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??OBJECTIVE To develop an HPLC-ELSD method for determination of MPEG-DSPE and HSPC in doxorubicin hydrochloride liposome injection. METHODS The column was Waters Symmetry 300 C₁₈(4.6 mm??150 mm, 5 ??m;pore size:300). The mobile phase was methonal-tetrahydrofuran-0.17 mol??L^-1 ammonium acetate(93??6??1). The flow rate was 1.0 mL??min^-1. The column temperature was 25 ?? and the injection volume was 10 ??L. The ELSD conditions were as follows: Alltech 2000ES ELSD detector; drift temperature: 110 ??; rate: 2.6 L??min^-1. RESULTS This method had good specificity. The linear ranges of the calibration curves for MPEG-DSPE and HSPC were 0.03-0.48 mg??mL^-1 (r=0.999 8) and 0.1-1.0 mg??mL^-1 (r = 0.999 8), respectively. The average recovery rates of MPEG-DSPE and HSPC were 100.0% (n=3??3) and 101.0% (n=3??3), respectively. The LODs of MPEG-DSPE and HSPC were 13 and 52 ng, respectively. The repeatability and intermediate precision of MPEG-DSPE were 0.9% (n=5) and ??1.9%(n=3), respectively. The repeatability and intermediate precision of HSPC were 1.1%(n=5) and ??1.3%(n=3) , respectively. CONCLUSION The established method is accurate, reliable, repeatable and suitable for the determination of MPEG-DSPE and HSPC in doxorubicin hydrochloride liposome injection.     

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??OBJECTIVE To develop a method for the determination of iridoid,phenylpropanoid glycosides,and organic acids in Scrophulariae Radix from different habitats and commercial herbs by UPLC-QTRAP-MS/MS. METHODS The analysis was carried out on a BDS HYPERSIL C₁₈ column (4.6 mm×250 mm,5 μm) with elution by mobile phase of acetonitrile-water at a flow rate of 1.0 mL·min^-1. The column temperature was maintained at 35 ??. The target compounds were analyzed by the negative ion multiple reaction monitoring (MRM) mode. RESULTS Twelve multiple constituents showed good linearity (r>0.999 4) in the range of the tested concentration. The average recoveries of the 12 components were 99.59%-101.24% with relative standard deviations of 0.93%-1.60%. CONCLUSION The established method is accurate and precise,which provides a reliable and effective technique for the quality evaluation of Scrophulariae Radix.     

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??OBJECTIVE To establish an HPLC-MS/MS method for the determination of melatonin in human plasma. METHODS The plasma samples were extracted with ethyl acetate, using melatonin-D7 as the internal standard (IS). Then the ethyl acetate layer was evaporated in vacuum concentrator. Dried samples were redissolved in mobile phase (0.1% formic acid-methanol= 56:44, V/V), vortexed and centrifuged. The redissolved solution was transferred to an auto sampler vial and the supernatant was injected to the HPLC-MS/MS system. The MS/MS analysis was carried out in positive ionization mode by multiple reactions monitoring (MRM) at m/z 233.1??174.1 for melatonin and m/z 240.2??178.0 for IS, respectively. RESULTS The calibration curve of melatonin in human plasma was linear over the concentration range of 0.020 00-30.00 ng??mL^{- 1}. The lower limit of quantitation was 0.020 00 ng??mL^{- 1}. The RSDs of within-day and between-day were less than 15%. The extraction recoveries were between 59.0%-65.0%. The matrix effects were between 95.5%-98.9%. CONCLUSION The method is proved to be convenient, sensitive and accurate. It can be applied to study the pharmacokinetics of melatonin prolonged-release tablets in healthy Chinese volunteers.     

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??OBJECTIVE To establish an HPLC-ELSD method for determination of mesaconine.METHODS Gemini C₁₈ 110A column (4.6 mm??150 mm,5 ??m) was adopted with the mobile phase consisting of methanol-0.1%TFA (25:75) at the flow rate of 0.6 mL??min^-1. The detector was Alltech ELSD 2000ES, with the drift tube temperature of 90 ?? and the airflow velocity of 2.3 L??min^-1. RESULTS Under this conditions, mesaconine and the related substances were well separated. The linear range of mesaconine was 0.40-0.60 mg??mL^-1 (r=0.999 6). RSD of injection precision and repeatability test were 0.04% (n=5) and 0.7% (n=6), the intra day and inter day precision RSD were 0.4% and 0.6%, respectively. The average recovery was 101.0% (RSD=1.1%, n=9). CONCLUSION The established method was specific, simple, accurate, and suitable for determination of mesaconine.     

