高效液相色谱/二极管阵列检测/质谱/质谱(HPLC/DAD/MS2)联用在板蓝根注射液成分鉴定中的应用

用HPLC/DAD/MS2联用仪,鉴定了板蓝根注射液中含有的核苷类物质。色谱条件:Luna C18 色谱柱(250 mm×4.6 mm i.d., 5 μm);甲醇-水梯度洗脱,流量0.8 mL/min; 柱温25 ℃。用二极管阵列检测器记录各个色谱峰的紫外吸收光谱,色谱检测波长254 nm。质谱条件:电喷雾离子源(ESI);正离子检测;扫描范围m/z 50-800。记录质量色谱图和各个色谱峰的一级、二级质谱图,并对质谱结果进行解析,通过与对照品比较,确定板蓝根注射液样品中含有腺苷、鸟苷、尿苷、胞苷及腺     

HPLC／UV法对银鲫肌肉组织中甲苯咪唑及其代谢产物残留的同时测定

建立了一种用高效液相色谱同时测定银鲫肌肉组织中甲苯咪唑(MBZ)及其代谢物氨基甲苯咪唑和羟基甲苯咪唑的方法.银鲫肌肉组织用乙酸乙酯提取,萃取物旋转蒸发至干后用1 mL二甲基甲酰胺-0.05 mol/L磷酸盐缓冲液(体积比3 ∶ 7)定容.色谱条件:Waters symmetry C18反相色谱柱(250 mm×4.6 mm,5 μm);流动相:乙腈-0.05 mol/L磷酸二氢铵溶液(体积比33 ∶ 67);流速:0.8 mL/min;检测波长为298 nm;检测温度为室温.在10 ～120 μg/kg添加水平,MBZ、MBZ-NH2、MBZ-OH的回收率分别为81% ～86%、71% ～75%、86% ～93%.MBZ的检出限为2 μg/kg,MBZ-OH和MBZ-NH2检出限均为3 μg/kg.     

高效液相色谱-质谱法分析菊芋叶中的绿原酸类化合物

建立了菊芋叶中绿原酸类化合物的高效液相色谱-紫外检测-质谱(HPLC-UV-MS)定性分析方法。液相色谱条件:Inertsil ODS-3色谱柱(250 mm×4.6 mm,5 μm);甲醇和水(含1%乙酸)梯度洗脱,流量1.0 mL/min;柱温35 ℃;检测波长327 nm。质谱条件:Thermo公司TSQ三级四极杆质谱仪;电喷雾电离(ESI)接口;负离子检出模式。采用该方法得到了菊芋叶提取物的紫外检测的色谱图、负离子监测的总离子流图以及相应色谱峰的紫外光谱图和一级、二级质谱图,对其进行解析,鉴别出菊芋叶中的7个绿原酸类成分。该方法简便、快速、灵敏度高,可以很好地对菊芋叶中的绿原酸类化合物进行定性分析。     

高效液相色谱法同时测定丹参提取物中4种成分的含量

建立反相高效液相色谱法同时测定丹参醇提物和超临界提取物中原儿茶醛、丹酚酸B、隐丹参酮和丹参酮ⅡA含量的方法.采用RP-HPLC梯度洗脱的方法进行测定,色谱条件为:Agilent C18柱(5 μm,4.6×250 mm);以1%醋酸乙腈-1%醋酸水为流动相进行梯度洗脱;检测波长:0～25 min (280 nm),25～60 min (270 nm);流速0.5 mL/min;柱温:35 ℃.测定了丹参醇提取及超临界提取物中以上4种成分的含量;4种成分线性关系均良好(r>0.9995),平均加样回收率均大于95.0%,RSD均小于3.0%.该方法一次进样,可以同时测定丹参中水溶性成分原儿茶醛和丹酚酸B、脂溶性成分隐丹参酮和丹参酮ⅡA的含量.     

苯酚氧化羰基化反应产物的高效液相色谱分析

采用高效液相色谱法分析了苯酚氧化羰基化反应产物,确定了色谱条件:KromasilTM C18色谱柱(150 mm×4.6 mm,5μm),检测波长254 nm,流动相V(甲醇):V(水)=65:35,流速0.6 mL/min.对碳酸二苯酯(DPC)和苯酚进行了定量分析,DPC和苯酚的外标曲线相关系数分别为0.99966...     

