二乙基二硫代氨基甲酸钠柱前衍生化HPLC法测一捻金中朱砂的含量

目的建立用高效液相色谱测定一捻金中朱砂含量的方法。方法以二乙基二硫代氨基甲酸钠(NaDDC)为衍生化试剂,样品经消解、衍生化和有机溶剂提取后测定。采用高效液相色谱法,使用C18柱,流动相为甲醇-水(含0.02 mmoL.L-1的NaDDC)(87:13);检测波长272 nm;流速1 mL.min-1,柱温35℃。结果 Hg2+在0.1052~0.6312μg范围内线性关系良好,平均加样回收率99.23%。结论本法结果准确,便于操作,可作为一捻金的质量控制方法之一。     

朱砂中汞的二乙基二硫代氨基甲酸盐柱前衍生化HPLC测定

建立了柱前衍生化HPLC法测定朱砂中的汞含量。以二乙基二硫代氨基甲酸钠(NaDDC)为衍生化试剂,样品经消解、衍生化和有机溶剂提取后测定。采用C18色谱柱,流动相为甲醇-水(含0.022mmol/LNaDDC)(90∶10),检测波长为272nm。结果Hg2 在8.4～42.0μg/ml浓度范围内线性关系良好,检测限为4.2ng,平均回收率为99.9%,RSD为2.2%。     

柱前衍生化HPLC法测小儿金丹片中朱砂的含量

目的：建立柱前衍生化HPLC法测定小儿金丹片中朱砂的含量。方法：以二乙基二硫代氨基甲酸钠（NaDDC）为衍生化试剂,样品经消解、衍生化和有机溶剂提取后测定。采用HPLC法,使用C18柱,流动相为甲醇-水（含0．02mmol·L^-1的NaDDC）（87：13）;检测波长272nm;流速1．0ml·min^-1,柱温35℃。结果：Hg^2＋在0．10～0．63μg范围内线性关系良好,r=0．9992,平均加样回收率98．4％（RSD为0．40％,n=9）。结论：本法结果准确,便于操作,可作为小儿金丹片的质量控制方法之一。     

微透析-柱前衍生化高效液相色谱法研究丙戊酸在大鼠血-脑双位点的药动学

目的研究大鼠腹腔注射丙戊酸钠后游离丙戊酸在大鼠血液和脑内的药动学。方法采用血-脑双位点微透析技术结合柱前衍生化高效液相色谱法,测定给药后240 min内各时间段大鼠血液透析样品和脑部海马区细胞外液透析样品中丙戊酸的浓度。数据采用DAS 3.0软件计算药代动力学参数。结果大鼠海马区细胞外液中游离丙戊酸达峰时间显著滞后于其在血液中的达峰时间,海马区细胞外液中游离丙戊酸的C_(max)和AUC_(0~∞)均明显低于血液中的响应值。结论本文建立的微透析技术结合柱前衍生化HPLC法可以成功用于丙戊酸在大鼠血-脑双位点的药动学研究。     

紫杉醇固体脂质纳米粒大鼠体内药动学

目的研究紫杉醇固体脂质纳米粒在大鼠体内的药动学。方法10只健康大鼠,雌雄各半,分为2组,分别口服给药紫杉醇固体脂质纳米粒和紫杉醇乳剂30 mg.kg-1,在设计的时间点从颈静脉取血,采用RP-HPLC测定紫杉醇在全血中的药物浓度,药动学参数用3P97软件进行处理。结果大鼠口服给药后,紫杉醇固体脂质纳米粒和乳剂的tm ax分别为3.133 h和1.627 h,MRT分别为10.362 h和3.297 h,mρax分别为1.512 2 mg.L-1和0.718 9 mg.L-1。结论固体脂质纳米粒能够显著改善大鼠体内紫杉醇的药动学行为,有利于其更好地发挥抗肿瘤作用。     

HPLC法测定人血浆中顺铂的浓度及其药动学研究

目的:建立测定非小细胞肺癌患者静脉滴注小剂量顺铂后血浆中药物浓度的方法,并分析其药动学特点。方法:15名非小细胞肺癌化疗患者静脉滴注小剂量顺铂,动态采集患者给药前及末次给药后96h内的血浆样本,以高效液相色谱(HPLC)法测定顺铂的血药浓度,DAS软件计算药动学参数。结果:顺铂血药浓度在0.16～4mg.L-1范围内线性关系良好,相对回收率为96.60%～113.95%,日内、日间RSD均在10%以内。顺铂的药动学模型为开放二室模型,药动学参数分别为:消除半衰期(t1/2β)(62.89±10.84)h,表观分布容积(V)(13.39±4.86)L,清除率(CL)(1.42±0.57)L.h-1,药-时曲线下面积(AUC)(74.24±43.83)mg.h.L-1。结论:HPLC法测定顺铂血药浓度,操作简单易行,重现性好,符合血药浓度监测及人体内药动学研究要求。     

