柱切换液相色谱法快速定量检测参附注射液中乌头碱类生物碱和人参皂苷

利用柱切换液相色谱,建立了参附注射液中苯甲酰新乌头原碱、苯甲酰乌头原碱、苯甲酰次乌头原碱、新乌头碱、次乌头碱和乌头碱6种乌头碱类生物碱,以及Rg1、Re、Rf、Rb1、Rc、Ro、Rb2、Rb3、Rd 9种人参皂苷的分析方法。首先利用强阳离子交换的在线固相小柱选择性富集和净化样品中生物碱类成分,优化了色谱条件;并采用EC－C18柱作为人参皂苷的分析柱,通过优化实验条件,结合柱切换方式,去除了样品中辅料等大极性基质成分对色谱柱的污染,实现了生物碱分析和人参皂苷分析的自动切换。结果显示,样品中的生物碱和人参皂苷分离良好,线性相关系数（r2）均大于0.999,连续进样精密度的相对标准偏差（RSD） < 2.0%,重复性的RSD < 2.0%;其中6种生物碱的平均回收率为95.1%～98.6%,检出限为4.0～8.2 ng/mL;9种人参皂苷的平均回收率为91.7%～104%。所构建的基于柱切换液相色谱技术的在线固相萃取方法能够有效去除样品中的基质干扰,快速完成参附注射液中3种单酯型生物碱和9种人参皂苷的快速定量,同时也可对3种双酯型生物碱进行限量检测,可应用于药物的质量评价。     

RP-HPLC法测定黑顺片中6种酯型生物碱的含量

建立黑顺片中6种酯型生物碱含量的高效液相测定方法。采用岛津C18色谱柱(250 mm×4.6 mm,5μm),以乙腈–四氢呋喃(体积比为25∶15)为流动相A,以0.1 mol/L乙酸铵溶液(每1 000 m L加冰乙酸0.5 m L)为流动相B梯度洗脱,流量为1.0 ml/min,检测波长为235 nm,柱温为35℃。苯甲酰新乌头原碱、苯甲酰乌头原碱、苯甲酰次乌头原碱、新乌头碱、次乌头碱、乌头碱进样量分别在0.222~4.430,0.060~1.119,0.144~2.878,0.015~0.297,0.025~0.496,0.011~0.229μg范围内与色谱峰面积线性呈良好的线性关系,相关系数r均大于0.999 5,检出限分别为2.10,2.70,2.20,2.60,1.60,3.10 ng,样品在12 h内稳定。苯甲酰新乌头原碱、苯甲酰乌头原碱、苯甲酰次乌头原碱、新乌头碱、次乌头碱、乌头碱的加标回收率分别为98.7%~102.2%,98.6%~102.3%,99.0%~101.9%,102.0%~97.3%,102.1%~98.3%,95.7%~103.5%,测定结果的相对标准偏差分别为2.5%,2.5%,2.2%,2.3%,2.2%,1.7%(n=6)。该方法简便,测定结果准确、可靠,可为黑顺片的质量评价和质量控制提供参考。     

硼亲和吸附剂的制备及其对苯甲酰脲类农药的萃取性能

以3-丙烯酰胺苯硼酸（APB）为单体,二乙烯基苯（DVB）为交联剂,“原位”聚合制备了聚（3-丙烯酰胺苯硼酸-二乙烯基苯）多孔硼亲和整体材料并作为搅拌饼固相萃取（SCSE-APBDVB）的萃取介质。以5种苯甲酰脲农药为目标化合物,详细考察了萃取过程中解吸溶剂、样品基底中pH值以及离子强度、萃取和解吸时间等实验条件对萃取效率的影响。在此基础上,与高效液相色谱-二极管阵列检测器联用建立了环境水样和果汁样品中苯甲酰脲农药残留的测定方法。在最佳条件下,在水样和果汁样品中,5种目标化合物的检出限（LOD,S/N=3）分别在0.055~0.11 μg/L和0.095~0.31 μg/L之间,所建立的方法具有理想的日内和日间重现性（RSD值均小于9.0%）。在对实际环境水样和果汁样品的测定中,不同加标浓度苯甲酰脲的回收率为75.6%~109%。研究表明,由于所制备吸附剂与目标化合物存在B-N配位作用、氢键和疏水等多种作用力,因此SCSE-APBDVB可对苯甲酰脲农药进行有效萃取,所建立的分析方法具有简便、灵敏和环境友好等特点。     

