Many diseases are closely related to the level of Fe3 + which mainly stored in the liver of human body, and mitochondria is the dominant organelle for iron metabolism. In this work, based on a glycocluster strategy, three novel dual-cascade targeting (hepatocyte and mitochondria) fluorescent probes were designed to monitor Fe3+ in hepatic mitochondria with good selectivity and sensitivity. Low toxic glycosylated probes exhibited obvious hepatic-targeting capacity in vitro and in vivo, since the galactose residue of probes confer highly affinity to asialoglycoprotein receptor (ASGPR) specifically expressed on the membrane of hepatocytes. Moreover, the hepatic targeting ability presented a trend of TP-Gal-1 < TP-Gal-2 < TP-Gal-3, which may be ascribed to a "cluster effect" of galactose-recognition toward ASGPR. Besides, the aimed probes displayed favorable mitochondria-targeting capacity under the assistance of electropositive triphenylphosphine. Overall, these galactosylated probes with dual-cascade targeting capacity provided promising detected tools for Fe3 + in hepatocyte mitochondria, which is greatly important for precise diagnosis and insight into liver mitochondrial Fe3 + -related diseases.

目的 建立以特征离子为目标的鉴别蛇胆汁的方法,并用于检查蛇胆川贝液.方法采用超高效液相色谱-四极杆-飞行时间质谱(UPLC-Q-TOF-MS)测定蛇胆汁和其他动物胆汁的质谱数据,并进行主成分分析(PCA),找出蛇胆汁的特征成分.再用高效液相色谱-串联质谱(HPLC-MS-MS)的多反应检测(MRM)定性和验证特征成分.针对蛇胆川贝液处方中的药材,制订供试品制备方法,并将蛇胆汁的MRM条件应用于蛇胆川贝液的检测中.结果 UPLC-Q-TOF-MS的液相方法条件为:甲醇(A)-10 mmol·L-1乙酸铵溶液(B),梯度洗脱(0～5 min,30％→50％A;5～20 min,50％→ 90％A;20～25 min,90％A;25～26 min,90％→30％A;26～30 min,30％A);质谱条件为:电喷雾电离源(ESI-),干燥气温度:220℃,干燥气流速8L·min-1,雾化气压力2bar,毛细管电压3500 V,喷嘴电压500 V,扫描范围50～1000,扫描频率1Hz.共找出两个蛇胆汁的专属性成分,分别为牛磺胆酸硫酸酯(SOTC)和牛磺去氧胆酸硫酸酯(SOTDC).建立了 SOTC和SOTDC的HPLC-MS-MSMRM方法,条件为:甲醇(A)-10mmol·L-1乙酸铵溶液(B),梯度洗脱(0～10 min,40％→50％A;10～25 min,50％→80％A;25～25.5 min,80％→40％A;25.5～30 min,40％A),质谱条件为:电喷雾电离源(ESI-),SOTC的离子对为594.2 → 514.3,594.2→ 124.0;SOTDC 为 578.2 → 498.3,578.2→124.0.将建立的方法应用于 41 批蛇胆川贝液制剂中,发现40批样品均含有蛇胆汁.结论 该方法可有效鉴别蛇胆汁和检查蛇胆川贝液中是否含蛇胆汁.

壮医萌芽于先秦时期,经历朝历代发展形成了自己独特的理论体系,其中"三道两路"理论是壮医生理病理观,对临床辨病和制方用药影响深远."龙路"为"两路"之一,意为运行血液的通路.能够通畅"龙路",保持人体气血运行平衡的药物,称为龙路药.通过论述龙路药的传统功效和促凝血、降血糖、降血压、抗心衰、抗血栓等现代药理作用之间的关系,为壮医药的临床应用提供一定理论支撑.