乳糖化-去甲斑蝥素纳米粒在大鼠体内的药动学研究

目的:研究乳糖化-去甲斑蝥素(lactosyl-norcantharidin,Lac-NCTD)水溶液及纳米粒2种制剂在大鼠体内药动学特征。方法:建立测定大鼠血浆中Lac-NCTD的HPLC检测法,并用3P97软件进行药动学拟合。结果:Lac-NCTD能与血浆中其他成分较好分离,在0.1～100μg/mL范围内线性关系良好。与Lac-NCTD水溶液相比,Lac-NCTD纳米粒的AUC及t1/2β显著增加,分别为Lac-NCTD水溶液的8.96倍和1.68倍,清除率显著降低(P<0.01)。结论:与水溶液相比,Lac-NCTD制备成纳米粒制剂后能显著改变药物在大鼠体内的药动学特征,延长了药物在体内的循环时间,有利于更好地发挥抗肿瘤效果。     

顺铂用于非小细胞肺癌化疗的药动学

目的:比较不同顺铂给药方案治疗非小细胞肺癌的顺铂血药浓度及药动学参数变化.方法:16名非小细胞肺癌化疗患者使用顺铂联合长春瑞滨方案治疗,静脉滴注给药,根据顺铂用药方案不同分为高剂量组与低剂量组.高剂量组用药方案:第1,2天顺铂60 mg·m-2,第1,8天长春瑞滨25 mg·m-2低剂量组用药方案:第1～5天顺铂20 mg·m-2,第1,8天长春瑞滨25 mg·m-2.动态采集患者给药前及末次给药后96 h内血样本,高效液相色谱法测定给药后顺铂血药浓度,3P97软件计算药动学参数.结果:从0～96 h,高剂量组血药浓度显著高于低剂量组(P<0.05),但差异随时间延长逐渐缩小.除AUC外,两组间其他药动学参数差异无显著性,但个体间变异大,变异系教为12.7%～99%.结论:结论AUC值对于顺铂个体化方案设计具有重要参考价值.     

柱前衍生化HPLC测定大鼠血浆中的盐酸美金刚胺

目的 建立测定大鼠血浆中盐酸美金刚胺浓度的方法.方法 采用柱前衍生化HPLC-UV法,色谱柱为Luna C_(18)柱(150 mm ×4.6 mm,5 μm),流动相为甲醇-水-三乙胺(70:30:0.5,磷酸调pH7),流速为1.0 mL·min-1,检测波长为225nm;以盐酸金刚烷胺为内标,采用液-液萃取法,经衍生化后进样测定.结果 大鼠血浆中的内源性成分对美金刚胺的测定无干扰;美金刚胺0.14～9.20μg·mL~(-1)与峰面积线性关系良好(r=0.9950),低、中、高浓度样品的平均提取同收率分别为83.46%、80.34%、82.77%,平均方法回收率分别为112.01%、99.96%、99.10%,日内RSD分别为10.24%、8.97%、6.94%,日间RSD分别为12.10%、8.77%、4.60%.结论 所建方法灵敏、准确,可为盐酸美金刚胺的体内研究提供参考.     

HPLC柱前衍生法测定妥布霉素

方法 对妥布霉素HPLC测定方法进行了研究。目 的 选用2,4- 二硝基氟苯(DNFB)柱前衍生化,以Agilent Hypersil ODS为色谱柱,0.25%三羟甲基氨基甲烷溶液-乙腈-0.5mol·L-1硫酸溶液(40∶59∶1)为流动相,流速为1.2ml·min -1,检测波长365nm。结果 在10～100μg范围内峰面积与浓度呈良好的线性关系,r＝0.9999。平均回收率为100.9%,RSD为1.5%。结论 本方法简便准确,重 现性好,可用于妥布霉素的质量控制。     

A sensitive assay for clavulanic acid and sulbactam in biological fluids by high-performance liquid chromatography and precolumn derivatization

Precolumn derivatization procedures using 1,2,4-triazole for the detection and quantitation of sulbactam and clavulanic acid spiked into urine and blood serum at trace levels have been developed. Sulbactam and clavulanic acid produced derivatives which absorbed maximally at 325 and 315 nm, respectively. The methods allow the detection of clavulanic acid and sulbactam down to 0.05 micrograms ml-1 in serum and 0.5 micrograms ml-1 in urine. The relative standard deviation for five replicate analyses of sulbactam and clavulanic acid at a concentration of 20 micrograms ml-1 in serum and urine ranged from 2-6%. In further HPLC experiments with sulbactam in phosphate buffer solution, ampicillin was found as a contaminant (0.5% by mass) in the sulbactam sample provided. The significance of this finding is discussed.     

