HPLC法测定安宫牛黄栓中体外培育牛黄含量

目的:用HPLC法测定安宫牛黄栓中体外培育牛黄含量。方法:样品经二氯甲烷超声提取后,采用HPLC法测定安宫牛黄栓中胆红素的含量,使用C18色谱柱,乙腈—1%醋酸溶液(95:5)为流动相,检测波长450nm。结果:胆红素线性范围0.0638～1.276μg,平均回收率99.89%。结论:样品处理方法合理,方法学考察符合定量要求,可用于安宫牛黄栓中胆红素的含量测定。     

高效液相色谱法测定浓缩苹果汁中棒曲霉素

建立了高效液相色谱法 (HPLC)测定浓缩苹果汁中棒曲霉素含量的方法。用乙酸乙酯提取浓缩苹果汁中棒曲霉素 ,碳酸钠水溶液净化提取液 ,蒸干乙酸乙酯层 ,其残留物用醋酸缓冲液溶解。棒曲霉素的含量测定采用HypersilODS (5 μm ,2 5 0mm×4 6mmi d)色谱柱 ,流动相V(乙腈 ) :V(醋酸盐缓冲液 ) :V(水 ) =5 :3:2 ,流速 1ml min ,检测波长 2 76nm。结果表明 :棒曲霉素浓度与峰面积有很好的线性关系 ;用HPLC测定浓缩苹果汁中棒曲霉素的含量可操作性强、准确度高 ,棒曲霉素回收率高达 99 2 %,重现性好 (RSD为 0 4 %) ;灵敏度高最小检出量为 10 μg L。     

高效液相法测定香菇菇柄麦角甾醇的含量

采用高效液相色谱法(High performance liquid chromatography,HPLC)建立了测定菇柄麦角甾醇的含量测定方法。确定提取过程中皂化剂的种类和醇碱比后,将样品皂化,萃取后蒸干溶剂,乙醇定容测定。采用Phe-nomenex-C18色谱柱,V(流动相甲醇)∶V(水)=98∶2,流速1.0 mL/min,检测波长282 nm。结果表明:麦角甾醇线性回归方程为Y=9E+9×106X-8919.9(X:质量浓度,mg/mL),R2=0.998 9,0.01~0.30 mg/mL范围内线性关系良好,回收率为97.31%~101.95%。与紫外分光光度法所测结果比较,HPLC法测定菇柄中麦角甾醇含量灵敏、快速、准确,适用菇柄中麦角甾醇的含量测定。     

近红外光谱法快速测定白芍中芍药苷含量

本研究旨在应用近红外光谱法建立一种白芍药材中芍药苷含量的快速测定方法。利用HPLC测定样品中芍药苷含量,并以其作为参考值,运用偏最小二乘法(PLS)建立芍药苷含量与近红外光谱之间的多元校正模型,对未知样品进行含量预测。结果表明,所建芍药苷定量分析模型的相关系数(R2)、内部交叉验证均方差(RMSECV)、校正均方差(RMSEC)分别为0.99395、0.33068、0.0563;经内部验证,模型的预测均方差(RMSEP)和平均回收率分别为0.0756和100.07%。该方法操作简便,无污染,结果准确可靠,可用于白芍中芍药苷含量的快速测定。     

近红外光谱法快速测定木香中木香烃内酯与去氢木香内酯的含量

本文主要研究近红外光谱法在快速测定木香药材中木香烃内酯与去氢木香内酯含量的应用。采用近红外漫反射光谱法采集木香的近红外光谱,以HPLC测量值为参考值,运用偏最小二乘法(PLS)建立木香烃内酯与去氢木香内酯含量的定量模型,并用未知样品验证该模型。结果表明所建定量模型的校正集内部交叉验证相关系数(R2)、校正均方差(RMSEC)和内部交叉验证均方差(RMSECV)分别为0.9783、0.161和0.374;经外部验证的预测相关系数(r2)和预测均方差(RMSEP)分别为0.9546和0.162。该方法操作简便,测定快速,结果准确,无污染,可用于木香药材中木香烃内酯与去氢木香内酯含量的快速测定。     