目的:建立13个厂家111批消渴灵片中黄连的6个生物碱、丹皮酚等多指标性成分含量测定方法,并结合主成分分析及聚类分析等化学计量学方法对其进行质量评价研究.方法:采用HPLC-UV法建立6个黄连生物碱类成分(非洲防己碱、表小檗碱、盐酸药根碱、盐酸黄连碱、盐酸巴马汀、盐酸小檗碱)、丹皮酚的含量测定方法及通过Chempatten化学计量学软件对不同生产企业的消渴灵片进行系统比较和评价.结果:建立消渴灵片的含量测定方法,测定了其中7个成分的含量,各个成分均能达到基线分离,进样量分别在6.05～249、9.29～226、5.94～195、11.0～283 ng、9.94～267、55.4～1 847、23.576～589.410 ng范围内与峰面积呈良好的线性关系,平均回收率分别为90.0％、84.8％、91.9％、90.3％、93.1％、93.0％、92.3％,符合方法验证要求.同时,化学计量统计体现了所建立的含量测定方法可以对不同生产企业生产的样品进行区分.结论:本实验所建立的多指标含量测定方法简单易行,准确性、稳定性和重复性良好,综合主成分分析和聚类分析,可为消渴灵片质量标准提供参考.

目的:建立同时测定罗汉茶中新落新妇苷、落新妇苷、新异落新妇苷、异落新妇苷、槲皮苷和黄杞苷含量的方法,并进行多元统计分析.方法:采用高效液相色谱-一测多评法.色谱柱为Phenomenex SuperLu C18,流动相为乙腈-0.1％甲酸溶液(19:81,V/V),检测波长分别为254 nm(新落新妇苷、落新妇苷、新异落新妇苷、异落新妇苷、黄杞苷)、291 nm(槲皮苷),流速为1.0 mL/min,柱温为30℃,进样量为10μL.以落新妇苷为内参物,计算其他5种成分的相对校正因子,再根据相对校正因子计算各成分的含量,并与外标法进行比较;采用SPSS 22.0软件进行聚类分析和主成分分析.结果:新落新妇苷、落新妇苷、新异落新妇苷、异落新妇苷、槲皮苷和黄杞苷检测进样量的线性范围分别为0.007～0.311、0.871～18.184、0.002～0.119、0.052～1.251、0.105～2.202、0.020～2.319μg(r>0.999);精密度、重复性、稳定性(24 h)试验的RSD均小于3％;平均加样回收率分别为97.32％、94.89％、97.15％、96.90％、97.52％、97.53％(RSD为1.09％～2.60％,n=6).新落新妇苷、新异落新妇苷、异落新妇苷、槲皮苷、黄杞苷相对于落新妇苷的平均相对校正因子分别为1.2526、1.1983、0.9586、0.8071、1.1381.一测多评法测得新落新妇苷、新异落新妇苷、异落新妇苷、槲皮苷、黄杞苷等5种成分的含量分别为0.3942～2.0672、0.1391～0.8047、2.8648～8.5548、4.5812～11.3711、1.0289～13.4015 mg/g;外标法测得新落新妇苷、落新妇苷、新异落新妇苷、异落新妇苷、槲皮苷和黄杞苷等6种成分的含量分别为0.3672～1.9253、46.3611～126.3421、0.1381～0.7988、2.9662～8.8578、4.6425～11.5233、0.9706～12.6419 mg/g;两种方法含量测定结果的相对误差均不高于3.55％.聚类分析结果显示,9批罗汉茶样品可聚为两类,S8聚为一类、其余聚为一类.主成分分析结果显示,前2个主成分的累计方差贡献率为84.745％,分类结果与前者一致.结论:所建高效液相色谱-一测多评法准确、可行,重复性良好,可用于罗汉茶中6种黄酮类成分的同时测定,并可为其质量控制提供参考.

目的:用电感耦合等离子体质谱(ICP-MS)对薄荷中铬(Cr)、锰(Mn)、钴(Co)、镍(Ni)、铜(Cu)、砷(As)、硒(Se)、钼(Mo)、镉(Cd)、钡(Ba)、镝(Dy)、铊(Tl)、铅(Pb)含量的不确定度评定.方法:电感耦合等离子体质谱法测定其含量,并对测定方法流程分析,确定不确定度来源,计算合成不确定度.结果:各元素的扩展不确定度分别为Cr 0.31mg/kg、Mn 4.8 mg/kg、Co 0.052 mg/kg、Ni 0.14 mg/kg、Cu 1.16 mg/kg、As 0.14 mg/kg、Se 0.028 mg/kg、Mo 0.024 mg/kg、Cd0.0076 mg/kg、Ba0.97 mg/kg、Dy0.018 mg/kg、Tl0.0038 mg/kg、Pb0.35 mg/kg.结论:该方法用于ICP-MS测定薄荷中13种元素的含量并评价其不确定度,提供了重金属及有害物质的不确定度的重要参考.

