NP-HPLC测定阿敏-额尔敦中士的宁的含量

目的:建立蒙药阿敏-额尔敦中士宁含量测定方法.方法:采用正相高效液相色谱法.色谱柱为JapanInertsil(SIL 100A-5μm,4.6mm×150mm),流动相为正己烷-二氯甲烷-甲醇-浓氨试液(47.5:47.5:5:0.35),流速1mL·min-1,柱温:30℃,检测波长为254nm.结果:士的宁进样量在0.0918～0.459μg范围内呈良好的线性关系(r=0.9996),平均加样回收率为98.12%,RSD为0.87%.结论:采用正相高效液相色谱法测定士宁含量,方法简便、灵敏、准确,可作为蒙药阿敏-额尔敦的质量控制方法.     

NP-HPLC法测定二氯二茂钛原料药的含量及有关物质

目的:建立正相高效液相色谱法测定有机金属抗癌原料药二氯二茂钛的含量及有关物质。方法:采用 Shimadzu LC-10A 液相色谱系统,色谱条件:以氰基 Lichrosorb—CN(4 mm×250 mm,5 μm)为色谱柱,以正己烷-二氯甲烷(50:50)为流动相,流速:1.0 mL·min~(-1),紫外检测波长:254 nm,柱温:25℃。以ω-溴代苯乙酮为内标测定二氯二茂钛原料药的含量;以自身对照法测定有关物质。结果:样品中待测组分与内标物分离良好,线性范围为2.5～50μg·mL~(-1),回归方程为 Y=0.0710X 0.0562,r=0.9999(n=7);日内和日间精密度 RSD 均小于1.0%,最低检测限为2 ng;方法的平均回收率(n=9)为99.9%。结论:方法简便、快速,结果准确,专属性强,灵敏度高,可同时测定有机金属类抗癌原料药二氯二茂钛的含量及有关物质。     

反相离子对高效液相色谱法测定人血浆中阿昔洛韦浓度及药代动力学

建立了用反相离子对高效液相色谱法测定人血浆中阿昔洛韦浓度的方法。色谱柱采用ＡｌｌｔｅｃｈＣ＿１８分析柱，以０．０４ｍｏｌ／Ｌ氯化钠－甲醇－ＩＰＲ－Ｂ７离子对色谱试剂混合液（１００：１５：０．６；ｖ／ｖ）为流动相。检测波长为２５４ｎｍ，肾上腺素为内标。血浆样品经高氯酸沉淀蛋白后直接进样测定。阿昔洛韦浓度在４０～１６００ｎｇ／ｍｌ范围内线性良好，ｒ＝０．９９９２．测定含阿苷洛韦１６０ｎｇ／ｍｌ的血浆样品，其日内（ｎ＝７）及日间的ＲＳＤ分别为４．７％和３．０％。平均回收率为９８．２±５．３％。测定了１０名健康志愿者单次口服阿昔洛韦片剂４００ｍｇ后不同时间的血药浓度并计算了有关的药代动力学参数。     

两种高效液相色谱法测定血清中茶碱浓度方法的评价

本文从经济、实用的角度出发,建立并比较了在反相(A法)和正相(B法)色谱系统下测定血清中茶碱浓度的两种高效液相色谱法(HPLC)。A法采用了ZORBAXODS色谱柱,甲醇、醋酸盐缓冲液为流动相;B法采用ZORBAXSIL色谱柱,正已烷、氯仿、甲醇三元混合溶剂为流动相,均以β-羟乙基茶碱为内标、275nm为检测波长。测定结果表明,两法线性范围均为2.5～40.Oμg/ml,最低检测浓度为2.0μg/ml,并且均有良好的相关性(r=0.999)。目内、目间变异小于6.36％,A法和B法的回收率分别为100.1±3.68％和99.1±4.33％,分析时间分别为11min和7min,血清内源物和部分临床常用药物不干扰两法测定。两法的测定结果相关性良好,回归方程为 =0.9681× 0.1504(r=0.999,n=5)。经临床的茶碱样品测定表明,两法简便灵敏,快速实用,能满足临床日常监测血药浓度水平的需要。     

Arachidonic and eicosapentaenoic acid metabolism by human CYP1A1: highly stereoselective formation of 17(R),18(S)-epoxyeicosatetraenoic acid