NIRS快速测定河南产不同加工方法野菊花中木犀草素的含量

本文利用近红外光谱技术(NIRS)快速测定河南产不同加工方法野菊花药材中木犀草素的含量。采用3种加工方法(直接晒干、蒸后晒干和炒后晒干)对河南新郑、三门峡和信阳三个不同产地的野菊花药材进行炮制加工,采用高效液相色谱法(HPLC)测定野菊花药材中木犀草素的含量,采集野菊花药材的NIRS图谱,运用偏最小二乘法(PLS)建立河南不同产区不同加工方法野菊花药材中木犀草素的定量分析模型。结果表明所建立的木犀草素定量分析模型的内部交叉验证相关系数(R2)、校正均方差(RMSEC)、预测均方差(RMSEP)分别为0.94525、0.0142、0.0428;验证集样品的木犀草素含量近红外预测值的平均相对偏差为2.95%,预测性能良好。该方法测定高效、快捷、准确度高,可用于河南产不同加工方法的野菊花药材中木犀草素含量的快速测定。     

威灵仙药材中三萜苷元齐墩果酸含量的HPLC测定

为探索可用于威灵仙药材质量控制的三萜苷元齐墩果酸HPLC含量测定方法,用HPLC法测定了威灵仙药材中齐墩果酸含量。色谱柱:Kromasil C18(4.6×150 mm,5μm),检测波长:220 nm,流动相:甲醇-水(95:5),流速:0.5 mL/min。齐墩果酸在0.004~0.015 mg范围内线性关系良好,回归方程为:Y=437954101.73X 144588.26(R=0.9991),平均回收率为:98.33%,RSD=1.76%。样品中齐墩果酸含量分别为:东北铁线莲0.2281%;威灵仙0.1260%、棉团铁线莲0.1851%。本方法快速、准确、灵敏度高、重现性好,适用于对威灵仙中齐墩果酸的含量进行测定。     

饲料中添加胆酸钠对大口黑鲈生长及糖代谢的影响

为探究胆酸(Cholic acid, CA)作为饲料添加剂对大口黑鲈(Micropterus salmoides)生长及糖代谢的影响, 实验以饲料中添加300 mg/kg胆酸钠(Sodium cholate, CAS)作为胆酸钠组, 以不添加胆酸钠作为对照组。在饲养8周后, 分析胆酸钠对大口黑鲈生长性能、肠道菌群、糖代谢及与糖代谢相关酶的活性和基因表达的影响。结果显示: 与对照组相比, 胆酸钠组中大口黑鲈的生长指数和体成分的变化均没有显著差异; 胆酸钠组中大口黑鲈的肠道菌群组成无显著差异; 胆酸钠组中肝糖原含量和肝中糖原合成酶(Glycogen synthase, GCS)的活性显著增加, 肝中糖原分解酶糖原磷酸化酶a(Glycogen phosphorylase a, GPa)活性无显著变化, 而胆酸钠组中肌糖原含量、肌肉GCS与GPa的活性无显著差异; 胆酸钠显著促进肝中糖异生途径中果糖-1,6-二磷酸酶(Fructose-1,6-bisphosphatase, FBPase)和葡萄糖-6-磷酸酶(Glucose-6-phosphatase, G6Pase)基因的表达; 胆酸钠显著降低肝中糖酵解基因丙酮酸激酶(Pyruvate kinase, PK)和肌肉中己糖激酶(Hexokinase, HK)基因的表达; 同时, 研究还发现饲料中胆酸钠的添加可以显著降低肝中胆汁酸受体法尼醇受体(Farnesoid X receptor, FXR)基因的表达量, 而不改变肠道FXR的表达量。研究表明: 在饲料中添加300 mg/kg胆酸钠可以促进大口黑鲈肝脏糖异生, 抑制肝脏和肌肉糖酵解, 并促进鱼体肝糖原的合成。这些糖代谢的变化与肠道菌群没有直接关系, 但可能与肝中FXR的表达量降低有关。     

