高效液相色谱法测定扑热息痛药代动力学参数

采用反相高效液相色谱法测定扑热息痛在健康人体内的药代动力学参数。Zorbax ODS色谱柱,流动相为甲醇一水一冰醋酸(20:75:5),茶碱为内标,检测波长245nm,线性范围0.5～25μg/mL,r=0.9987,回收率101.58%,日内、日间测定 RSD 均小于10%,血浆中扑热息痛最低检测浓度小于0.1μg/mL。     

HPLC及SEP-PAK C_(18)小预柱法测定复方甘草片中吗啡的含量

本文介绍了用SEP—PAK C_(18)小预柱固相萃取技术和反相高效液相色谱法测定复方甘草片中微量吗啡含量的方法。对固相萃取条件进行了研究,选择出以10％甲醇溶液为淋洗溶液,以70％甲醇溶液为洗脱溶液。这样基本消除了干扰组分对测定的影响,色谱图背景吸收小,基线平稳。色谱柱为μBondapak C_(18)柱,以0.1mol/L NaH_2PO_4溶液—甲醇(5:1)为流动相,检测波长为280nm。平均回收率为(101.2±1.5)％,RSD小于2％。该方法简便、快速、准确,重现性好,可用于药品质量控制。     

均匀设计优选提取条件测定沙棘粉中儿茶素含量

目的:筛选优化沙棘粉中儿茶素的提取方法,并用HPLC测定其含量。方法:以均匀设计法优化提取条件,采用Agilent ZORBAX SB-C18柱,以甲醇—乙酸溶液(用乙酸调p H值至3.2)(25:75)为流动相,流速1 m L/min;柱温25℃,检测波长280nm。结果:优选出的最佳提取条件:甲醇浓度为74.0%,水浴温度为70℃,超声时间为30min。儿茶素在0.50~10.0μg/m L浓度范围内线性良好,相关系数(r)为0.9999,平均回收率为97.25%(RSD=1.77%)。结论:本试验建立的含量测定方法操作简便、重现性好且具有较强的专属性。     

乌龙茶中咖啡因的高效液相色谱测定法

五种乌龙茶中的咖啡因以ZBX50010—86方法提取和净化后,用高效液相色谱测定。色谱柱为NOVA PAK~(TM)C18柱,流动相为0.02mol/LNH4Ac溶液(pH=4.0)—甲醇(75+25)混合液,紫外检测波长230nm。方法的测定低限为0.004mg/mL,相对标准偏差为1.56％。     

水产品中甲基睾丸酮残留ELISA测定

建立了养殖水产品中甲基睾丸酮(MET)残留ELISA测定方法,利用甲醇提取,过酸性氧化铝柱去杂质后进行ELSIA分析。结果:方法最低检测限为10.0μg/kg,样品中添加浓度为10μg/kg、20μg/kg和50μg/kg 3个梯度水平平均回收率范围86.3-96.5%,RSD为4.39-5.78%。结论:该方法适合水产品中MET残留快速测定。     

HPLC法同时测定季铵盐中氯化苄和苯甲醇

本文提出同时测定季铵盐苄基氯化铵中氯化苄、苯甲醇的新方法。采用高效液相色谱法,以C18(4.6mm×250mm,5μm)为分离柱;磷酸二氢盐—甲醇进行梯度洗脱;检测波长为210nm;柱温为25℃;流速为1.0m L/min。结果表明氯化苄在6.5~130μg/m L(r=0.9998),苯甲醇在6.25~125μg/m L(r=0.9999)浓度范围内呈良好的线性关系,氯化苄和苯甲醇的回收率分别为95.00~102.00%和102.86~105.71%,检出限分别为3.0μg/g、1.5μg/g。该方法操作简单,未用到有毒的试剂,准确性高,是季铵盐原料质量控制的理想方法。     

