特非那定片有关物质的检测方法研究

目的建立特非那定片的有关物质高效液相色谱检查法（HPLC）。方法采用HPLC法,使用HypersilPhenyl2（250x4．6mm,5p,m）色谱柱,流动相为缓冲液．乙腈．三乙胺（650：350：3;检测波长为220hm;流速为1．5mL·min-1;柱温为室温。结果该色谱条件下,杂质与主成分能完全分离,重复性试验RSD为0．27％（n=6）,最低检测量为0．005μg,有关物质含量均为0．3％。结论该方法简便、准确、灵敏度高,可用于特非那定片中杂质含量的测定。     

HPLC和络合-UV法测定特非那丁颗粒剂的含量

目的:用HPLC和电荷转移络合UV法分别测定特非那丁颗粒剂的含量,并对测定结果进行比较。方法:HPLC法,色谱柱:SpherisorbC8( 1 50×4 .6mm ,5μm) ;流动相:甲醇0 .1mol/L三乙胺磷酸缓冲液( 80∶2 0 ) ,检测波长2 3 5nm。电荷转移络合UV法,利用碘与TFN在氯仿中形成电荷转移络合物的原理,在紫外2 94nm的波长处测定TFN的含量。结果:两法测定的线性范围和平均回收率分别为:HPLC法1 50～1 50 0 μg/ml,99.4 0 % (RSD0.92%) ;络合UV法2～1 2 μg/ml,99.4 5% (RSD0.67%)。结论:两种测定方法准确可靠,并无显著性差异。     

HPLC法测定小儿氨酚黄那敏片中马来酸氯苯那敏含量及含量均匀度

目的：建立HPLC法测定小儿氨酚黄那敏片马来酸氯苯那敏含量及含量均匀度。方法：采用HPLC法,色谱柱：KromasilC18（250mm×4.6mm,5μm）;流动相：乙腈-0.05mol/L磷酸氢二钠（含0.02%三乙胺,用磷酸调pH为3.5）（20：80）;流速：1.0ml/min;检测波长：260nm。柱温：30℃。结果：马来酸氯苯那敏检测浓度的线性范围为0.811～8.12μg/ml（r=0.9996）。平均回收率为99.21%,RSD为0.25%。结论：方法灵敏,可靠,简单可行,可用于该制剂的马来酸氯苯那敏含量及含量均匀度测定。     

3种方法测定伪麻黄碱血药浓度的比较研究

目的 建立并比较测定伪麻黄碱血药浓度的HPLC法、柱前衍生-HPLC法和LC/MS/MS法,为体内低浓度伪麻黄碱的分析提供更为简便、准确的方法.方法 摸索并优化血浆前处理条件、衍生化条件、色谱条件、质谱条件等.结果 3种方法均具有较好的专属性、线性、精密度、回收率和稳定性,可准确、灵敏地检测生物样本中的伪麻黄碱.3种方...     

LC-MS/MS方法测定人血浆中盐酸非那吡啶的浓度

目的 建立LC-MS/MS法测定人血浆中盐酸非那吡啶浓度.方法 色谱柱为Capcell PAK C18,流动相为甲醇-10 mmol乙酸铵(85:15,pH =4),流速为0.15 mL·min-1,电喷雾离子源(ESI),正离子方式检测,多反应监测(MRM).结果 盐酸非那吡啶的线性范围为0.2～50.0 ng·mL-1,日内、间差RSD均≤11.5％,提取回收率为≥60.2％.结论 该方法灵敏、准确、快速,可用于人血浆盐酸非那吡啶浓度的测定.     

两种方法测定复方氨基酸注射液(18AA-Ⅴ)中亚硫酸氢钠含量

目的:建立高效液相色谱(HPLC)法和比色法两种方法用于测定复方氨基酸注射液(18AA-Ⅴ)中的抗氧剂亚硫酸氢钠,并比较两种方法是否存在显著性差异.方法:HPLC法色谱条件为采用Shoedx Sugar SH1011色谱柱(300 mm×8 mm,6 μm),以0.04 mol/L磷酸溶液为流动相,流速0.8 ml/min,柱温为30 ℃,检测波长为200 nm;比色法原理为亚硫酸氢钠在酸性条件下与甲醛和碱性品红可生成紫红色络合物,通过测定该络合物在556 nm波长处的吸光度,可计算出亚硫酸氢钠的含量.结果:HPLC法亚硫酸氢钠在41.8~313.9 μg/ml范围内峰面积与浓度呈良好线性关系(r=0.999 5),平均回收率为102.4%,RSD为0.65%(n=9);比色法亚硫酸氢钠在0.8~2.5 μg/ml浓度范围内呈良好的线性关系(r=0.999 0),平均回收率为99.7%,RSD为0.83%(n=9).结论:两种方法均可用于测定复方氨基酸注射液(18AA-Ⅴ)中的抗氧剂亚硫酸氢钠,且无显著性差异,但HPLC法操作更简便,适合批量样品的测定.     

