HPLC法测定注射用甲氨蝶呤含量方法的改进

目的:改进注射用甲氨蝶呤(MTX)含量测定的高效液相色谱法。方法:以内标法代替《中国药典》采用的外标法进行注射用MTX的含量测定。色谱柱为InertsilC_8,流动相为乙腈-水-冰醋酸-三乙胺(14:86:0.5:0.3),紫外检测波长为306nm,流速为0.8mL·min~(-1),以甲硝唑为内标,并与外标法结果比较。结果:MTX检测浓度线性范围为0.52～3.64μg·mL~(-1)(r=0.9997),平均回收率为99.77%,RSD=0.42%。2种方法含量测定结果相似。结论:改进后的方法简便、准确、重复性好,可有效地控制MTX的质量。     

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.     

高效液相色谱法测定注射用法罗培南在健康人血浆和尿中的浓度

目的:建立测定人血浆和尿中注射用法罗培南浓度的高效液相色谱(HPLC)法。方法:血浆样品经10%三氯醋酸沉淀蛋白,尿样直接稀释,以替硝唑为内标,采用乙腈:0.05 mol·L~(-1)磷酸二氢钾缓冲液(pH 3.5,16:84,V:V)为流动相,经Zorbax XDB-C_8柱分离,318 nm波长检测。结果:法罗培南的血浆样品和尿样线性范围分别为0.1～50 mg·L~(-1)和0.5～250 mg·L~(-1),提取回收率分别为51.2%～54.9%和99.6%～104%,日内、日间精密度(RSD)均低于9.14%。结论:本方法准确性好、操作简单,可满足药动学研究的要求。     

胶束液相色谱法测定山莨菪碱的血药浓度和药代动力学参数

用胶束液相色谱技术以十二烷基硫酸钠(SDS)为表面活性剂,正丙醇为改性剂,硫酸阿托品为内标,用甲醇初步沉淀蛋白后,不经萃取,直接注射血浆样品,成功地测定了山茛菪碱的体内血药浓度;进行了健康受试者肌注给药的药代动力学研究。方法准确、灵敏、简便,可供进一步研究山茛菪碱及其生物利用度等测定血药浓度,并可用于临床监测。     

高效液相色谱法检测血清中的高三尖杉酯碱

目的：利用高效液相色谱法测定血清中的高三尖杉酯碱的含量。方法：以内标法为基础,取１ｍＬ血清标本加入内标三尖杉酯碱,经二氯甲烷２次重复提取,减压蒸干,残渣用流动相溶解,进样。色谱条件：以Ｃ１８反相柱为分析柱,流动相组成为０１ｍｏｌ·Ｌ－１甲酸铵－甲醇（２∶１）,流速为１０ｍＬ·ｍｉｎ－１,在紫外检测器波长２９０ｎｍ处进行检测。结果：方法学检查：最低检测限２ｎｇ·ｍＬ－１,在４～１５００ｎｇ·ｍＬ－１范围内呈线性,回收率９１３％（ｎ＝８）,批内和批间的相对标准偏差分别为１２％（ｎ＝８）和３６％（ｎ＝４）。结论：对临床化疗常与之配伍使用的３种药物,进行干扰实验,未发现干扰峰。     

高效液相色谱法测定人体血浆中吡喹酮及相对生物利用度

目的建立高效液相色谱法测定血浆中吡喹酮浓度。方法血浆样品沉淀后经NUCLEODUR100-5C18柱分离,流动相为乙腈-水-四氢呋喃=51：47：2（v/v/v）。24名健康志愿者采用自身对照随机交叉试验设计,分别单剂量口服吡喹酮片参比制剂（R）或受试制剂（T）1．2g后测定两者相对生物利用度。结果血浆中吡喹酮在25～3200μg·L^-1线性良好,最低定量浓度为25μg·L^-1;方法学回收率为104．3％～119．9％（n=15）;精密度（RSD）实验结果表明日内、日间变异均〈6．9％（n=15）。与R相比,T的相对生物利用度为104．0％±24．8％。结论该法简单,准确度高,灵敏度好;方差分析结果表明T与R的主要药动学参数之间无显著差异,2制剂生物等效。     