HPLC-UV法测定人血浆及尿中多尼培南

建立HPLC-UV法测定人血浆及尿中多尼培南(DP)的质量浓度,用于DP人体药代动力学研究.采用UltimateXB-C18(150 mmn×4.6 mm×3 μm)色谱柱,血浆及尿样测定用流动相分别为10 mmol/L醋酸钠∶乙腈∶三乙胺(体积比94∶6∶0.1,冰乙酸调pH为4.51)和8 mmol/L醋酸钠∶乙腈∶三乙胺(体积比95.5∶4.5∶0.1,冰乙酸调pH值为4.99),流速均为1 mL/min,检测波长为297 nm.血浆样品经乙睛沉淀蛋白,再用二氯甲烷反洗后取上清液进样,以美罗培南(MRP)做内标;尿样直接用水稀释后进样,外标法定量.血药浓度在0.062 5～200 μg/mL范围内与峰面积线性关系良好,定量限为0.0625 μg/mL,批内精密度在1.0％～6.0％之间,批间精密度在3.0％～6.7％之间,方法回收率在92.4％～104.5％之间,预处理回收率在91.2％～103.8％之间.尿药浓度在0.625～4 800 μg/mL范围内与峰面积线性关系良好,定量限为0.625 μg/mL,批内精密度在0.7％～4.0％之间,批间精密度在1.1％～5.3％之间,方法回收率在94.1％～106.7％之间.该方法具有简便、灵敏、准确等特点,适用于人血浆及尿中多尼培南质量浓度的测定.     

梯度洗脱HPLC法测定利培酮的含量

目的:建立测定利培酮含量的梯度洗脱高效液相色谱(HPLC)法。方法:色谱柱为C18柱(4.6mm×100mm,3μm),流动相A为乙酸铵缓冲液(0.2mol/L,PH6.5)-甲醇-水(100∶150∶750),流动相B为乙酸铵缓冲液(0.2mol/L,PH6.5)-甲醇-水(100∶850∶50),检测波长为275nm,流速1.5ml/min,柱温35℃。结果:利培酮质量浓度在0.5～1.5mg/mL范围内与峰面积呈良好的线性关系(r=0.9998)。结论:梯度洗脱HPLC法测定利培酮的含量,方法准确、专属性强、重现性好,适用于利培酮的含量测定。     

高效液相色谱法测定茶皂苷的含量及其血药浓度

目的:建立了高效液相色谱法对茶皂苷含量及其血药浓度的测定.方法:色谱柱为Hypersil ODS(25mm,4.6*250mm),流动相为甲醇-水(90:10),流速为0.5ml/min,检测波长为215nm.结果:茶皂苷在0.16～1.28mg/mL的浓度范围内,茶皂苷的浓度与主峰面积具有良好的线性关系,回归方程:y...     

等度高效液相色谱测定大豆异黄酮中大豆甙元和金雀异黄酮

本文介绍了等度高效液相色谱(HPLC)测定大豆异黄酮中二种主要成份:大豆甙元(Daidzein)和金雀异黄酮(Genistein)的方法。采用Atlantis C18色谱柱和EasyGuard C18,保护柱;以甲醇:0·1%醋酸(pH3·11)=51·5:48·5(v/v)为流动相;检测波长λ=254nm;流速1mL/min。在测定范围内(10-200ng)峰面积与质量浓度线性关系良好。大豆甙元和金雀异黄酮的相关系数分别为0·9984,0·9997·两组份回收率为97·0—102·92%·     

Differences in the Method by Which Plasma Is Separated from Whole Blood Influences Amphotericin B Plasma Recovery and Distribution Following Amphotericin B Lipid Complex Incubation Within Whole Blood

A previous investigation suggested that the use of plasma as the biological fluid for measurement of amphotericin B (AmpB) concentrations greatly underestimates the concentrations of AmpB in the total blood circulation following amphotericin B lipid complex (ABLC) administration to humans. The purpose of this study was to determine if differences in the method used to obtain plasma from whole blood influences the percentage of AmpB recovered in plasma following ABLC incubation in whole blood. ABLC (5 μg AmpB/ml; peak blood concentration observed in rabbits following intravenous bolus of ABLC at a dose of 1 mg/kg) was incubated in whole blood for 5 min at 25°C. These conditions were used to mimic the sample retrieval conditions used when blood is obtained from animals and human patients. Following incubation, plasma was obtained from whole blood using five different methods: (A) Whole blood was centrifuged for 5 min at 23°C, and the plasma was separated; (B) whole blood was stored at 4°C for 18 h, and the plasma was separated by gravity; (C) whole blood was stored at 23°C for 18 h, and the plasma was separated by gravity; (D) whole blood was stored at 37°C for 18 h in a water bath, and the plasma was separated by gravity; and (E) whole blood was stored at 30°C for 18 h in a water bath, and the plasma was separated by gravity. All samples were protected from light throughout the duration of the experiment. AmpB concentration in each plasma sample was determined by high-performance liquid chromatography (HPLC) using an external calibration curve. The whole blood : plasma Amp B concentration ratio and the percentage of AmpB partitioned into plasma following incubation of ABLC in whole blood for each plasma separation procedure was as follows: (A) 6.5 : 1 blood : plasma AmpB concentration ratio, 15.4% ± 1.6% AmpB in plasma; (B) 2.98 : 1 blood : plasma AmpB concentration ratio, 33.6% ± 7.7% AmpB in plasma; (C) 1.5 : 1 blood : plasma AmpB concentration ratio, 67.6% ± 10.3% AmpB in plasma; (D) 1.5 : 1 blood : plasma concentration ratio, 68.1% ± 1.1% AmpB in plasma; and (E) 1.2 : 1 blood : plasma AmpB concentration ratio; 83.4% ± 5.5% AmpB in plasma. These findings suggest that when measurement of AmpB in plasma is required following ABLC administration, incubation of whole blood at 30°C for 18 h appears to be the most effective method.     

