Determination of Sparfloxacin in Human Urine by Reversed-Phase High Performance Liquid Chromatography With Nitrous Acid and Hydroiodic Pre-Column Derivatization

Sparfloxacin can be oxidized by nitrous acid, then react with hydroiodic acid to form a fluorescent derivative. Based on this, a reversed-phase high performance liquid chromatographic pre-column derivatization new method is described for the determination of sparfloxacin in human urine. The linear range is 0.05 mg/L to 4.0 mg/L, the recoveries are 91.5%similar to 95.7% and the RSD is 1.2%similar to4.2%. The results showed that this method is suitable for the determination of sparfloxacin in human urine.

Determination of 18 Kinds of Amino Acids in Fresh Tea Leaves by HPLC Coupled with Pre-column Derivatization

A rapid and accurate quantitative method of high performance liquid chromatography( HPLC) with fluorescence detector has been developed for the analysis of 18 kinds of amino acids in fresh tea leaves. The samples were minced and mixed,and extracted with ultra pure water at 90℃ for 20 min. The 6-aminoquinolyl N-hydroxy-succinimidyl carbamate( AQC) was used as pre-column derivatization reagent. Gradient HPLC separation was performed on a C_(18) column( Symmetry C_(18),3. 9 mm × 15 cm,4 μm). Good linearity between concentrations and peak areas was achieved in the concentration range of 5. 0-250 μmol/L for 18 kinds of amino acids. The method was validated by the analysis of five replicates. The 18 kinds of amino acid standards were spiked in fresh tea leaf samples and the average recovery rate was 86. 25%-109. 05% with relative standard deviations( n = 5) ranging from 6. 03% to 10. 56%. The limit of detection( LOD) for the analytes was0. 05-1. 27 μmol/L. The method was successfully applied to the analysis of the 18 kinds of amino acids in fresh tea leaves from east Dongting and west Dongting mountains in Suzhou. The results indicate that the method is simple,rapid,precise and reliable.

Enantioresolution of a Series of Chiral Benzyl Alcohols by HPLC on a Dinitrobenzoylphenylglycine Stationary Phase after Achiral Pre-Column Derivatization

High performance liquid chromatography method for the separation of a series of chiral benzyl alcohols on N-(3,5-dinitrobenzoyl)-D-phenylglycine stationary phase (Macherey Nagel, Chiral-2) after pre-column achiral derivatization was developed. Cheap and easy available aromatic acid chlorides were used as derivatization agents. Good to excellent separations of the enantiomers were achieved in all cases in relatively short analytical runs. It was shown that the enantiorecognition depends on the substituents both in the starting alcohol and in the acid chloride. The method presents an efficient alternative to the direct analyses on polysaccharide and cyclodextrine-derived stationary phases.

Determination of Lignin in Marine Sediment Using Alkaline Cupric Oxide Oxidation-Solid Phase Extraction-on-Column Derivatization-Gas Chromatography

Lignin serves as one of the most important molecular fossils for tracing Terrestrial Organic Matters (TOMs) in marine environment. Extraction and derivatization of lignin oxidation products (LOPs) are crucial for accurate quantification of lignin in marine sediment. Here we report a modification of the conventional alkaline cupric oxide (CuO) oxidation method, the modification consisting in a solid phase extraction (SPE) and a novel on-column derivatization being employed for better efficiency and reproducibility. In spiking blanks, recoveries with SPE for the LOPs are between 77.84% and 99.57% with relative standard deviations (RSDs) ranging from 0.57% to 8.04% (n=3), while those with traditional liquid-liquid extraction (LLE) are from 44.52% to 86.16% with RSDs being from 0.53% to 13.14% (n=3). Moreover, the reproducibility is greatly improved with SPE, with less solvent consumption and shorter processing time. The average efficiency of on-column derivatization for LOPs is 100.8% ± 0.68%, which is significantly higher than those of in-vial or in-syringe derivatization, thus resulting in still less consumption of derivatizing reagents. Lignin in the surface sediments sampled from the south of Yangtze River estuary, China, was determined with the established method. Recoveries of 72.66% to 85.99% with standard deviation less than 0.01mg/10g dry weight are obtained except for p-hydroxyben-zaldehyde. The lignin content Σ8 (produced from 10 g dry sediment) in the research area is between 0.231 and 0.587 mg. S/V and C/V ratios (1.028 ± 0.433 and 0.192 ± 0.066, respectively) indicate that the TOMs in this region are originated from a mixture of woody and nonwoody angiosperm plants; the high values of (Ad/Al) v suggest that the TOMs has been highly degraded.