重组人粒细胞集落刺激因子成品蛋白质含量测定方法的研究

目的研究重组人粒细胞集落刺激因子 (rhG CSF)成品蛋白质含量测定方法。方法用三氯醋酸 Lowry法和高效液相色谱外标法测定rhG CSF成品蛋白质含量。结果两种方法均能有效排除rhG CSF成品中甘露醇的干扰 ,与经典的Lowry法相比 ,蛋白质的回收率均大于 96 % ,RSD均 <5 %。结论三氯醋酸 Lowry法和高效液相色谱外标法均可用于测定rhG CSF成品的蛋白质含量 ,高效液相色谱法操作更简便 ,更准确。     

2种测定N-乙酰半胱氨酸含量的方法比较

目的:比较2种测定N-乙酰半胱氨酸(NAC)原料含量的方法。方法:采用容量法和高效液相色谱(HPLC)法对NAC原料进行含量测定。结果:容量法测定结果偏高,HPLC法NAC检测浓度在0.1080～1.2690mg·mL-1范围内与峰面积积分值线性关系良好(r=0.9995);低、中、高平均加样回收率分别为99.77%、100.20%、100.20%,RSD=0.48%,含量测定结果较容量法低。结论:容量法测NAC含量操作简单,方法快速;HPLC法能够比较直观、真实的反映样品中所含有的杂质,与容量法比较,含量偏低。     

Rapid determination of serum melatonin by ESI-MS-MS with direct sample injection

This paper describes a rapid, simple and sensitive analytical method for the quantitative determination of melatonin in human serum by ESI-MS-MS with direct serum sample injection and on-line extraction. The method uses N-acetyltryptamine as the internal standard. It has high specificity and sensitivity for serum melatonin analysis. The internal calibration curve shows a wide linear range from 0.500 to 200 ng/ml with a correlation coefficient, R(2) > 0.999. The limit of quantitation is 0.500 ng/ml and the limit of detection is 0.100 ng/ml with 10-microl sample injection. The recoveries of serum melatonin at three levels are approximately 70%. The intra-assay precision (n = 5) is between 0.8 and 2.0% and the inter-assay precision (n = 3) is between 1.5 and 5.9% over the calibration range. This method has a total analysis time of less than 9 min. It can be used for the measurement of melatonin in human blood.     

高效液相色谱法测定人血浆中阿莫地喹的浓度(英文)

目的:建立高效液相色谱法测定人血浆中阿莫地喹的浓度。方法:分析柱采用KromasilC18(150mm×4.6mm,5μm),流动相为甲醇∶水∶三乙胺∶磷酸∶(21∶77.5∶1∶0.5),流速为1.0mL·min-1,检测波长为294nm,内标为羟氯喹。结果:阿莫地喹和羟氯喹的保留时间分别为5.82min,8.56min。该法在10～1000μg·L-1浓度范围内有良好的线性关系(r=0.9998,n=9),最低检测限为5μg·L-1,提取回收率为75.5%～82.7%,方法回收率为97.0%～104.8%。日内精密度的RSD<6.0%,日间精密度的RSD<7.5%。结论:该法简单,灵敏适合于阿莫地喹的药动学研究。     

Evaluation of sample work-up methods and internal standards for the determination of catecholamines in urine by HPLC with electrochemical detection

Two sample work-up methods: (I) one consisting of adsorption of the catecholamines onto alumina followed by ion pair extraction and (II) another consisting of isolation by cation exchange and subsequent adsorption onto alumina, have been evaluated for the assay of urinary catecholamines by means of HPLC with electrochemical detection. With the aim of achieving high precision, two internal standards, i.e. dihydroxybenzylamine and epinine, have been compared. The results indicate that clean HPLC chromatograms are obtained with both work-up methods and that the highest precision (RSD < 4%) is achieved with method II and with epinine as internal standard, whereas the lowest precision is obtained with method I and with dihydroxybenzylamine.     

