内标对比标准曲线法与标准加入对比法的误差分析

目的对内标对比标准曲线法与标准加入对比法的原理与误差进行分析。方法以相对峰面积为指标 ,利用偏微分原理对二种方法进行误差分析。结果对于内标对比标准曲线法在Ai/Ar=0 3～ 3 0区间 ,标准曲线斜率B(b)较大而可获得较高灵敏度。Ai/Ar=a +b时 ,结果误差较小 ;样品相对峰面积越大及其相对测量误差越小 ,以及a越小时 ,分析误差越小。对于标准加入对比法得到了标准加入曲线不通过原点时的定量计算公式 ;当B与b的差值越小 ,或横轴截距 -A/B与 -a/b差值越大 ,以及测定相对峰面积的误差越小时 ,可使分析相对误差越小。结论两种方法在CE定量分析中应用可行 ,而且特别适合进样不能准确控制的色谱定量分析     

毛细管电泳叠加对比法测定复方降压片中5组分含量

目的建立复方降压片 (二方 )的高效毛细管电泳多组分同时定量方法。方法用高效毛细管电泳叠加对比法 ,采用石英毛细管 ( 6 5cm× 75 μmI.D .)有效长度 5 0cm ,5 0mmol/L硼砂溶液为背景电解质 ,重力进样 1 0s(高度 7cm) ,检测温度 ( 2 4± 1 )℃ ,紫外检测波长 2 4 6nm。结果测定复方降压片中维生素B1、磷酸氯喹、氢氯噻嗪、维生素B6和芦丁 5组分的相对峰面积的RSD不大于2 4 % ,回收率为 96 8%～ 1 0 4 2 % ,标示百分含量均在 96 7%～ 1 0 3 5 %。结论该法可同时分离测定复方降压片中 5个组分     

毛细管电泳法测定复方降压片中5组分含量

目的建立复方降压片(二方)的高效毛细管电泳5组分同时定量方法.方法用高效毛细管电泳内标对比法,采用石英毛细管(65 cm×75 μm,I.D.)有效长度50 cm,50 mmol·L-1硼砂溶液为背景电解质,重力进样10 s(高度7 cm),检测温度(23±1)℃,紫外检测波长246 m.结果测定复方降压片(二方)中维生素B1、磷酸氯喹、氢氯噻嗪、维生素B6和芦丁5组分的相对峰面积的RSD≤4.1%,回收率为94.6%～105.3%,标示百分含量均在96.1%～106.4%.结论该法可同时分离测定复方降压片中5个组分.     

复方甘草片的毛细管电泳指纹图谱研究

目的 建立复方甘草片的毛细管电泳指纹图谱。方法 以50 mmol·L-1硼砂溶液(含10％甲醇,v/v)提取样品,用毛细管区带电泳法,紫外检测波长228 nm,电压14 kV,以50mmol·L-1硼砂溶液为背景电解质,氢氯噻嗪为参照物(s),对6个厂家各10批以上复方甘草片进行指纹图谱研究。以20个峰的迁移时间、峰面积、相对迁移时间和相对峰面积分别构成表征样品性质的4个向量,以不同批指纹向量与标准对照指纹向量间的相关系数和夹角余弦为测度评价指纹的相似性,并以所建立的CE指纹图谱检验另外1个厂家样品为合格。结果 所建立的CE指纹图谱具有稳定、重现的特点。结论 该指纹图谱可控制复方甘草片的质量。     

非水毛细管电泳法同时测定苦参栓中苦参碱和氧化苦参碱的含量

目的:建立简单、快速、灵敏的能同时分离测定苦参栓中苦参碱和氧化苦参碱的非水毛细管电泳方法。方法:采用非水毛细管电泳法,缓冲体系70 mmol.L-1乙酸铵-7.0%醋酸-10%乙腈的甲醇液,熔融石英毛细管(50μm×50 cm),分离电压25 kV,检测波长205 nm,阳极进样。结果:苦参碱和氧化苦参碱在1.00～500μg.mL-1与峰面积线性关系良好。迁移时间的日内相对标准偏差分别为0.09%和0.59%,日间相对标准偏差分别为0.63%和1.9%;峰面积的日内相对标准偏差分别为0.45%和4.9%,日间相对标准偏差分别为2.3%和5.6%。平均回收率分别为96.63%和97.65%。结论:该方法简单、快速、准确、重复性好,可用于苦参栓的质量控制。20123214Chin Hosp Pharm J2012     