High－Performance Liquid Chromatographic Determination Of N－Nitrosoamines by Pre－Column Fluorescence Derivatization with 9－Phenanthrene Chloride

A sensitive high-performance chromatographic method for the detection of N-nitrosoamines with pre-column fluorescence derivatization has been developed.Eight representative N-nitrosoamines, which were separated on a reversed-phase C18column with methanol - Water-triethylamine as eluent, were used as model compounds to optimize the derivathation and the chromatographic conditions. The relative standard deviations(n=7) at an analytical concentration of 4×10-6 mol/L are less than 5%. The detection limits (signal-to-noise ratio=3) for the N-nitrosoamines compounds are in the range of 0.02～0. 14 nmol per injection volume(10μl).

柱前衍生高效液相荧光检测法测定复方氨基酸注射液中甲硫氨酸亚砜的含量

目的建立测定复方氨基酸注射液中甲硫氨酸亚砜质量分数的高效液相色谱法。方法采用柱前衍生HPLCFLD法,色谱柱为Agilent Poroshell 120 EC-C18柱(3.0 mm×100 mm,2.7μm)或效能相当的色谱柱,以乙酸钠四氢呋喃溶液为流动相A,乙酸钠溶液-乙腈-甲醇(体积比20∶40∶40)为流动相B,流速为1.0 mL/min,采用荧光检测器,激发波长为233 nm,发射波长为441 nm。结果甲硫氨酸亚砜质量浓度在0.02605~2.605μg/mL范围内线性关系良好(r=0.9996),检测限为1.3×10^(-3)ng,平均回收率为100.1%,RSD为0.3%。结论建立的方法操作简单、准确、灵敏度高,适用于复方氨基酸注射液中甲硫氨酸亚砜的测定。

不同产地及加热方式文冠果仁中的氨基酸成分分析

目的 分析不同产地的文冠果仁氨基酸成分及加热方式对氨基酸的影响。方法 采用ZORBAX Eclipse Plus C18柱(4.6 mm×250 mm,5μm),样品以异硫氰酸苯酯进行柱前衍生化,以乙腈-甲醇-水(3∶1∶1)为流动相A,0.1 mol·L^(-1)的醋酸钠溶液(p H 6.5)-乙腈(93∶7)为流动相B,柱温35℃,检测波长254 nm。结果 17种氨基酸在1.806~225.200μg·m L^(-1)与峰面积呈良好的线性关系,各产地必需氨基酸、药用氨基酸、苦味氨基酸和儿童必需氨基酸占总氨基酸的含量分别为14.40%~18.99%、64.06%~73.79%、14.82%~17.09%、23.53%~29.30%,北方地区的文冠果仁氨基酸含量、必需氨基酸含量和药用氨基酸含量均比较高,文冠果仁经过加热处理后各类氨基酸均有所提高。结论 不同产地文冠果仁的氨基酸在含量和营养价值上具有差异,北方地区的文冠果仁氨基酸营养和药用价值较高,文冠果仁经过一定的加热后食用营养价值和药用价值得到提高。

柱前衍生化液质联用法测定氯乙酰氯试剂中的乙酰氯和二氯乙酰氯

建立了柱前衍生化超高效液相色谱-三重四级杆联用(UPLC-MS/MS)法同时测定酰化试剂氯乙酰氯中的杂质乙酰氯和二氯乙酰氯的含量。以苯胺作衍生化试剂,采用Poroshell 120 Aq-C18(150 mm×2.1 mm, 2.7μm)为色谱柱,0.1%(V/V)甲酸水和乙腈为流动相,梯度洗脱,流速为0.3 mL/min。在电喷雾离子源(ESI),正离子监测下,采用多反应监测扫描(MRM)模式。乙酰氯和二氯乙酰氯分别在0.5~50 ng/mL和5~500 ng/mL范围内线性关系良好(R^(2)≥0.998 6),定量限为0.5和5 ng/mL,检出限为0.2和2 ng/mL;回收率(n=3)为90.8%~103.1%和92.5%~108.7%,RSD为1.4%~6.6%和0.9%~3.8%。衍生化产物室温放置24 h内稳定。方法准确可靠,简便高效,可用于氯乙酰氯中乙酰氯和二氯乙酰氯的同时检测。