战骨药材及其复方制剂的指纹图谱分析及牡荆素含量测定

建立战骨药材及其复方制剂的HPLC指纹图谱分析及牡荆素的HPLC含量测定方法,并考察牡荆素在跌打生骨胶囊制备过程中的含量变化。采用Waters Xbridge C18(4.6 mm×250 mm,5μm)色谱柱;流动相为乙腈-0.5%冰醋酸水溶液,梯度洗脱;流速为1.0mL/min;检测波长为350 nm;柱温:25oC。结果表明牡荆素在0.16~0.80μg范围内线性关系良好(R2=0.9996);10批战骨药材的指纹图谱有19个共有特征峰,其峰面积占总峰面积的90%以上。每克药材含牡荆素在15.8~30.2μg之间。制备胶囊的稠膏及内容物颗粒未检出牡荆素。表明本实验建立的HPLC方法可用于测定战骨药材中牡荆素的含量,而不适用于其复方制剂中牡荆素的含量测定。     

毛萼香茶菜二萜化合物的高效液相色谱定量分析

本文报道用HPLC分离测定毛萼香茶菜(Rabdosia eriocalyx)中的二萜化合物。总二萜提取物在zorbax ODS柱上,用甲醇-水(70:30)作流动相,毛萼晶乙、毛萼晶丙等五个化合物在18分钟内能很好分离。以odonicin为内标,用峰面积比测定各组份的含量。结果符合一般含量测定的要求,此法可作为香茶菜二萜化合物一类生物活性物质的分析及发现新二萜化合物的前导性有效筛选手段。     

固相萃取-反相高效液相色谱法分析粗裂地钱中的黄酮类化合物

建立了粗裂地钱中四种黄酮类化合物的固相萃取高效液相色谱分析方法。样品经95%乙醇溶液提取后,用C18固相萃取小柱预处理,采用甲醇∶乙腈∶0.5%乙酸(150∶100∶150,V/V)为流动相,C18色谱柱(150 mm×4.6 mm,5μm)进行分离与测定。低、中、高浓度下方法的回收率为89.70%～97.95%,RSD小于6%。结果表明,该法简便快速,节省溶剂,分析结果准确可靠,重复性好,可用于粗裂地钱中四种黄酮类化合物的含量分析。     

Enantioseparation and determination of flumequine enantiomers in multiple food matrices with chiral liquid chromatography coupled with tandem mass spectrometry

The present work firstly described the enantioseparation and determination of flumequine enantiomers in milk, yogurt, chicken, beef, egg, and honey samples by chiral liquid chromatography‐tandem mass spectrometry. The enantioseparation was performed under reversed‐phase conditions on a Chiralpak IC column at 20°C. The effects of chiral stationary phase, mobile phase components, and column temperature on the separation of flumequine enantiomers have been studied in detail. Target compounds were extracted from six different matrices with individual extraction procedure followed by cleanup using Cleanert C18 solid phase extraction cartridge. Good linearity (R²>0.9913) was obtained over the concentration range of 0.125 to 12.5 ng g^‐1 for each enantiomer in matrix‐matched standard calibration curves. The limits of detection and limits of quantification of two flumequine enantiomers were 0.015‐0.024 and 0.045‐0.063 ng g^‐1, respectively. The average recoveries of the targeted compounds varied from 82.3 to 110.5%, with relative standard deviation less than 11.7%. The method was successfully applied to the determination of flumequine enantiomers in multiple food matrices, providing a reliable method for evaluating the potential risk in animal productions.     