目的 对二维液相色谱应用的发展与原理进行概述.方法 介绍了二维液相色谱仪的构造与二维液相色谱技术的应用范围,着重描述了该技术在中药定性和定量中的研究进展.结果和结论 展望了在中药质量控制中二维液相色谱的应用前景.

目的 建立一种丸剂中非法添加环丙沙星的快速检测方法.方法 通过19F-核磁共振定量(19F-qNMR)法对添加入当归苦参丸中的环丙沙星进行检测,以4,4'-二氟二苯甲酮为内标,以氘代DMSO为溶剂,采集δ-120.3处环丙沙星信号和δ-104.8处内标信号面积,计算环丙沙星的含量.结果 以环丙沙星和内标信号面积比为纵坐标,环丙沙星和内标质量比为横坐标,得到线性回归方程为:Y=3.384 1X+0.114 8,R2=0.992,环丙沙星在5～30 mmol/L具有良好的线性,检出限为7.798×10-2mg/mL,定量限为0.260 mg/mL.建立的19F-qNMR法精密度、重复性、稳定性、加样回收率均符合检测要求.结论 所建立的19F-qNMR法样品制备简单,检测时间短,灵敏度高,适合对丸剂中非法添加的环丙沙星进行含量测定.

[Objectives] To establish a HPLC method for the simultaneous determination of 14 characteristic components in Gegen Qinlian tablets and the content of Gegen Qinlian tablets in 11 manufacturers. [Methods] A Inertsil ODS-3（250 mm×4.6 mm, 5 μm） reversed-phase high performance liquid chromatography column was used with acetonitrile-0.02 mol/L ammonium acetate+0.03% triethylamine solution（adjusted with glacial acetic acid to pH of 4.3） as the mobile phase for gradient elution at a flow rate of 1.0 mL/min. The determination wavelength of 8 components（puerarin, daidzin, liquiritin, baicalin, wogonoside, daidzein, ammonium glycyrrhizinate, baicalein） was 250 nm; the determination wavelength of wogonoside was 280 nm; the determination wavelength of 5 components（epierberine, jatrorrhizine hydrochloride, coptisine, palmatine hydrochloride and berberine hydrochloride） was 346 nm. [Results] There was a good linear relationship between the injection volume of puerarin, daidzin, liquiritin, baicalin, wogonoside, epierberine, jatrorrhizine hydrochloride, coptisine, daidzein, palmatine hydrochloride, ammonium glycyrrhizinate, berberine hydrochloride, baicalein and wogonin and peak area（r＞0.999 0） in the range of 146.26-585 0.24, 24.13-965.04, 18.45-738.00, 79.18-316 7.32, 9.57-382.80, 4.76-190.40, 2.57-102.80, 13.41-536.40, 10.60-424.00, 11.33-453.22, 12.08-483.20, 46.73-186 9.25, 20.28-811.20, 12.11-484.50 ng, respectively; the average recovery rates（n=6） were 2.18%, 1.79%, 1.81%, 1.68%, 2.27%, 2.13%, 1.96%, 1.07%, 0.93%, 0.61%, 2.92%, 0.77%, 2.79% and 0.62%, respectively; the precision, repeatability and stability were good, and the RSD was less than 3%. This method was used to determine 16 batches of Gegen Qinlian tablets produced by 11 enterprises. Wogonoside was not detected in a lot of samples, but 14 components were detected for other enterprises, but the content was different. [Conclusions] The method was accurate and feasible and could be used for the overall quality control of this variety.

A new dual hepatocyte-targeting fluorescent probe HPL-1, which can precisely distinguish tumorous pH from physiological pH, was developed. The OFF-ON switch of HPL-1 can be triggered via pH-induced structural change of the lactam group of the rhodamine moiety from closed-ring to open-ring. Our results showed that the phosphate group of HPL-1 is beneficial to its accumulation in liver cells, and combination of the phosphate and galactose units could synergistically increase the hepatocyte-targeting capacity. HPL-1 could selectively distinguish hepatoma cells from other tissue cells, and precisely distinguish cancerous liver cells from normal liver cells. Compared with other reported probes, HPL-1 not only enable a simple and convenient detection method, but also has good hepatocyte-targeting capacity and precise recognition capacity of tumors under weak acid micro-environment, which opens new avenues for precise diagnosis and treatment of hepatocellular carcinoma.