药品标准中HPLC法色谱条件的优化与系统适用性试验的重要性

本文通过三例药品标准中的HPLC法试验,对HPLC法色谱条件的优化和系统适应性试验的重要性进行了讨论.提出了HPLC法作为含量测定方法和有关物质的检查方法,必须对色谱条件进行优化筛选并确立一个操作性强、判断明确的系统适应性实验方法,才能确保方法的重现性和准确性.     

HPLC法测定复方醋酸氟轻松酊中水杨酸和间苯二酚的含量

目的建立一种用HPLC法,同时测定复方醋酸氟轻松酊中水杨酸和间苯二酚含量的方法。方法采用C18色谱柱,流动相为甲醇-0.02mol/L磷酸二氢钾溶液(以0.1mol/L氢氧化钠溶液调节pH值至5.8),检测波长为230nm。结果水杨酸、间苯二酚线性范围分别在0.01～0.1mg/mL和0.02～0.2mg/mL的浓度范围内均呈现出良好的线性关系。结论该法具有简便、准确等特点。     

HPLC法测定盐酸环丙沙星含量方法的建立

目的：建立盐酸环丙沙星含量测定的HPLC法。方法：以Diamonsil^TM(钻石）C18为固定相，0.05mol/l枸橼酸溶液－乙腈（82：18）为流动相，在检测波长277nm条件下测定环丙沙星含量，并与药典方法进行比较。结果：在建立的色谱条件下，在浓度5－30μg/ml范围内线性良好，且与其杂质能完全分离，相关系数为0．9985。结论：本法准确、方便、可靠。     

高效液相色谱法测定保济丸中厚朴酚与和厚朴酚的含量

目的：测定保济丸中厚朴酚与和厚朴酚的含量，并对两个生产厂家的样品进行含量比较。方法：采用高效液相色谱(HPLC)法、C18柱，以乙腈—水—冰醋酸(60：40：2)为流动相，检测波长为294nm。结果：厚朴酚浓度在0．1061—1．6976μg／mL和厚朴酚浓度在0.0559～0.8944μg／mL内与峰面积线性关系良好，加样回收率分别为100．21％，98．83％。结论：HPLC法简便，测定结果准确、重现性好，可用于保济丸的质量控制。     

Characterization and quantitative determination of impurities in piperaquine phosphate by HPLC and LC/MS/MS

Four impurities in piperaquine phosphate bulk drug substance were detected by a newly developed gradient reverse phase high performance liquid chromatographic (HPLC) method. These impurities were identified by LC/MS/MS. The structures of impurities were confirmed by spectroscopic studies (NMR and IR) conducted using synthesized authentic compounds. The synthesized reference samples of the impurity compounds were used for the quantitative HPLC determination. The system suitability of HPLC analysis established the validity of the separation. The method was validated according to ICH guidelines with respect to specificity, precision, accuracy and linearity. Forced degradation studies were also performed for piperaquine phosphate bulk drug samples to demonstrate the stability indicating power of the newly developed HPLC method.     

Isolation and characterization of a potential process related impurity of phenazopyridine HCl by preparative HPLC followed by MS–MS and 2D-NMR spectroscopy

During the process development of phenazopyridine HCl bulk drug, a potential impurity was detected in the routine impurity profiles by HPLC. Using MS–MS and multidimensional NMR techniques, the trace level impurity was unambiguously identified to be 3-phenyl-5-phenylazo-pyridine-2,6-diamine after its isolation from phenazopyridine HCl by semi-preparative HPLC. The formation of the impurity was discussed. To our knowledge, it is a novel impurity not reported elsewhere.     