Determination of clozapine and its metabolite, norclozapine in various biological matrices using high-performance liquid chromatography

Purpose. To develop and validate an HPLC method for the quantitation of clozapine and its metabolite, norclozapine in human plasma, rat plasma, and the various human plasma lipoprotein fractions. Methods. Using liquid-liquid extraction, clozapine, and norclozapine are extracted from the biological matrix with MTBE. After concentration, the residue was reconstituted in 1mM HCl and injected on to a C6 Phenyl column (3μm, 2.0×150 mm). Mobile phase was 10mM Ammonium Acetate, pH 5—Acetonitrile—Methanol (5:3:2, v/v/v). Loxapine served as the internal standard. Absorbance was measured at 254 nm. Results. Quantitation limits for clozapine and norclozapine was 100 ng/mL and 50 ng/mL, respectively. Recovery for both clozapine and norclozapine was near 100%. Percent accuracy for intraday variability in human plasma, rat plasma, and human TRL, HDL, LDL, and LPDP lipoprotein fraction was between 92 to 113% for both analytes. Intraday precision for the same matrices was less than 9% CV for both analytes. Percent accuracy and precision for interday variability in human plasma was 97 to 103% and less than 10% CV, respectively. Samples were stabile in the autosampler for 80 h at 10°C and on the benchtop for 2 h. It should be noted, Clozapine-N-oxide, which is a known metabolite of Clozapine, was not determined since it is not clinically active. Conclusions. This method is a simple, fast and robust HPLC assay for the determination of clozapine and norclozapine in various matrices and lipoprotein fractions.     

高效液相色谱法测定妥布霉素的含量

本文采用高效液相色谱法测定妥布霉素的含量,以Waters Nova-pak C_(18)为固定相,水—冰醋酸—甲醇—乙腈(40:5:30:25 v/v)配制的庚烷碘酸钠溶液(0.02mol/L)为流动相,在330nm处测定,线性范围0.1～0.6mg/ml,r=0.9988。方法简便、快速,重现性良好。     

Determination of Clozapine and its Metabolite,Norclozapine in Various Biological Matrices Using High-Performance Liquid Chromatography

Purpose. To develop and validate an HPLC method for the quantitation of clozapine and its metabolite, norclozapine in human plasma, rat plasma, and the various human plasma lipoprotein fractions. Methods. Using liquid-liquid extraction, clozapine, and norclozapine are extracted from the biological matrix with MTBE. After concentration, the residue was reconstituted in 1mM HCl and injected on to a C6 Phenyl column (3μm, 2.0×150 mm). Mobile phase was 10mM Ammonium Acetate, pH 5—Acetonitrile—Methanol (5:3:2, v/v/v). Loxapine served as the internal standard. Absorbance was measured at 254 nm. Results. Quantitation limits for clozapine and norclozapine was 100 ng/mL and 50 ng/mL, respectively. Recovery for both clozapine and norclozapine was near 100%. Percent accuracy for intraday variability in human plasma, rat plasma, and human TRL, HDL, LDL, and LPDP lipoprotein fraction was between 92 to 113% for both analytes. Intraday precision for the same matrices was less than 9% CV for both analytes. Percent accuracy and precision for interday variability in human plasma was 97 to 103% and less than 10% CV, respectively. Samples were stabile in the autosampler for 80 h at 10°C and on the benchtop for 2 h. It should be noted, Clozapine-N-oxide, which is a known metabolite of Clozapine, was not determined since it is not clinically active. Conclusions. This method is a simple, fast and robust HPLC assay for the determination of clozapine and norclozapine in various matrices and lipoprotein fractions.     