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??OBJECTIVE To establish an analysis method for identification of gelatine ingredients in Chinese patent medicines. METHODS Trypsin was used for hydrolysis of gelatins. The characteristic getalins analyses were identified by rapid resolution liquid chromatography (RRLC) coupled to triple quadruple mass spectrometry (QQQ-MS). Trypsin was used for hydrolysis of gelatins.RESULTS The established present method was specific, precise and reliable. Gelatine ingredients as prescribed wereas detected for several samples. However, instead of gelatin ingredients as prescribed, unprescribed gelatine ingredients were as detected for some samples. CONCLUSION The methodology validation is performed and the RESULTS are satisfied. The method can be used for identification of donkey-hide gelatin, oxhide gelatin, deer-horn glue and tortoise-shell glue in Chinese patent medicines, and provides a scientific reference for the study of quality control method of gelatin ingredients in Chinese Patent Medicines.     

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??OBJECTIVE To identify rapidly the chemical constituents in Tanreqing injection and Tanreqing capsules by ultra-performance liquid chromatography with quadrupole-time-of-flight mass spectrometry (UPLC/Q-TOF-MS^E).METHODS The separation was performed on a Waters Acquity UPLC BEH C₁₈ column (1.0 mm??100 mm, 1.7 ??m) with acetonitrile-0.1% formic acid as mobile phase in gradient elution. ESI ion source was employed in negative ion mode. The differences of chemical compositions between the two preparations and the sources of these compounds were illustrated based on their retention time, accurate mass measurements and the mass fragments by comparison with those in the literature or database and the reference standards.RESULTS A total of 111 compounds including 13 unknown components were identified or tentatively characterized. Among these compounds, 14 were derived from Scutellariae Radix (SR) intermediate, 36 were from Bear Bile Powder(BBP) intermediate, 7 were from Caprae Hircus Cornu(CHC) intermediate, 34 were from Lonicerae japonicae Flos(LJF) intermediate and 22 were from Forsythiae Fructus(FF) intermediate. Moreover, quinic acid and rutin were simultaneously detected in LJF and FF intermediates, 28 constituents were unambiguously confirmed by their reference standards. However, 71 compounds were observed both in injection and capsules, while 24 compounds were only found in Tanreqing injection and 16 compounds only in the capsules.CONCLUSION The differences of chemical constituents between Tanreqing injection and capsules are effectively characterized by UPLC/Q-TOF-MS^E method, which will facilitate the quality control of the two preparations.     

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??OBJECTIVE To establish an HPLC-HRMS/MS method for analyzing the structures and sources of the related substances in etimicin sulfate. METHODS The chromatographic separation was achieved on a broad pH range column with a basic elution system composed of[H₂O-ammonia-glacial acid(96??3.6??0.4)]-methanol(70??30). Twenty percent of the eluent was detected under positive electrospray ionization(ESI) by Q-Exactive mass spectrometry. The fragmentation pathways were elucidated according to the HRMS and HRMS/MS fragmentation of etimicin and some known compounds. Then the unknown related substances were identified by analyzing their HRMS and HRMS/MS fragmentation with the help of the rule. RESULTS Sixty-five related substances were detected by the HPLC-HRMS/MS method in the two samples from different companies,among which 38 were detected in the product of company A, 59 in that of company B, and 32 were detected for both companies. Fifty-four related substances were identified or deduced, and the other 11 were not able to be identified due to limited information. Based on the significant difference of impurity spectra between the two enterprises, the key parameters of the synthesis process were analyzed. CONCLUSION An HPLC-HRMS/MS method, which showes excellent accuracy, is developed to identify the related substances in etimicin sulfate. The resources and structures of the related substances are analyzed, which will be helpful to the process optimization.     

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??OBJECTIVE To establish new category identification models of Rhei Radix et Rhizoma for authenticity and species prediction. METHODS The samples were extracted by ultrasonic with methanol, separated by UPLC and detected by DAD. Data were processed and screened by MPP software to reveal the variation of components of Rhei Radix et Rhizoma. The category prediction models were established to distinguish authentic from unauthentic Rhei Radix et Rhizoma and three species of authentic Rhubarb. RESULTS These models were confirmed to prensent prediction accuracy of 100%. Fifteen samples were predicted and classified with confidence level of greater than 0.57. CONCLUSION The prediction model is simple, reliable and accurate. It is suitable for the identification and quality evaluation of different species of Rhei Radix et Rhizoma crude drugs and decoction pieces.
     