高效液相色谱-程序波长紫外检测法同时测定血浆中的色氨酸及其代谢物

建立了一种高效液相色谱-程序波长紫外检测法同时测定血浆中色氨酸(Trp)及其主要代谢产物犬尿氨酸(Kyn)和5-羟色胺(5-HT)。以茶碱为内标(IS),采用BDS-Hypersil-C8柱(150 mm×4.6 mm, 5 μm)分离。流动相为10 mmol/L醋酸钠缓冲液(pH 4.5)-乙腈(94:6, v/v),流速为0.6 mL/min;柱温为25 ℃;紫外检测波长设定: Kyn和IS为360 nm, 5-HT为220 nm, Trp为302 nm。3种物质的平均回收率为87%～113%;线性范围分别为3.97～400 μmol/L(Trp), 0.421～20.2 μmol/L(Kyn), 4.36～980 nmol/L(5-HT);检出限分别为0.134 μmol/L(Trp), 0.0160 μmol/L(Kyn), 2.03 nmol/L(5-HT)。利用该方法对15例抑郁症患者和15例健康志愿者的血浆进行测定,结果表明两组间Trp的代谢存在显著的差异。     

固相萃取-高效液相色谱法测定啤酒中的黄腐酚

采用Waters Sep-Pak C18固相萃取小柱对啤酒样品进行分离纯化,建立了啤酒中黄腐酚的固相萃取-高效液相色谱检测方法。选用色谱柱Zorbax Eclipse XDB-C18柱(250 mm×4.6 mm,5 μm),以甲醇和0.1%甲酸水溶液为流动相进行梯度洗脱,柱温25 ℃,流速0.4 mL/min,检测波长370 nm。在此条件下,黄腐酚分离良好且无杂质峰干扰,在0.5～500 μg/L的范围内线性关系良好(r21),在高、中、低浓度下的加标回收率为91.21%～95.58%,相对标准偏差小于2%。方法的检出限为0.24 μg/L,定量限为0.80 μg/L。该方法简便快速、结果准确、重现性好,是检测啤酒中黄腐酚含量的有效方法。     

高效液相色谱法测定复方甘草口服溶液中8种防腐剂

建立高效液相色谱法测定复方甘草口服溶液中苯甲醇、苯酚、苯甲酸、山梨酸、对羟基苯甲酸甲酯、对羟基苯甲酸乙酯、对羟基苯甲酸丙酯和对羟基苯甲酸丁酯8种防腐剂。样品经体积分数为50%的甲醇溶液稀释后,以SHISEIDO CAPCELL MGⅡC₁₈色谱柱(250 mm ×4.6 mm,5 μm)为分离柱,以乙腈–0.02 mol/L乙酸铵溶液(用冰乙酸调至pH 5.0)为流动相,梯度洗脱,流量为1.0 mL/min,检测波长分别为211 nm (苯甲醇、苯酚)、225 nm (苯甲酸)、256 nm (山梨酸、对羟基苯甲酸甲酯、对羟基苯甲酸乙酯、对羟基苯甲酸丙酯和对羟基苯甲酸丁酯),柱温为30℃。苯甲醇的质量浓度在8.0～160.0 μg/mL范围内,山梨酸的质量浓度在3.0～150.0 μg/mL范围内,苯酚、苯甲酸的质量浓度在3.0～300.0 μg/mL范围内,对羟基苯甲酸甲酯、对羟基苯甲酸乙酯、对羟基苯甲酸丙酯、对羟基苯甲酸丁酯的质量浓度在0.5～50.0 μg/mL范围内与色谱峰面积线性关系良好,相关系数均不小于0.999 7,检出限为0.01～2.0μg/mL...     

