蘑菇中甲氨基阿维菌素苯甲酸盐的固相萃取-高效液相色谱荧光分析方法。蔬菜样品用乙酸乙酯提取,提取液旋转浓缩近干后用少量乙酸乙酯溶解,再经PRS固相萃取(SPE)柱净化,洗脱液经氮气吹干后用氮甲基咪唑和三氟乙酸酐衍生,衍生物用高效液相色谱分析,采用外标法定量。在添加浓度1.0～20.0 μg/kg范围内,平均添加回收率为78.6%～84.9%,日内相对标准偏差(RSD)为2.7%～6.0%,日间RSD为3.1%～8.9%,检出限为0.10 μg/kg。甲氨基阿维菌素苯甲酸盐衍生物在0.002～0.10 mg/L范围内呈良好的线性关系,相关系数为0.9999。     

Determination of ochratoxin A in Portuguese rice samples by high performance liquid chromatography with fluorescence detection

A sensitive and accurate analytical method for the determination of ochratoxin A (OTA) in rice, based on extraction with phosphate-buffered saline/methanol, an immunoaffinity column (IAC) for clean-up, and high performance liquid chromatography with fluorescence detection (HPLC-FD), is described. The limit of quantification of the proposed method was 0.05 g kg^–1. Recovery of OTA from rice samples spiked at 0.05 g kg^–1 was 92%, with a within-day RSD of 5.4%. The proposed method was applied to 42 rice samples from Portugal and the presence of OTA was found in six samples at concentrations ranging from 0.09 to 3.52 g kg^–1. The identification of OTA was confirmed by methyl ester derivatization and then HPLC analysis. The daily intake of OTA by the Portuguese population was also estimated.     

柱前衍生高效液相色谱-荧光检测法测定血浆中同型半胱氨酸

建立了一种柱前衍生高效液相色谱-荧光检测法用于测定血浆中同型半胱氨酸(Hcy)。使用三(2-羧乙基)膦盐酸盐(TCEP)为还原剂,N-(1-芘)马来酰亚胺(NPM)为衍生剂进行样品预处理,Agilent Hypersil C-18柱(250 mm×4.0 mm, 5 μm)进行分离,流动相为15 mmol/L醋酸钠-乙腈-混合酸(300 mL水中含1 mL醋酸和1 mL磷酸)混合溶液,采用梯度洗脱,荧光检测激发波长为330 nm,发射波长为380 nm。Hcy的回收率为(102.08±4.94)%。线性范围为0.500～100 μmol/L,检出限（以信噪比为3计）为0.016 μmol/L。日内与日间相对标准偏差均小于5%。利用该方法对7例高血压患者和7例健康志愿者的血浆进行了测定,结果表明两组间的Hcy含量存在显著的差异(p＜0.05)。本方法简单、快速、灵敏、特异,适用于血浆Hcy的临床定量测定。     

Analysis of zearalenone and α-zearalenol in animal feed using high-performance liquid chromatography

Different extraction and clean-up techniques used before HPLC analysis were compared in order to obtain a reliable method for the quantitative determination of zearalenone (ZEA) and α-zearalenol (α-ZOL) in animal feed. Immunoaffinity clean-up was compared to C₁₈ and Florisil column clean-up. Extracted samples were analysed by reversed-phase HPLC with fluorescence detection (λ_ex=274 nm, λ_em=440 nm). A mobile phase of acetonitrile:water (50:50 (v/v)) and a flow-rate of 1.0 ml min⁻¹ resulted in a good separation between ZEA and α-ZOL. Using immunoaffinity clean-up the linear range was between 25 and 600 μg kg⁻¹ for ZEA and α-ZOL in maize. Intra-laboratory coefficients of variation (CV) (under repeatability conditions) were 9.16% for ZEA and 2.18% for α-ZOL. Recoveries for spiked ZEA and α-ZOL samples ranged from 89 to 110% with CVs between 5.2 and 11.2% (under within-laboratory reproducibility conditions). Using C₁₈ and Florisil solid-phase clean-up, matrix interference was too high. Therefore, naturally contaminated animal feed samples were analysed using the developed HPLC method coupled to the immunoaffinity clean-up.     

柱前衍生反相高效液相色谱-荧光检测法测定大鼠血浆中的奈替米星

建立了一种简便、灵敏的氯甲酸芴甲酯(FMOC-Cl)柱前衍生反相高效液相色谱-荧光检测血浆中奈替米星的新方法,同时研究了其药代动力学。对色谱条件进行了优化,采用ZORBAX Eclipse XDB-C8柱(150 mm×4.6 mm,5 μm),流动相为乙腈-水(体积比为85:15),流速为1.0 mL/min,荧光检测激发波长为265 nm,发射波长为315 nm,得到奈替米星的平均加标回收率为96.62%～100.84%(n=3),对奈替米星检测的线性范围为0.045～8.88 mg/L,相关系数为0.9993,方法的日内与日间精密度分别低于3%与3.5%,最低检出限(S/N=3)与定量限(以3倍检出限计)分别为0.01和0.03 mg/L。方法简便、快速、灵敏,样品用量少(30 μL奈替米星血浆溶液已能满足该药含量的测定以及药物代谢的研究),为大鼠体内奈替米星的药代动力学研究提供了可靠的分析手段。     

