高效液相色谱法测定大鼠组织中双酚A和4-壬基酚浓度

建立了大鼠组织中双酚A和4-壬基酚的提取和含量测定方法。大鼠组织样品经甲醇-乙酸铵缓冲液匀浆、 正己烷-乙醚混合溶剂提取、氮气吹干后用流动相溶解,以乙腈-0.01 mol/L乙酸铵缓冲液(pH 4.5)(体积比为75∶25)为流动相,经C18色谱柱分离,在激发波长227 nm、发射波长313 nm下进行荧光检测。大鼠心、脑、肝和肾脏组织样品中,双酚A的检出限为3.2~4.6 ng/g,4-壬基酚的检出限为11.8~15.6 ng/g;日内检测精密度为0.89%~4.50%,日间检测精密度为3.10%~12     

Polyamide as an efficient sorbent for simultaneous interference-free determination of three Sudan dyes in saffron and urine using high-performance liquid chromatography-ultra violet detection北大核心CSCD

With polyamide( PA)as an efficient sorbent for solid phase extraction( SPE)of Sudan dyes II,III and Red 7B from saffron and urine,their determination by HPLC was performed. The optimum conditions for SPE were achieved using 7 mL methanol/water( 1:9,v/v,pH 7)as the washing solvent and 3 mL tetrahydrofu-ran for elution. Good clean-up and high( above 90%)recoveries were observed for all the analytes. The opti-mized mobile phase composition for HPLC analysis of these compounds was methanol-water( 70:30,v/v). The SPE parameters,such as the maximum loading capacity and breakthrough volume,were also determined for each analyte. The limits of detection( LODs),limits of quantification( LOQs),linear ranges and recoveries for the analytes were 4. 6-6. 6 μg/L,13. 0-19. 8 μg/L,13. 0-5 000 μg/L( r2> 0. 99)and 92. 5% -113. 4%,respec-tively. The precisions( RSDs)of the overall analytical procedure,estimated by five replicate measurements for Sudan II,III and Red 7B in saffron and urine samples were 2. 3%,1. 8% and 3. 6%,respectively. The developed method is simple and successful in the application to the determination of Sudan dyes in saffron and urine sam-ples with HPLC coupled with UV detection.     

使用限进介质色谱柱的柱切换高效液相色谱法直接进样测定大鼠血浆中舒必利

提出一种直接进样测定大鼠血浆中舒必利浓度的高效液相色谱方法,使用限进介质色谱柱作为预柱在线去除血浆蛋白后,将舒必利通过柱切换转移到分析柱中进行分析。限进介质色谱柱为CAPCELLPAKMFSCX阳离子交换柱（20×4．0mmi．d．,5μm）,分析柱为Kromasil C18柱（150×4．6mm i．d．,5μm）,限进介质柱预分离时流动相为PH=6．88的50mmol／L磷酸盐缓冲液乙腈（100：5,V／V）,切换后分析流动相为PH=6．83的50mmol／L磷酸盐缓冲液-乙腈（100：10,V／V）。流速均为1mL／min,检测波长为240nm。该方法检出限为17ng／mL,定量限为50ng／mL。舒必利在50～1400ng／mL之间线性良好（r=0．9997）,高中低浓度的日内、日间相对标准偏差分别为1．5％～4．2％及2．0％～5．2％,方法回收率为98．8％～104．1％．     

Normal phase high performance liquid chromatography for determination of paclitaxel incorporated in a lipophilic polymer matrix

