指示稳定性的高效液相色谱方法和强制降解研究片剂型马西替坦中的杂质（英文）

Macitentan (MAC) is a pulmonary arterial hypertension (PAH) drug marketed as a tablet and often has stability issues in the final dosage form. Quantitative determination of MAC and its associated impurities in tablet dosage form has not been previously reported. This study quantified impurities present in Macitentan tablets using a binary solvent-based gradient elution method using reversed phase-high performance liquid chromatography.The developed method w as validated per International Conference on Harmonization (ICH) guidelines and the drug product w as subjected to forced degradation studies to evaluate stability. The developed method efficiently separated the drug and impurities (48 min) w ithout interference from solvents,excipients,or other impurities. The developed method met all guidelines in all characteristics w ith recoveries ranging from 85%-115%,linearity w ith r 2≥0. 996 6,and substantial robustness. The stability-indicating nature of the method w as evaluated using stressed conditions (hydrolysis:1 N HCl at 80℃/15 min; 1 N NaOH at 25℃/45 min; humidity stress (90%relative humidity) at 25℃for 24 h,oxidation:at 6%(v/v) H2O2,80℃/15 min,thermolysis:at105℃/16 h and photolysis:UV light at 200 Wh/m2; Fluorescent light at 1. 2 million luxh). Forced degradation experiments show ed that the developed method w as effective for impurity profiling. All stressed samples w ere assayed and mass balance w as 96%. Forced degradation results indicated that MAC tablets w ere sensitive to hydrolysis (acid and alkali) and thermal conditions. The developed method is suitable for both assay and impurity determination,w hich is applicable to the pharmaceutical industry.     

反相高效液相色谱表征无定型聚芳醚酮(砜)成环解聚产物

建立了一种反相高效液相色谱表征无定型聚芳醚酮(砜)成环解聚产物的新方法。采用岛津VP-ODSC18反相色谱柱(150mm×4.6mm;5μm,i.d.),以四氢呋喃/水为流动相,流速1.0mL/min,梯度淋洗方式;254nm紫外检测,由此方法可将全部同系物组分有效地分离并可准确给出产物中各环状低聚体组分的百分含量。分析结果表明,酚酞聚醚酮、酚酞聚醚砜、双酚A聚砜经成环解聚反应高产率得到环状聚合体,最高成环率达97.57%。通过与激光质谱的比较证实了HPLC方法更适合于对成环解聚产物体系的表征。     

超高效液相色谱-串联质谱法同时测定肿瘤细胞mRNA中12种正常核苷和修饰核苷（英文）

建立了一种肿瘤细胞mRNA中12种正常核苷和修饰核苷的分析方法。肿瘤细胞样品经总RNA提取、mRNA纯化和mRNA水解后,加入8-溴鸟苷(8Br-G)为内标,以0.1%甲酸水溶液和0.1%甲酸甲醇溶液为流动相,采用Agilent RRHD SB-C₁₈(2.1 mm×50 mm,1.8μm)色谱柱进行分离,电喷雾电离源正离子扫描(ESI⁺),多反应监测(MRM)模式检测。该方法符合方法学验证要求,12种正常核苷和修饰核苷在一定浓度范围内线性关系良好,相关系数(r)均大于0.99,定量下限(LOQ)为0.17～3.13 ng/mL,检出限(LOD)为0.08～1.56 ng/mL。将建立的方法应用于人鼻咽癌5-8F细胞系、人宫颈癌Hela细胞系和人胚肾上皮293T细胞系,成功地检测出多种修饰核苷水平。该方法细胞用量少(每样品2×10⁶个),分析时间短(每样品6 min),灵敏度高,可为癌症研究和临床诊断等提供一种有效的分析方法。     

高效液相色谱-电感耦合等离子体质谱联用技术测定皮革中Cr(Ⅵ)

