衍生化-高效液相色谱法测定西索米星

采用反相高效液相色谱法，以2，4-二硝基氟苯为衍生化试剂。其线性范围为10mg/L-500mg/L；检出限为5.0mg/L；相对标准偏差为2.3%。方法适用于西索米星的产品质量控制。     

Determination of lysinoalanine by high performance liquid chromatography

This work suggests an HPLC method for qualitative and quantitative determination of N^ε(2-amino-2-carboxyethyl)-L -lysine (LAL). LAL was released from total hydrolysates of various proteins of animal origin and derivatized with dansyl chloride. The performance of two different columns, Spherisorb 3S TG and μ-Bondapack C₁₈, was compared; better resolution and quantitative response were obtained with the former. The mobile phase was a mixture of 0.01 M phosphate buffer (pH 7) and acetonitrile. Linear response and quantitative repeatability were tested for both detectors used (UV-Vis set at 254 nm; fluorimetric set at λ_ex(max) = 360 nm and λ_em(max) = 525 nm). For LAL standard the minimum detectable amount was 0.05 ng, whereas for LAL in actual samples the amount was 0.5 ng (40 μg/g of analyzed proteins). Good analytical repeatability was obtained, resulting in CV % of 4.7 and 3.8 for UV and fluorimetric detectors, respectively. LAL recovery was determined using both detectors; the values obtained were 94 % (fluorimetric) and 92 % (UV). Greater noise levels were observed with the fluorimetric detector and its higher sensitivity could not, therefore, be fully utilized. The highest amounts of LAL were found in the casein (2816 μg/g) and cooked albumin (615 μg/g) samples.     

Determination and purification of metallothioneins by high performance liquid chromatography.

A rapid, reproducible and sensitive high performance liquid chromatography (HPLC) method for the determination and purification of metallothionein-I (MT-I) and metallothionein-II (MT-II) in mouse and rabbit livers has been developed. Methallothioneins (MTs) were separated by an HPLC anion exchange column, eluted through a linear gradient of Tris buffer and the peak containing MTs was determined by atomic absorption spectrophotometry. Furthermore, the content of MT-I or MT-II was calculated by protein peak area in a short time (about 20 min). The sample to be tested was homogenized, centrifuged and saturated by cadmium. MT-I and MT-II were eluted at 15.9 and 19.3 min, respectively. The following mouse liver cytosols were tested: controls, Cd-injected samples and 60Co-irradiated samples. A detection limit of 5 micrograms/g liver was established for this method. We have analysed more than 100 biological samples and obtained satisfactory results.     

高效液相色谱法分析硫双灭多威

研究了用高效液相色谱分析杀虫剂硫双灭多威的方法。在反相ＯＤＳ柱上,用甲醇－水作流动相,紫外检测器检测,以邻苯二甲酸二甲酯作内标定量。方法快速、准确,重现性好,线性范围宽,变异系数为０．１８％,回收率为９９．８９％,色谱分析周期仅为８ｍｉｎ。     

Determination of cortisol in guinea pig urine by high performance thin-layer chromatography, high performance liquid chromatography, and thin-layer chromatography/radioimmunoassay

Summary In order to collect urinary samples from unrestrained guinea pigs, animals were kept in their familiar home cages with wood shavings for bedding. Cortisol was removed from shavings by a simple washing step, and an attempt was made to measure its concentrations by high performance thin-layer chromatography (HPTLC), high performance liquid chromatography (HPLC), or thin layer chromatography/radioimmunoassay (TLC-RIA). After intramuscular administration of 25 mg cortisol, cortisol excretion increased from about 20–30 μg/day to 400–500 μg/day (HPTLC: 531 μg/day, HPLC: 493 μg/day; TLC-RIA: 394 μg/day). Similarly, the treatment of the animals with 20 IU ACTH resulted in an augmented cortisol excretion, with mean values of 294 μg/day (HPTLC), 256 μg/day (HPLC) and 143 μg/day (TLC-RIA), respectively. The present study shows, for the first time, that cortisol excretion in unrestrained laboratory animals can be determined. Whilst the cortisol values measured by HPTLC and HPLC agree, the amounts measured by TLC-RIA were significantly lower. These differences are probably due to the presence of substances in urine or shavings which interfere with the radioimmunological determination. Hence, cortisol should be determined either by HPTLC or HPLC. Beside having a desirable specificity, these methods are more suited than TLC/RIA for steroid analysis since they confer the possibility of measuring additional steroids (e.g. precursors and/or metabolites of cortisol) in a single urine extract. This is especially the case for the HPTLC method since substances can be transformed into fluorescent derivatives.     

