毛细管电泳叠加对比法的误差分析

目的对毛细管电泳叠加对比法的原理和误差理论进行探索.方法以相对峰面积为定量指标,利用全微分原理进行误差分析,并以极值误差理论对方法进行误差预测.结果当叠加样品相对峰面积增大3～10倍时,结果的相对误差较小,在实验的设计上也比较容易实现;样品相对峰面积的相对误差和称量叠加对照品的相对误差是决定结果误差的主要因素;若称量误差＜0.5%,相对峰面积的测定误差在1%以内,则分析结果的相对误差＜2%,其极值相对误差＜3.5%.结论毛细管电泳叠加对比法是一种简便、准确的定量方法.     

应用误差叠加原理评价色谱内标法

本文应用误差叠加原理,对药物分析中的内标定量法进行了评价。五个实际例子说明,内标法改善分析方法精密度的条件是C(?)。作者提出了在药物分析中所用内标法的条件。本文表明内标法除了其优点外,还存在一些局限性。     

毛细管区带电泳法测定麻黄中麻黄碱和伪麻黄碱的含量

目的建立测定麻黄中麻黄碱和伪麻黄碱含量的毛细管区带电泳（CZE）法。方法以200 mmol.L-1硼砂-500 mmol.L-1硼酸溶液（1∶1,v/v）含2 mg.mL-1庚烷磺酸钠和0.4%（v/v）乙腈为背景电解质（BGE）,石英毛细管（75 cm×75μm）,有效分离长度60 cm,运行电压15 kV,紫外检测波长210 nm,重力进样20 s（高度10 cm）,以苯甲醇作内标,对麻黄中的麻黄碱和伪麻黄碱进行定量分析,以两者含量为参量对10批麻黄进行系统聚类分析并进行质量评价。结果麻黄碱和伪麻黄碱的相对峰面积（Ai/Ar）与各自的质量浓度C（mg.mL-1）呈良好的线性关系,两者的平均回收率分别为100.4%和100.9%。结论该法准确可靠,操作便捷,可用于麻黄中麻黄碱和伪麻黄碱的含量测定。     

31例卵巢恶性肿瘤的B超与临床病理对比分析

目的探讨女性卵巢恶性肿瘤的B超声像特征及临床病理诊断价值。方法回顾性分析经手术病理确诊的31例女性卵巢恶性肿瘤的B超与临床病理对比。结果肿瘤体积越小恶性发生率越高;肿瘤体积越大良性发生概率反而越高。但是,<5 cm的肿瘤不论囊肿还是实性肿瘤,均存在恶性肿瘤的危险。结论 B超对卵巢肿瘤类型诊断具有很高的诊断价值。直径5¹0 cm的卵巢囊肿建议均施切除术。     

应用误差叠加原理评价色谱内标法

本文应用误差叠加原理,对药物分析中的内标定量法进行了评价。五个实际例子说明,内标法改善分析方法精密度的条件是C_V;s<2rC_V,i。作者提出了在药物分析中所用内标法的条件。本文表明内标法除了其优点外,还存在一些局限性。     