A normal phase (NP) high performance liquid chromatography (HPLC) method was developed for analysis of paclitaxel incorporated in poly(sebacic-co-ricinoleic acid), a lipophilic polymer matrix utilized for preparation of an injectable formulation for the localized delivery of paclitaxel. Thin layer chromatography experiments revealed that separation of paclitaxel from the polymer is dependent on the eluting strength (solvent strength) of the mobile phase. The HPLC system consists of a Purospher STRAR Si analytical HPLC column (5 microm, 250mm x 4mm, Merck), and 1-2.5% (v/v) methanol in dichloromethane as the mobile phase. Detection was by UV absorbance at 240 and 254 nm. The effect of the mobile phase composition on paclitaxel retention, peak shape and column efficiency, and the influence of the sample loading on the shape of the paclitaxel peak were studied. The mobile phases used for the chromatography consisted of 1.5% (v/v) methanol in dichloromethane. Paclitaxel was determined in the formulation and in the samples from degradation studies using UV detection at a wavelength of 254 nm. UV detection at 240 nm has advantages for following polymer matrix degradation products due to higher detector response at this wavelength. The utility of the proposed NP HPLC approach was demonstrated by assessment of intra- and inter-batch content uniformity, and by the determination of paclitaxel content after 7 and 60 days exposure of the paclitaxel-loaded polymer matrix to in vitro and in vivo degradation.     

HPLC analysis and pharmacokinetic study of quercitrin and isoquercitrin in rat plasma after administration of Hypericum japonicum thunb. extract

A simple HPLC method was developed for determination of quercitrin and isoquercitrin in rat plasma. Reversed-phase HPLC was employed for the quantitative analysis using kaempferol-3-O-beta-D-glucopyranoside-7-O-alpha-L-rhamnoside as an internal standard. Following extraction from the plasma samples with ethyl acetate-isopropanol (95:5, v/v), these two compounds were successfully separated on a Luna C(18) column (250 x 4.6 mm, 5 microm) with isocratic elution of acetonitrile-0.5% aqueous acetic acid (17:83, v/v) as the mobile phase. The flow-rate was set at 1 mL/min and the eluent was detected at 350 nm for both quercitrin and isoquercitrin. The method was linear over the studied ranges of 50-6000 and 50-5000 ng/mL for quercitrin and isoquercitrin, respectively. The intra- and inter-day precisions of the analysis were better than 13.1 and 13.2%, respectively. The lower limits of quantitation for quercitrin and isoquercitrin in plasma were both of 50 ng/mL. The mean extraction recoveries were 73 and 61% for quercitrin and isoquercitrin, respectively. The validated method was successfully applied to pharmacokinetic studies of the two analytes in rat plasma after the oral administration of Hypericum japonicum thunb. ethanol extract.     

Simultaneous determination of phenylglyoxylic acid, mandelic acid, styrene glycol and hippuric acid in primary culture of rat hepatocytes incubate by high-performance liquid chromatography

A simple HPLC method for the simultaneous determination of phenylglyoxylic acid (PGA), mandelic acid (MA), styrene glycol (SG) and hippuric acid (HA) in cell culture medium was developed. Analysis was performed on a C(18) column with a mobile phase composed of methanol-potassium dihydrogen phosphate (pH 2.5; 10 mM; 10:90, v/v) at 220 nm. The flow-rate of mobile phase was set at 0.5 mL/min. The mean absolute recoveries of PGA, MA, SG and HA were 95.9, 98.4, 98.0 and 97.1%, respectively. The inter-day and intra-day precisions, determined at three concentration levels, were less than 10% of RSD. The limits of quantification for PGA, MA, SG and HA were 13.2, 13.1, 14.5 and 11.2 microM with RSD less than 20%. The limits of detection for PGA, MA, SG and HA were 4.6, 4.6, 5.1 and 3.9 microM, respectively. The method was successfully applied to study the stereoselective metabolism of SG and MA in primary culture of rat hepatocytes. The results show that there is stereoselective metabolism for both of MA and SG in primary culture of rat hepatocytes. The extent of biotransformation from S-MA to PGA is significantly greater than that from the R enantiomer and the main metabolites are PGA and HA for S-SG and R-SG, respectively.     