Determination of paraquat in marijuana by reversed-phase paired-ion high performance liquid chromatography.

A sensitive and selective method is described for the quantitative determination of paraquat in marijuana. Paraquat is extracted from finely ground plant material with hydrochloric acid with sonification, and the resulting acidic solution is extracted with chloroform:isopropanol (9:1) and evaporated to dryness. The residue is reconstituted with aqueous phosphate buffer pH 7.0; the solution is passed through a C-18 SEP-PAK TM and is analyzed with high performance liquid chromatography, using a reversed-phase column and an "ion pairing" reagent in the mobile phase. The recovery of paraquat in laboratory-spiked material varied from 90-97%. Results obtained with confiscated, field-sprayed marijuana by the procedure described were in excellent agreement with those obtained with a well-established ultraviolet procedure. The calculated limit of detection with this method is 2 ng of paraquat.     

32种氧化型染料的高效液相色谱定量及高效液相色谱-串联质谱确证方法北大核心CSCD

染发类产品中氧化型染料种类多,实际样品测定时干扰多,建立染发类产品中多种常用染料的测定方法,为该类产品的有效监管提供技术手段十分必要。该研究根据染料使用频率分组,采用能够屏蔽硅羟基和金属离子影响的C_(18)柱,优化了《化妆品安全技术规范》(2015年版)中32种染料的高效液相色谱法(HPLC)并建立了高效液相色谱-串联质谱(HPLC-MS/MS)确证方法。样品以10 g/L亚硫酸氢钠水溶液为抗氧化剂,用无水乙醇-水(1∶1,v/v)混合溶液冰浴超声提取10 min。HPLC方法采用甲醇、乙腈和磷酸盐缓冲液为流动相分两个液相色谱条件进行梯度洗脱分离,于280 nm波长下检测,其中一个HPLC条件中的相互干扰组分均在另一个HPLC条件下完全分离,避免了实际样品检测时组分间的干扰,并排除了32种以外的其他15种常用染料的干扰。HPLC-MS/MS方法分别采用5 mmol/L乙酸铵水溶液-乙腈和5 mmol/L乙酸水溶液-乙腈为正离子和负离子模式下的流动相,电喷雾离子模式下用多反应监测(MRM)模式进行定性和定量分析。HPLC和HPLC-MS/MS两个方法中,日内精密度和48 h内稳定性的相对标准偏差(RSD)<10%,回收率为82.6%~114.9%(RSD<10%)。HPLC方法中32种染料在大约10~500 mg/L范围内线性关系良好(r^(2)>0.99),检出限为9.7~40.1μg/g;HPLC-MS/MS方法中氢醌线性范围为2.0~79.7 mg/L,检出限为8.0μg/g,其他组分线性范围约为0.1~4 mg/L,检出限为0.01~0.4μg/g。采用HPLC、HPLC-MS/MS两个方法和《化妆品安全技术规范》方法同时测定实际样品,共检出16种染料,检出含量范围为58~25160μg/g。3个方法检测结果的RSD为1.9%~10.1%。该研究增加了HPLC-MS/MS确证方法,适应化妆品法定检验中的未知物确认程序;方法简便快速,结果准确,专属性强,具有较好的通用性和可操作性。     

电合成产物苯甲醛的高效液相色谱法测定

建立了一种用高效液相色谱法(HPLC)测定电合成产物中苯甲醛的方法.采用Kronmsil C(18)色谱柱(5μm,250 mm×4.6 mm I.d.),流动相为甲醇,检测波长为250 nm,外标法定量.苯甲醛的质量浓度在2.0-70.0 mg/L范围内线性关系良好,其相关系数r=0.9998(n=3);将本法应用于电合成产物中苯甲醛的测定,其加标回收率为93.0%～97.0%,相对标准偏差为0.05%～1.2%.     