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??OBJECTIVE To rapidly identify the related substances in doxycycline hyclate tablets and investigate the possible degradation pathways of doxycycline hyclate solution by 2D-LC-IT-TOF/MS. METHODS Firstly,the chromatography was carried out using the 1stD InertSustain^TM C₁₈(4.6 mm??150 mm,5 ??m) column and a mobile phase containing a mixture of buffer solution(0.25 mol??L^-1 ammonium acetate solution-0.1 mol??L^-1 ethyldiaminetetraacetic acid disodium: triethyiamine=100??10??1)-acetonitrile(85??15). Solution pH was adjusted to 8.8 with glacial acetic acid. Then a InertSustain^TM C₁₈(2.1 mm??50 mm,2 ??m) column was used with 0.1% formic acid-water as mobile A and 0.1% formic acid-acetonitrile as mobile B in a gradient elution mode in 2ndD. Electrospray ionization (ESI) source was tested in both positive and negative ion modes. Nebulized gas flow was 1.5 L?? min^-1. Dry gas flow was 10 L?? min^-1. The desolvation tube temperature was kept at 200 ??. Related substances were characterized according to multi-level MS behaviors. The 2D-LC-IT-TOF/MS method was employed to identify the structures of impurities in forced degradation solutions and illuminate the degradation pathways of doxycycline hyclate solution. RESULTS A total of eight related substances were detected in doxycycline hyclate tablets. Four of them had a content of more than 0.1%, and their structures were identified to be 4-epidoxycycline, metacycline, ??-epidoxycycline and 2-acetyl-2-decarbamoyldoxycycline. The solution of doxycycline hyclate easily degraded under alkaline condition and generated an open loop compound taken off the keto group. The solution was sensitive to heat, and generated 4-epidoxycycline with a little amount of 4-epi-6-epidoxycycline; but the solution was stable under illumination and acidic condition. CONCLUSION The established method is suitable for rapid identification of impurities in doxycycline hyclate, which can be applied as a useful analytical tool for quality control and drug process optimization of doxycycline hyclate.     

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??OBJECTIVE To establish an HPLC-MS/MS method for determination of ASC-J9 in rat blood. METHODS After liquid-liquid extraction, ASC-J9 was separated on a Symmetry C₁₈colume, with mobile phase of acetonitrile-water containing 0.1% formic acid and 10 mmol??L^-1 ammonium formate.The flow rate was 1.0 mL??min^-1, mass shunt was 0.4 mL??min^-1, and column temperature was main tained at 35 ??. Quantification was performed in positive ion multiple-reaction-monitoring(MRM) mode. RESULTS The calibration curve of ASC-J9 had good linearity in the concentration range of 3.54-1 180 ng??mL^-1. The extraction recovery rate was within 83.19% to 87.27%,and the intra-day and inter-day RSDs were both less than 8.89%. CONCLUSION This method is specific, sensitive and suitable for determination of ASC-J9 in rat blood.
     

超声循环法提取连钱草总黄酮

目的:在超声条件下寻找提取连钱草总黄酮的优化条件。方法：采用正交设计法,以超声功率、超声时间、提取温度、溶剂体积分数为因素,每个因素3个水平,选择L9(3⁴)正交表,用紫外分光光度法测定总黄酮的含量做为评价指标。结果：最佳提取条件为超声功率800 W,超声时间90(45,45) min, 提取温度40 ℃,溶剂浓度65%。最佳提取条件下总黄酮的质量分数为5.228%。结论：以乙醇为溶剂,采用超声循环提取技术提取连钱草中总黄酮,提取温度低,时间短,提取效率高,具有广阔前景。     

连钱草中黄酮、有机酸类成分的HPLC指纹图谱

目的:建立连钱草药材中黄酮、有机酸类成分的HPLC指纹图谱分析方法,为整体控制和评价连钱草药材的质量提供依据。方法:采用大连依利特Hypersil BDS C18(4.6 mm×250 mm,5μm),流动相为甲醇-0.1%磷酸梯度洗脱,流速1.0mL·min-1,检测波长330 nm。结果:建立了连钱草药材黄酮、有机酸类成分的HPLC指纹图谱,标定了17个共有峰,其精密度、稳定性、重复性均符合指纹图谱技术要求(RSD<5%),13批连钱草药材的相似度在0.713～0.987。结论:该指纹图谱方法简便、准确、重复性好,可作为连钱草药材质量控制的依据。     