Determination of 4-nonylphenol and 4-octylphenol in human blood samples by high-performance liquid chromatography with multi-electrode electrochemical coulometric-array detection

Alkylphenols can affect human health because they disrupt the endocrine system. In this study, an analytical method for determining trace amounts of 4-nonylphenol (NP) and 4-octylphenol (OP) in human blood samples was developed. Reversed-phase HPLC with multi-electrode electrochemical coulometric-array detection was used for the determination of NP and OP in plasma and serum samples prepared with a solid-phase extraction method. The separation was achieved using an isocratic mobile phase of 0.7% phosphoric acid-acetonitrile with a C18 reversed phase column. The detection limits of NP and OP were 1.0 and 0.5 ng ml-1, respectively. The recoveries of NP and OP added to human plasma samples were above 70.0% with a relative standard deviation of less than 15.5%. The method was found to be applicable to the determination of NP and OP in various human blood samples such as serum and plasma.     

High-performance liquid chromatography with fluorescence detection for the simultaneous determination of 3,4-methylenedioxymethamphetamine, methamphetamine and their metabolites in human hair using DIB-Cl as a label

This paper describes a highly sensitive HPLC method for the simultaneous determination of 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxyamphetamine (MDA), amphetamine (AP) and methamphetamine (MP) in human hair samples. The amphetamines investigated were derivatized with the fluorescent reagent, DIB-Cl to yield highly fluorescent DIB-derivatives, which were then analyzed by HPLC with fluorescence detection at excitation and emission wavelengths of 325 nm and 430 nm, respectively. The separation was achieved on an ODS column with an isocratic mobile phase composed of acetonitrile-methanol-water (30:40:30, v/v/v). The limits of detection for the four compounds obtained by the proposed method ranged from 11 to 200 pg/mg. The method was successfully applied to the determination of MDMA and MDA in hair samples obtained from MDMA abuser.     

Analysis of tetramethylpyrazine in Ephedrae herba by gas chromatography-mass spectrometry and high-performance liquid chromatography

A simple and reliable HPLC method was developed for the determination of 2,3,5,6-tetramethylpyrazine (TMP) in Ephedrae herba. Further identification of TMP was achieved using GC-MS. The mobile phase used was methanol-water-35% acetic acid (35:65:0.5, v/v/v) at a flow-rate of 0.8 ml/min. The detection wavelength was set at 290 nm. The linear range of the peak area calibration curve of TMP was 2.64-264 mg/l (r=0.9987) and the recovery for TMP in Ephedrae herba extracts was 101.1-106.9%. The relative standard deviations of retention time and peak area were 0.18 and 1.5% (n=6), respectively. The detection limit of TMP was 0.03 mg/l. The contents of TMP in Ephedrae herba could easily be determined within 10 min.     

色谱指纹图谱在苹果酒质量评价中的应用

采用反相高效液相色谱-电化学检测法研究了14种苹果酒样品的指纹图谱。以标准品绿原酸进行定位,通过对图谱分析和相对保留时间计算,确定了8个共有峰。根据共有峰的峰面积用相关系数法和向量夹角余弦法计算相似度,两种方法的计算结果一致。实验结果表明同一厂家生产的苹果酒相似度较好。该法为苹果酒的产品分析提供了有效的微观信息,为苹果酒的质量控制、新产品的研发以及苹果酒行业标准的制定提供一种可行思路。     

高效液相色谱-二极管阵列检测器同时测定化妆品中的九种染料及中间体

建立了用高效液相色谱-二极管阵列检测器(HPLC-PDA)同时检测9种染料及中间体的系统方法。首先采用超声提取的方法处理样品,对提取溶剂和提取时间进行了选择,确定用甲醇-0.01 mol/L 乙酸铵(体积比为2∶1)作提取溶剂,超声提取20 min。然后,采用C18柱,以甲醇-0.01 mol/L乙酸铵(pH 6.2)为流动相梯度洗脱,用PDA检测。以保留时间定性,并以紫外吸收光谱图辅助定性,以外标法定量。定量检测波长为230 nm,15 min内可对9种目标物同时进行测定,且各化合物都达到基线分离(分离度大于1.5)。经测定,该方法的平均回收率(n=8)为81.0%～105.6%,相对标准偏差（RSD）为0.8%～4.9%,检出限(以信噪比为3计)为0.1～2 μg。该方法简单、快速,能有效提取和分离测定化妆品中9种染料及中间体。将该方法用于实际化妆品样品的检测,结果令人满意。     