高效液相色谱法测定水性涂料中的酚类防霉剂

建立了简便、准确的高效液相色谱法检测涂料样品中五氯苯酚(PCP)、四氯苯酚(TeCP)和邻苯基苯酚(OPP)。选用C18柱为分离柱,以甲醇-10 mmol/LNH4Ac为流动相进行梯度洗脱,二极管阵列检测器检测,检测波长为220 nm。在该色谱条件下,PCP、TeCP、OPP分析线性范围分别为0.02~200 mg/L,0.02~200 mg/L,0.04~200 mg/L;最低检测浓度分别为0.01、0.01、0.02 mg/L,分别相当于涂料中的质量分数为5.0×10-4%,5.0×10-4%,1.0×10-3%;方法加标回收率为96.6%~103.6%,相对标准偏差为0.98~4.3%(n=7)。本法能满足涂料中PCP、TeCP、OPP的分析测定。     

反相高效液相色谱法同时测定对苯氧基苯酚和对氯苯酚

建立了一种同时测定对苯氧基苯酚和对氯苯酚的反相高效液相色谱方法。采用Diamonsil(钻石)C18(250 mm×4.6 mm i.d.,5μm)色谱柱,以甲醇-水(V(甲醇):V(水)=8∶2)为流动相,在波长为230 nm处进行检测。对苯氧基苯酚和对氯苯酚在20~230μg/mL范围内,峰面积与质量浓度呈良好的线性关系,相对标准偏差RSD分别为0.26%、0.53%,回收率在99.0%~101%之间。方法可用于对苯氧基苯酚的合成工艺研究及成品质量控制。     

Measurement of N-methyl-2-pyridone-5-carboxamide in urine by high performance liquid chromatography

We have previously described a simple and reproducible method for the measurement of nicotinamide and its major metabolite N-methyl-2-pyridone-5-carboxamide (2-pyr) in human plasma. We now describe a low-cost high-throughput method for measurement of urinary 2-pyr, and demonstrate that Isolute C18 bulk can replace use of the column to clean up the samples prior to injection into the HPLC apparatus. Using a standard curve together with an internal standard for each sample, with mean recovery of 2-pyr greater than 95%, the assay has proved reproducible, with considerable savings in cost and time. The principal advantages of this method are the rapid column clean up of samples prior to injection and the simple but effective methodology.     

牛肉中阿莫西林残留量的高效液相色谱分析

建立了反相高效液相色谱分析牛肉样品中阿莫西林残留量的方法.对动物组织样品采用磷酸盐缓冲溶液提取、三氯乙酸沉淀蛋白、固相萃取净化和富集.研究了样品在C18柱上的保留行为和阿莫西林的特征谱图.待测物用外标定法定量分析,标准曲线的线性范围为5.0～500 μg/L,相关系数为0.99934.动物组织样品中阿莫西林的加标回收率为77.3%～97.4%,相对标准偏差为1.2%～13.5%,检出限低于10 μg/kg.     

高效液相色谱法测定乳制品中的双氰胺

建立了乳制品中双氰胺的高效液相色谱(HPLC)检测方法。样品经乙腈提取后采用HPLC测定。优化后的色谱条件:XBridge Amide柱(250 mm×4.6 mm, 3.5 μm),流动相为乙腈和水(体积比为90:10,含0.2%甲酸),流速为1.0 mL/min,检测波长为218 nm。方法在0.5~50 mg/L范围内线性关系良好,相关系数(r²)为0.9999,回收率在96.7%~101.0%之间,RSD为4.5%~4.9%(n=6),检出限为0.2 mg/kg(S/N=3),定量限为0.5 mg/kg(S/N=10)。该方法简单、准确、灵敏度高,适用于各种乳制品中双氰胺的检测。     