Oral bioavailability and intestinal absorption of candesartan cilexetil: role of naringin as P-glycoprotein inhibitor

Objective: The aim of the study is to explore the pharmacokinetic behavior of candesartan solid dispersions prepared by different pharmaceutical interventions using P-gp inhibitor in rabbits to validate the effectiveness of naringin as a pharmaceutical excipient in enhancing the oral delivery of lipophilic candesartan cilexetil.

Methods: Male albino rabbits (1–1.5?kg) were orally administered pure CAN suspensions and various candesartan solid dispersions (10?mg/kg) with and without naringin (15?mg/kg) and blood samples were collected at specified time points. CAN plasma samples were measured using HPLC.

Key findings: After oral dosing of pure CAN suspension, the mean AUC0-8?h was found to be 0.14?±?0.09?μgh/ml which was increased significantly, i.e. 0.52?±?0.13?μgh/ml with freeze-dried solid dispersions in the presence of naringin (p?p?Conclusion: These results are quite stimulating for further development of a clinically useful oral formulation of candesartan cilexetil based on P-gp inhibition using naringin, a natural flavonoid as a pharmaceutical excipient.     

Evaluation of bioequivalence of two formulations containing 100 milligrams of aceclofenac

The bioequivalence of two oral formulations containing aceclofenac 100 mg was determined in 24 healthy Indian male volunteers. The study was designed as a single dose, fasting, two-period two-sequence crossover study with a washout period of 1 week. The content of aceclofenac in plasma was determined by a validated HPLC method with UV detection. The preparations were compared using the parameters area under the plasma concentration-time curve (AUC_0-t), area under the plasma concentration-time curve from zero to infinity (AUC_0-∞), peak plasma concentration (C_max), and time to reach peak plasma concentration (t_max). No statistically significant difference was observed between the logarithmic transformed AUC_0-∞ and C_max values of the two preparations. The 90% confidence interval for the ratio of the logarithmic transformed AUC_0-t, AUC_0-∞, and C_max were within the bioequivalence limit of 0.80-1.25.     

A Stability Indicating Method for Dipyridamole

An HPLC method for the assay and chromatographic purity assessment of dipyridamole raw material and capsule product was developed. A mobile phase composing of methanol: aq 200 mM pentane sulphonic acid, sodium salt, [70:30 v/v], had triethylamine added [2ml/L] and was then adjusted to pH 3.0 with phosphoric acid. The mobile phase was pumped through an octadecylsilated-silica column, being held at 60° C, at a flow rate of 1.5ml/min. Detection was made at 288nm.

The resolution of degradation products, along with precision and accuracy were investigated for the system.     

Enantiomeric Separation and Quantitative Determination of Atenolol in Tablets by Chiral High-Performance Liquid Chromatography

The separation and quantitative determination of atenolol isomers by chiral high-performance liquid chromatography (HPLC) are described. Atenolol isomers were separated using a Chiralcel OD® column (250 × 4.6 mm,10 μm); the mobile phase was hexane-ethanol-diethylamine (75:25:0.1 v/v/v); ultraviolet detection was at 276 nm; and flow rate was 0.7 ml/min. The coefficient of variation and average recovery of (R)-isomer were 0.60% and 100.37%, respectively, for sample A and 0.69% and 100.63%, respectively, for sample B. The coefficient of variation and average recovery of (S)-isomer were 0.59% and 100.33%, respectively, for sample A and 0.63% and 99.78%, respectively, for sample B.     

Enantiomeric separation and quantitative determination of atenolol in tablets by chiral high-performance liquid chromatography

The separation and quantitative determination of atenolol isomers by chiral high-performance liquid chromatography (HPLC) are described. Atenolol isomers were separated using a Chiralcel OD® column (250 × 4.6 mm,10 μm); the mobile phase was hexane-ethanol-diethylamine (75:25:0.1 v/v/v); ultraviolet detection was at 276 nm; and flow rate was 0.7 ml/min. The coefficient of variation and average recovery of (R)-isomer were 0.60% and 100.37%, respectively, for sample A and 0.69% and 100.63%, respectively, for sample B. The coefficient of variation and average recovery of (S)-isomer were 0.59% and 100.33%, respectively, for sample A and 0.63% and 99.78%, respectively, for sample B.