以HPLC法测定马来酸氯苯那敏含量的方法中马来酸峰及氯苯那敏峰定位的问题

目的：探讨在《国家药品标准化学药品地方标准上升国家标准》收载品种中采用HPLC方法测定马来酸氯苯那敏的含量时,所采集的HPLC图谱中,出现的马来酸峰与氯苯那敏峰保留位置如何确定的问题。方法：采用高效液相色谱法,色谱柱：Waters C18（4．6mm×150mm,5μm）,江苏淮阴C18（4．6mm×250mm,5μm）;流动相：庚烷磺酸钠溶液（冰醋酸调pH值至3．3）-甲醇-乙腈（25：30：18）;检测波长262nm。结果：在本文收集的试验条件下所采集的色谱图中,马来酸氯苯那敏对照品均出现了前后两个峰,马来酸峰与氯苯那敏峰保留位置可能在不同的色谱条件下有明显变化。结论：色谱图中马来酸峰与氯苯那敏峰保留位置应加以确定,以防止对马来酸氯苯那敏对照品纯度产生疑虑。     

Simultaneous Determination of Contents of Three Active Components in Jiejia Tincture by HPLC Method

The objective of the study was to determine the contents of three active components in Jiejia tincture by establishing HPLC method. Test articles were prepared by ultrasonic extraction. Separation was performed using a Kromasil C18 (250 mm × 4.6 mm, 5 µm) chromatographic column, and gradient elution was performed with acetonitrile-0.3% phosphoric acid solution as the mobile phase at a volumetric flow rate of 0.80 mL/min. The contents of catechin, baicalin and berberine in Jiejia tincture were determined at the wavelength of 276 nm and a column temperature of 30 □. The results revealed that catechin showed a good linear relationship at the range of 100∼800 µg/mL (r=0.9997); baicalin showed a good linear relationship at the range of 15∼120 µg/mL (r=0.9996), and berberine at the range of 7∼56 µg/mL (r=0.9995). Their average recovery rates were 99.67% (RSD 1.01%, n=6), 98.7% (RSD 1.93%, n=6) and 100.5% (RSD 2.88%, n=6) respectively. The study concluded that the high-performance liquid chromatography established in this study was simple, accurate and reproducible, and can also be used in the determination of catechin, baicalin and berberine contents in Jiejia tincture.     

HPLC法同时测定人血浆中帕罗西汀和喹硫平浓度

目的 建立同时测定人血浆中帕罗西汀和喹硫平浓度的高效液相色谱( HPLC)法.方法 采用HPLC法进行测定:Inertsil ODS-C18柱(4.6×150mm,5μm)为色谱柱,流动相为乙腈-磷酸二氢钠(0.05 mol/L磷酸二氢钠)(39∶61),流速为1.0mL/m in,检测波长为210nm,柱温为40℃,以乙酸乙酯-二氯甲烷(体积比75∶25)为萃取剂.结果 帕罗西汀在10～640μ g/L、喹硫平在20 ～ 1200μg/L范围内峰面积与浓度呈良好线性关系,检测限分别为5μg/L,8μg/L.结论 该方法简单、快速、灵敏、准确,可用于临床帕罗西汀与喹硫平的血药浓度监测和药动学研究.     

HPLC法测定不同产地附子中主要成分的含量

目的 考察不同产地的西南附子中新乌头原碱,次乌头原碱和乌头原碱的含量。方法 采用HPLC进行测定,色谱柱为Diamonsil C18 柱（150 mm×4.6 mm,5 μm）;流动相A为四氢呋喃-乙腈（15∶25）;流动相B为醋酸铵溶液（0.1 mol/L）,检测波长λ=235 nm;流速1 ml/min;柱温25 ℃。结果 不同产地的西南附子样品中生物碱类成分的含量有着明显差异,新乌头碱的含量以贵州小河的最高为0.17%,贵州兴义的最低为0.06%;次乌头碱的含量以四川江油的最高为0.28%,贵州花江的最低为0.13%;乌头碱的含量以云南腾冲与贵州水口寺的最高为0.04%,四川江油与贵州龙里的最低为0.01%。结论 西南附子中三种乌头碱的总含量以四川江油和云南腾冲产者最高。另外,HPLC测定法简单、易行,可为附子的产区扩大研究提供科学依据。     

高效凝胶色谱法测定多花黄精多糖分子量与分子量分布

目的：建立多花黄精多糖的分子量与分子量分布分析方法。方法：应用高效凝胶渗透色谱法，色谱柱为Shodex OHPak SB-803HQ（8mm×300mm），流动相为0．71％硫酸钠溶液（内含0．02％叠氮钠），柱温：35℃，示差折光检测器（检测器温度35℃），流速0．5ml·min^-1。结果：根据建立的方法测定，得到多糖的高效凝胶色谱图，计算出多花黄精多糖的重均分子量及其分布。结论：所用方法简便、快速、准确，可用于多花黄精多糖的质量控制。     