本文建立了高效液相色谱-电感耦合等离子体质谱(HPLC-ICP-MS)法测定皮革中Cr(Ⅵ)含量。采用Dionex AS19色谱柱,以NH4NO3(pH=7.4)为流动相,在流速为1.0mL/min的条件下,可以很好地分离Cr(Ⅲ)与Cr(Ⅵ)。在色谱进样量为100μL,质谱采用He碰撞池模式下,对Cr(Ⅵ)的检出限达到了0.02μg/L;2.0、20.0μg/L两个加标水平的回收率分别为93.5%~97.0%和97.8%~102.3%,方法精密度优于4.1%,可以满足测定要求。应用该方法测定了各类皮革产品中的Cr(Ⅵ)。     

4-(2-苯并噻唑偶氮)间苯二酚反相高效液相色谱分离和测定钒(Ⅴ)、铜(Ⅱ)、铁(Ⅲ)、钴(Ⅱ)和镍(Ⅱ)

以4-(2-苯并噻唑偶氮)间苯二酚作柱前衍生试剂,CLC-C₈为固定相,甲醇-水(68:32,V/V)含2×10^-3mol/LTBA·Br为流动相,在波长550nm处光度检测.可分离和测定Cu(Ⅱ)、V(Ⅴ)、Co(Ⅱ)、Fe(Ⅲ)和Ni(Ⅱ)。检出限分别为2.5×10^-1ngCu、9×10^-2ngV、1.3×10^-1ngCo、1.4×10^-1ng Fe和2.9×10^-1ng Ni.方法应用于保证纯及分析纯盐酸试样分析,获得满意的结果.     

苯并(a)芘的高效液相色谱荧光分析法的研究Ⅰ

本文对苯并(a)芘的荧光性质进行了研究,建立了苯并(a)芘的高效液相色谱分离荧光检测法.并用于环境水样分析,结果令人满意。     

凝胶渗透色谱净化-高效液相色谱法测定润滑脂中苯并(a)芘含量

提出了高效液相色谱法测定润滑脂中苯并(a)芘含量的方法。样品用环己烷-乙酸乙酯(1+1)混合溶剂提取,所得提取液须经凝胶色谱净化大分子干扰物质。采用高效液相色谱分离后,用荧光检测器进行测定。苯并(a)芘的质量浓度在5.00～200μg.L-1范围内与其峰面积呈线性关系,方法检出限(3S/N)为3μg.L-1。在3个标准加入水平下进行了回收试验,所得回收率在85.6%～97.8%之间,相对标准偏差(n=6)小于3.0%。     

反相高效液相色谱法测定光诱导反应产物N-(α-萘基)咔唑和N-(β-萘基)咔唑含量

报道了以Ｗａｔｅｒｓ ＢｏｎｄａｐａｋＴＭ，ＯＤＳ—Ｃ１８为色谱柱，甲醇：四氢呋喃：水（７２：６：２２，Ｖ／Ｖ为 流动相，在ＵＶ２５４ｎｍ为测定波长的条件下，利用外标法定量，建立了反相高效液相色谱法测 定光诱导反应粗产物中Ｎ－（α－萘基）咔唑和Ｎ－（β－萘基）咔唑的方法。为光诱导自由基链式亲 核取代反应（ＳＲＮ１）反应机理的探讨提供了可靠的理论依据。     

超高效液相色谱-串联质谱法测定纺织助剂中双(氢化牛脂基)二甲基季铵化合物

建立了快速测定纺织助剂中双(氢化牛脂基)二甲基季铵化合物(DHTDMAC)的超高效液相色谱-串联质谱(UPLC-MS/MS)分析方法.样品经5%(v/v)甲酸甲醇超声波提取、稀释后,采用Eclipse Plus C18柱(50 mm×2.1 mm, 1.8 μm)分离,甲醇和0.1%(v/v)甲酸水溶液为流动相梯度洗脱,电喷雾离子源正离子多反应监测模式进行定性和定量分析.实验结果表明,DHTDMAC在10~280 μg/L范围内与峰面积呈良好的线性关系,相关系数(r²)为0.9991;以不低于3倍信噪比计算方法检出限(LOD, S/N=3)为3 mg/kg;方法定量限(LOQ, S/N=10)为10 mg/kg.3种纺织助剂基质中DHTDMAC在3个不同加标水平的平均回收率为97.2%~108.3%,相对标准偏差(RSD)为1.5%~4.6%.该方法灵敏度高、操作简便、快速准确,适用于纺织助剂中DHTDMAC的分析测定.     

