An improved high-performance liquid chromatographic method with a solid-phase extraction for the determination of piperacillin and tazobactam: application to pharmacokinetic study of different dosage in Chinese healthy volunteers

An improved HPLC method was developed for the determination of piperacillin and tazobactam in human plasma and pharmacokinetic study in Chinese healthy volunteers. Piperacillin and tazobactam in human plasma were extracted by solid-phase extraction and separated on a C(18) column and detected at 220 nm. The mobile phase for piepracillin consisted of 0.01 mol/L sodium dihydrogen phosphate (pH = 4.65) and acetonitrile (71:29, v/v), and that for tazobactam was 0.05 mol/L sodium dihydrogen phosphate (pH = 4.45) and methanol (90:10, v/v). The method was linear in the range 0.25-320.00 microg/mL for piperacillin (r(2) = 0.995) and 0.25-64.00 microg/mL for tazobactam (r(2) = 0.994). The lower limit of quantification of both compounds was 0.25 microg/mL. The intra- and inter-day precisions of piperacillin and tazobactam at three concentrations were all less than 9.2% and accuracies were within the range 97.0-108.0%. The method was used to investigate the pharmacokinetics of piperacillin and tazobactam in 12 volunteers who were intravenously given a dosage of 1.25, 2.50 and 3.75 g in three periods. The results showed that piperacillin sodium-tazobactom sodium (4:1) for injection in Chinese people fits linear dynamics, and the administred dosage can be adjusted with therapeutic effect.     

Simultaneous determination of rifampicin and sulbactam in mouse plasma by high-performance liquid chromatography

A simple and rapid high-performance liquid chromatographic (HPLC) method with ultraviolet detection has been developed and validated for the simultaneous determination of rifampicin and sulbactam in mouse plasma. Plasma samples were deproteinized with acetonitrile and separated by HPLC on a RP-18 (125 x 4 mm, 5 microm) column and gradient elution with potassium dihydrogen phosphate solution (pH 4.5; 50 mm) and acetonitrile at a flow-rate of 1.0 mL/min. Rifampicin and sulbactam were monitored at 230 nm and confirmed by means of their UV spectra using a diode-array detector. The method was linear at plasma levels from 1 to 100 microg/mL for rifampicin and from 5 to 200 microg/mL for sulbactam. The limits of quantification were 0.6 microg/mL for rifampicin and 4.2 microg/mL for sulbactam. The intra- and inter-day precisions of the method (RSD) were lower than 5% for both compounds. Average recoveries of rifampicin and sulbactam from mice plasma were 98.2 and 89.3%, respectively. The developed method was successfully applied to the determination of the pharmacokinetic profile of both compounds in mice.     

高效液相色谱指纹图谱法分析南方红豆杉药材的氯仿提取物

建立了南方红豆杉药材氯仿提取物的高效液相色谱(HPLC)指纹图谱分析方法。采用Eurospher 100 C18色谱柱(250 mm×4 mm, 5 μm),以甲醇和水为流动相进行梯度洗脱,流速为1 mL/min,检测波长为232 nm,柱温为30 ℃。以10-脱乙酰巴卡亭III(10-DABIII)为参照物,在相同的色谱条件下测定了10批不同产地的南方红豆杉药材氯仿提取物的指纹图谱,获得了11个共有指纹峰,并利用主成分分析法(PCA)对指纹图谱进行统计分析。结果表明南方红豆杉药材的质量与种植区域有关。该方法稳定、可靠,可用于南方红豆杉药材的质量控制。     

A solid-phase extraction method for high-performance liquid chromatographic determination of salvianolic acid B in rabbit plasma: application to pharmacokinetic study

A sensitive solid-phase extraction/high-performance liquid chromatographic method with ultraviolet detection was established for the analysis of salvianolic acid B in rabbit plasma. The analyte was separated on a reversed-phase column with trifluoroacetic acid-methanol-acetonitrile (70:10:20, v/v/v) as mobile phase at a flow rate of 1 mL/min, and ultraviolet detection at 315 nm. The calibration curve for salvianolic acid B was linear over the range 35-1400 microg/L with coefficients of correlation >0.999. The inter-day and intra-day precisions of analysis were <15%, and assay accuracy ranged from 95.3 to 109.1%. This method is suitable for determining salvianolic acid B in plasma and thus investigating the pharmacokinetics of salvianolic acid B.     