基于HPLC研究柏子仁中黄曲霉毒素的污染状况与风险分析

目的基于HPLC研究柏子仁中4种黄曲霉毒素的污染情况,并进行风险分析,评估柏子仁的用药安全。方法采用Agilent Eclipse Plus C 18(4.6×250 mm 5μm)为色谱柱,柱后光化学衍生法检测,以甲醇∶乙腈∶水(35∶10∶55)为流动相,流速1.0 mL·min^(-1);柱温:40℃;荧光检测器检测。结果黄曲霉毒素B 1、B 2、G 1、G 2分别在0.35~20.8μg·L^(-1)、0.13~7.6μg·L^(-1)、0.36~21.6μg·L^(-1)、0.13~7.6μg·L^(-1)范围内呈良好的线性关系,黄曲霉毒素B 1、B 2、G 1、G 2平均回收率分别为90.6%、83.46%、87.84%、86.58%,相对偏差分别为2.9%、3.6%、3.6%、4.7%。23批柏子仁中有14批检出黄曲霉毒素B 1和总量,残留量分别在1~4μg·kg^(-1)和1~5μg·kg^(-1)之间,均符合规定,但检出率高达61%,存在安全隐患。结论该方法简单、准确、方便,能有效评价柏子仁中黄曲霉毒素的污染情况。警示柏子仁易受黄曲霉毒素的污染,需加强柏子仁监管力度和不定期的专项抽检,降低安全风险,以确保临床用药安全。

HPLC-柱后衍生荧光法测定水产品中河豚毒素含量结果的不确定度评定

通过建立HPLC-柱后衍生荧光法测定水产品中河豚毒素含量的不确定度评定方法,量化分析测定结果的不确定度分量,并对各分量相对贡献进行比较。结果表明:测定结果的不确定度主要来源于标准品的校准过程、样品称量、实验过程移取及定容、样品重复实验误差、前处理过程中的回收率不同等。其中校准不确定度中标准溶液配制的不确定度贡献最多,称样量不确定度最小;当水产品中河豚毒素的含量为90.6μg/kg时,其扩展不确定度为0.69μg/kg(包含因子k=2)。

高效液相色谱法测定罗望子胶的单糖组成

采用柱前衍生化高效液相色谱法,对罗望子胶中主要单糖组分进行检测。选用Diamonsil-plus C 18-A^(*)色谱柱,用磷酸盐缓冲液-乙腈溶液流动相进行等度洗脱,流速设置1.0 mL/min,柱温35℃,进样量5μL,检测波长245 nm。罗望子胶中主要单糖组分葡萄糖、木糖和半乳糖的比例为52.05%、20.03%和12.17%。数据在各自浓度范围内线性关系良好(R≥0.9992),加样回收率在100.33%~101.34%之间,RSD小于2.0%,检测限在0.0005~0.0009 mg/L之间。综上,该检测方法适用于罗望子多糖胶中单糖组分测定。

氧化型染发剂中过氧化氢含量检测方法研究

简述过氧化氢在氧化型染发剂中的作用原理及对产品质量的影响,对氧化型染发剂中过氧化氢含量主要检测方法——化学滴定分析法(高锰酸钾法)、柱前衍生高效液相色谱法、柱后衍生高效液相色谱法、鲁米诺化学发光法及荧光光谱法进行对比分析。结果表明:柱后衍生高效液相色谱法具有较高的自动化、较宽的线性范围、较低的检测限、较稳定的回收率,优于其他三种检测方法,适合大批量快速检测,但此法操作相对复杂、分析成本较高;化学滴定分析法以其具有的操作简单、精密度低、分析成本低等优点,在染发剂中过氧化氢常量分析中仍然占据着重要地位。

湖南地区特色发酵食品黄曲霉毒素B_(1)污染情况分析

随机抽取湖南地区特色发酵食品辣酱、豆瓣酱、豆豉共218批次,对其中的黄曲霉毒素B_(1)含量进行测定,并分析此3类食品中黄曲霉毒素B_(1)的污染情况,同时对黄曲霉毒素B1的测定方法进行比较分析及选择,最后对相关产品质量的提高,提出有效预防控制措施。