近红外光谱法快速测定附子中6个生物碱的含量

应用近红外光谱(NIRS)技术结合偏最小二乘(PLS)和最小二乘支持向量机(LS-SVM)建立了附子中多指标成分的快速无损检测方法。选取38批样品建立了同时测定附子样品中6种成分含量的高效液相色谱(HPLC)方法;通过采集附子样品的NIRS图,分别采用PLS和LS-SVM建立了各个成分HPLC测定值与NIRS图的定量校正模型。所建立的苯甲酰新乌头原碱、苯甲酰乌头原碱、苯甲酰次乌头原碱、新乌头碱、次乌头碱、乌头碱、单酯型生物碱总量和双酯型生物碱总量LS-SVM模型的相对预测偏差(RPD)分别为3.3、3.2、4.1、7.7、8.8、7.6、4.0和8.6;验证集相关系数(rpre)分别为0.9486、0.9475、0.9668、0.9909、0.9946、0.9969、0.9669和0.9927,且LS-SVM模型优于PLS模型,说明NIRS模型验证集与HPLC测定值具有良好的非线性关系,模型预测效果良好。采用NIRS技术结合LS-SVM模型可以快速对附子中的上述6个生物碱含量以及单酯型生物碱总量和双酯型生物碱总量进行检测,方法操作简便,对控制附子中的生物碱含量具有一定的指导作用。     

大孔树脂对麻黄细辛附子汤生物碱成分静态吸附及解吸附性能研究

采用高效液相色谱法(HPLC)测定麻黄细辛附子(MXF)汤复方水提液中盐酸麻黄碱、盐酸伪麻黄碱、苯甲酰新乌头原碱、苯甲酰乌头原碱、苯甲酰次乌头原碱含量,考察了4种大孔树脂(HPD–100,HPD–722,HPD–400,HPD–600型)对5种生物碱的静态吸附、解吸附性能,筛选出分离纯化麻黄细辛附子汤中生物碱的最佳树脂。结果显示,HPD–722型树脂对5种生物碱的吸附过程稳定且吸附量大,具有较高的解吸附率(87.26%),优于其它3种树脂。HPD–722型大孔树脂适用于富集麻黄细辛附子汤中的生物碱类化合物。     

基于UPLC/Q-TOF-MS分析附子半夏配伍相反的物质基础

利用超高效液相色谱-飞行时间质谱联用技术(UPLC/Q-TOF-MS)分析附子半夏药对配伍相反的物质基础,从化学成分层次阐释其配伍相反机制.基于UPLC/Q-TOF-MS建立附子半夏药对配伍后生物碱类成分的化学指纹图谱,通过主成分分析法和正交偏最小二乘判别法分析药对配伍在合煎过程中的生物碱类成分的含量变化,找出差异变化显著的化学成分.结果表明正离子模式时附子半夏药对合煎液中次乌头碱,中乌头碱,乌头碱,去氧乌头碱,10-OH-中乌头碱,10-OH-乌头碱等的含量明显增高,而中乌头原碱,去乙酸中乌头原碱,去乙酸次乌头原碱,苯甲酰乌头原碱,苯甲酰次乌头原碱,10-OH-苯甲酰中乌头原碱等含量降低.附子半夏药对配伍应用时双酯型二萜生物碱的含量明显增高,而单酯型二萜生物碱的含量明显降低,这可能是附子半夏药对配伍相反作用的物质基础.     

自制离子液体固相微萃取涂层分析人体尿液中的五氯酚

建立了顶空固相微萃取与气相色谱法（HS-SPME-GC）测定人体尿液中五氯酚（PCP）的新方法。 采用溶胶-凝胶法,加入自制的离子液体键合固相微萃取涂层,优化了萃取温度、萃取时间、pH值、离子强度及解吸时间。 结果表明,样品中加入3 g NaCl,溶液的pH值为2,并以一定速度搅拌的条件下,在80 ℃顶空萃取50 min,300 ℃下解吸5 min,方法的检测限为5.0 ng/L,线性范围为0.05～100 μg/L,相对标准偏差(RSD)为5.9%,加标回收率为106.6%。     