高效毛细管电泳法测定人血浆育亨宾浓度

目的 建立非水毛细管电泳法(NCAE) 测定人血浆育亨宾浓度的新方法 . 方法 运用非水毛细管电泳方法 , 熔融石英毛细管(75 μm ×50 cm) , 缓冲液为30 mmol•L 1醋酸铵的甲醇 乙腈液(4：1, V/V, 含1%冰醋酸), 检测波长：220 nm , 分离电压：25 kV, 柱温25 ℃, 用0.45 μm微孔滤膜过滤后进样, 压力进样：50 kPa×5 s, 采用峰面积内标法定量. 结果方法 最低检测浓度为0.01 μg•mL 1, 线性范围0.054 8～5.480 0 μg•mL 1, r＝0.999 1 , 线性关系良好. 日内RSD 为2.81%～5.22%, 日间RSD 为4.72%～6.52%. 结论该方法 测定人血清中育亨宾含量快速、准确和灵敏.     

氢核磁共振定量法测定恩替卡韦

目的 建立氢核磁共振定量法测定恩替卡韦含量的方法.方法 以氘代DMSO为溶剂,恩替卡韦为样品,1,4-二硝基苯为内标测定氢核磁共振谱,通过比较样品定量峰与内标物质响应峰面积,计算恩替卡韦的量.结果 同一样品在相同条件下测定6次,定量峰与内标响应峰比值的RSD值为1.1%;平行配制3份样品,恩替卡韦质量分数93.8%,RSD值为0.70%,与质量平衡法测定结果93.8%一致.结论 氢核磁共振定量法与质量平衡法相比样品用量少,不需要对照品,测定快速准确,是一种有效地化学样品含量测定手段.     

尿激酶中分子组分比2种测定方法的建立和比较研究

目的:建立尿激酶中分子组分比测定的分子排阻色谱法与十二烷基硫酸钠毛细管电泳法,并对2种方法的方法学验证与样品测定结果比较。方法:分子排阻色谱法采用TSKgel G2000SWXL色谱柱(7.8mm×300 mm,5μm),以0.1 mol·L^-1磷酸二氢钠缓冲液(pH 3.0)为流动相,流速0.5 mL·min^-1,柱温35℃,检测波长280 nm,进样量50μL;十二烷基硫酸钠毛细管电泳法采用未涂层熔融石英毛细管(50μm×30.2cm,有效长度20.2 cm),分离电压为15 kV,柱温25℃,检测波长214 nm,进样端为正极,5 kV压力进样20 s。结果:通过对分子排阻色谱法与十二烷基硫酸钠毛细管电泳法的方法学验证与样品测定结果的比较,证明两种方法测定结果基本一致。分子排阻色谱法:高分子量与低分子量尿激酶质量浓度在0.05~5.1mg·mL^-1范围内呈良好的线性关系;检测下限分别为1.5μg·mL^-1与5.1μg·mL^-1,定量下限分别为5.1μg·mL^-1与16.9μg·m L^-1;8批样品中高分子量与低分子量尿激酶相对含量范围分别为87.0%~96.4%与3.6%~13.0%。十二烷基硫酸钠毛细管电泳法:高分子量与低分子量尿激酶质量浓度在0.1~5.2 mg·mL^-1范围内呈良好的线性关系;检测下限分别为2.5μg·mL^-1与51.8μg·mL^-1,定量下限分别为7.5μg·mL^-1与155.4μg·m L^-1;8批样品中高分子量与低分子量尿激酶相对含量范围分别为88.2%~97.3%与2.7%~11.8%。结论:分子排阻色谱法与十二烷基硫酸钠毛细管电泳法都适用于尿激酶中分子组分比的测定并互为补充。     