The mechanisms and regioselectivities of Fe(0)-catalyzed alkylation of aromatic compounds with alkenes were explored by DFT calculations. Our calculations show that these systems tend to undergo anti-Markovnikov addition. The influence of steric effects on the reaction mechanism has been investigated. The results indicate that the reaction is more likely to provide the linear product with dissociation of the 1PMe3 ligand. The reductive elimination is the rate-determining step of the whole process, such that the electrostatic interactions of the reaction site and the steric hindrance of the trimethylsilyl groups are favorable for the anti-Markovnikov rather than the Markovnikov addition pathway. Our calculations provide insights into the regioselective origin of the alkylation of aromatic compounds mediated by Fe(PMe3)4.

A series of novel low-toxic hepatoma cell–targeting lipid materials were designed and synthesized, in which monogalactose, digalactose, and galactose-biotin were used as targeting moieties and hydrophilic heads while stearate was used as hydrophobic tail (Mono-Gal-ST, Di-Gal-ST, and Gal-Biotin-ST). The corresponding galactose-biotin-modified liposomes (Mono-Gal-LPs, Di-Gal-LPs, and Gal-Biotin-LPs) and conventional liposomes (LPs) were prepared. These galactose-biotin-modified liposomes can distinguish hepatoma cells from other tissue cells owing to the recognition of asialoglycoprotein receptor by galactose group. Moreover, the ability of liposomes to distinguish hepatoma cells from normal hepatocytes follows a trend of LPs < Mono-Gal-LPs < Di-Gal-LPs < Gal-Biotin-LPs, which is attributed to the cluster glycoside effect and the synergistic effect of galactose and biotin. In addition, the endocytosis of these galactose-biotin-modified liposomes were competitively inhibited by galactose, further confirming these liposomes entered hepatoma cells via asialoglycoprotein receptor–mediated pathway.

葛根芩连制剂源于《伤寒论》所载的葛根芩连汤.文章比较分析了五种不同剂型葛根芩连制剂(包括片剂、微丸、胶囊、颗粒及口服液)的质量标准、各药味的定性鉴别以及含量测定,总结了葛根芩连制剂在解热、抑菌及糖尿病治疗的临床应用及联用药效,以供中医药创新研发及临床用药参考.

A novel method for the dynamic determination of cetirizine dihydrochloride in urine using surface enhanced Raman scattering (SERS) has been developed. Comparison of SERS activities between gold and silver colloid was made based on the analysis of the transmission electron micrographs and the SERS spectra, revealing that silver colloid is much more efficient on the signal enhancement performance. The primary sites of the adsorption on the nanoparticle surface were represented by the feature peaks of piperazine at ca. 1050cm(-1) and phenyl rings at ca. 1630cm(-1), respectively. The best signal response was extracted at pH 7 at the Cl-:Ag+ molar ratio of 2:1. The matrix effect was eliminated by subtracting the spectral contribution of the blank urine from the sample spectra. Sample preparation procedures were minimized. Quantification could be simply accomplished on the predicted versus actual concentration curve, within the Raman shift range from 500 to 2500cm(-1). Neither modeling nor complicate calculation was required. The method was shown to be specific and applicable for drug urine concentration monitoring in situ.