UHPLC指纹图谱结合化学计量学评价不同产地显脉旋覆花花序的质量

目的建立不同产地显脉旋覆花Inula nervosa Wall.花序的超高效液相色谱(UHPLC)指纹图谱,用于显脉旋覆花花序质量的评价。方法采用Agilent Poroshell 120 C18色谱柱(100 mm×3.0 mm,2.7μm);以0.1%甲酸水(A)-甲醇(B)为流动相,梯度洗脱;流速:0.3 mL·min^-1,检测波长:254 nm。对建立的指纹图谱的主要特征峰进行确认,采用相似度评价软件,结合主成分分析(PCA)方法,对21个批次药材进行质量评价。结果建立了显脉旋覆花花序的UHPLC指纹图谱,用对照品标定了9个峰,相似度在0.9以上,21批药材亲缘关系较近。PCA可直观地把4个不同产地的样品进行分类。结论该方法专属性强,重现性良好,可为显脉旋覆花花序的鉴别及质量评价提供依据。     

紫外光谱结合化学计量学区分不同产地川东獐牙菜

目的:分析鉴别4个产地川东獐牙菜,并建立预测模型,预测产地区分准确性。方法:光谱数据导入UVProbe2.34,比较不同产地相同部位的紫外光谱图,将原始光谱数据以及经过8点平滑、一阶求导和二阶求导后的数据导入SIMCA-P11.5,进行主成分分析(PCA),比较三维得分图的产地鉴别效果。结果:主成分分析中以叶的原始数据以及8点平滑处理数据鉴别效果最佳,主成分累计贡献率均为98.8%,其余预处理方式无法取得较好的鉴别效果可能与主成分数累计值有关(一阶求导为83.9%,二阶求导为47.3%)。根部数据能将重庆、湖北的样品和湖南样品分开,但重庆和湖北的样品无法区分。建立偏最小二乘判别分析(PLS-DA)模型,检测鉴别模型的可靠性,并为预测更多产地的区分提供依据。将验证集带入训练集建立的模型进行偏最小二乘判别分析,能区分产地,证明该模型产地鉴别效果可行。PLS-DA中训练集的预测值和真实值相关系数为0.985,其评估均方差(RMSEE)为0.159,验证集导入训练集后其预测值与真实值的相关系数为0.927,预测均方差(RMSEP)为0.327,RMSEE与RMSEP两者相近,且都0.500,该模型的预测可靠性高。结论:运用紫外光谱结合主成分分析和偏最小二乘判别分析能够较好的鉴别不同产地川东獐牙菜,构建模型预测效果较好,加入未知产地样品也能较好区分。     

UPLC结合化学计量学方法的白及指纹图谱分析

目的:建立不同产地白及药材UPLC指纹图谱,为建立快速而高效的白及药材整体质量控制方法提供实验依据。方法:采用UPLC-PDA检测法,ACQUITY UPLC BEH C_(18)色谱柱(2.1 mm×150 mm,1.7μm),流动相乙腈-水溶液梯度洗脱,柱温45℃,检测波长280 nm,流速0.3 m L·min~(-1),建立白及药材指纹图谱;通过相似度评价,结合聚类分析和主成分分析,对不同批次的白及药材进行质量评价。结果:建立的白及药材UPLC指纹图谱方法确定了20个共有峰,并对9个共有峰进行了明确化学成分指认;43批白及药材的相似度在0.540~0.942;通过聚类分析可大致聚成2类;PCA大致聚成4类,并发现指纹图谱中6个造成43批次白及药材差异性的化合物。结论:白及的UPLC指纹图谱的构建和化学模式的识别为药材质量控制提供更全面的参考。     

HS-SPME-GC-MS结合化学计量法对不同产地艾叶药材挥发性成分的比较分析

目的:建立艾叶挥发性成分的快速分析方法,研究不同产地艾叶挥发性成分的含量及分布特征。方法:采用顶空固相微萃取-气相色谱-质谱(HS-SPME-GC-MS)联用技术,结合安捷伦化学工作站对不同产地的艾叶中挥发性化学成分进行定性分析,以峰面积归一化法计算各组分的相对含量,并通过化学计量法主成分分析和聚类分析对艾叶中的挥发性成分含量进行分析。结果:初步鉴定出84种化合物,主要为酮类、烯类、醇类化合物。不同产地艾叶挥发性成分有一定差异,采用主成分分析及聚类分析法能有效区分不同产地的艾叶,其中湖北蕲春产的艾叶品质最好,其次为湖南广布、河南汤阴的艾叶质量较佳,聚类分析结果将不同产地艾叶分为5类。结论:此方法稳定可靠,适用于艾叶挥发性成分的快速分析,并为艾叶挥发性成分的质量评价提供一定的科学依据。     