毛细管电泳法对肝素钠注射液中杂质的测定

目的:在2008年国家食品药品监督管理局颁布的毛细管电泳法分析肝素钠杂质的补充检验方法的基础上优化毛细管电泳色谱条件,对全国评价性抽验肝素钠注射液样品进行测定。方法:采用毛细管电泳法,缓冲液为pH 3.0的400 mmol.L-1三羟甲基氨基甲烷(Tris)溶液;分离电压为-25 kV;检测波长为200 nm;进样压力3.447 kPa,进样时间为4 s;毛细管温度:25℃。结果:肝素主峰与多硫酸软骨素峰峰之间分离度为1.0,与硫酸皮肤素之间分离度为2.0,10批供试品的杂质含量结果与离子色谱法结果基本一致。结论:优化后的色谱条件使肝素与硫酸皮肤素、多硫酸软骨素的分离度增加,该方法使肝素钠样品中的杂质定性、定量更为准确。     

毛细管电泳法测定盐酸洛美沙星片的含量

目的 建立毛细管电泳法(HPCE)测定盐酸洛荑沙星片的舍量.方法 以0.2mol/L NaH2PO4-H3PO4缓冲液(pH4.0)为背景电解质,50era×75iμm未涂层空心毛细管柱为分离通道,检测波长为282nm,运行电压25kV,压力进样(0.5psi×3s),柱温25℃,采用峰面积外标法定量.结果 在选定的电泳条件下,洛美沙星的浓度在50.00-500.00mg/L范围内与峰面积呈良好的线性关系(r=0.9995);低、中、高三种浓度(n=3)的平均回收率分别为99.94%、99.94%与96.25%;方法的检测限(S/N=3)为5.00mg/L;日内和日间峰迁移时间的RSD分别2.17%、1.13%,日内和日间峰面积的RSD分别为3.94%、1.97%.结论 本法试剂用量少、简便、快速、准确,可用于洛美沙星的质量检控.     

The determination of a degradation product in clidinium bromide drug substance by capillary electrophoresis with indirect UV detection

A capillary electrophoresis (CE) method utilizing indirect ultraviolet (UV) detection was developed for the determination of a non-UV absorbing degradation product, Ro 5–5172, in clidinium bromide drug substance. The electrophoresis buffer consisted of sodium phosphate and benzyltrimethylammonium bromide. Rinsing the capillary with sodium hydroxide followed by water then fresh capillary electrophoresis buffer was found to significantly improve the reproducibility of the migration times of the analytes. To further improve run-to-run reproducibility, an internal marker was used to account for differences in injection volumes and migration times between runs. The precision of the method was found to be less than 1% relative standard deviation for the migration time ratio and peak area ratio of Ro 5–5172 to the internal standard. The method was found to be linear for 0.05–1% Ro 5–5172 with respect to a 10 mg ml⁻¹ sample preparation. The limit of detection was found to be less than 0.01% Ro 5–5172. Results obtained for the analysis of a clidinium bromide drug substance lot using this CE method and a thin layer chromatography method were compared and found to be in agreement.     

Quantitative analysis of polymorphs in binary and multi-component powder mixtures by near-infrared reflectance spectroscopy

Near-infrared reflectance spectroscopy was employed to quantify polymorphs in binary and multi-component powder mixtures. Sulfamethoxazole (SMZ) forms I and II were used as model polymorphs for this study. The instrument reproducibility, method error, precision, and limits of detection and quantification of the method were assessed. Physical mixtures of the polymorph pair were made by weight, ranging from 0 to 100% SMZ form I in II. Near-infrared spectra of the powder samples contained in glass vials were obtained over the wavelength region of 1100-2500 nm. A calibration plot was constructed by plotting SMZ form I weight percent against a ratio of second derivative values of log(1/R') (where R' is the relative reflectance) versus wavelength. The coefficients of determination, R(2), were generally greater than 0.9997 and standard errors were low for all the systems. Instrument error was assessed by analyzing a sample 10 times without perturbation. Method error was assessed in the same manner except the sample was re-mixed between analyses. A precision study was conducted by analyzing aliquots from a larger homogeneous sample. Limits of detection (LOD) and quantification (LOQ) were determined from the standard deviation of the response of the blank samples (100% SMZ form II, undiluted or diluted with 60% lactose). These limits were subsequently validated with independent samples. The results show that polymorphs can be quantified in binary and multi-component mixtures in the 2% polymorph composition range. These studies indicate that NIRS is a precise and accurate quantitative tool for determination of polymorphs in the solid-state, is comparable to other characterization techniques, and is more convenient to use than many other methods.     