A simple RP-HPLC method for the simultaneous determination of curcumin and its prodrug, curcumin didecanoate, in rat plasma and the application to pharmacokinetic study

A high-performance liquid chromatographic (HPLC) method has been developed for the simultaneous determination of curcumin and its prodrug, curcumin didecanoate (CurDD), in rat plasma. The analytes were extracted by ethyl acetate following the addition of sodium dodecyl sulfate, and separated on a reverse-phase C(18) column using a gradient mobile phase system of acetonitrile-tetrahydrofuran-water containing 0.1% formic acid. Detection by UV absorption at 425 nm gave a lower limit of quantitation (LLOQ) of 5 and 10 ng/mL for curcumin and CurDD in 50 μL of plasma, respectively. Intra- and inter-day precisions of quality control samples except those at LLOQ were within 15% for curcumin and CurDD, respectively, and the accuracies for both compounds were between 93.9 and 108%. The method was successfully applied to determine plasma concentration-time curves of curcumin and CurDD in rats following intravenous (i.v.) administration of curcumin or CurDD at doses of 1 mg/kg (calculated as curcumin). The results suggested that i.v. dosed CurDD provided sustained plasma levels of curcumin.     

Determination of sodium monofluoroacetate (1080) in biological samples as its 4-bromomethyl-7-methoxycoumarin derivative by RP-HPLC

A high-performance liquid chromatography (HPLC) method with fluorescence detection is described for the determination of sodium monofluoroacetate (MFA-Na) in biological samples. 4-Bromomethyl-7-methoxycoumarin is used as a derivatization reagent and reacted with MFA-Na to form 7-methoxy-4-methylenecoumarin monofluoroacetate for HPLC analysis. Chromatographic separation is performed on a Hewlett Packard RP-18 column using methanol-water (60:40, v/v) as the mobile phase. A fluorescent detector is employed with the excitation and emission wavelengths as 319 nm and 390 nm, respectively. The novel method yields a good linear relationship when the concentration of MFA-Na is within 1 and 500 nmol/mL (r = 0.9996). The detection limit is 50 pmol/mL. The established method is applied to determine MFA-Na in biological samples. The recovery rates of MFA-Na are between 81% and 88%, and the relative standard deviations are less than 5%. The method shows good sensitivity and selectivity for the determination of MFA-Na in biological samples.     

Simplified procedure for the determination of sotalol in plasma by high-performance liquid chromatography

A simple, specific and rapid reversed-phase high-performance liquid chromatographic (HPLC) procedure for sotalol determination is described requiring small plasma volumes. The high recovery of sotalol from plasma and the high precision of measurement obviate the need for an internal standard. Plasma samples (300 microliters) were deproteinised with 50 microliters of 70% (w/w) perchloric acid in disposable glass tubes. After vortex-mixing and centrifugation, 30 microliters of 4 M K2HPO4 were added followed by gentle shaking. A 20-microliters aliquot was then injected (by autosampler) for HPLC analysis. Chromatography was performed on a glass-lined 250 mm x 4 mm 5-micron C18 steel column. The mobile phase was 6% (v/v) acetonitrile in 0.08 M KH2PO4 buffer (pH 4.6). The flow-rate was 0.8 ml/min. Detection was by fluorescence with excitation and emission wavelengths at 235 and 310 nm, respectively. The retention time for sotalol was 7.1 min. Calibration was linear from 0.16 to 10 micrograms/ml in plasma (r greater than 0.999 for detector response to sotalol). The minimum concentration for quantitation was 0.08 micrograms/ml [within assay coefficient of variation (C.V.) less than 5%]. Recovery was near quantitative (greater than 98%) and replicate (intra-assay precision was less than 5% C.V.). Analysis of samples (n = 10) at concentrations of 0.42 and 4.2 micrograms/ml gave mean values of 0.44 and 4.3 micrograms/ml, respectively. The inter-assay C.V. values were 4.5 and 2.2%, respectively. Other clinically used antiarrhythmic drugs did not interfere. This assay can be performed using other commercial C18 analytical columns by suitable adjustment of mobile phase flow-rate and acetonitrile composition.     