应用HPLC-DAD/MS技术评价中药天麻的质量

分析比较不同产地及不同炮制方法的天麻药材质量。天麻药材指纹图谱HPLC-DAD分析条件如下：Zorbax XDB C₁₈色谱柱;流动相为乙腈-0.1%醋酸水梯度洗脱,检测波长为270 nm。MS分析条件：采用ESI电离源,负离子检测模式。天麻药材含量测定HPLC条件：流动相为乙腈-0.1%醋酸水等度洗脱,其余均同天麻药材指纹图谱的液相分析条件。结果表明,不同产地天麻药材具有相似的指纹图谱,原药材直接冻干的炮制方法较其他方法具有较高的相似度;指认了天麻药材指纹图谱中8个主要的色谱峰;指纹图谱相似度的分析结果与主要成分定量分析的结果基本一致。本文所建立的HPLC-DAD/MS分析方法能够对天麻药材进行全面的质量评价。     

Simultaneous determination of zidovudine and lamivudine in human serum using HPLC with tandem mass spectrometry

A method employing high performance liquid chromatography (HPLC) with tandem mass spectrometry (MS) has been developed and validated for the simultaneous determination of clinically relevant levels of zidovudine (AZT) and lamivudine (3TC) in human serum. The method incorporates a fully automated ultrafiltration sample preparation step that replaces the solid-phase extraction step typically used for HPLC with UV detection. The calibration range of the dual-analyte LC-MS/MS method is 2.5–2500 and 2.5–5000 ng ml⁻¹ for AZT and 3TC, respectively, using 0.25 ml of human serum. The lower limit of quantification was 2.5 ng ml⁻¹ for each analyte, with a chromatographic run time of approximately 6 min. Overall accuracy, expressed as bias, and inter- and intra-assay precision are <±7 and <10% for AZT, and <±5 and <12.1% for 3TC over the full concentration ranges. A cross-validation study demonstrated that the LC-MS/MS method afforded equivalent results to established methods consisting of a radioimmuno-assay for AZT and an HPLC-UV method for 3TC. Moreover, the LC-MS/MS was more sensitive, allowed markedly higher-throughput, and required smaller sample volumes (for 3TC only). The validated method has been used to support post-marketing clinical studies for Combivir™ — a combination tablet containing AZT and 3TC.     

高效液相色谱法与容量分析法测定发酵液中L-赤藓酮糖含量的比较

目的：建立高效液相色谱法（ HPLC）测定发酵液中L-赤藓酮糖含量的方法,并与测定还原糖常用的容量分析法（ VA）斐林试剂滴定法进行了比较。方法 HPLC法的色谱条件为：色谱柱：Lichrospher 5-NH2（4．6 mm ×250 mm）;柱温：32℃;流动相：甲醇—水（85∶15,V／V）;流速：1．0 mL· min－1。用紫外检测器室温下检测L-赤藓酮糖,检测波长为277 nm,检测器温度为35℃。结果所得L-赤藓酮糖的线性范围为1．00～100．00 g· L－1,R2为0．9999,最低检测限为0．50 g· L－1,最低定量限为0．85 g· L－1。 F检验和t检验结果显示：在各自的线性范围内, VA法与HPLC法在95％的置信区间上存在显著性差异。结论与VA法相比,HPLC法精密度好,准确性高,不受发酵液中其它组分干扰,更适用于测定发酵液中L-赤藓酮糖的含量。     

色谱法测定人血浆中万古霉素的浓度

目的：建立HPLC法和液相色谱-串联质谱（LC-MS/MS）法检测人血浆中万古霉素的浓度,并分别与FPIA法进行比较。方法：HPLC法：100μL血浆样品加入等体积10%的高氯酸沉淀,振荡离心后取上清液进样。采用Agilent Eclipse XDB-C18（250 mm×4.6 mm,5μm）,柱温为35℃,流动相为0.05 mol·L-1 KH2PO4（pH=3.2）-甲醇（74∶26,v/v）,流速为1 mL·min-1,检测波长为230 nm。 LC-MS/MS法：100μL血浆样品用300μL乙腈沉淀,振荡离心后取上清液进样。色谱柱为Agilent Eclipse XDB-C18（100 mm ×2.1 mm,3.5μm）,柱温为40℃,流动相为水（含0.1%甲酸）-乙腈（90∶10,v/v）,流速为0.20 mL·min-1。采用电喷雾化离子源（ESI）,多离子反应模式（MRM）检测,万古霉素和内标去甲万古霉素的监测离子对分别为m/z 725.0→144.0和718.5→144.0。采用建立的HPLC法和LC-MS/MS法测定95份临床样本,并与FPIA法进行相关性和测定方法的偏倚分析。结果：HPLC法和LC-MS/MS法测定万古霉素的线性范围分别为1.2~96μg·mL-1和0.4~96μg·mL-1,低、中、高三种浓度质控品日内和日间相对标准差（RSD）均〈15%。两种方法与FPIA法有很强的相关性（r=0.9621,P〈0.0001和r=0.9466,P〈0.0001）,无明显偏倚。结论：建立的HPLC和LC-MS/MS法快速、灵敏、准确,样本测定结果与FPIA法无显著差异,适用于万古霉素的常规血药浓度监测及人体药物代谢动力学研究。     