Human cytochrome P450 1A1 (CYP1A1) and human NADPH-cytochrome P450 reductase were expressed and purified from Spodoptera frugiperda insect cells. A reconstituted enzymatically active system metabolized polyunsaturated fatty acids such as arachidonic (AA) and eicosapentaenoic acid (EPA). CYP1A1 was an AA hydroxylase which oxidizes this substrate at a rate of 650+/-10 pmol/min/nmol CYP1A1, with over 90% of metabolites accounted for by hydroxylation products and with 19-OH-AA as major product. Epoxyeicosatrienoic acid (EET), mainly 14,15-EET, accounted for about 7% of total metabolites. Unlike rat CYP1A1, the human enzyme exhibited no 20-OH-AA as product. In contrast, with EPA as substrate CYP1A1 was mainly an epoxygenase, oxidizing with over 68% of total metabolites EPA to 17(R),18(S)-epoxyeicosatetraenoic acid (17(R),18(S)-EETeTr). 19-OH-EPA accounted for about 31% of total metabolites. Significantly, the 17,18-olefinic bond of EPA was epoxidized to 17(R),18(S)-EETeTr with nearly absolute regio- and stereoselectivity. Molecular modeling analyses provided rationale for high efficiency of AA hydroxylation at C(19) and its gradual decrease down to C(14), as well as for the limited EPA 17(S),18(R) epoxidation due to unfavorable enzyme-substrate interactions. The absence of omega-hydroxylation for both substrates is not due to steric factors, but probably a consequence of different reactivities of omega and (omega-1) carbons for hydrogen abstraction. It is suggested that the capacity of human CYP1A1 to metabolize AA and EPA and its inducibility by polycyclic aromatic hydrocarbons may affect the production of physiologically active metabolites, in particular, in the cardiovascular system and other extrahepatic tissues including lung.     

NP—HPLC与RP—HPLC两种方法测定制马钱子中士的宁含量

目的比较NP—HPLC与RP—HPLC两种测定制马钱子中士的宁含量的方法。方法按照2000版和2005版《中国药舆》，采用NP—HPLC与RP—HPLC两种方法。结果加样回收率考察中NP-HPLC法平均回收率为97．16％，RSD=0．71％（n=6）；RP—HPLC法平均回收率为98．13％，RSD=0．45％（n=6）。结论此二种方法都准确可靠，RP—HPLC法更为合适。     

盐酸帕洛诺司琼注射液中异构体含量的NP-HPLC测定

建立了正相高效液相色谱(NP-HPLC)法测定盐酸帕洛诺司琼注射液中(R,S)-异构体的含量.样品经氧氧化钠碱化后,再用二氯甲烷提取.用Inertsil SIL-100A色谱柱,二氯甲烷-甲醇-氨水(448:50:2)为流动相,检测波长261 nm.采用加校正因子的主成分自身对照法计算(R,S)-异构体的含量.     

正相高效液相色谱法测定人血浆中的氢氯噻嗪及药动学研究

 目的：建立正相高效液相色谱法测定人血浆中的氢氯噻嗪(HCTZ)浓度，研究正常人po后的药动学。方法：色谱柱采用YWG-SiO2柱，以氯仿-甲醇-冰乙酸(500∶50∶6)为流动相，紫外λ/nm 272检测。血浆样品在酸性条件下用乙酸乙酯提取吹干浓缩后进样。对10名健康志愿者po复方卡托普利片(含HCTZ₁₈mg)后不同时间的血药浓度进行测定。结果：方法的平均回收率为94.7%。血浆最低检测限5 ng/ml。正常人体内HCTZ消除相平均T_1/2为2.8 h;T_PK为2.5 h;c_max为97 ng/ml。结论：方法能满足HCTZ血药浓度监测和药动学研究的需要。     

手性固定相NP-HPLC法拆分比卡鲁胺对映体

建立了正相色谱法拆分比卡鲁胺对映体.考察了色谱柱、流动相中异丙醇浓度、流速和温度对对映体拆分的影响,确定拆分条件如下:采用Chiralpak AD-H手性固定相,以正己烷-异丙醇(70:30)为流动相,检测波长270 nm,流速0.6 ml/min,柱温20℃.在此条件下,比卡鲁胺对映体分离完全,分离度为7.0.