高效液相色谱法测定化妆品中α-、β-熊果苷及烟酰胺

建立了美白类化妆品中熊果苷的两种光学异构体α-熊果苷和β-熊果苷及烟酰胺的高效液相色谱检测方法。样品用氯化钠水溶液-氯仿(2:1, v/v)进行萃取。固定相为依利特ODS-BP柱(200 mm×4.6 mm, 5 μm),流动相为甲醇-水(10:90, v/v),柱温为25 ℃,检测波长为220 nm,流速为0.5 mL/min,进样量为20 μL。在上述条件下α-熊果苷、β-熊果苷和烟酰胺的质量浓度依次在0.07~50, 0.06~50, 0.05~50 mg/L时与色谱峰面积之间的线性关系良好,相对标准偏差(n=7)分别为1.65%、1.73%和1.33%。将该方法用于化妆品的检测,回收率为91.7%～109.6%。该法简便、快速、准确,可用于化妆品美白成分的测定。     

Fast HPLC method using ion-pair and hydrophilic interaction liquid chromatography for determination of phenylephrine in pharmaceutical formulations

A rapid procedure based on a direct extraction and HPLC determination with fluorescence detection of phenylephrine in pharmaceutical sachets that include a large excess of paracetamol (65 + 1, w/w), ascorbic acid (5 + 1, w/w), and other excipients (aspartame and sucrose) was developed and validated. The final optimized chromatographic method for ion-pair chromatography used an XTerra RP18 column, 3 microm particle size, 50 x 3.0 mm id. The mobile phase consisted of a mixture of acetonitrile and buffer (10 mM sodium octane-1-sulfonate, adjusted with H3PO4 to pH 2.2; 200 + 800, v/v), with a constant flow rate of 0.3 mL/min. The separation was carried out at 30 degrees C, and the injection volume was 3 microL. Fluorescence detection was performed at excitation and emission wavelengths of 275 and 310 nm, respectively. The mobile phase parameters, such as the organic solvent fraction (acetonitrile) in mobile phase as an organic modifier, the concentration of sodium octane-1-sulfonate as a counter-ion, temperature, and pH of mobile phase, were studied. As an alternative to ion-pair chromatography, hydrophilic interaction liquid chromatography (HILIC) was investigated using a Luna HILIC column, 3 microm, 100 x 4.6 mm id. The mobile phase consisted of acetonitrile and buffer (5 mM potassium dihydrogen phosphate, adjusted with H3PO4 to pH 2.5; 750 + 250, v/v) at a flow rate of 0.8 mL/min. The separation was carried out at 25 degrees C, and the injection volume was 5 microL. The proposed method has an advantage of a very simple sample pretreatment, and is much faster than the currently utilized HPLC methods using gradient elution and UV detection. Commercial samples of sachets were successfully analyzed by the proposed HPLC method.     