天冬酰胺合成酶B抑制剂高效液相色谱筛选方法的建立与应用

建立了一种快速、高效测定天冬酰胺合成酶B(AS-B)酶活性的反相高效液相色谱法(RP-HPLC)。酶反应体系中的氨基酸经2,4-二硝基氟苯(DNFB)柱前衍生,通过RP-HPLC测定酶反应体系前后底物及产物的变化来分析酶的活性。采用的色谱柱为Agilent C18柱(250 mm×4.6 mm,5 μm),以50 mmol/L醋酸钠缓冲液(pH 6.2)-乙腈(体积比为15:85)为流动相,流速为1.0 mL/min,柱温为30 ℃,检测波长365 nm,于6 min内实现了各组分的基线分离。通过该方法测定反应动力学参数进行AS-B的抑制定量分析。将已知AS-B抑制剂L-谷氨酸-γ-甲酯作用于酶反应体系,测得的抑制剂的抑制常数与文献值相接近,证明该方法可用于AS-B抑制剂的筛选。     

Ultra-high performance liquid chromatographic method for the determination of polycyclic aromatic hydrocarbons in a passive environmental sampler

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous compounds released in the environment by different sources. The aim of the present work was to validate a solid‐phase extraction (SPE) and a rapid ultra‐high performance liquid chromatographic (UHPLC) method for the analysis of PAHs in a passive environmental sampler, namely a Dacron® (the commercial name of a synthetic fiber based on polyethylene terephthalate) textile. The elution temperature was optimized to improve the resolution of early‐eluted compounds, namely acenaphthene (Ac) and fluorene (F). The UHPLC method lasts about 10 min and showed good linearity for all the 16 PAHs considered, with regression coefficients over 0.99. Recoveries, limits of detection (LODs), and limits of quantification (LOQs) of the SPE method were well within the performance criteria fixed by the Regulation n. 836/2011, namely 0.3 and 0.9 μg/kg, respectively.     

剖析液相色谱柱技术的复杂状况(英文)

 Column packings continue to evolve as the needs of users for high efficiency, high resolution and highly sensitive high performance liquid chromatographic (HPLC) analysis drive further developments. In comparing and contrasting modern HPLC columns technologies, diameters of column packings and particle materials are covered. Some products and applications of modern HPLC columns are provided. Future directions in packing developments are predicted in this introductory article.     

五味子提取物高效液相色谱分析方法的优化

针对五味子乙醇提取物的复杂体系,借助于复杂样品分析系统软件(CSASS),根据组分在4次简单线性梯度下42个峰的保留时间,快速准确地计算出各组分的液相色谱保留参数a,c值和峰形参数σ,W1/2。借助这些参数,对五味子色谱的分离情况进行高精度仿真预测。在此基础上,应用移动重叠分辨分离图和谱图仿真技术,发展了计算机辅助的五味子提取物的高效液相色谱全局优化方法。在优化条件下,五味子提取物的高效液相色谱分析可在40 min内完成,且常量成分和部分低含量成分都能够得到较好的分离。所建立的方法已成功地用于五味子中化合物保留时间及峰形的预测,并在此基础上对其色谱分离条件进行优化。     

高效液相色谱法同时测定化妆品中的10种合成着色剂

建立了高效液相色谱同时测定化妆品中颜料橙5、酸性黄36、颜料红53、酸性紫49、罗丹明B、溶剂蓝35、苏丹红Ⅱ、苏丹红Ⅳ、分散黄3和颜料红57等10种合成着色剂的方法。用四氢呋喃(THF)、二甲基亚砜(DMSO)和甲醇对样品进行分步超声辅助萃取、离心净化后,在Eclipse XDB-C₁₈(150 mm×4.6 mm, 5 μm)色谱柱上分离。用乙腈-0.02 mol/L乙酸铵溶液(用乙酸调pH至4.60)作为流动相进行梯度洗脱,二极管阵列检测器(DAD)检测。在0.5~20.0 mg/L范围内,10种着色剂的峰面积与质量浓度呈线性关系,相关系数(r)大于0.999;定量限(LOQ)为10~20 mg/kg。在3个添加水平的回收率均在92.9%~108.8%之间,相对标准偏差(RSD)为0.5%~6.1%(n=6)。该方法简便、快速、灵敏,适合油状、粉状和膏状化妆品中禁限用着色剂的定量检测。     

百合知母汤的高效液相色谱指纹图谱及其与组方药味的相关性

研究和建立了百合知母汤的高效液相色谱(HPLC)指纹图谱,为研究百合知母汤的药效物质基础及配伍变化提供了手段。采用Agela Venusil XBP-C18色谱柱(250 mm × 4.6 mm,5 μm),以乙腈和0.1%甲酸为流动相二元梯度洗脱,流速1 mL/min,检测波长315 nm,柱温25 ℃。以芒果苷为参照物,在相同的色谱条件下测定了10批不同产地的百合与知母制备的百合知母汤的指纹图谱,获得了16个共有指纹峰,通过与对照品的保留时间及紫外光谱比较,标定了5-羟甲基糠醛(5-HMF)、新芒果苷、芒果苷、异芒果苷、王百合苷B的出峰位置。该方法得到的百合知母汤的指纹图谱特征性和重现性较好,方法稳定、可靠,可以为百合知母汤的质量控制提供参考。通过实验归属了百合知母汤指纹图谱中的主要色谱峰,并确定了煎煮过程中的主要变化成分为5-HMF。     