Validated HPLC method for determination of chlorzoxazone in human serum and its application in a clinical pharmacokinetic study

A high performance liquid chromatographic (HPLC) method for the determination of chloroxazone in human serum using phenacetin as internal standard (IS) is described. Protein precipitation is used for preparation of the sample. A mobile phase consisting of acetonitrile and 0.5% acetic acid in water mixture (40:60 v/v) was used at a flow rate of 1 ml/min on a C18 column. The eluate was monitored using an UV/VIS detector set at 287 nm. Ratio of peak area of analyte to IS was used for quantification of serum samples. The absolute recovery was greater than 96% over a concentration range of 1 to 100 micrograms/ml and the limit of quantitation was 0.05 microgram/ml. The intra-day relative standard deviation (RSD) measured at 1, 10, 50, and 100 micrograms/ml ranged from 0.9 to 5.1%. The inter-day RSD ranged from 0.6 to 3.0%. The method is simple, sensitive and has been successfully used in pharmacokinetic study conducted in healthy human volunteers.     

高效液相色谱法测定益泼丰的含量

本文报道了ＨＰＬＣ内标法加校正因子测定法测定益泼丰原料药中益泼丰的含量。选ＯＤＳ色谱柱，以醋酸曲安缩松为内标物，甲醇：水（７２：２８）为流动相。益泼丰对照品与内标物的量之比对其色谱峰面积比作图，线性关系良好，相关系数ｒ＝０．９９９８，平均回收率为９９．８％。相对标准差ＲＳＤ为０．４％（ｎ＝１５）．平均校正因子ｆ＝０．２７６４，ＲＳＤ＝０６％（ｎ＝５）．方法简便，快速，准确。     

藏药八味獐牙菜丸的质量标准研究

目的：建立测定藏药八味獐牙菜丸中盐酸小檗碱含量的方法。方法：采用薄层色谱法和高效液相色谱法。结果：建立了八味獐牙菜丸中兔耳草和小檗皮的薄层色谱鉴别方法,并采用HPLC测定了该藏药制剂中小檗皮的有效成分盐酸小檗碱的含量,其回归方程为：y=4629102.46x-136034.70,r=0.9995,盐酸小檗碱的平均回收率为97.30%,RSD=1.57%（n=6）。结论：该方法测定快速、重现性好,可用于八味獐牙菜丸的质量控制。     

高效液相色谱法测定甲氧氯普胺片含量

本文为甲氧氯普胺片中甲氧氯普胺的HPLC测定法。用苯巴比妥钠作内标物。色谱柱CLC-C_8,检测波长214nm,流动相由磷酸盐缓冲液(0.01mol/L)—乙腈(1∶1)组成,并用磷酸溶液调至pH值5左右,供试品和内标物能在短时间内完全出峰,且达到较好的分离。该法专一,简便,灵敏度高,和中国药典(1985)方法比较,结果一致,适用于常规分析,更适宜药厂多批次的含量测定。     

高效液相色谱法测定人血浆中甲氨蝶呤浓度

目的:建立高效液相色谱测定人血浆中甲氨蝶呤（MTX）浓度的方法。方法:以茶碱（Theophyline）为内标,色谱柱为Nova-pak C₁₈柱（150 mm×3.9 mm,4μm）,流动相:乙腈-甲醇-0.05 mol·L^-1磷酸二氢钾缓冲液pH 6.5（6:2:92）,流速:1.0 ml·min^-1,检测波长303 nm及254 nm,柱温25℃。结果:甲氨蝶呤浓度在0.05～10.0 mg·L^-1范围内线性良好,r=0.999 6。绝对回收率为66.72%,相对回收率为98.35%,日内、日间RSD均<10%。结论:该法快速,简便,灵敏度高,适用于临床常规血药浓度监测。     

高效液相色谱法测定人血浆中头孢克肟浓度

目的 :建立以高效液相色谱法测定头孢克肟血药浓度的方法。方法 :以DiscoveryC18(250mm×4 6mm ,5μm )为分析柱 ,甲醇 -醋酸盐缓冲液 -三乙胺 (28∶72∶0 5)为流动相 ,检测波长为286nm ,流速为1 0ml/min ,头孢拉定为内标物 ,测定头孢克肟血药浓度。结果 :头孢克肟在0 1～5 0μg/ml检测浓度范围内呈良好线性关系 (r=0 9995) ;高、中、低3种浓度的日间RSD≤6 69 %、日内RSD≤6 10 % ;相对回收率为 (96 63±3 17) %。结论 :该方法操作简便、灵敏、准确 ,适用于临床头孢克肟的血药浓度测定及药动学研究     