1-(2-噻唑偶氮)-2-萘酚螯合物反相高效液相色谱法分离测定铜(Ⅱ)、铁(Ⅱ)和镍(Ⅱ)

以１－（２－噻唑偶氮）－２－萘酚（ＴＡＮ）作柱前显色剂，于ＯＤＳ柱上，用内含０．１ｍｏｌ／ＬＬｉＣｌ，５×１０－６ｍｏｌ／Ｌ ＴＡＮ和ＨＡｃ－ＮＨ４Ａｃ缓冲溶液（ｐＨ ５．５）的甲醇－水溶液（８０：２０，Ｖ／Ｖ）作流动相，流速为０．６ｍＬ／ｍｉｎ，并以紫外－可见检测器于５９０ｎｍ处进行检测，发展了一种ＲＰ－ＨＰＬＣ法同时分离测定铜（Ⅱ）、铁（Ⅱ）、镍（Ⅱ）的方法，方法灵敏度高，对于铜、铁、镍的检测限分别为１μｇ／Ｌ， ２ μｇ／Ｌ和 ０．４ μｇ／Ｌ。用于实际样品测定，结果满意。     

Development and validation of a reversed-phase liquid chromatographic method for analysis of demeclocycline and related impurities

A simple, robust, and rapid reversedphase high-performance liquid chromatographic method for the analysis of demeclocycline and its impurities is described. Chromatographic separations were achieved on a Symmetry Shield RP8 (75 mm × 4.6 mm, 3.5 μm) column kept at 40°C. The mobile phase was a gradient mixture of acetonitrile, 0.06 M sodium edetate (pH 7.5), 0.06 M tetrapropylammonium hydrogen sulphate (pH 7.5) and water, A (2:35:35:28 v/v/v/v) and B (30:35:35:0 v/v/v/v) pumped at a flow rate of 1 mL/min. UV detection was performed at 280 nm. The developed method was validated according to the ICH guidelines for specificity, limit of detection, limit of quantification, linearity, precision, and robustness. An experimental design was applied for robustness study. Results show that the peak shape, chromatographic resolution between the impurities, and the total analysis time are satisfactory and better than previous methods. The method has been applied for the analysis of commercial demeclocycline bulk samples available on the market.     

Development and validation of an improved liquid chromatographic method for the analysis of oxytetracycline

Summary An improved LC method is described for the separation of oxytetracycline and its impurities. The separation is much better than that obtained with official pharmacopoeia methods. The method uses XTerra RP-18, 5 μm (25cm×4.6 mm I.D.), a silica-based stationary phase with methyl end-capping, claimed to reduce silanol activity. The column temperature is set at 30°C and a UV detection is performed at 280 nm. Mobile phase containing acetonitrile −0.25 M tetrabutylammonium hydrogen sulfate pH 7.5−0.25 M ethylenediaminetetraacetic acid pH 7.5-water (115:360:160:365,v/v/v/v) is used at a flow rate of 1.0 mL.min⁻¹, to separate the impurities present in oxytetracycline base. A central composite experimental design is used to optimize the separation. A second isocratic method with higher content of acetonitrile is needed to separate the more retained impurities present only in oxytetracycline hydrochloride. The method is robust and shows good selectivity, repeatability, linearity and sensitivity.     