纳米胶束载体聚乙二醇单甲醚聚乳酸嵌段共聚物中细菌内毒素动态显色定量方法的建立

目的: 建立纳米胶束载体聚乙二醇单甲醚聚乳酸嵌段共聚物细菌内毒素动态显色定量方法。 方法: 首先建立动态显色法测定细菌内毒素的标准曲线,并验证标准曲线的可靠性,考察样品对细菌内毒素检查的干扰情况,确定样品的最大无干扰浓度,随后验证方法的精密性和准确性,并进行初步应用。 结果: 标准曲线可靠性验证中,相关系数r＞0.99,标准曲线可靠。供试品用水稀释并添加内毒素标准品至供试品终浓度0.625mg.ml^-1(稀释40倍)及内毒素终浓度为0.25 Eu.ml^-1时,测定其回收率接近100%,供试品在该浓度对内毒素实验的干扰作用较小。方法的精密度和准确度验证中,标准曲线各浓度点测量结果的变异系数均＜7%,加入各浓度的内毒素标准品的测定结果的变异系数均＜5%,回收率在94%-112%之间。另外,日常检测3批供试品的内毒素含量均小于限值,回收率在97%-105%之间。结论: 动态显色法可用于定量检测纳米胶束载体聚乙二醇单甲醚聚乳酸嵌段共聚物的细菌内毒素。     

Reproducibility of standard preparation in digoxin radioimmunoassay in plasma and serum

We studied the reproducibility of standard preparations in digoxin radioimmunoassay in a randomized trial using serum and plasma as matrices. The errors expressed relative to the observed counts per minute (cpm) attributable to each of the procedures involved in the preparation of standards were as follows: preparation of stock solutions and dilutions, 1.3%; addition of diluted solutions to the medium, 1.2%; and residual error due to the assay procedure, 3.7%. No error caused by mixing, portioning, and storage was detected. Heparinized plasma gave lower cpm values than serum at 4.0 ng/ml (p less than 0.01), an effect that would give over- or underestimations by about 5% at that level, depending on the medium used. This suggests that standard and sample matrices should be similar. Our procedure for preparing the standards seems to be reasonably reliable; this is necessary for satisfactory monitoring of patients on digitalis therapy.     

Capillary electrophoretic method for determination of protease inhibitor indinavir sulfate used in human immunodeficiency virus therapy

A simple, fast and reliable capillary electrophoresis (CE) method for determination of indinavir sulfate, a potent protease inhibitor used in human immunodeficiency virus (HIV) therapy, in commercial and simulated capsule formulations is described. The analysis was performed in a 75microm i.d. uncoated fused-silica capillary with 27cm length (effective length of 19.4cm) using a 20mmoll-1 phosphate buffer at pH 2.52. Samples were injected hydrodynamically by applying 0.5psi pressure during 2s. The applied voltage was 28kV. Direct UV detection at 214nm led to an adequate sensitivity without interference from sample excipients and known impurities. For quantitative purposes, diazepam was used as internal standard. Under optimized conditions, the migration times for indinavir sulfate and diazepam were 1.06 and 1.66min, respectively. Analytical curve of peak area ratios versus concentration in the range of 20.0-100.0microg/ml gave a coefficient of correlation of 0.9992, establishing the method linearity. The limits of detection and quantitation were 4.61 and 14.0microg/ml, respectively. The within-day precision expressed as relative standard deviation was <1.5% for 10 consecutive sample injections. An average recovery of 100.81+/-0.56% at three concentration levels was obtained. Based on the performance characteristics, the proposed methodology was found suitable for the determination of indinavir sulfate in capsule formulations, presenting additional advantages inherent to the CE technology, such as low consumption of reagents and column endurance.     