Objective:To establish an HPLC method for simultaneous determination of fourteen index components in Gegen Qinlian tablets and perform the quantitative analysis on Gegen Qinlian tablets from eleven pharmaceutical companies.Methods:RP-HPLC separation was carried on an Inertsil ODS-3 column (250 mm × 4.6 mm,5 μm)with the mobile phase consisting of acetonitrile and ammonium acetate+0.03％ triethylamine aq.(adjusting pH to 4.3 by acetic acid)with a gradient elution at the flow rate of 1.0 mL· min-1.The detection wavelength was set at 250 nm for puerarin,daidzin,liquiritin,baicalin,wogonoside,daidzein,ammonium glycyrrhizinate and baicalein,at 280 nm for wogonin,and at 346 nm for epierberine,jatrorrhizine hydrochloride,coptisine,palmatine hydrochloride and berberine hydrochloride.Results:Good linear correlations (r＞0.999 0) were found in the range of 146.26-5 850.24 ng,24.13-965.04 ng,18.45-738.00 ng,79.18-3 167.32 ng,9.57-382.80 ng,4.76-190.40 ng,2.57-102.80 ng,13.41-536.40 ng,10.60-424.00 ng,11.33-453.22 ng,12.08-483.20 ng,46.73-1 869.25 ng,20.28-811.20 ng,12.11-484.50 ng for puerarin,daidzin,liquiritin,baicalin,wogonoside,epierberine,jatrorrhizine hydrochloride,coptisine,daidzein,palmatine hydrochloride,ammonium glycyrrhizinate,berberine hydrochloride,baicalein and wogonin,respectively.The average recoveries (n-6) of the fourteen components were 2.18％、1.79％、1.81％、1.68％、2.27％、2.13％、1.96％、1.07％、0.93％、0.61％、2.92％、0.77％、2.79％、0.62％,respectively.The accuracy,precision and stability were satisfactory with RSD values less than 3％.Sixteen batches of Gegen Qinlian tablets from eleven pharmaceutical companies were investigated.Wogonoside was not founded in one batch and all the fourteen components were detected in the rest samples but their contents varied.Conclusion:The established method was accurate and feasible,which could be applied in comprehensive quality control of Gegen Qinlian tablets.

Objective To establish method for determination of dye auramine O in Gegen Qinlian tablets by LC-MS/MS.Methods Agilent Eclipse plus C18(2.1×100 mm, 1.8μm) was used. A gradient elution program of acetonitrile-0.01 mol·L-1 ammonium acetate was adopted at the lfow rate of 0.3 mL· min-1. The column temperature was maintained at 35℃ and the sample size was 2 μL. The analyses were detected by an electrospray ionization tandem mass spectrometry with dynamic MRM.Results The results of veriifcation of methodology showed the calibration curve of auramine O was good linear in the range of 0.093 4~1.869 pg (r =0.999 9), and the recovery was 94.38% (RSD=3.60%, n=9).ConclusionThe method was convenient, accurate with good precision, reproducibility and stability which has good linear relationship. It can be used for the determination the content of dye auramine O in Gegen Qinlian tablets.

目的:建立葛根芩连片中可能掺入的染色剂含量测定方法,并采用LC-MS/MS方法进行专属性确证.方法:采用HPLC法测定染色剂含量,使用Kromasil C18(4.6 mm×250 mm,5μm)色谱柱,以乙腈-0.05 mol·L-1乙酸铵溶液(含0.5％冰乙酸)为流动相,梯度洗脱,流速1.0 mL· min-1,检测波长为420 nm和500 nm,柱温35℃.用LC-MS/MS进行确证,以乙腈-0.01 mol· L-1乙酸铵溶液为流动相,梯度洗脱;离子源为电喷雾电离源(ESI),扫描模式为多反应监测(MRM).结果:建立了HPLC法检测葛根芩连片中可能含有的10种染色剂含量的方法.方法学验证结果表明,柠檬黄、胭脂红、日落黄、亮黄、酸性红73、甲基橙、金橙Ⅱ、金胺O、赤藓红、碱性橙Ⅱ的线性范围分别为1.498 6～49.955 0、0.672 7～22.423 5、1.497 8～49.925 0、0.223 5～7.451 6、0.447 0～14.900 0、0.398 0～13.267 8、0.497 0～16.566 7、0.190 6～6.353 2、0.695 3～23.1764和0.119 3～3.977 6μg;加样回收率(n=6)分别为97.2％(RSD=0.86％)、99.1％(RSD=0.48％)、98.6％(RSD=0.86％)、101.0％(RSD=1.2％)、98.2％(RSD=0.89％)、98.3％(RSD=1.0％)、97.5％(RSD=0.63％)、96.3％(RSD=0.40％)、97.8％(RSD=1.0％)和96.3％(RSD=0.96％).在对11个厂家123批样品的检验中,有5个厂家33批样品检出金胺O,检出率为26.8％,含量在0.013～83.51μg·g-1之间,其中1批样品同时检出金胺O和碱性橙Ⅱ.用LC-MS/MS进行专属验证,33批出现HPLC阳性峰样品均检出相应的金胺O和碱性橙Ⅱ.结论:本方法准确、便捷,具有良好的精密度、重现性和稳定性,线性关系良好,可用于葛根芩连片的质量控制.