基于色差原理分析不同产地桑白皮有效成分含量与颜色的相关性

目的:研究桑白皮药材有效成分含量与颜色的相关性,并对不同产地样品进行指纹图谱和聚类分析,为该药材质量控制及品质评价提供依据。方法:分别采用高效液相色谱法(HPLC)和盐酸-镁粉反应比色法测定桑白皮中桑根皮素和总黄酮的含量;肉眼观察桑白皮颜色,采用色差仪测定其色度值(L*、a*、b*)并计算总色差值(E*ab);利用SPSS 21.0软件对桑根皮素和总黄酮的含量与各颜色指标(L*、a*、b*、E*ab)进行Pearson相关性分析。采用HPLC法建立3个不同产地共20批桑白皮药材的指纹图谱,并进行相似度分析。采用SPSS 21.0软件对20批样品进行K-均值聚类分析(基于桑根皮素、总黄酮的含量及颜色指标)和系统聚类分析(基于指纹图谱共有峰相对峰面积)。结果:桑白皮中桑根皮素和总黄酮的平均含量分别为0.0960~0.6186 mg/g、0.48%~1.51%,其各颜色指标均呈显著相关(P<0.01)。其中,L*、b*、E*ab值越小,a*值越大,则桑根皮素含量越高;L*、b*、E*ab值越大,a*值越小,则总黄酮含量越高。20批药材样品的指纹图谱与对照图谱的相似度为0.883~0.983;标定了13个共有峰,并指认1号峰为绿原酸。K-均值聚类分析结果显示,20批样品可聚为2类,Ⅰ类以安徽产样品为主,其桑根皮素含量较高、总黄酮含量偏低,颜色较暗、偏黄棕色;Ⅱ类以四川、贵州产样品为主,其桑根皮素含量偏低、总黄酮含量较高,颜色较明亮、偏黄白色。系统聚类分析结果与其较为一致。结论:桑白皮颜色与其桑根皮素、总黄酮含量密切相关,颜色偏黄棕色的桑白皮中桑根皮素含量较高,颜色偏黄白色的桑白皮中总黄酮含量较高。     

LC-MS法快速测定人血浆中格列吡嗪

目的 建立快速测定人血浆中格列吡嗪的HPLC-ESI-MS法,测定志愿者口服格列吡嗪片剂10 mg后的血药浓度,并对受试制剂和参比制剂的生物等效性进行评价.方法 血浆样品以甲醇沉淀蛋白,直接进行HPLC-ESI-MS分析,色谱柱为Lichrospher C18(5 μm,250 mm×4.6mm),流动相为10mmol/L乙酸铵缓冲液-甲醇(22:78,V/V),内标为格列本脲,检测离子分别为m/z444(格列吡嗪)、m/z492(内标),裂解电压为120 V.20名健康志愿者交叉口服供试片和参比片,计算主要药动学参数及相对生物利用度,以判断生物等效性.结果 在10.04～2008 ng/ml范围内格列吡嗪与内标的峰面积比值与浓度呈良好的线性关系.受试制剂及参比制剂的消除半衰期分别为(3.46±0.52)小时和(3.56±0.54)小时,达峰时间分别为(2.7±0.7)小时和(2.6±0.8)小时,达峰浓度分别为(734.72±210.34)ng/ml和(743.57±239.77)ng/ml.以AUC0-16计算的受试制剂的相对生物利用度为(96.1%±11.5%).结论 本实验建立的分析方法快速、灵敏、准确、简便.统计学结果表明两种制剂生物等效.     

外标一点法在人体甲氨蝶呤血浆浓度测定中的应用

目的：建立甲氨蝶呤（ MTX）血浆浓度的高效液相色谱测定方法。方法：利用外标一点法检测甲氨蝶呤血浆浓度,并与标准曲线法进行对比。结果：使用高效液相色谱法测定甲氨蝶呤血浆浓度时,标准曲线法在0.05～5.00μg／ml范围内药物浓度与峰面积呈现良好的线性关系,回收率、相对标准偏差符合测定要求;外标一点法在0.36～0.65μg／ml范围内药物浓度与峰面积呈现良好的线性关系,回收率、相对标准偏差符合要求。2种方法对患者服药44 h后甲氨蝶呤血浆浓度测定结果一致,差异无统计学意义（ P＞0.05）。结论：外标一点法简单、快速,精密度和重复性较好,适用于临床甲氨蝶呤血浆浓度测定。     