高效液相色谱-质谱法测定发酵液中的喷司他丁

建立了测定发酵液中喷司他丁含量的反相高效液相色谱-质谱分析方法。采用的色谱条件: 色谱柱为Hypersil ODS2柱(250 mm×4.6 mm, 5 μm);流动相为甲醇/乙腈/10 mmol/L乙酸铵(pH 7.6)(2.5/2.5/95, v/v/v),流速为1.0 mL/min;检测波长为280 nm;柱温为40 ℃;进样量为10 μL。喷司他丁在1.0～100 mg/L范围内有良好的线性关系,相关系数为0.9999。该方法精密度好,稳定性高,能简便、快速、准确地测定发酵液中喷司他丁的含量。     

固相萃取/反相高效液相色谱法对血清中胰岛素的测定

采用固相萃取纯化血清样品,荧光胺柱前衍生,建立了反相高效液相色谱/荧光检测血清中胰岛素的方法.色谱条件:LunaC5柱(100 mm×4.6 mm,5 μm)分离,流动相为乙腈-Tris缓冲溶液(10 mmol/L,pH 8.5)梯度洗脱,流速:1 mL/min.激发波长(λex)273 nm,发射波长(λem)476 nm,进样量20 μL.结果表明:胰岛素在10 ～800 μg/L范围内峰面积和质量浓度呈良好的线性关系,相关系数r为0.997 1;检出限为5 μg/L.日内、日间精密度分别为1.5%、1.0%,回收率为92%.     

固相萃取-微乳液相色谱法测定环境水体中的3种酚类化合物

建立了固相萃取-微乳液相色谱法同时测定环境水体中的苯酚、双酚A (BPA)、2,4-二氯苯酚3种酚类化合物的检测方法。水样加酸酸化后,经C18固相萃取小柱富集净化,用微乳液相色谱法测定3种目标物的含量。在Inertsil C18色谱柱(150 mm×4.6 mm, 5 μm)上以微乳(3.0%十二烷基硫酸钠(SDS)-6.0%正丁醇-0.8%正庚烷-90.2%(水+0.5%HAc))和乙腈作为流动相进行梯度洗脱,流速1.0 mL/min,检测波长280 nm。结果表明,苯酚、双酚A、2,4-二氯苯酚的检出限(S/N=3)依次为0.74、8.0、8.0 μg/L,线性范围在0.1~10 mg/L范围内,相关系数(r)均大于0.999。将3种酚类化合物定量加到空白水样中,苯酚、双酚A、2,4-二氯苯酚的加标回收率分别为82.7%、87.8%、82.6%,其RSD均小于5%(n=6)。对环境水样的酚类化合物分析也取得了良好的加标回收率,其值均在85.7%~113.2%之间。结果表明,该方法准确可靠、灵敏度高,适用于环境水体中酚类化合物的检测。     

A 13C NMR study of the methylol derivatives of 2,4′‐ and 4,4′‐dihydroxydiphenylmethanes found in resol phenol–formaldehyde resins

A total of 13 of the 16 possible methylol derivatives of 2,4′‐ and 4,4′‐dihydroxydiphenylmethane have been synthesized, isolated, and identified. These compounds are found as intermediates in the cure process of resol phenol–formaldehyde (PF) resins. Analysis of the ¹³C NMR spectra (in acetone‐d₆) of these compounds provided a way to evaluate the seven methylolphenol ring types (methylol derivatives of 2‐hydroxyphenyl and 4‐hydroxyphenyl rings) found in typical resol PF resins using the ipso carbon region from 150 to 160 ppm. A simple diagnostic test was developed using the chemical shift values of the methylol methylene carbon atoms to identify the presence of intermediates containing either a 2‐hydroxyphenyl or a 4‐hydroxyphenyl ring. Using these data it is now possible to analyze the major components in extracted prepreg PF resins. Copyright © 2002 John Wiley & Sons, Ltd.     

整体柱高效液相色谱法测定草甘膦原药中甲醛含量

建立了衍生化-高效液相色谱法对草甘膦原药中痕量甲醛含量进行测定的方法。样品中残留甲醛经超声波水浴提取，与2，4-二硝基苯肼衍生反应，生成的2，4-二硝基苯腙用反相整体柱色谱进行快速分离，在360nm紫外波长下检测，外标法定量。该分析方法在2．0～200mg/L浓度范围呈良好线性，添加回收率在88％～105％之间，相对标准偏差小于5％。样品中甲醛的最小检测浓度为0．5mg/kg。比较了整体色谱柱和常规C18反相柱分离效果，表明整体色谱柱可在1．5min内实现衍生化产物的快速分离并进行定性定量，同时发现相对于常规柱，采用整体柱提高了检测灵敏度约10倍。     