Determination of octreotide and assessment of matrix effects in human plasma using ultra high performance liquid chromatography-tandem mass spectrometry

A selective UHPLC-MS/MS method for determination of the therapeutic peptide octreotide in human plasma was developed and validated. This assay used a UHPLC C(18) column with 1.7 μm particle size for efficient separation and an ion-exchange SPE for selective extraction. Octreotide and its labeled internal standard, [(13)C(6)Phe(3)] octreotide, were extracted from human plasma using a simple Oasis? WCX μElution SPE method and analyzed with a total chromatographic run time of 7.5 min. Matrix effects were studied during method development by direct monitoring of representative phospholipids. On-line removal of phospholipids using column switching and pre-column back-flushing was carried out to trap and remove any residual phospholipid matrix interferences. The UHPLC column provided baseline separation between the analyte and matrix peaks. The chromatographic conditions yielded optimal retention and excellent peak shape for both the analyte and internal standard. The assay was linear in the concentration range of 0.025-25.0 ng/ml, inter- and intra-assay precision and accuracy were within 6.1% and ±1.93%, respectively. Recovery was ～73%. Post-extraction addition experiments showed that matrix effects were less than 4%. This method for octreotide in human plasma has been validated and utilized to support of clinical pharmacokinetic studies.     

High-performance liquid chromatographic assay of formycin A in plasma after solid-phase extraction

A new, simple and accurate high-performance liquid chromatography (HPLC) method for the determination of formycin A in plasma is presented. The samples were chromatographed on a LiChrosorb RP-18 column after purification using a Bakerbond SPE column. The mobile phase was methanol–0.067 M phosphate buffer, pH 4.20 (1:4, v/v) containing 0.005 M sodium hexanesulfonate. Azathioprine was applied as an internal standard. UV detection was carried out at 293 nm. The method was tested for linearity (over the range 0.1–9.0 μg/ml). The recovery was 91.89% (mean). The described method has been successfully applied to the quantitative determination of formycin A in plasma and should be useful for clinical and bioavailability investigations.     

Determination by liquid chromatography-mass spectrometry of clomiphene isomers in the plasma of patients undergoing treatment for the induction of ovulation

A rapid, sensitive and selective LC-MS method is described for the simultaneous determination of zuclomiphene and enclomiphene in plasma from patients undergoing treatment with clomiphene citrate for the induction of ovulation. Samples spiked with N-didesmethyltamoxifen, the internal standard, were extracted into methyl tertiary butyl ether. The compounds were separated on a Luna C(18) analytical column, and a mobile phase of methanol-water (70:30 v/v) containing 0.05% trifluoroacetic acid at a flow rate of 1ml/min. The limits of determination were 35pg/ml and 7pg/ml for zu- and enclomiphene, respectively. Within-day coefficients of variation ranged from 2.1% to 7.2%.     

固相萃取-气相色谱法测定水和废水中四种氯苯类有机污染物

研究了用固相萃取技术结合氢火焰气相色谱法测定水和废水中四种氯苯类有机污染物。通过实验比较Sdex C18,Sep-Park Vac Silica,Bond Elut CARBON,Bond Elut SI和Bond Elut PLEXA五种SPE小柱对四种氯苯类的萃取效率,发现Dionex的Sdex C18柱有很好的回收率。系统研究了最佳萃取条件,甲醇洗脱体积为4.0 mL,且洗脱液不可通过氮吹浓缩;四种氯苯类化合物除氯苯外穿透体积都在1.0 L以上,而氯苯的穿透体积为300 mL,表明Sdex C18柱对二氯苯和三氯苯有很强的吸附性。在最佳萃取和测定条件下,方法线性范围为20.0~400μg/L,检出限为0.383~0.635μg/L,完全满足日常环境监测分析要求。     

Enantiomeric separation of tramadol and its active metabolite in human plasma by chiral high-performance liquid chromatography: application to pharmacokinetic studies