Development and validation of a stability-indicating column high-performance liquid chromatographic assay method for determination of nebivolol in tablet formulation

A simple, precise, and accurate isocratic reversed-phase (RP) stability-indicating column high-performance liquid chromatographic (HPLC) assay method was developed and validated for determination of nebivolol in solid pharmaceutical dosage forms. Isocratic RP-HPLC separation was achieved on a Phenomenex Luna C8 (2) column (250 mm x 4.6 mm id, 5 microm particle size) using mobile phase composed of acetonitrile-pH 3.5 phosphate buffer (35 + 65, v/v) at a flow rate of 1.0 mL/min, and detection was performed at 280 nm using a photodiode array detector. The drug was subjected to oxidation, hydrolysis, photolysis, and heat to apply stress conditions. The method was validated for specificity, linearity, precision, accuracy, robustness, and solution stability. The method was linear in the drug concentration range of 40-160 microg/mL with a correlation coefficient of 0.9999. The repeatability relative standard deviation (RSD) for 6 samples was 0.69%, and the intermediate precision (RSD) for 6 samples was 1.39%. The accuracy (recovery) was between 98.57 and 99.55%. Degradation products produced as a result of stress studies did not interfere with detection of nebivolol, and the assay can thus be considered stability-indicating.     

超高效液相色谱-静电场轨道阱高分辨质谱法测定硝苯地平中痕量基因毒性杂质

建立了测定硝苯地平中基因毒性杂质2、6和12的超高效液相色谱-静电场轨道阱高分辨质谱法(UHPLC-Orbitrap HRMS).样品以甲醇为溶剂,提取后直接进样分析.采用ACE EXCELTM 3 C18-AR色谱柱(150 mm×4.6 mm,3μm)分离,流动相为甲醇-0.1％甲酸水(65:35,v/v),等度洗...     

高效液相色谱法测定头孢他啶的含量及杂质

采用高效液相色谱法测定了头孢他啶的含量及杂质。以Alltima C18色谱柱(250 mm×4.6 mm,5 μm)为分离柱,以乙腈和磷酸盐缓冲溶液(pH 3.9)分别为流动相A和流动相B进行梯度洗脱,流速1.3 mL/min,柱温35 ℃,紫外检测波长255 nm。从头孢他啶药物中共分出14个杂质,且14个杂质间具有良好的分离度。头孢他啶在0.267~1069 μg/mL范围内与峰面积具有良好的线性关系(r=1.0000);其定量限(S/N=10)和最低检出限(S/N=3)分别为3.1 ng和0.93 ng。3个浓度的日内测定值的相对标准偏差(RSD）为0.72%(n=3),日间测定值的RSD为0.91%(n=3)。头孢他啶溶液在4 ℃避光条件下放置24 h保持稳定。本方法与欧洲/英国药典和日本药局方的方法比较,具有分离杂质数量多、分离度好的优点。     

Impurity profile of macitentan in tablet dosage form using a stability-indicating high performance liquid chromatography method and forced degradation study

Macitentan (MAC) is a pulmonary arterial hypertension (PAH) drug marketed as a tablet and often has stability issues in the final dosage form. Quantitative determination of MAC and its associated impurities in tablet dosage form has not been previously reported. This study quantified impurities present in Macitentan tablets using a binary solvent-based gradient elution method using reversed phase-high performance liquid chromatography. The developed method was validated per International Conference on Harmonization (ICH) guidelines and the drug product was subjected to forced degradation studies to evaluate stability. The developed method efficiently separated the drug and impurities (48 min) without interference from solvents, excipients, or other impurities. The developed method met all guidelines in all characteristics with recoveries ranging from 85%-115%, linearity with r² ≥ 0.9966, and substantial robustness. The stability-indicating nature of the method was evaluated using stressed conditions (hydrolysis:1 N HCl at 80℃/15 min; 1 N NaOH at 25℃/45 min; humidity stress (90% relative humidity) at 25℃ for 24 h, oxidation:at 6% (v/v) H₂O₂, 80℃/15 min, thermolysis:at 105℃/16 h and photolysis:UV light at 200 Wh/m²; Fluorescent light at 1.2 million luxh). Forced degradation experiments showed that the developed method was effective for impurity profiling. All stressed samples were assayed and mass balance was>96%. Forced degradation results indicated that MAC tablets were sensitive to hydrolysis (acid and alkali) and thermal conditions. The developed method is suitable for both assay and impurity determination, which is applicable to the pharmaceutical industry.     