柱前衍生化高效液相色谱法测定2-去氧葡萄糖的含量

目的建立柱前衍生化高效液相色谱法测定2-去氧葡萄糖(2-DG)含量的方法。方法采用1-苯基-3-甲基-5-吡唑酮(PMP)为柱前衍生化试剂,将2-DG在碱性条件下衍生化后直接进样测定。分离柱为HypersilODS2色谱柱(4.6 mm×250 mm,5μm),流动相为100 mmol·L-1醋酸铵缓冲液(pH 5.5)-乙腈(78∶22),流速1.0 mL·min-1,波长249 nm。结果 2-DG在19.68~393.6μg·mL-1浓度范围内与峰面积具有良好的线性关系(r=0.9997);其定量限(S/N=10)和最低检出限(S/N=3)分别为7.8和3.1 ng;平均回收率为101.21%,RSD 0.63%。结论该方法简便实用、检测灵敏度高、测定结果准确,适用于2-DG的质量控制。     

High-performance micellar liquid chromatography determination of sulphonamides in pharmaceuticals after azodye precolumn derivatization

A chromatographic procedure with precolumn derivatization to form the N-(1-naphthyl)ethylenediamine dihydrochloride azodyes is proposed for the analysis of several sulphonamides (sodium sulphacetamide, sulphadiazine, sulphaguanidine, sulphamerazine, sulphamethizole, sulphamethoxazole, sulphanilamide and sulphathiazole) in pharmaceutical preparations (tablets, pills, capsules, suspensions and drops). The separation is performed with a 0.05 M sodium dodecyl sulphate/2.4% pentanol eluent at pH 7. The precolumn derivatization improved the resolution in the chromatograms and increased the selectivity in the determination of mixtures of sulphonamides and in preparations where other drugs were present. The derivatization reaction was readily performed in a micellar medium of SDS at pH 1, leading to a rapid and simple procedure. The recoveries were in the 97–104% range with relative standard deviations usually below 3%.     

柱前衍生化HPLC法分析沙参多糖中单糖组成

目的建立北沙参多糖中单糖的组成和含量的检测方法。方法以三氟乙酸水解沙参多糖,水解产物加入衍生化试剂1-苯基-3-甲基-5-吡唑酮(PMP)生成衍生化产物,采用RP-HPLC法,色谱柱:C18色谱柱(250 mm×4.6 mm,5μm),流动相:体积分数为19%的乙腈溶液-醋酸铵缓冲液(醋酸铵-冰醋酸,pH5.5,体积比为100∶1),等度洗脱,流速:1.0 mL.min-1,检测器:紫外检测器,检测波长:250 nm,柱温:35℃。结果沙参多糖由D-甘露糖、D-鼠李糖、半乳糖醛酸、D-葡萄糖、D-半乳糖、L-(+)-阿拉伯糖等8种单糖(有2种未鉴定出的单糖)组成。D-甘露糖、D-鼠李糖、半乳糖醛酸、D-葡萄糖、D-半乳糖、L-(+)-阿拉伯糖的线性分别为4~80μmol.L-1(r=0.999 0)、12~240μmol.L-1(r=0.999 0)、5.6~1.12×103μmol.L-1(r=0.999 4)、1.54×103~30.80×103μmol.L-1(r=0.999 0)、80.0~1.6×103μmol.L-1(r=0.999 0)、64.00~1.28×103μmo.lL-1(r=0.9992);平均加样回收率分别为94.0%、88.5%、93.9%、95.9%、89.2%、90.8%。含量分别为0.22%、0.61%、8.55%、72.88%、4.79%、2.71%。结论该方法可以准确的测定出北沙参多糖中含有的各种单糖和含量。     

柱前衍生液相色谱法与柱后衍生液相色谱法测定疫苗中游离甲醛残留量的对比研究

目的:建立柱前衍生液相色谱法与柱后衍生液相色谱法测定疫苗中游离甲醛残留量,并考察2种方法测定结果的一致性程度。方法:柱前衍生液相色谱法色谱条件：岛津LC-20AT液相色谱仪(SPD-20A紫外检测器),采用Kromasil 100-5-C₁₈(250 mm×4.6 mm)色谱柱,流动相为60%乙腈溶液,流速0.8mL·min^-1,柱温40℃,测定波长360 nm。柱后衍生液相色谱法色谱条件：岛津LC-20AT液相色谱仪(SPD-M20A二极管阵列检测器和vector衍生装置),采用纳谱AQ-C₁₈(250 mm×4.6 mm)色谱柱,流动相为0.2%(V/V)磷酸溶液,流速1.0 mL·min^-1,柱温25℃,检测波长412 nm;衍生溶液为醋酸缓冲液,流速0.5 mL·min^-1,温度100℃。分别对2种方法的精密度、重复性、加样回收率等项目进行考察,并对测定结果进行显著性检验(F检验和t检验)。结果:柱前衍生液相色谱法线性范围为0.025～100μg·mL^-1     