Determination of urinary free cortisol by HPLC (high performance liquid chromatography) and UV detection

Using basal isocratic equipment of HPLC and only one step extraction the determination of urinary free cortisol is available in non specialized laboratory. Results are displayed about reliability and specificity of the method. An extraction coefficient of 97.8 per cent, a good practicability in connection with the use of C18 column chromatography and a reagent flow of 0.5 ml/min. enable a low cost of this determination. Reference values are established for a working adult population.     

Direct Radioimmunoassay of Rat Urinary Kallikrein: Its Application to the Determination of Active and Inactive Kallikrein Concentration after HPLC Analysis

Abstract

Rat urinary kallikrein (RUK) was purified to apparent homogeneity by a three-step procedure and antibodies were raised in rabbits. Renal kallikrein exists as an active and an inactive form. A specific radioimmunoassay (RIA) was developped to measure directly the total kallikrein. The antibody used in this radioimmunoassay recognized both forms. No cross reactivity was detected with trypsin, esterase A or human urine. When iodinated rat urinary kallikrein was used, the detection range was between 0.125 and 16 ng with 6.5% intraassay variation and 8.1% between assay variation. Intrarenal kallikrein was measured in renal tissue after homogeneisation and solubilisation. Correlations between this RIA and the kininogenase activity or the amidolytic activity were highly significant. Since kallikrein exists as an inactive precursor the direct measurement of the total immunoreactive protein differs from activities determinations. An HPLC ion exchange system has been developped to separate active and inactive forms directly from urine, with a recovery of 79 ± 11%. This procedure permits measurement of inactive forms. Rat urine contains as much inactive kallikrein as active kallikrein.     

Determination of the histamine metabolite tele-methylimidazoleacetic acid and of creatinine in urine by the same HPLC system

OBJECTIVE AND DESIGN: To develop an HPLC method with UV detection for determination of tele-methylimidazoleacetic acid (t-MelmAA) in urine of man or animals, which is reliable, simple and less expensive than existing GC/MS techniques. METHOD: The elaborated procedure enables separation of t-MelmAA from its pros-isomer (p-MelmAA) as well as from imidazoleacetic acid. As an internal standard tele-ethylimidazoleacetic acid (t-EtlmAA) is used. The acids, after being converted to their stable isopropylesters, are extracted at pH 6.0-6.5. The further separation prior to HPLC utilises a small cellulose phosphate column. The HPLC system is isocratic with a C18 column and mobile phase consisting of an aqueous solution of SDS at pH 3.5 mixed with acetonitrile (65:35). An advantage of this system is that it can be used to determine the urine creatinine concentration to express excreted t-MelmAA in mmol/mol creatinine. RESULTS AND CONCLUSIONS: The method applicability is demonstrated in repeated studies in mastocytosis patients. The broad range of excretion values, from normal up to a high level (0.9 to 30 mmol/mol creatinine), indicates that it can be satisfactorily used for evaluation of histamine turnover under various clinical conditions. The method appears to be a good alternative to GC/MS based ones.     

高效液相色谱法测定血清碘海醇浓度方法的建立和应用

目的 建立利用高效液相色谱法(HPLC)测定血清碘海醇浓度的方法,为碘海醇血浆清除率临床应用及推广提供方法学支持.方法 设定色谱条件,对HPLC测定血清碘海醇浓度的方法进行方法学验证.对33例慢性肾脏病(CKD)儿童同时进行碘海醇清除率与99mTc-DTPA血浆清除率测定,并采用最新Schwartz公式估算肾小球滤过率(eGFR).以99mTc-DTPA血浆清除率为参照,分析碘海醇清除率及eGFR与参照方法的相关性和一致性.结果 在设定HPLC色谱条件下,碘海醇浓度在10～1 000 mg/L范围内线性良好(R2 =0.9998).批间测量和批内测量相对标准偏差(RSD％)＜5％,平均回收率(R％)＞96％.碘海醇血浆清除率、eGFR与99mTc-DTPA血浆清除率的相关系数(r)分别为0.95和0.63,偏差分别为(6.24±10.81)mL/(min·1.73 m2)和(12.11±22.96) mL/(min·1.73 m2).结论 建立了测定血清碘海醇浓度的HPLC法;初步探索并验证了碘海醇血浆清除率评价儿童肾功能的准确、可行.