不同产地金银花药材的UPLC指纹图谱分析

目的： 建立不同产地金银花药材的超高效液相特征性指纹图谱,为有效控制和科学评价金银花药材整体质量提供依据。 方法： 采用Agilent C₁₈ 色谱柱（2.1 mm×50 mm,1.8 μm）,流动相乙腈-0.2%磷酸水,以0.4 mL·min^-1的流速进行梯度洗脱,检测波长238 nm,柱温30 ℃。 结果： 在21 min内得到金银花药材的指纹图谱,对其中5个色谱峰进行了初步归属,并对14批药材样品进行了分析,其相似度为0.915~0.987。 结论： UPLC指纹图谱方法较HPLC大大缩短了分析时间,可用于金银花药材的质量评价。     

基于指纹图谱和化学计量法评价不同产地黑顺片的质量

目的基于指纹图谱和化学计量学对3个主产区(四川江油、四川布拖和云南)的35批黑顺片进行质量评价。方法采用HPLC建立黑顺片的指纹图谱和多组分含量的测定方法,色谱条件为AlltimaTMC18色谱柱(4.6 mm×250 mm,5μm);柱温:20℃;以乙腈-四氢呋喃(25∶15)为流动相A,0.1 mol·L^-1乙酸铵溶液为流动相B,梯度洗脱;检测波长:235 nm。根据指纹图谱确定黑顺片的共有峰,分别采用相似度评价、聚类分析和主成分分析对不同产地样品进行系统比较和评价。结果35批不同产地黑顺片样品指纹图谱的相似度均>0.900。35批样品可聚为3类,11批江油产样品聚为一类,布拖和10批云南产样品聚为一类,2批云南产样品聚为一类。2个主成分因子的累计方差贡献率为92.586%。结论四川江油产黑顺片与四川布拖、云南产存在一定差异性,可为黑顺片的质量评价提供参考。     

GC-MS联合化学计量学方法分析鱼腥草不同部位挥发油

目的对鱼腥草各部位挥发油进行分析,为鱼腥草药用部位提供科学依据。方法采用水蒸气蒸馏提取鱼腥草挥发油,气相色谱质谱联用仪对挥发油组分进行鉴定,采用峰面积归一化法测定各组分含量,结合主成分分析、马氏距离和相似度评价等方法探求各部位之间的异同。结果根、茎、叶和花四部位共鉴定出35种化合物,其中共有化合物24种,各部位挥发油组分基本一致;各组份含量存在较大差异,甲基正壬酮含量最高;根和其它三个部位差异大,茎和叶之间的差异小。结论今后鱼腥草用药时应充分考虑各部位挥发油成分的差异。     

UPLC测定不同产地菟丝子药材中6种黄酮类成分的含量

目的:采用UPLC建立同时测定菟丝子药材中金丝桃苷、异槲皮苷、紫云英苷、槲皮素、山柰酚、异鼠李素6种黄酮类成分含量的方法。方法:以内蒙古产菟丝子为代表建立菟丝子药材中6种黄酮类成分的定量方法,用此方法测定12批不同产地菟丝子药材中6种黄酮类成分的含量。色谱条件为Agilent Eclipse Plus C_(18)色谱柱(2.1 mm×50 mm,1.8μm),流动相乙腈-0.1%甲酸水梯度洗脱,流速0.21 m L·min~(-1),柱温30℃,检测波长360 nm。结果:金丝桃苷、异槲皮苷、紫云英苷、槲皮素、山柰酚、异鼠李素分别在0.046 8~0.748 8μg(r=0.999 9),0.024 2~0.387 2μg(r=0.999 9),0.088 2~1.411 2μg(r=0.999 9),0.002 3~0.037 0μg(r=0.999 7),0.028 8~0.460 8μg(r=0.999 6)和0.002 3~0.037 0μg(r=0.999 4)线性关系良好,检出限分别为0.292 5,0.302 5,0.367 5,0.289 0,0.360 0,0.286 0 mg·L~(-1),定量限分别为0.877 5,1.210 0,1.470 0,1.012 0,1.260 0,1.215 5 mg·L~(-1)。平均回收率分别为99.95%,99.78%,99.53%,99.38%,100.03%,99.35%,RSD分别为1.0%,1.2%,1.0%,1.4%,1.1%,0.5%。12批来自不同产地的菟丝子6种黄酮类成分含量差异较大。结论:该方法专属性强、重复性好、快速、稳定、可控,为科学评价及有效控制菟丝子药材的质量提供依据。