采用ZTC1＋1澄清剂法、水醇法制备扶正解毒口服液和克喘口服液的研究

扶正解毒口服液和克喘口服液是由中国中医研究院广安门医院研制生产的院内制剂,多年来临床应用疗效确切。原工艺中一直采用水醇法沉淀除杂质的制备方法。本实验采用ZTC1＋1澄清剂沉淀法的新工艺方法制备扶正解毒口服液、克喘口服液并与水醇法制备工艺进行了比较。 1实验材料 　　黄芪、党参、枸杞子、藤梨根、麻黄、石膏、紫苏子、莱菔子、葶苈子等十九味中药;ZTC1＋1澄清剂;何首乌对照药材,麻黄碱对照品;GC－105管式高速离心机;苯、氯仿、乙醚、丙酮、甲醇、正丁醇、无水乙醇等化学试剂均为分析纯。 2制备方法 2.1水醇法制备工艺 　　扶正解毒口服液：取浓缩液1份,加入95％乙醇使含醇量达到70％,搅匀,静置48小时,滤过,滤液回收乙醇至无醇味,加入甜叶菊甙,调整体积至规定量,搅匀,冷藏48小时,滤过,滤液高速离心(16000r/min),调PH至规定值,灌封于10ml安瓿中,灭菌即得样品1a。 　　克喘口服液：制法同扶正解毒口服液,得样品2a。 2.2ZTC1＋1澄清剂法制备工艺 　　扶正解毒口服液：取另一份浓缩液,加入ZTC1＋1澄清剂预处理剂A,煮沸20分钟,然后加预处理剂B,静置12小时,滤过,滤液煮沸20分钟灭酶。然后加入ZTC1＋1澄清剂Ⅳ型B组及A组,静置12小时,滤过,滤液高速离心(16000r/min),加入甜叶菊甙,调pH至规定值,调整体积至规定量,搅匀,灌封于10ml安瓿中,灭菌即得样品1b。 　　克喘口服液：取另一份浓缩液,加入ZTC1＋1澄清剂Ⅳ型B组及A组,静置12小时,滤过,滤液高速离心(16000r/min),加入甜叶菊甙,调pH至规定值,调整体积至规定量,搅匀,灌封于10ml安瓿中,灭菌即得样品2b。 3稳定性观察 　　将上述两种工艺制成的扶正解毒口服液样品1a、1b,克喘口服液样品2a、2b分别置于室温下放置3个月,观察其澄明度、色泽变化,测定其相对密度、pH值。两种工艺制备的样品均稳定、澄明,不混浊,色泽相近,无明显变化。 4质量鉴别 4.1扶正解毒口服液 4.1.1供试品溶液的配制：分别取样品1a、1b各30ml,加乙酸乙酯30ml振摇提取,提取液浓缩至1ml,作为供试品溶液1a、1b。 4.1.2对照品溶液配制：取何首乌对照药材1g,加甲醇10ml,加热回流30分钟,滤过,滤液浓缩至1ml,作为对照品溶液。 4.1.3样品的薄层色谱分析：按照薄层色谱法(中国药典2000年版一部附录ⅥB)试验。供试品色谱中,在与对照品药材色谱相应的位置上,显桔黄色的荧光斑点,各斑点大小相近。见图1。 4.2克喘口服液 4.2.1供试品溶液的配制：分别取样品2a、2b各20ml,加浓氨试液1ml,用乙醚振摇提取二次,每次20ml,合并乙醚提取液,加盐酸乙醇溶液(1→20)1ml,摇匀,蒸干,残渣加无水乙醇1ml使溶解,作为供试品溶液2a、2b。     

程序升温法研究药物稳定性的试验误差

通过电子计算机模拟程序升温加速试验数据,从理论上探讨了各种程序升温法的试验误差,并与恒温法作了对比。结果表明,药物降解程度越大,试验温度差越大,平均温度越接近室温,抽样频率越高,试验结果误差就越小。在药物降解程度和加热温度相同的条件下,指数程序升温法的试验误差明显小于线性升温、倒数升温和对数升温法。恒温法的试验结果误差小于指数升温法,但所需的加热时间约为指数升温法的5倍。     