高效液相色谱法同时检测化妆品中12种直接染料

建立了高效液相色谱分离检测化妆品中12种直接染料的方法。采用XB-C18色谱柱分离,以乙腈-0.02 mol/L乙酸铵水溶液（pH 9.0）为流动相梯度洗脱,二极管阵列检测器检测。结果表明,12种直接染料在线性范围内具有良好的线性关系,相关系数均> 0.999;方法的检出限为0.017~1.7 μ g/g,回收率为85.6%~113.4%,相对标准偏差为1.0%~7.7%;日内和日间精密度分别为0.4%~6.5%和0.2%~7.5%。该法简单快速,灵敏度高,定量准确可靠,适用于化妆品中直接染料的日常检测。     

仿刺参中皂苷类成分的高效液相色谱指纹图谱

利用高效液相色谱法建立了仿刺参皂苷类成分的指纹图谱,为仿刺参的质量控制提供了新的方法。采用固相萃取制备供试品溶液,选用Zorbox SB-C18色谱柱(250 mm×4.6 mm, 5 μm),以乙腈-0.1%磷酸水溶液为流动相进行梯度洗脱,检测波长为205 nm,柱温30 ℃。分析了不同产地的10批仿刺参样品,采用国家药典委员会推荐的“中药色谱指纹图谱相似度评价系统(2004 A版)”处理谱图,确定了6个共有峰。计算了10个样本间的指纹图谱相似度,所得相似度计算结果均大于0.97。该方法具有良好的稳定性和重现性,可用于仿刺参的质量控制。     

HPLC测定一次性塑料用品中邻苯二甲酸酯类增塑剂

本文用高压、液体色谱柱Liehmspher C-18(250mm×4.6mmID,5μm),以乙腈-水为流动相,流速为1.0mL/min,线性梯度乙腈从70%到100%,采用质谱检测器对邻苯二甲酸酯进行定性鉴定.邻苯二甲酸二甲脂(DMP)、邻苯二甲酸二乙酯(DEP)、邻苯二甲酸二丁酯(DBP)和邻苯二甲酸二甲酸辛酯(DOP)浓度分别为1.1～89mg/L,0.9～74mg/L,0.9～71mg/L和0.9～68mg/L时线性关系良好,平均回收率分别为83%,89%,86%,87%(n=5).建立了准确度和灵敏度高,方便快捷有效的一次性塑料用品中邻苯二甲酸酯类增塑剂的高效液相色谱定量分析方法.     

A high-performance liquid chromatographic method for determination of flecainide in human plasma using fluorescence detection

Summary A high performance liquid chromatographic method for the determination of flecainide in serum has been developed. The analysis is performed on a microparticulate silica column. The eluate is monitored by fluorescence detection at an excitation wavelength of 300nm and an emission wavelength of 370nm. No sources of interference were identified and a coefficient of variation of less than 8% was observed on repeated flecainide determinations. The method has a good reproducibility, specificity and accuracy, and can be applied in therapeutic drug monitoring of flecainide in patients.     

高效液相色谱法同时检测化妆品中38种限用着色剂

建立了高效液相色谱法同时测定化妆品中颜料红4等38种限用着色剂的检测方法。样品经四氢呋喃、甲醇、乙酸铵水溶液混合溶剂超声提取后,经离心、氮吹和复溶,于Agilent zorbax SB-Aq色谱柱(150 mm×3.0 mm, 3.5 μm)上进行反相液相色谱分离,以乙腈和含0.075%(v/v)甲酸的30 mmol/L乙酸铵溶液为流动相进行梯度洗脱。选择254、416、484、514、590和620 nm作为检测波长进行定量分析。结果表明:在1~10 mg/L的质量浓度范围内,38种限用着色剂的色谱峰面积与质量浓度呈线性相关,相关系数(r)大于0.999, LOQ值为5~50 μg/g。在100 μg/g和500 μg/g两个加标水平下,各目标化合物回收率均在93.2%~107.6%之间,相对标准偏差(RSD)小于10%(n=6)。该方法简便、快速、灵敏度高、重现性好,适合于化妆品中限用着色剂的定性与定量检测。     