固相萃取RP-HPLC测定小鼠肝脏组织中的多沙唑嗪

研究建立了小鼠肝脏组织中多沙唑嗪的反相高效液相色谱测定方法. 肝组织样品经过匀浆、提取、C18固相萃取小柱富集净化后, 在YMC C18色谱柱(4.6 mm i.d.×250 mm, 5 μm)上, 以V(甲醇)∶V(0.02 mol/L KH2PO4) ＝70∶30, pH 3.0为流动相, 流速0.6 mL/min, 检测波长246 nm对多沙唑嗪进行测定. 结果表明, 肝脏组织中的多沙唑嗪在0.5～10 μg/mL范围内与峰面积呈良好线性关系. 平均回收率为91.0%, RSD为3.3%. 检出限为1 ng. 方法操作简便, 重现性好, 适用于肝脏组织中多沙唑嗪药物的浓度测定及代谢研究.     

用高效液相色谱法测定灯盏花中槲皮素的含量

分别采用溶剂回流提取和超声提取两种方法对灯盏花样品中的槲皮素进行提取,优化了提取条件,并用高效液相色谱进行了定量分析,采用扫描电镜分析了提取样品的表面形貌.结果表明:两种方法的提取率分别为0.048 7mg/g和0.042 3mg/g,溶剂回流的提取效果略优于超声提取;回流提取时的高温有利于提取物的溶出,而超声提取节能、快速.与此同时,采用两种方法提取的样品表面都受到明显的侵蚀,且回流提取时的机械搅拌促进样品的破碎,这些都有利于槲皮素的溶出.     

高效液相色谱法同时测定中药材虎掌南星的核苷类成分

建立了高效液相色谱同时测定中药材虎掌南星中核苷类活性成分(腺嘌呤、次黄嘌呤、黄嘌呤、尿苷、胸腺嘧啶、腺苷、鸟苷)含量的方法。以Lichrospher C18色谱柱(150 mm×4.6 mm, 5 μm)分离,以乙腈-水(含0.1%甲酸)为流动相,梯度洗脱,在254 nm下检测,腺嘌呤、次黄嘌呤、黄嘌呤、尿苷及鸟苷分别在1.6～50 mg/L范围内、胸腺嘧啶和腺苷分别在1.2～40 mg/L范围内的线性关系良好,相关系数均大于0.9995,加标回收率为98.9%～101.2%,相对标准偏差均小于3%。方法学考察结果显示符合含量测定要求,并应用于不同产地虎掌南星的测定。该方法操作简便、快速,结果可靠,重现性好,可作为虎掌南星质量评价的参考依据。     

高效液相色谱法测定化妆品中的7种色素

建立了用高效液相色谱-二极管阵列检测法( HPLC-DAD)同时检测化妆品中7种着色剂(胭脂红Ponceau)、萘酚黄(Naphthol Yellow S Hydrate)、日落黄( Food Yellow 3)、酸性红33( Acid Red 33)、诱惑红(Allura Red AC)、酸性红13(Acid Red...     

高效液相色谱法测定食品中氟啶脲的残留量

建立了高效液相色谱测定食品中氟啶脲残留量的方法.样品中的氟啶脲经正己烷或乙腈提取,弗罗里硅土净化后,以乙腈-水( 85:15,v/v)混合溶液为流动相,经C18色谱柱分离,紫外检测器(260 nm)测定.结果表明:氟啶脲在0 05 ～2.0 mg/L范围内线性良好(相关系数为0 999 8),定量限(以信噪比为10计)...     

高效液相色谱法测定酒花α-酸含量

介绍反相液相色谱测定酒花和酒花浸膏中的α-酸含量的方法,反相色谱分离α-酸,外标法定量。方法具有样品处理简单、快速、定量准确和重复的特点。酒花和酒花浸膏的α-酸测定相对偏差分别为2.02％和1.78％。