PMP柱前衍生反相高效液相色谱—紫外检测法结合离子色谱—脉冲安倍法测定不同产地泽泻多糖的单糖组成

本文建立了1-苯基-3-甲基-5-吡唑啉酮(PMP)柱前衍生高效液相色谱-紫外检测法结合离子色谱-脉冲安倍法测定泽泻中10种单糖含量的测定方法,并比较了不同产地泽泻多糖中的单糖组成差异。经测定泽泻多糖中含有葡萄糖、半乳糖、半乳糖醛酸和阿拉伯糖,其中葡萄糖含量最高,说明泽泻多糖是以葡聚糖为主的杂多糖。不同产地泽泻多糖中葡萄糖、半乳糖、半乳糖醛酸含量差异显著(P<0.05),葡萄糖含量由高到低依次为广泽泻(166.940~264.724 mg/g)、川泽泻(82.647~155.019 mg/g)和建泽泻(51.148~97.746 mg/g);广泽泻多糖中半乳糖含量(3.511~3.840 mg/g)显著高于川泽泻(1.193~1.797 mg/g,P<0.01)和建泽泻(低于定量限);仅有广泽泻中半乳糖醛酸含量(1.342~1.823 mg/g)高于方法定量限。广泽泻中葡萄糖、半乳糖、半乳糖醛酸摩尔比为60.13∶1∶0.40,川泽泻中葡萄糖、半乳糖摩尔比为1545.05∶1,建泽泻多糖中几乎都是葡萄糖,葡萄糖占比在上述产地多糖中依次增加,三种单糖的杂合程度依次减弱。研究结果将为泽泻多糖的结构研究和质量控制奠定基础。

PMP柱前衍生HPLC法测定八月瓜果皮多糖中单糖组成

为建立同时测定八月瓜果皮中6种单糖组分的色谱分离方法,采用1-苯基-3-甲基-5-吡唑啉酮(1-phenyl-3-methyl-5-pyrazolone,PMP)柱前衍生和高效液相色谱法测定八月瓜果皮中6种单糖组分。色谱柱为SHIMADZU-C18(4.6 mm×250 mm,5μm),流动相为乙腈(A)-0.05 mol/L甲酸铵溶液(B)梯度洗脱(0~10 min:18%A;10~20 min:18%~20%A;20~40 min:20%~23%A;40~60 min:23%A),柱温30℃,流速1 m L/min,检测波长245 nm。结果表明,所建立的PMP-HPLC柱前衍生化法可准确地测定八月瓜果皮中的D-甘露糖、L-鼠李糖、D-半乳糖醛酸、D-葡萄糖、D-半乳糖和D-阿拉伯糖含量,6种单糖成分在各自质量浓度范围内线性关系良好(R2≥0.998 3),加样回收率为92.58%~100.02%。10批八月瓜果皮样品中均检出6种单糖,D-甘露糖、L-鼠李糖、D-半乳糖醛酸、D-葡萄糖、D-半乳糖和D-阿拉伯糖平均物质的量比为1.18∶1.00∶23.25∶11.73∶3.51∶5.88,其中D-半乳糖醛酸含量为51.84~212.84 mg/g,D-葡萄糖含量为22.44~73.23 mg/g,D-阿拉伯糖含量为6.14~60.01 mg/g,是八月瓜果皮中的单糖主要成分。该试验所建立的方法操作简便、重复性好和准确度高,适用于八月瓜果皮多糖中单糖成分的分析。

地龙药材中黄曲霉毒素B_(1)、B_(2)、G_(1)、G_(2)的含量检测

目的建立超声萃取-免疫亲和柱净化-柱后光化学衍生法高效液相色谱法同时测定地龙药材中黄曲霉毒素B_(1)、B_(2)、G_(1)、G_(2)含量的测定方法。方法样品粉碎过120目筛后,采用70%甲醇超声处理30min,经免疫亲和柱净化、高效液相色谱分离、光化学柱后衍生,通过荧光检测器测定4种黄曲霉毒素的含量。结果线性范分别为:黄曲霉毒素B_(1)0.0104~0.0520ng(r=0.9999)、黄曲霉毒素B_(2)0.0038~0.0190ng(r=0.9998)、黄曲霉毒素G_(1)0.0108~0.0540ng(r=0.9998)、黄曲霉毒素G_(2)0.0038~0.0190ng(r=0.9998),线性关系良好。检出限分别为0.43、0.15、0.43、0.15μg·kg^(-1)。回收率在90.42~99.47%之间,RSD≤3.2%(n=6)。结论该方法操作简便,灵敏度高、重复性好、结果准确,可用于地龙中黄曲霉毒素含量的测定。

基于柱前衍生化GC-MS技术探究紫苏叶与白苏叶的质量差异

为比较紫苏叶与白苏叶化学成分差异,采用柱前衍生化气相色谱-质谱联用技术(GC-MS)对紫苏叶与白苏叶的氨基酸、有机酸、糖类、酚酸类等多种成分进行了定性和定量分析。进一步结合主成分分析(principal component analysis,PCA)和正交偏最小二乘判别分析(orthogonal partial least squares discrimination analysis,OPLS-DA)等多元统计分析方法筛选得到8个显著性差异化合物,分别为D-阿拉伯糖醇、D-果糖、D-半乳糖、D-葡糖醇、D-山梨糖醇、咖啡酸、麦芽糖、迷迭香酸,其中D-果糖、D-葡糖醇、咖啡酸、麦芽糖、迷迭香酸在紫苏叶中的相对含量均显著高于白苏叶。上述结果表明紫苏叶与白苏叶中的多种类型非挥发性成分含量存在显著差异,同时为不同种质紫苏叶的应用与开发提供了数据参考。