A sensitive and stereoselective high-performance liquid chromatographic assay for the quantitative determination of the analgesic tramadol and O-demethyltramadol, an active metabolite, is described in this work. Ketamine was used as internal standard. The assay involved a single tert-butymethylether extraction and liquid chromatography analysis with fluorescence detection. Chromatography was performed at 20 degrees C on a Chiracel OD-R column containing cellulose tris-(3,5-dimethylphenylcarbamate) as stationary phase, preceded by an achiral end-capped C18 column. The mobile phase was a mixture of phosphate buffer (containing sodium perchlorate (0.2 M) and triethylamine (0.09 M) adjusted to pH 6) and acetonitrile (80:20). The method developed was validated. The limit of quantitation of each enantiomer of tramadol and its active metabolite by this method was 0.5 ng/mL; only 0.5 mL of the plasma sample was required for the determination. The calibration curve was linear from 0.5 to 750 ng/mL for tramadol enantiomers, and from 0.5 to 500 ng/mL for O-demethyltramadol enantiomers. Intra and interday precision [coefficient of variation (CV)] did not exceed 10%. Mean recoveries of 95.95 and 97.87% for (+)R,R- and (-)S,S-tramadol and 97.70 and 98.79% for (+)R,R- and (-)S,S-O-demethyltramadol with CVs < 2.15% were obtained. Applicability of the method was demonstrated by a pharmacokinetic study in normal volunteers who received 100 mg of tramadol by the intravenous route.     

Determination of omeprazole in human plasma by high-performance liquid chromatography

A new method for the determination of omeprazole in human plasma was developed. Omeprazole was extracted from plasma with toluene-isoamylalcohol (95:5, v/v), the organic phase was evaporated, dissolved in the mobile phase and injected into a reversed-phase C₁₈ column. Flunitrazepam was used as an internal standard. The mobile phase consisted of 47% methanol and 53% of 0.1 M dipotassium hydrogenphosphate, pH 7.8. The spectrophotometric detection was performed at 302 nm. Limit of quantitation was 9.7 ng/ml and the calibration curve was linear up to 1240 ng/ml.     

Simultaneous quantitation of five flavonoid glycosides in Herba Epimedii by high-performance liquid chromatography-tandem mass spectrometry

A rapid and accurate reversed-phase liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method has been developed and validated for the quantitative determination of five flavonoid glycosides, icariin, epimedin A, epimedin B, epimedin C and hyperin in Herba Epimedii. Chromatographic separations were performed using a C(18) narrow-bore HPLC column; a mixture of an aqueous solution of ammonium formate (pH 4.0) and acetonitrile was used as the mobile phase, with compounds detected in the positive ion mode with multiple-reaction monitoring using a triple-quadrupole mass spectrometer equipped with an electrospray ionisation interface. This method for the determination of the reported flavonoid glycosides was accurate and reproducible, with a lower limit of quantication of 0.5 microg/mL. The standard calibration curves for the above-mentioned compounds were linear (r(2) > 0.998) over the concentration range 0.5-10.0 microg/mL. The relative standard deviations for intra- and inter-day precision over the concentration range for the flavonoid glycosides were lower than 7.8% with accuracy between 90.1 and 111.0%. The established method was successfully applied to the quality assessment of samples of Herba Epimedii collected from Korea and China.     

Liquid chromatographic determination and depletion profile of oxytetracycline in milk after repeated intramuscular administration in sheep

A simple, rapid and specific ion-pair liquid chromatographic method for the routine determination of the marker residue of oxytetracycline in sheep milk, at levels as low as 20 microg/kg, has been developed. Milk samples were acidified and extracted with acetonitrile. The extracts were purified by treatment with ammonium sulphate and concentrated into diluted phosphoric acid. Separation was carried out isocratically on a Nucleosil C(18) column using a mobile phase that contained both positively and negatively charged pairing ions. The in-house validated method gave overall recoveries and overall relative standard deviations better than 86% and 4.6%, respectively. The method was successfully applied to study the depletion of oxytetracycline in sheep milk and to estimate the withdrawal period after intramuscular administration of a commercial oxytetracycline formulation.