以离子液体为流动相添加剂的高效液相色谱法测定钩藤药材中钩藤碱和异钩藤碱

采用以离子液体为流动相添加剂的高效液相色谱法测定钩藤药材中钩藤碱和异钩藤碱的含量。钩藤药材样品0.532g用60%(体积分数)甲醇溶液10mL在25℃下超声提取30min。过滤后的续滤液以C18色谱柱为分离柱,以16.7 mmol·L~(-1)磷酸二氢钾缓冲溶液(pH 3.0,20mmol·L~(-1) 1-己基-3-甲基咪唑四氟硼酸盐离子液体作为添加剂)为流动相,在检测波长245nm处进行测定。钩藤碱与异钩藤碱的质量浓度均在1.56~100mg·L~(-1)内与其对应的峰面积呈线性关系,检出限(3S/N)依次为0.914,0.194 mg·L~(-1)。方法用于钩藤药材样品的分析,回收率为98.0%~101%,测定值的相对标准偏差(n=6)为0.60%~1.8%。     

人血清白蛋白生物色谱柱的性能研究及应用于铁棒锤中活性成分的筛选

以大孔硅胶为基质,采用羰基咪唑法合成了人血清白蛋白(HSA)生物色谱填料。详细评价了该填料对药物的分离性能,温度和pH对药物保留的影响。结果表明,该种生物色谱填料的分离性能优良,可以在短时间内基线分离4种药物分子。用HSA色谱柱对铁棒锤中有效成分进行了分离,通过LC-MS分析结果显示,该色谱柱可以分离出其中4种活性成分(脱乙酰去氧乌头碱、苯甲酰脱氧乌头碱、乌头原碱、16-O-去甲基乌头次碱)。     

复方中药四逆汤中乌头碱类二萜生物碱的电喷雾串联质谱研究

利用电喷雾串联质谱方法不经柱分离而直接分析复方中药四逆汤中二萜生物碱的组成,乌头碱类二萜生物碱在质谱条件下形成质子化分子,一种生物碱对应一个分子离子峰.检测到了苯甲酰单酯型、双酯型和脂类生物碱等18种二萜生物碱,其中苯甲酰中乌头原碱等单酯型生物碱是四逆汤中的主要生物碱成分.     

中空纤维支载离子液体液液微萃取法检测环境水体中的三氯生

三氯生(5 Chloro-2-(2,4-dichlorophenoxy) phenol,TCS)是一种新型环境水体污染物,具有潜在的生态与健康风险,因此发展合适的分析方法来检测水环境中这类化合物极其必要.本研究以1-辛基-3-甲基咪唑六氟磷酸离子液体( [C8 MIM][PF6])为萃取剂,基于中空纤维的离子液体液液微萃取方法,结合HPLC/UV用于环境水样中TCS的分析测定;通过对各参数(萃取剂、供体相的体积、供体相pH值、离子强度、萃取时间等)的优化在最优条件下(样品相体积为50 mL,pH值2,盐浓度为0.2 mol/L,200 r/min振荡萃取8 h),获得了较高的富集倍数(907倍)、较低的检出限(0.05 μg/L,RSD=7.4％,n=6)和较好的线性范围(0.1～100 μg/L);以4种环境水样加标实验对方法的准确性进行评估,其回收率可达94.2％～108.5％(RSD=5.5％～8.0％,n=6);本方法可广泛应用于环境水体中痕量TCS的分析检测.     

顶空液相微萃取-气相色谱法测定盐酸雷尼替丁中二氯甲烷和三氯甲烷的残留量

采用顶空液相微萃取与气相色谱联用技术测定雷尼替丁中二氯甲烷和三氯甲烷的残留量。自制了萃取液保护装置。考察了萃取溶剂的种类、萃取时间、萃取温度、萃取液的体积对二氯甲烷和三氯甲烷萃取效果的影响。以正十三烷为萃取剂,在60 ℃下萃取30 min,萃取液滴体积2 μL。二氯甲烷含量在1～10 μg/g范围内与色谱峰高呈线性关系,相关系数(r2)为0.9733;三氯甲烷含量在1～10 μg/g范围内与色谱峰高呈线性关系,r2为0.9724。二氯甲烷和三氯甲烷的最低检出限分别为0.0273 μg/g和0.0410 μg/g,加标回收率分别为93.6%～102%和98.1%～103%。方法简便易行,测定结果准确。     