Determination Of Formaldehyde And Acetaldehyde Associated To Atmospheric Aerosols By HPLC

Abstract

A rapid new analytical protocol was developed for the determination of formaldehyde and acetaldehyde associated to atmospheric particulate matter, at ng/m³ levels. The aerosols were collected on glass fiber filters (8″×10″) at face velocities ranging from 15 m/min to 23 m/min. Aliquots of 15.4 cm² were sonicated, for 20 min, with 5.0 mL of 0,01% 2,4-dinitrophenylhydrazine (DNPH), 1 % phosphoric acid. The liquid phase was then filtered and the separation and quantification of the corresponding 2,4-dinitrophenylhidrazone (DNPHo) derivatives carried out by reverse phase HPLC. Acetonitrile:water (57:43, v/v) as mobile phase at 1.0 mL/min and absorbance detection at 350 nm and 365 nm for, respectively, formaldehyde-DNPHo (0.04 AUFS) and acetaldehyde-DNPHo (0.01 AUFS) were used. The precision for four different aliquots, from a 8″×10″ glass fiber filter, were under 0.04% for formaldehyde and 14.16 % for acetaldehyde. In Salvador, Bahia, Brazil, formaldehyde and acetaldehyde were determined, respectively, in the range of 6.8 ng/m³ to 27.3 ng/m³ and 9.1 ng/m³ to 54.6 ng/m³.     

柱前衍生-萃取阻断反应-高效液相色谱法测定化妆品中游离甲醛

建立了柱前衍生化-萃取阻断反应-高效液相色谱(HPLC)测定化妆品中甲醛的方法。化妆品中甲醛检测的难点是: 甲醛缓释剂类防腐剂在衍生过程中释放甲醛,影响游离甲醛的准确测定。以2,4-二硝基苯肼(DNPH)乙腈溶液-磷酸盐缓冲液(pH 2)(1:1, v/v)为提取溶液,于室温下快速衍生2 min后,立即加入二氯甲烷萃取,阻断衍生反应,经乙腈稀释后进行HPLC测定。以Agilent C18柱(250 mm×4.6 mm, 5 μm)为分离柱,乙腈-水(60:40, v/v)为流动相,流速为1.0 mL/min,于355 nm波长下检测。在洗发水、乳液、膏霜、洗手液、牙膏、指甲油、粉饼中分别添加50、100、500、1000 μg/g 4个浓度水平的甲醛,其回收率为81%～106%,相对标准偏差(n=6)＜5.0%。方法的定量限(以信噪比(S/N)＞10计)为50 μg/g。该方法快速、简便、重现性好,且可以有效避免甲醛缓释剂类防腐剂分解释放甲醛,适用于化妆品中游离甲醛的测定。     

高效液相色谱法定量测定中药千层塔提取物中的石杉碱甲

建立了高效液相色谱快速定量测定中药千层塔提取物中石杉碱甲含量的分析方法。千层塔提取物经甲醇/水/甲酸(10/90/0.2, v/v/v)提取并定容后,过滤膜后直接分析。色谱分离选用XCharge C18色谱柱(150 mm×4.6 mm, 5 μm),以水(含0.1%三氟乙酸)和乙腈(含0.09%三氟乙酸)为流动相进行梯度洗脱,流速为2 mL/min,于310 nm波长下检测,可在10 min内完成石杉碱甲的快速分离分析。结果表明,石杉碱甲在2.12～106 mg/L范围内线性关系良好(相关系数为0.9999);平均加标回收率为102.34%,相对标准偏差(RSD)为0.46%;日内及日间精密度均小于2%,满足定量要求。该方法简便、快速,结果可靠,重现性好,可作为千层塔提取物质量评价的依据。     

Effects of production parameters on the structure of resol type phenolic resin/layered silicate nanocomposites