Precision of internal standard method in HPLC analysis

The internal standard methods are known to compensate for the errors from sample preparation and injection into an analytical instrument. However, recent HPLC apparatuses have injectors of excellent repeatability and it is dubious whether the cancellation of injection error can lead to substantial improvement in the precision of analysis. This paper answers the above question experimentally and theoretically. The HPLC analysis of butylscopolamine bromide is taken as an example. The relative standard deviations (RSD) of measurements in the internal standard method and absolute calibration curve method are compared and the advantages of these methods are discussed. The measurement RSD is shown to be well estimated by the (function of mutual information) (FUMI) theory without repeating measurements. This report also demonstrates simple equations for calculating the measurement RSD at an arbitrary concentration of analyte and for selecting the better method between the internal standard method and absolute calibration curve method under specific experimental conditions.     

GC一测多评法测定小儿健脾贴膏中丁香酚、异龙脑和龙脑的含量

目的建立测定小儿健脾贴膏中丁香酚、异龙脑和龙脑3种成分含量的一测多评法。方法采用GC法,以萘为内标物、丁香酚为内参物,建立该成分与异龙脑和龙脑的相对校正因子,用该校正因子进行3种有效成分的含量测定,与内标法计算值进行比较。结果一测多评法计算值和内标法测得的异龙脑和龙脑的百分含量相对误差均小于2.0%。结论 GC一测多评法用于小儿健脾贴膏中丁香酚、异龙脑和龙脑3个有效成分的含量测定可行、准确。     

Direct quantification in bioanalytical LC-MS/MS using internal calibration via analyte/stable isotope ratio

The possibility to rationalize and simplify bioanalysis, without compromising the analytical quality, by omitting the calibration curves was studied. Using mass spectrometry (MS) and a stable isotope labeled internal standard it was possible to get equally good results by calculating the results directly from the analyte/internal standard area ratio and a predetermined response factor as by the traditional way, using a calibration curve run at the same occasion. To be able to use this simplified quantification method, that we call internal calibration, in its most simple form there are some prerequisites that must be considered: (1) The relative response should not be concentration dependent. (2) The relative response should be constant between batches/days. (3) The level of analyte in the internal standard should not be detectable. (4) There should be no influence from naturally occurring isotopes of the analyte on the internal standard peak area. A bioanalytical LC-MS/MS method for a research compound was validated both with and without calibration curves and no significant differences were found regarding precision and accuracy. It was shown that all four prerequisites above were fulfilled. Validation data were very good for the whole concentration range, 0.010-30 micromol/L. Long-term data for QC samples showed excellent precision and accuracy.     

内标多控法测定枳实中黄酮类成分及其不确定度评定研究

目的建立HPLC内标多控法测定枳实药材中黄酮类成分的方法,并对测定结果进行不确定度评定。方法以柚皮苷为内标,建立柚皮苷与橙皮苷、新橙皮苷的相对校正因子,计算枳实中黄酮类成分的含量,再根据数学模型,对测定过程中引入的不确定度进行评估,由此计算测定结果不确定度。结果采用本方法测定枳实中柚皮苷与橙皮苷、新橙皮苷含量的扩展不确定度分别为W×2.94%、W×3.49%、W×3.34%。结论建立的不确定度评定法可为内标多控法测定中药有效成分的不确定度分析提供参考。     

Development and validation of an accurate HPLC method for the quantitative determination of picroside II in tablets

A simple, sensitive and accurate high performance liquid chromatographic method (HPLC) with UV detection was developed and validated to determine picroside II in a new tablet formulation with paeoniflorin as internal standard. Chromatographic separation was achieved on an Agilent XDB C18 column (250 x 4.6 mm I.D., 5 microm) using a mobile phase consisting of acetonitrile-water-acetic acid (18:82:0.4, v/v/v) at a flow rate of 1.0 ml/min. The UV detection wavelength was set at 265 nm. Linear calibration curves were obtained in the concentration range of 0.10-100 microg/ml with the limit of quantification (LOQ) of 0.10 microg/ml. The within- and between-run precisions in terms of % relative standard deviation (RSD) were lower than 5.7% and 6.3%, respectively. The accuracy in terms of % relative error (RE) ranged from -2.3% to 5.0%. This validated method was successfully applied to the determination of the content of picroside II in a new tablet formulation.     