Derivatization following hollow‐fiber microextraction with tetramethylammonium acetate as a dual‐function reagent for the determination of benzoic acid and sorbic acid by GC

Derivatization at the injection port following hollow‐fiber‐based liquid–liquid–liquid microextraction with tetramethylammonium acetate as a dual‐function reagent, i.e. an acceptor and derivatization reagent, for the determination of benzoic acid (BA) and sorbic acid (SA) in real samples by GC was developed. BA and SA were extracted from aqueous samples to an organic phase impregnated into the pores of the hollow fiber wall, and then back‐extracted to the acceptor solution located inside the lumen of the hollow fiber. Upon injection, the extracted analytes were quantitatively derivatized to their methyl esters with tetramethylammonium acetate in the GC injection port. Several parameters related to the derivatization and extraction efficiency were optimized. The linearity was satisfactory over a concentration range of 0.1–50 mg/L with r > 0.993 for both analytes. The LODs were 2.0 μg/L for SA and 20 μg/L for BA. The recoveries (83–116%) and precisions (RSDs of 1.2–11.4% (n = 3)) were examined by analyzing real spiked samples. The enrichment factors of BA and SA were 300 and 425. The results demonstrated that this is a simple, rapid, accurate, and sensitive method for the determination of BA and SA in various samples.     

Determination of Lignans in Schisandra sphenanthera and Schisandra chinensis Using Ionic Liquid-based Ultrasonic-assisted Extraction and High-performance Liquid Chromatography

A green, rapid and precise sample pretreatment technique, IL-based UAE(ionic liquid-based ultrasonic-assisted extraction), was coupled with high-performance liquid chromatographic separation to identify the main effective components in Schisandra sphenanthera(S. sphenanthera) and Schisandra chinensis(S. chinensis) including schisantherin A, schisandrin A, and deoxyschizandrin. Four different types of ionic liquids have been investigated, finally[C₆MIM] [BF₄] was used as the extraction solvent. A powder form of S. sphenanthera and S. chinensis was mixed with the[C₆MIM] [BF₄] to produce a suspension. This suspension was ultrasonically extracted in a water bath at room temperature. Several of the process parameters were optimized, including the type of ionic liquid used and its volume, the sample amount, the size of the sample particle, the extraction time, etc. HPLC calibration curves were established for all the analytes and proved to be linear(r>0.9999). The lowest detection level for schisandrin A was 0.12 μg/mL, for schisantherin A was 0.08 μg/mL, and for deoxyschizandrin was 0.10 μg/mL. The recoveries of the target compounds were from 74.19% to 109.33%. The standard deviations for detection were generally no more than 6.31%. In contrast to conventional extraction methods, the IL-based UAE did not involve volatile organic volatile solvents, and the analysis time, required sample and solvent volumes were also lower than those of the conventional techniques.     

Liquid chromatography-electrochemical detection for studying the effects of tetrahydrobiopterin on monoamine neurotransmitters in rat striatum

BH4 is the natural cofactor of two important amino acid hydroxylases: tyrosine hydroxylase, the rate-limiting enzyme in the biosynthesis of dopamine from tyrosine, and tryptophan hydroxylase, the rate-limiting enzyme in the biosynthesis of serotonin from …     

三相中空纤维膜液相微萃取-高效液相色谱法测定水中痕量双酚A

建立了三相中空纤维膜液相微萃取-高效液相色谱(HF-LPME-HPLC)方法,用于分析测定水中痕量双酚A的含量.设计了三相中空纤维膜液相微萃取系统,优化的HP-LPME最佳萃取条件为:萃取剂为正辛醇,接受相NaOH浓度为0.09 mol/L,样品溶液pH=4.0,NaC1加入量为30 g/L,搅拌速度为900 r/min,萃取时间为60 min.萃取后取20 μL接受相进行色谱分析.在最佳萃取条件下,方法的线性范围为0.5～200 μg/L(r＞ 0.999),检出限(信噪比为3)为0.2 μg/L;富集因子为241;方法RSD＜3.2％ (n=3).在实际环境水样中添加5,20和50μg/L的双酚A标准物质,加标平均回收率为92.8％～101.9％.表明本方法可用于水中痕量双酚A的快速准确测定.     