高效液相色谱法测定癫痫患儿血清中左乙拉西坦的浓度

目的建立测定癫痫患儿血清中左乙拉西坦药物浓度的高效液相色谱(HPLC)法。方法用乙酸乙酯从血清中按液液萃取法提取左乙拉西坦和内标α-溴苯乙酮,经氮气恒流吹干富集浓缩,流动相复溶后20μL进样,0.4%磷酸-乙腈(92:8,V/V)为流动相,流速:1.0 mL·min~(-1),柱温35℃,以RP-C_(18)色谱分析柱(150 mm×3.9 mm,5μm)在210 nm波长处测定,记录左乙拉西坦与内标的峰面积比值并拟合浓度工作曲线。结果左乙拉西坦和内标峰面积比与左乙拉西坦浓度在0.234.～60.00 mg·L~(-1)的范围内线性关系良好,Y=0.065 14 X-0.004 7,r=0.999 5,最低定量限为0.234 mg·L~(-1)(S/N=4.8)。方法平均绝对回收率为(89.18±3.57)%,日间和日内RSD分别为(5.84±2.15)%和(5.48±2.61)%,冻融稳定性RSD<5%。该方法能完全分离临床可能合并所用药物的干扰(分离度R均大于1.5),采用单盲法对方法学质量控制结果RSD<4%。结论该方法经方法学验证符合血清样品的测定要求,可用于临床血药浓度测定和药动学研究。     

Validation of a simple bioanalytical method for the determination of DRF-6196, a novel oxazolidinone, in mouse plasma: application to a single-dose pharmacokinetic study

An isocratic simple, specific, sensitive and reproducible high performance liquid chromatography (HPLC) method was developed and validated for the estimation of DRF-6196, a novel oxazolidinone in mouse plasma. This method involves a simple liquid/liquid extraction of DRF-6196 and the internal standard (IS; chlorzoxazone, CAS 95-25-0) from plasma into dichloromethane/ethyl acetate mixture that was evaporated under nitrogen. The HPLC analysis was carried out on an Inertsil ODS 2 column using 0.01 mol/L potassium dihydorgen ortho phosphate (pH 3.2) and acetonitrile (65:35, v/v) as mobile phase. The eluate was monitored using an UV detector set at 266 nm. Ratio of peak area of analyte to IS was used for quantification of plasma samples. The retention time of DRF-6196 and IS were 8.2 and 11.1 min, respectively. The assay was linear (r2 > 0.999) in the concentration range 0.1-50 microg/ml. Absolute recovery for analyte and IS was > 94 % from mouse plasma. The lower limit of quantification (LLOQ) of DRF-6196 was 0.1 microg/ml. The inter- and intra-day precision in the measurement of quality control (QC) samples, 0.1, 0.3, 15.0 and 40.0 microg/ml, were in the range 3.64 to 9.51 % relative standard deviation (RSD) and 0.92 to 6.23 % RSD, respectively. Accuracy in the measurement of QC samples was in the range 88.15 to 106.05 % of the nominal values. The analyte and IS were stable in the stability studies viz., benchtop, autosampler and freeze/thaw cycles. The stability of DRF-6196 was established for 1 month at -80 degrees C. The assay method was successfully applied to a pharmacokinetic study of DRF-6196 in mice.     

高效液相色谱法测定烧伤酊中黄芩苷的含量

目的建立烧伤酊中黄芩苷的高效液相色谱测定方法。方法采用Hypersil-ODS C18柱,流动相为：甲醇-水-冰醋酸（50∶50∶1）,检测波长：274 nm,流速：1.0 ml.min-1。结果黄芩苷对照品在30.096~150.48 g.L-1浓度范围内具有良好的线性关系,回归方程为：Y=14633X＋1096.9（r=0.99998,n=5）。平均加样回收率为98.84%（RSD=1.62%,n=5）。结论本方法简便快捷,结果准确,并具有良好的重复性,可用于本制剂的含量测定和质量控制。