一点法与工作曲线法两种外标法选择依据的探讨

目的 :根据试验测定的标准直线 y=kx+b,如何根据直线方程中系数 k、b及进样量选择使用外标一点法还是用工作曲线法进行测定。方法 :使用数学推导出偏差计算公式并通过测定芍药苷的量进行验证。结果 :根据数学推导得因使用不同外标法而引起的偏差公式为 ε=bkx0 +b× (1- x0x1) (x0 ,x1 分别为对照品及样品进样量 )。结论 :高效液相色谱法 (气相色谱法 ) ,允许使用不同外标法引起偏差为 1% ,当标准直线方程中系数符合 bkx0 +b ≤ 3.5 %时 ,或薄层色谱扫描法 ,允许使用不同外标法引起偏差为 3% ,当标准直线方程中系数符合 bkx0 +b ≤ 10 .5 %时 ,样品进样量至少为对照品进样量的 80 %～ 140 %范围内 ,可用外标一点法代替工作曲线法进行定量测定     

A simple and fast method for quantification of ertapenem using meropenem as internal standard in human plasma in a clinical setting

A high-performance liquid chromatography method for the determination of ertapenem in human plasma using ultraviolet detection (300 nm) was developed and validated. The method involved a simple organic protein precipitation that guarantees rapid sample preparation. By using meropenem as internal standard, it also guarantees improved precision (relative standard deviation <3.70) and accuracy (deviation from true value <6.58%). The method was linear from 0.59 to 150 microg/mL and reproducible. In conclusion, this method allows the measurement of ertapenem in human plasma and may be used for both clinical routine applications and pharmacokinetic studies.     

Determination of bromazepam in plasma with an internal standard by gas-liquid chromatography.

A gaschromatographic assay method is described for the determination of Ro 5-3350 (bromazepam) using Ro 5-4547 (methylbromazepam) as internal standard. After alkaline ether extraction the bromazepam and methylbromazepam obtained from sulfuric acid reextraction are hydrolyzed to ABBP and to MABBP, respectively. After neutralization, the bromo-pyridine-benzophenones are extracted with ether and dissolved in hexane after evaporation of the ether. Under the described gaschromatographic conditions it was found that MABBP and ABBP have retention times of 10.5 and 12.5 min, respectively. The limit of sensitivity is situated at 5 ng/ml of plasma. The specificity is satisfactory since the metabolite which might have interfered (hydroxy-3-bromazepam) appears only at very low concentrations in the blood. The linearity of the calibration curve was confirmed for plasma concentrations up to 100 ng/ml.     

斜率校正“一测多评法”同时测定杜仲中京尼平苷酸、绿原酸、京尼平苷和松脂素二葡萄糖苷的含量

目的:建立斜率校正"一测多评法"同时测定杜仲中京尼平苷酸(GPA)、绿原酸(CHA)、京尼平苷(GP)、松脂素二葡萄糖苷(PDG)含量的方法。方法:以HPLC法含量测定为基础, CHA为对照,计算GPA、GP、PDG的斜率相对校正因子,并比较斜率校正"一测多评法"与外标法两种结果相关性,从而验证斜率校正"一测多评法"的准确性,最终建立测定杜仲多种活性成分的斜率校正"一测多评法"。结果:在一定的线性范围内,GPA、GP、PDG的斜率相对校正因子分别为:0.7628,0.4786,0.7112,重复性良好。斜率校正"一测多评法"计算的含量值与外标法实测值相关性较好,56批杜仲样本中GPA、GP、PDG含量结果的相关系数均大于0.99。结论:建立的斜率校正"一测多评法"准确性高,可用于杜仲中GPA、CHA、GP、PDG活性成分的含量测定。