高效液相色谱法同时测定化妆品中的呋喃妥因和呋喃唑酮

建立了一种同时检测化妆品中呋喃妥因和呋喃唑酮的高效液相色谱法(HPLC)。乳液、膏霜、化妆水、散粉、唇膏类等不同类型的化妆品样品由乙腈-甲醇(体积比为1∶1)混合溶液超声提取后进行HPLC分析。采用Kromasil C18色谱柱,以乙腈-0.4%乙酸溶液(体积比为30∶70)为流动相,流速1.0 mL/min,采用二极管阵列检测器检测,检测波长为365 nm;采用外标法定量,呋喃妥因和呋喃唑酮检测的线性范围为0.1～20 mg/L,相关系数为0.9999,最低检出限为1.2 mg/kg;在0.2,1.0,10.0 mg/L加标水平下,平均回收率为88.0%～104.6%,相对标准偏差为0.5%～4.8%。该方法快速准确,可用于化妆品中呋喃妥因和呋喃唑酮的同时测定。     

鸡蛋中三聚氰胺的液相色谱快速测定

建立了鸡蛋中三聚氰胺的液相色谱快速检测方法。样品经1%三氯乙酸溶液超声提取,磺基水杨酸和乙腈沉淀蛋白,高速离心分离,上清液过滤后直接由高效液相色谱分析。在优化的条件下,样品的色谱分析时间仅为10 min;三聚氰胺标准品在0.05～20 mg/L范围内,线性相关系数为0.9999;方法定量限为0.10 mg/kg (S/N＞10);1.0,2.0,4.0,6.0 mg/kg 4个加标水平测试,回收率在94.2%～107%之间;6次平行测试的相对标准偏差(RSD)为1.53%～2.06%。该方法前处理简单、分析速度快、灵敏度高,满足鸡蛋中三聚氰胺检验工作的快速测定要求。     

色谱柱选择对23种防腐剂测定结果的影响

以化妆品中23种防腐剂检测方法为例,探讨色谱柱选择对液相色谱方法测定结果的影响。参照《化妆品安全技术规范》甲基异噻唑啉酮等23个组分的检验方法,在2台不同的高效液相色谱仪上用15款不同品牌、型号的C₁₈色谱柱检测23种防腐剂,计算色谱峰的理论塔板数和分离度,对23种组分的分离效果进行分析,并应用USP (United States Pharmacopeia)数据库和PQRI (Product Quality Research Institute)数据库等2种等效色谱柱选择方法,对不同色谱柱的分离效果及等效性进行评价和预测。实验结果表明,15款色谱柱对23种防腐剂的分离效果差异显著,仅有2款色谱柱可以实现23种组分的完全分离。USP和PQRI数据库中2种等效色谱柱选择方法均无法预测出合适的等效色谱柱,对23种防腐剂的液相色谱分析参考价值均较小。色谱柱是影响23种防腐剂液相色谱法测定结果准确性的关键因素,有关实验室在应用该方法时,应考虑色谱柱选择性差异。化妆品基质复杂,如何在现有研究成果的基础上,开发色谱柱的筛选和预测评价体系,进而指导实际样品的分离是下一步研究的重点、难点。建议有关部门在制修订检测方法时,注重色谱柱的耐用性考察,完善系统适应性指标,细化色谱柱分类和增加描述信息,指导色谱柱的合理选择,从而规避由于色谱柱使用过程中选择依据缺失而导致测定结果不准确的风险。     

运用色谱指纹图谱与化学计量学方法对灵芝进行分类

采用95%乙醇为提取溶剂,运用高效液相色谱(HPLC)指纹图谱技术与化学计量学方法,对11个不同灵芝菌株子实体进行分类。通过相似度分析分别获得提取样品指纹图谱的13个共有峰及每个样品之间的相似度;以相对共有峰面积为分析参数,运用化学计量学方法包括聚类分析(HCA)、主成分分析(PCA)及判别分析(DA)对其进行分类,结果分为紫芝、赤芝和美国大灵芝3类。实验结果表明,用化学计量学的方法对灵芝样品的指纹图谱数据进行分析,是一种可用于其分类的科学方法。     

高效液相色谱法同时测定厚朴温中胶囊中的7种有效成分

建立了同时测定厚朴温中胶囊中山姜素、甘草酸、和厚朴酚、小豆蔻明、木香烃内酯、去氢木香内酯及厚朴酚含量的反相高效液相色谱法。固定相为Scienhome C18柱(250 mm×4.6 mm,5 μm),流动相为甲醇-乙腈-0.06%磷酸溶液(体积比为38∶27∶35),流速为1.0 mL/min,柱温为30 ℃,检测波长为235 nm。在上述条件下,山姜素、甘草酸、和厚朴酚、小豆蔻明、木香烃内酯、去氢木香内酯及厚朴酚的质量浓度分别在0.885～17.7,107～2140,8.85～17.7,1.035～20.7,4.85～97,5.9～118和17.5～350 mg/L时与色谱峰面积之间的线性关系良好;回收率分别为96.9%～101.1%,96.0%～100.5%,100.3%～100.8%,97.7%～101.4%,100.4%～102.3%,96.0%～102.3%和96.2%～100.6%。该方法简便、快速、准确,可用于厚朴温中胶囊的质量控制。