基于指纹图谱结合多元统计分析不同干燥方式蟾蜍肉氨基酸类成分

目的:建立不同干燥方式所得蟾蜍肉氨基酸类成分的指纹图谱及4个氨基酸的含量测定方法,比较不同干燥方式对蟾蜍肉品质及其氨基酸含量的影响。方法:采用异硫氰酸苯酯柱前衍生化-高效液相色谱法,建立热风干燥、冷冻干燥、真空干燥和阴干干燥4种方式所得蟾蜍肉干燥样品的指纹图谱,同时测定4个氨基酸含量,并进行相似度评价;运用SPSS 22.0软件进行聚类分析,运用SIMCA 14.1软件进行主成分分析(PCA)和偏最小二乘法-判别分析(PLS-DA)。结果:不同干燥方式得到的16批蟾蜍肉样品指纹图谱相似度结果均大于0.970,共匹配出10个共有峰,指认出缬氨酸、蛋氨酸、异亮氨酸、亮氨酸4个氨基酸;PCA提取出2个主成分,累积方差贡献率为87.95%;PLS-DA得出异亮氨酸、缬氨酸、亮氨酸、蛋氨酸的变量重要性投影(VIP)值均大于0.7,提示4个氨基酸可作为评价不同干燥方式所得蟾蜍肉品质的关键指标;阴干干燥、热风干燥、冷冻干燥和真空干燥4种方式所得蟾蜍肉氨基酸总质量分数测定值分别为7.92%、4.62%~5.37%、7.89%~9.02%和4.60%~9.24%。结论:所建立的指纹图谱及含量测定方法稳定可行,结合多元统计分析发现不同干燥方式对蟾蜍肉氨基酸含量产生较大影响,可为蟾蜍肉干燥方式和质量控制研究提供参考。

高效液相色谱法分析干旱胁迫下烟草中游离氨基酸的含量变化

建立了柱前衍生-高效液相色谱高灵敏、快速测定新鲜烟叶中21种游离氨基酸的新方法。新鲜烟叶样品经0.1 mol/L的盐酸超声提取30 min后,以6-氨基喹啉基-N-羟基琥珀酰亚胺基甲酸酯(AQC)作为衍生试剂,反向色谱柱分离,二极管阵列检测器测定。在最佳实验条件下,21种氨基酸在9 min内实现完全分离,在低、中、高3个浓度水平下的加标回收率在84.7%~104%之间,相对标准偏差(RSD)为1.8%~5.1%,定量限在0.02~0.10 mg/g之间。基于该方法,探讨了干旱胁迫下游离氨基酸含量的变化趋势。研究表明,在干旱胁迫下,总游离氨基酸、脯氨酸、天冬酰胺等含量明显升高,且大部分氨基酸在旺长期时变化最大,对明晰游离氨基酸参与烟草抗逆的调控机制提供了数据基础和技术支撑。

柱前手性衍生化-HPLC法测定科立内酯二醇对映异构体含量

基于柱前手性衍生化-HPLC法,以联苯-4-甲酰氯为衍生化试剂,采用硅胶表面涂覆有直链淀粉-三(s)-α-甲基苯基氨基甲酸酯手性色谱柱(AS-H,4.6 mm×250 mm,5μm),以正己烷-无水乙醇(V∶V=80∶20)为流动相,流速1 mL/min,柱温30℃,检测波长275 nm,外标法计算科立内酯二醇对映异构体超量值。结果显示,科立内酯二醇对映异构体衍生物分离度为2.8,(+)-科立内酯二醇衍生物在2.07~41.40μg/mL、(-)-科立内酯二醇衍生物在1.95~39.06μg/mL范围有良好线性关系(R2=0.9991、R2=0.9991)。(+)-科立内酯二醇衍生物低、中、高浓度平均加样回收率(RSD)分别为99.36%(0.51%)、99.24%(0.89%)、100.24%(0.65%);(-)-科立内酯二醇衍生物低、中、高浓度平均加样回收率(RSD)为99.74%(0.89%)、100.02%(0.31%)、98.81%(0.63%)。方法简便、快速,能准确测定科立内酯二醇对映异构体超量值,为前列腺素药物手性中间体提供新的分离和检测方法。