Comparison of high-performance liquid chromatography and capillary electrophoresis methods for quantitative determination of glycyrrhizinic acid in pharmaceutical preparations.

A high-performance liquid chromatographic (HPLC) and a capillary electrophoretic (CE) method have been developed for the determination of glycyrrhizinic acid in pharmaceuticals. The two methods have been validated and have been compared with respect to their suitability for the said purpose as well as in relation to requirements from the legal authorities. The HPLC method provide a repeatability of the quantitative analysis of glycyrrhizinic acid below 1% relative standard deviation (RSD) which makes the method suitable when the legal authorities requires the content to be within +/-5% of the declaration. The repeatability of the CE method is in the order of 3-5% RSD when using internal standardization. However, using internal standardization as well as peak normalisation (peak area/migration time) indicate that the RSD may be reduced to about 1%. The CE method is suitable as a stability indication method as the degradation product glycyrrhetinic acid can be determined simultaneously.     

Microdetermination of procainamide in human serum

An electron-capture GLC method to measure procainamide (0.1-1 microgram/sample) in human serum was developed. An internal standard, p-amino-N-[2-(dipropylamino)ethyl]benzamide, is added to the serum before the sample is alkalinized with pH 10.5 phosphate buffer and extracted with ethyl acetate. The ethyl acetate phase is evaporated to dryness, and the residue is reacted with pentafluoropropionic anhydride. N-Pentafluoropropionyl derivatives of the drug and the internal standard had retention times of 5 and 8 min, respectively, when chromatographed at 235 degrees on a 1-m (4-mm i.d.) glass column packed with 5% OV-17 (carrier gas flow of 40 ml/min). The coefficient of variation was less than 5% for spiked standards. Furthermore, N-acetylprocainamide added to samples did not interfere. One hundred and eighty-six samples from 16 patients receiving procainamide intravenously were assayed by this GLC procedure and by a standard colorimetric method. Linear regression analysis yielded a correlation coefficient of 0.985 (slope, 1.040; intercept, 0.015).     

动态显色法检测注射用重组人粒细胞巨噬细胞刺激因子中内毒素的含量

目的建立定量检测注射用重组人粒细胞巨噬细胞集落刺激因子（rhGM—CSF）中内毒素的质控方法。方法采用动态显色法,通过标准曲线的可靠性验证和干扰试验,建立检测rhGM—CSF中内毒素的定量方法。结果标准曲线可靠性验证中,曲线的R2均〉0．98,标准曲线成立;干扰试验显示,供试品复溶后稀释5倍后加入0．5EU／ml的标准内毒素作为供试品回收液,经检测其内毒素回收率均接近100％,该稀释倍数下,供试品浓度对内毒素实验的干扰作用较小;另外,日常检测2批供试品的内毒素含量均小于限值,与凝胶法检测结果一致。结论该动态显色法方法可用于rhGM—CSF中内毒素的定量检测。     

基于近红外光谱的疏血通注射液浸提过程总固体含量分析

目的 建立一种快速测定疏血通注射液浸提过程中总固体含量的近红外光谱法。方法 以水蛭和地龙2种药材浸提过程为例,采用偏最小二乘法(PLS)建立总固体含量的近红外光谱分析校正模型,实现对总固体含量的快速测定。结果 近红外光谱在一阶导数结合Karl Norris平滑滤波处理下,建模效果最佳。水蛭总固体含量校正集相关系数(R)为0.810 8,校正集和验证集预测误差均方根(RMSEC、RMSEP)分别为0.583和0.495,交叉验证误差均方根(RMSECV)为0.81,校正集和验证集相对偏差(RSEC、RSEP)分别为6.11%和5.25%;地龙总固体含量校正集R值为0.975 5,RMSEC和RMSEP分别为1.10和1.85,RMSECV为1.61,RSEC和RSEP分别为4.68%和7.80%。结论 近红外光谱可用于快速测定疏血通注射液浸提过程中总固体含量,有望推广应用于中药浸提过程的在线质量控制。