LC determination of glimepiride and its related impurities

Five impurities in glimepiride drug substance were detected and quantified using a simple isocratic reverse phase HPLC method. For the identification and characterization purpose these impurities were isolated from a crude reaction mixture of glimepiride using a normal phase HPLC system. Based on the spectroscopic data like NMR, FTIR, UV and MS these impurities were characterized and used as impurity standards for determining the relative response factor during the validation of the proposed isocratic reverse phase HPLC method. The chromatographic separation was achieved on a Phenomenex Luna C8 (2) 100 Å, 5 μm, 250 mm × 4.6 mm using a mobile phase consisting of phosphate buffer (pH 7.0)–acetonitrile–tetrahydrofuran (73:18:09, v/v/v) with UV detection at 228 nm and a flow rate of 1 ml/min. The column temperature was maintained at 35 °C through out the analysis. The method has been validated as per international guidelines on method validation and can be used for the routine quality control analysis of glimepiride as active pharmaceutical ingredient (API).     

脑栓通胶囊高效液相色谱指纹图谱质量控制方法研究

目的建立脑栓通胶囊HPLC指纹图谱,用于脑栓通胶囊质量控制。方法分别构建脑栓通胶囊HPLC指纹图谱A和HPLC指纹图谱B,共同检出复方中的全部5味药材成分。采用2种样品前处理方法（超声提取、超声提取后液液萃取纯化）制备脑栓通胶囊供试品溶液,使用Ultimate AQ-C18（150 mm×4.6 mm,3μm）色谱柱进行色谱分离。指纹图谱A色谱条件如下：以乙腈（A）-四氢呋喃（B）-0.05%磷酸溶液（C）为流动相,检测波长在0～53 min为254 nm,于53 min后将波长切换为275 nm,柱温20℃,流速1.1 mL min-1;指纹图谱B色谱条件如下：以乙腈（A）-0.05%磷酸溶液（B）为流动相,检测波长390 nm,柱温25℃,流速1.0 mL min-1。结果脑栓通胶囊指纹图谱A（可检出蒲黄、赤芍、天麻、漏芦4味药材）确定了27个共有峰,通过对照品对照、快速液相-三重串联四级杆质谱（RRLC/MS/MS）鉴定了其中10个色谱峰的化学成分;脑栓通胶囊指纹图谱B（可检出郁金药材）确定了5个共有峰,鉴定了其中1个色谱峰的化学成分。结论该方法具有良好的可行性、稳定性和重现性,为脑栓通胶囊的质量控制提供了科学依据。     

Chromatographic fingerprint analysis and characterization of furocoumarins in the roots of Angelica dahurica by HPLC/DAD/ESI-MSn technique

A high performance liquid chromatography-diode array detection-electrospray ionization tandem mass spectrometry (HPLC/DAD/ESI-MSn) method was used for the chromatographic fingerprint analysis and characterization of furocoumarins in the roots of Angelica dahurica for the first time. Nine "common peaks" were identified by comparing with the retention time, UV and MS spectra of reference furocoumarins. The software "Similarity Evaluation System for Chromatographic Fingerprint of TCM" was used to evaluate the similarities of 13 batches of Baizhi samples collected from Henan, Zhejiang, Sichuan and Anhui provinces of China. The results indicated that the samples from different batches had similar HPLC fingerprints, and the method could be applied for the quality control of the roots of Angelica dahurica. In addition, a total of 20 furocoumarins were identified or tentatively characterized by HPLC/DAD/ESI-MSn technique, and their fragmentation patterns in an electrospray ion trap mass spectrometer were also summarized.     

Estimation of impurity profiles of drugs and related materials: part XXI. HPLC/UV/MS study of the impurity profile of ethynodiol diacetate

The impurity profile of ethynodiol diacetate was investigated using the HPLC/UV/MS method. Using the slightly modified HPLC method of USP 24 two impurities, earlier isolated by preparative HPLC and investigated by NMR spectroscopy were separated and characterised. The mass spectra amended by the diode-array UV spectra supported the earlier found structures (E and Z isomers of 17-ethinyl-estr-4-ene-3β,17-diol-3-acetate-17-(3′-acetoxy-2′-butenoate). Another, hitherto not described impurity, 17-ethinyl-estr-4-ene-3β,17-diol-3-acetate-17-(3-oxo-butanoate) has also been separated and characterised by means of its mass spectrum, NMR and UV spectra.