柱切换技术-高效液相色谱法快速检测大鼠血浆中的普萘洛尔对映体

建立了快速检测大鼠血浆中普萘洛尔对映体浓度的柱切换-高效液相色谱法。将自制限进填料柱作为预处理柱,通过直接进样方式,使普萘洛尔对映体在预处理柱上保留,同时除去血浆中的蛋白质等大分子;再通过柱切换技术,使普萘洛尔对映体在键合型纤维素-三(3,5-二甲基苯基氨基甲酸酯)(Chiralcel OD-RH)分析柱上得到手性拆分。通过条件优化,确定切换前预处理流动相为硼酸盐缓冲液(pH 8.5)-甲醇(95:5, v/v),流速为1.0 mL/min;切换后分析流动相为异丙醇-乙醇-0.2 mmol/L硼酸盐缓冲液(pH 8.5)(30:30:40, v/v/v),流速为0.8 mL/min;切换时间为3 min;柱温为25 ℃;检测波长为293 nm。普萘洛尔两对映体在25～500 mg/L的质量浓度范围内具有良好的线性关系(r=0.9995), 3个加标水平(50、100、250 mg/L)的平均回收率为97.89%～101.56%,日内和日间精密度均小于5%。该方法简便、快速、灵敏、准确,适于血浆样本中手性药物的药代动力学研究。     

Simple and extractionless high-performance liquid chromatographic determination of rosiglitazone in human plasma and application to pharmacokinetics in humans

A simple and extractionless HPLC method using fluorescence detection was developed for the determination of rosiglitazone in human plasma. After deproteinization using perchloric acid the plasma samples were directly injected onto the HPLC system. The mobile phase was composed of acetonitrile (52%) and 20 mm ammonium acetate (48%, pH 7.5), and analysis was run at a flow rate of 0.2 mL/min with the detector operating at 247 nm for excitation wavelength and at 367 nm for emission wavelength, respectively. The method has a mean recovery of 97%, while the intra-day and inter-day precisions were all less than 7%. This method is simple, specific, sensitive and requires only a small plasma volume with short analytical time, and is suitable for the determination of plasma rosiglitazone in routine measurements for pharmacokinetic studies.     

Automated solid-phase extraction method for the determination of piperaquine in plasma by peak compression liquid chromatography

A validated bioanalytical method for the determination of piperaquine (PQ) in plasma by solid-phase extraction (SPE) and liquid chromatography (LC) using peak compression is presented. Protein is precipitated from plasma with acetonitrile-1% aqueous acetic acid (85:15, v/v). An internal standard (IS) is added to the samples before they are loaded onto a strong cation exchanger (Isolute PRS) SPE column. PQ and the IS are analyzed by LC on a Zorbax SB-CN column (250 x 4.0 mm) with the mobile phase acetonitrile-phosphate buffer [I = 0.1, pH 2.5 (12:88, v/v)] and UV detection at 345 nm. Trichloroacetic acid (TCA) is added to the samples prior to injection into the chromatography system. PQ elutes in a gradient of TCA, which enables peak compression of PQ and significantly higher peak efficiency as a result. The intraassay precision for plasma is determined to be 5.4% at 3.00 microM and 5.8% at 0.050 microM. The interassay precision for plasma is 1.3% at 3.00 microM and 10.0% at 0.050 microM. The lower limit of quantitation and the limit of detection are 0.025 and 0.005 microM, respectively.     

Organotin speciation in textile and plastics by microwave-assisted extraction HPLC-ESI-MS

The HPLC coupled with electrospray ionization mass spectrometry (ESI-MS) was applied for the determination of tributyltin chloride (TBT), triphenyltin chloride (TPhT), triethyltin chloride (TET) in textile and plastic samples. The separation was performed in the isocratic mode on ion exchange SCX column with a mobile phase containing methanol-ammonium acetate (50 mmol l(-1))-acetic acid (90/9.98/0.02, v/v/v). Sensitive detection of three organotin compounds by ESI-MS was performed on the basis of multiple reaction monitoring (MRM) mode. Limits of detection were 0.4, 20, 4 ng ml(-1) for TBT, TPhT, and TET, respectively. Microwave-assisted extraction (MAE) of organotin compounds was finished in 60% (v/v) methanol aqueous solution. Under the experimental conditions used, recoveries of three organotin compounds obtained for spiked textile and plastic samples are in the range of 55-95% and the R.S.D.s are 3-9%.