Polymer/layered silicate nanocomposites belong to one of the most promising group of materials of the past few decades and most probably for the near future. Following the pioneering works of Toyota Research Group in the 1980s, the interest on these materials increased rapidly and research is now being carried out world wide, using all kinds of polymers as base material.In this present study, the aim was to investigate the effects of several different production parameters; on the morphology of resol type phenol formaldehyde based layered silicate nanocomposites produced by mixing and casting. For this purpose; two different liquid resol type phenolic resins (PF76 and PF76TD), two different curing methods (heat cure route and acid cure route), two different montmorillonite clays (unmodified Cloisite Na+ and modified Rheospan), two different clay sources (Wyoming-USA and Tokat-Turkey), and five different clay amounts (0.5%, 1%, 1.5%, 3%, 10%) were used.XRD, SEM, TEM analyses and mechanical tests indicated that resol type phenolic resins lead to better structures when they were modified with ethylene glycol and cured by the use of an acidic curing agent. It was also observed that use of modified clay with no more than 1.5 wt% in the phenolic matrix lead to certain degree of exfoliation consequently better structure and higher mechanical performance.     

Transformation of dynamic DSC results into isothermal data for the curing kinetics study of the resol resins

Kinetics of thermosetting polymers curing is difficult to study by isothermal methods based on the differential scanning calorimetry (DSC) technique. The difficulty is due to the low sensitivity of the equipment for total reaction heat measurements during high temperature process. The aim of this paper is to display the equivalence between a dynamic model, the Ozawa method, and an isothermal isoconversional fit, which allows predicting the isothermal behavior of the resol resins cure through dynamic runs by DSC. In this work, lignin–phenol–formaldehyde and commercial phenol–formaldehyde resol resins were employed. In addition, the isothermal kinetic parameters for both resins were performed by means of transformation of the data obtained from the dynamic Ozawa method.     

HPLC-UV method development and validation for the determination of low level formaldehyde in a drug substance

A reversed-phase high-performance liquid chromatographic method (HPLC) with diode-array detection is developed and validated for the quantitative determination of formaldehyde in a drug substance. Formaldehyde (HCHO) is reacted with 2,4-dinitrophenylhydrazine (DNPH) to form a Schiff base (HCHO-DNPH derivatization product), which has an absorbing maximum (lambda max) at 360 nm. The HPLC method employs a C8, 3-microm particle size analytical column (150 mm x 4.6 mm), 15-microL injection volume, column temperature controlled at 30 degrees C, detection at 360 nm, and a water-acetonitrile (55:45, v/v) mobile phase at a flow rate of 1 mL/min. These conditions resolve the HCHO-DNPH product from unreacted DNPH, the drug substance and related impurities, as well as diluent peaks within 20 min. The retention time of the HCHO-DNPH product is approximately 6.4 min. The method is linear, accurate in the specified range (0.33-333 ppm), and robust based on analyte (HCHO-DNPH derivatization product) stability in standard and sample. Detection limit is 0.03 ng (0.1 ppm).     

Analysis of chlortetracycline by high performance liquid chromatography with postcolumn alkaline-induced fluorescence detection.

A high performance liquid chromatographic method for the analysis of chlortetracycline (CTC) using postcolumn fluorescence detection has been developed. After chromatographic separation of CTC on a polystyrene-divinylbenzene copolymer column, a highly fluorescent derivative isochlortetracycline (iso-CTC) was formed postcolumn in an on-line reaction coil with the addition of 25% NaOH (w/v). Chromatographic separation was achieved on a PRP-1 column, 15 cm x 4.6 mm, with 27:73 acetonitrile:0.2% perchloric acid (v/v), at 1.0 mL/min. Fluorescence derivatization was achieved by the on-line addition of 25% NaOH (w/v), at a flow rate of 0.2 mL/min, into the column eluant in a post-column reaction coil. The reaction coil was 9 m of teflon (1/16 in o.d., 0.3 mm i.d.) knitted into a six-sided coil. The fluorescent derivative was detected at lambda ex 355 nm and lambda em > 389 nm. Using this method after a simple sample cleanup, CTC can be detected in milk at 0.04 micrograms/mL, which is comparable to that obtained by microbiological assays. The detection method was linear between 0.02 micrograms/mL and 4 micrograms/mL. Because of the chromatographic separation, the method is more selective than microbiological assays and more sensitive than ultraviolet detection. With the chromatographic system described, the keto tautomeric forms of CTC and 4-epi-CTC are separated in a system which minimizes their formation on-column. In acidic aqueous organic solutions, the keto tautomer of CTC is the only product formed to any significant amount.