A modified assay for cyclosporin in blood using solid-phase extraction with high-performance liquid chromatography

A modified procedure for measuring cyclosporin in whole blood by high-performance liquid chromatography is described and evaluated for clinical use. Sample preparation uses solid-phase extraction cartridges that can be reused. Life of the reverse-phase analytical column exceeds 1,000 injections at 70 degrees C. Cyclosporins A and D (internal standard) elute after 5.6 and 7.6 min, respectively. Calibration plots are linear from 50 ng/ml to at least 2,000 ng/ml. Within-day and between-day imprecision is less than 9% (coefficient of variation). Minimum measurable concentration is 50 ng/ml.     

Determination of oligosaccharides and monosaccharides in Hakka rice wine by precolumn derivation high-performance liquid chromatography

This article presents a precolumn derivatization procedure with 1-phenyl-3-methyl-5-pyrazolone (PMP) reagent to detect oligosaccharides and monosaccharides in Hakka rice wine. The subsequent separation of the derivatized glucose–PMP also was performed using a mobile phase consisting of the molar ratio of acetonitrile to ammonium acetate buffer (0.1M) of 22:78 at a flow rate of 1.0 mL/min with the column temperature of 35°C, and the pH of ammonium acetate buffer at 5.5. The optimum derivation conditions were as follows: reaction temperature, 70°C; reaction time, 30 minutes; molar ratio of PMP to glucose, 10:1 (v/v); molar ratio of sodium hydroxide to glucose, 3:1 (v/v). The recovery rates were between 93.13% and 102.08% with relative standard deviation of 0.96–2.48%. The established method provides sufficient sensitivity with values of limit of detection of 0.09–0.26 mg/L and limit of quantification of 0.27–0.87 mg/L for determination of oligosaccharides and monosaccharides.     

Determination of catecholamines in rat tissues by high-performance liquid chromatography using a precolumn fluorescence labeling method

A high-performance liquid Chromatographic method using solid-phase dansylation on alumma for precolumn fluorescence label has been developed for the determination of Catecholamines (norepinephrine, epinephrine, and dopamine) in various rat tissues. After alumina treatment of the tissue homogenate, catecholamines adsorbed on the alumina were dansylated by solid-phase reaction. Both the excess reagent and fluorescent degradation products produced during dansylation were washed out from the alumina. Dansylated Catecholamines were eluted from the alumina and separated by reversed-phase high-performance liquid chromatography. The four catecholamine derivatives, including the internal standard, were separated within 17 min, and no major interfering peak could be detected on any chromatograms. The calibration graph showed a good linearity in a range of 10 to 500 pmol for each catecholamine per sample. This method was applied to different rat tissues, and both the recovery and the reproducibility for all samples proved to be satisfactory. The present study provides a simple, sensitive, and selective method useful for routine pharmacological experiments of the determination of Catecholamines.     

Determination of catecholamines in rat tissue by precolumn dansylation using micro high-performance liquid chromatography with fluorescence detection

A micro high-performance liquid chromatography (MHPLC)-precolumn dansylation procedure for determination of norepinephrine (NE), epinephrine (E), and dopamine (DO) in various kinds of rat tissue has been developed. After dansylation of the extracted catecholamines to get fluorescent derivatives, they were separated and determined using MHPLC equipment with a fluorimeter. The standard deviation of the method was ± 8% for NE, ± 4% for E, and ± 8% for DO. This relatively simple and rapid procedure for determination of catecholamines provides an efficient and reproducible method. Furthermore, the procedure is readily adaptable to routine pharmacological study of catecholamine metabolism in various kinds of rat tissue.     

Comparison of 9-fluorenylmethoxycarbonyl and 9-fluoreneacetyl-tagged silica-based derivatization reagents in high-performance liquid chromatography.

Two silica reagents based on a 4-hydroxy-3-nitrobenzoyl backbone were synthesized and characterized with 9-fluorenylmethoxycarbonyl (FMOC) and 9-fluoreneacetyl (FA) tags. These reagents were tested by derivatization of primary and secondary amines. Derivatization conditions such as temperature, time and triethylamine catalyst were tested. The FA-tagged silica reagent showed better performance than the FMOC-tagged silica reagent by a comparison of derivatization efficiencies, stabilities of reagents, and blank reagent interferences with derivatization. Finally, cadaverine and an aliphatic amine mixture were analysed using the FA-tagged reagent by pre-column, off-line derivatization and fluorescence detection.