目的:建立HPLC-柱后光化学衍生法测定蛤蚧定喘丸中黄曲霉毒素G2、G1、B2、B1的含量,并建立LC-MS/MS确证方法.方法:采用SPOLAR C18柱(4.6 mm×250 rnm,5μm)色谱柱,以甲醇(A)-乙腈(B)-水(C)为梯度洗脱流动相进行洗脱,流速1.1 mL·min-1,柱温35℃;采用柱后光化学衍生法,柱后光化学衍生器波长为254 nm;荧光检测器激发波长为365 nm,发射波长为450 nrn,增益值为15.用LC-MS/MS进行确证,流动相为水(0.1％甲酸+5 mmol·L-1甲酸铵)-乙腈,离子源:电喷雾电离源(ESI),扫描模式:多反应监测(MRM).结果:黄曲霉毒素G2、G1、B2、B1分别在0.001 18～0.029 6 ng(r=0.999 8)、0.004 18～0.104 6 ng(r=0.999 9)、0.001 23～0.030 8 ng(r=1.000 0)、0.004 267～0.106 7 ng(r=1.000 0)范围内线性关系良好;黄曲霉毒素G2、G1、B2、B1回收率(n=9)分别为101.2％(RSD=7.10％)、103.2％ (RSD=3.91％)、92.5％％(RSD=2.91％)和93.5％ (RSD=7.95％).结论:所建立的HPLC-柱后光化学衍生法操作简便、可靠,结果准确,灵敏度高,重现性好,可作为该产品中黄曲霉毒素检测质量控制的方法.

目的:建立离子色谱法测定血栓通注射液中氯化钠、枸橼酸钠的含量.方法:采用AS-11-HC离子交换色谱柱(250 mm×4 mm),AG-11-HC保护柱(50 mm ×4 mm),以35 mmol·L-1氢氧化钠溶液为流动相,等度洗脱,流速1.0 mL·min-1;阴离子抑制器,抑制电流90 mA;电导检测器,柱温30°;进样量100 μL.对建立的方法进行线性范围、准确性以及精密度验证.结果:标准溶液中氯离子在1.0 ～30.0 μg范围内,氯离子色谱峰与其浓度呈良好的线性关系(r=1.000,n=6),平均回收率为98.10％;标准溶液中枸橼酸根离子在0.5～4.0 μg范围内,枸橼酸根离子色谱峰与其浓度呈良好的线性关系(r=0.999 7,n=6),平均回收率为102.9％;结果5家企业13批样品均含有氯化钠,含量在7.407～8.705 mg· mL-1;3家企业7批样品含有枸橼酸钠,含量在0.319～1.157 mg·mL-1;2家企业均未检出枸橼酸钠.结论:本文设计方法经方法学验证可用于血栓通注射液中辅料氯化钠、枸橼酸钠的质量控制.

目的:建立银杏洋参胶囊中萜类内酯HPLC-ELSD定量测定方法.方法:色谱柱:SunfireTM C18柱(4.6 mm× 150 mm,5μm);流动相:甲醇-水(30:70);流速:1.0 mL·min-1;柱温:35℃,漂移管温度:110℃,气体流速:2.9L·min-1.结果:白果内酯、银杏内酯A、银杏内酯B和银杏内酯C分别在0.9333～7.778μg(r=0.9994)、0.9846～ 8.205 μg(r=0.9991)、0.9144 ～7.620μg(r=0.9992)和0.3027 ～ 2.5225μg(r=0.9991)进样量范围内线性关系良好,方法平均加样回收率分别为103.12％(RSD为1.81)、101.30％(RSD为2.93)、104.08％(RSD为4.41)和101.75％(RSD为1.00).结论:本方法快速、准确,可用于同时检测银杏洋参胶囊中的萜类内酯,可作为该产品质量控制的方法.