Comparison of ultra high performance supercritical fluid chromatography,ultra high performance liquid chromatography,and gas chromatography for the separation of synthetic cathinones

A comparison of ultra high performance supercritical fluid chromatography, ultra high performance liquid chromatography, and gas chromatography for the separation of synthetic cathinones has been conducted. Nine different mixtures of bath salts were analyzed in this study. The three different chromatographic techniques were examined using a general set of controlled synthetic cathinones as well as a variety of other synthetic cathinones that exist as positional isomers. Overall 35 different synthetic cathinones were analyzed. A variety of column types and chromatographic modes were examined for developing each separation. For the ultra high performance supercritical fluid chromatography separations, analyses were performed using a series of Torus and Trefoil columns with either ammonium formate or ammonium hydroxide as additives, and methanol, ethanol or isopropanol organic solvents as modifiers. Ultra high performance liquid chromatographic separations were performed in both reversed phase and hydrophilic interaction chromatographic modes using SPP C18 and SPP HILIC columns. Gas chromatography separations were performed using an Elite‐5MS capillary column. The orthogonality of ultra high performance supercritical fluid chromatography, ultra high performance liquid chromatography, and gas chromatography was examined using principal component analysis. For the best overall separation of synthetic cathinones, the use of ultra high performance supercritical fluid chromatography in combination with gas chromatography is recommended.     

超高效液相色谱-飞行时间质谱快速分析人参根中的皂甙化合物

建立了超高效液相色谱-飞行时间质谱快速分析人参根部提取物中的皂甙类化合物的方法。色谱柱为HSS T3超高效液相色谱柱(100 mm×2.1 mm, 1.8 μm);以15 mmol/L甲酸铵水溶液-乙腈为流动相,采用二元梯度洗脱的方式对人参主根的皂甙提取物进行分离。基于待测目标物的多级质谱碎片离子、精确质量等信息,结合9种人参皂甙标准化合物的多级质谱碎片离子质谱图,共鉴定出人参主根提取物中27种皂甙类化合物。在确定的条件下,以9种人参皂甙标样为研究对象,进行了全面的方法学考察,发现它们的线性范围分别为0.33～9.00 mg/L (Rg1), 0.11～9.00 mg/L (Re), 0.02～2.00 mg/L (Rf), 0.07～6.00 mg/L (Rg2), 0.04～3.00 mg/L (Rb1, Rb3), 0.22～6.00 mg/L (Rc), 0.04～9.00 mg/L (Rb2, Rd);在中等加标浓度时,经内标物峰面积校正的9种皂甙标准化合物的峰面积的相对标准偏差(RSD)不高于11.3%;低、中、高3个质量浓度加标水平的回收率范围分别为90%～100%、98%～104%及96%～103%;最低检出限为3.5～18.5 μg/L。该方法具有高分辨、快捷、简便、可靠等特点,并成功地应用于分析同一产地、不同生长时间的人参干燥主根中皂甙的差异。可以预计此方法可进一步应用于各种人参原料和制品中皂甙的快速测定。     

快速高效液相色谱分离技术在中药分析中的应用

中药化学成分分析是中药药效物质阐明及质量控制的关键问题之一。由于中药化学成分的复杂性,中药成分分析一直存在着分离难度大、分析时间长等问题。为此,针对这些问题发展起来的快速、高效液相色谱技术(主要包括超高压液相色谱、基于整体柱的高效液相色谱和高温液相色谱)备受关注。本文结合本实验室近年的研究,对该技术在中药复杂体系化学成分分析中的应用做简要综述,内容主要包括这些技术的原理、分离性能及其在中药复杂体系中的应用。