Ionic Liquid-based Microwave-assisted Liquid-liquid Microextraction and High Performance Liquid Chromatography Determination of Sulfonamides from Animal Oils

The authors performed ionic liquid-based microwave-assisted liquid-liquid microextraction(IL-based MALLME) coupled with high performance liquid chromatographic separation for the determination of 6 sulfonamides (SAs) from animal oils. The target analytes were extracted from animal oil samples with sodium hydroxide solution containing 1-butyl-3-methylimidazolium tetrafluoroborateand as the extraction solvent under microwave irradiation. The experimental parameters of the IL-based MALLME, including types of ILs, volume of IL, amount of ion-pairing agent(NH₄PF₆), pH value of sample solution, and extraction temperature and time were evaluated. The limits of detection and quantification obtained were in a range of 0.4―0.5 μg/kg and a range of 1.2―1.8 μg/kg, respectively. The accuracy of the method was evaluated by analyzing five spiked animal oil samples at two fortified levels(5 and 50 μg/kg), and the recoveries of SAs varied from 81.4% to 114.5% with relative standard deviations ranging from 0.8% to 9.0%.     

液相微萃取-高效液相色谱联用分析化妆品中痕量的雌激素

建立了液相微萃取-高效液相色谱联用(LPME-HPLC)测定爽肤水中痕量的雌三醇、雌二醇、炔雌醇和雌酮的分析方法,研究了萃取溶剂种类、接受相体积、搅拌速度、萃取时间等对萃取效率的影响。结果表明,该方法对4种雌激素的富集倍数可达到247~343倍,方法的线性范围为1~200 μg/L,检出限为0.4~1.0 μg/L,6次平行测定的相对标准偏差为3.6%~7.3%,爽肤水中的加标回收率为101.2%~114.9%。方法简单快速、灵敏度高、环境友好,满足痕量雌激素分析的要求。     

超声辅助离子液体分散液液微萃取/高效液相色谱法测定新型抗艾药ACC007

该文提出了一种基于超声辅助离子液体分散液液微萃取/高效液相色谱(HPLC)测定血清及药片中ACC007含量的新方法。在超声辅助下,无需分散剂即可将疏水性离子液体1-辛基-3-甲基咪唑六氟磷酸盐([C8mimPF6])形成的细小液滴分散于样品溶液中,从而有效萃取ACC007,萃取率在94.0%以上。实验对萃取剂种类、萃取剂用量、溶液pH值、萃取时间、冷却和离心时间等萃取条件进行了考察。在优化条件下,ACC007的线性范围为0.20~10.0μg/mL,检出限分别为0.062μg/mL(药片)和0.068μg/mL(血清)。采用该方法对药片和血清中ACC007进行测定,加标回收率为90.5%~103%,相对标准偏差为2.9%~5.1%,结果令人满意。     

Dispersive liquid–liquid microextraction followed by high-performance liquid chromatography for the determination of three carbamate pesticides in water samples

A simple, rapid and efficient method, dispersive liquid–liquid microextraction (DLLME) in conjunction with high-performance liquid chromatography (HPLC), has been developed for the determination of three carbamate pesticides (methomyl, carbofuran and carbaryl) in water samples. In this extraction process, a mixture of 35 µL chlorobenzene (extraction solvent) and 1.0 mL acetonitrile (disperser solvent) was rapidly injected into the 5.0 mL aqueous sample containing the analytes. After centrifuging (5 min at 4000 rpm), the fine droplets of chlorobenzene were sedimented in the bottom of the conical test tube. Sedimented phase (20 µL) was injected into the HPLC for analysis. Some important parameters, such as kind and volume of extraction and disperser solvent, extraction time and salt addition were investigated and optimised. Under the optimum extraction condition, the enrichment factors and extraction recoveries ranged from 148% to 189% and 74.2% to 94.4%, respectively. The methods yielded a linear range in the concentration from 1 to 1000 µg L^?1 for carbofuran and carbaryl, 5 to 1000 µg L^?1 for methomyl, and the limits of detection were 0.5, 0.9 and 0.1 µg L^?1, respectively. The relative standard deviations (RSD) for the extraction of 500 µg L^?1 carbamate pesticides were in the range of 1.8–4.6% (n = 6). This method could be successfully applied for the determination of carbamate pesticides in tap water, river water and rain water.