Determination of zearalenone in maize products by vortex‐assisted ionic‐liquid‐based dispersive liquid–liquid microextraction with high‐performance liquid chromatography

A novel method has been developed for the analysis of zearalenone in maize products by vortex‐assisted ionic‐liquid‐based dispersive liquid–liquid microextraction combined with HPLC and fluorescence detection. Maize samples were extracted with methanol/water (80:20, v/v) and the extraction solution was then used as the dispersive solvent in the microextraction procedure. The analyte was rapidly transmitted to a small volume of ionic liquid and was determined by HPLC. Various parameters affecting the recovery of the mycotoxin were investigated, such as the type and volume of the extraction solvent, the type and volume of the dispersive solvent, the pH of the aqueous phase, the salt addition, and the time of vortex and centrifugation. Under the optimal experimental conditions, a good linearity of the analyte was obtained in the range of 1.0–1000.0 μg/L with the correlation coefficient of 0.9998. The limit of detection (S/N = 3) and quantification (S/N = 10) were 0.3 and 1.0 μg/kg, and the mean recoveries ranged from 83.5 to 94.9%, with a relative standard deviation less than 5.0%. The proposed method was demonstrated to be simple, cheap, quick, and highly selective and was successfully applied to the determination of zearalenone in maize products.     

高效液相色谱荧光检测法测定药物中的氯乙酰氯

建立测定药物中氯乙酰氯含量的高效液相色谱–荧光检测方法。以吖啶酮乙酰肼为荧光标记试剂,对氯乙酰氯进行柱前衍生。在室温下反应15 min,衍生产率达到最大。衍生溶液在XDB–C18柱上,以水和乙腈为流动相进行分析,激发波长和发射波长分别为255 nm和429 nm。氯乙酰氯浓度在1~1 000 nmol/L范围内与色谱峰面积具有良好的线性关系,线性相关系数r=0.999 9。方法的检出限为0.35 nmol/L,仪器精密度和方法精密度分别为0.52%和0.67%(n=6)。样品加标回收率为92.5%~95.6%。该方法简单、准确,精密度良好,可用于测定药物中氯乙酰氯的残留量。     

Determination of atrasentan by high performance liquid chromatography with fluorescence detection in human plasma.

Atrasentan is an endothelin antagonist selective for the ET(A) receptor in development at Abbott Laboratories for the treatment of cardiovascular disease and cell proliferation disorders. A simple and sensitive chromatographic method for the determination of atrasentan in human plasma has been developed and validated. The analytical method involves acidification of the plasma samples with 0.3 N HCl prior to extraction with 1:1 (v:v) hexane/tert-butylmethylether. The organic extract was evaporated to dryness, reconstituted with 20:80 (v:v) acetonitrile/0.05 M K(2)HPO(4) and washed with 75:25 (v:v) hexane/tert-butylmethylether. The organic layer was discarded and the aqueous layer was injected into the HPLC. Atrasentan and internal standard (ABT-790) were separated from interference using a 250 x 4.6 mm, 5 microm, 120 A Phenomenex Spherisorb C(8) analytical column with a 50 x 4.6 mm, Alltech Absorbosphere 5 microm CN guard cartridge using a mobile phase consisting of 25:15:5:55 (v:v:v:v) acetonitrile/isopropanol/methanol/0.05 M K(2)HPO(4), pH 7.0, at a flow rate of 1.0 mL/min. Fluorescence detection was achieved using lambda(ex) 278 nm and lambda(em) 322 nm. For a 1.0 mL plasma sample volume, the limit of quantitation was approximately 200 pg/mL. The method was linear from 0.2 to 1300 ng/mL (r(2) = 0.9986). Inter- and intra-day assay RSD (n = 6) were less than 10%. Mean accuracy determinations showed the quality control samples to range between 94 and 99% of the theoretical concentration.     

高效液相色谱-柱后在线光化学衍生荧光检测法测定辣椒油中4种苏丹红染料

建立了在线光化学衍生、荧光检测、高效液相色谱(HPLC)测定辣椒油中苏丹红I、II、III和B的方法。以乙腈-水为流动相,采用梯度洗脱方式在SB-C18色谱柱上分离。用实验室自制的程序控制时间/光强光化学反应器作为在线衍生装置,优化了光衍生反应的条件和荧光检测条件。3种不同加标浓度下,辣椒油样品中4种苏丹红染料的加标回收率为81.3%～100.4%。加标水平为0.8 mg/kg下荧光信号强度的相对标准偏差(RSD,n=6)为2.6%～3.8%。苏丹红I、II、III和B的检出限(LOD)和定量限(LOQ)范围分别为0.009～0.054 mg/kg和0.030～0.181 mg/kg,优于传统的HPLC分离、二极管阵列检测器检测方法。该方法具有简单、灵敏、选择性好的特点,适用于食品样品中苏丹红的常规分析。