高效液相色谱法检测压敏胶及其制品中的防老剂D

建立了高效液相色谱法测定压敏胶及其制品中防老剂D的分析方法.对样品前处理和色谱分析条件进行了优化,以乙酸乙酯为溶剂,甲醇为沉淀剂,乙腈-水(90:10)为流动相,流速为0.5 mL/min,等度洗脱.采用C18色谱柱,二极管阵列检测器(DAD),高效液相色谱法测定,检测波长为310 nm,外标法定量.并建立了防老剂D的...     

丙泊酚微乳注射液血浆蛋白结合率的测定

采用Beagel犬血浆平衡透析法,高效液相色谱法测定丙泊酚微乳注射液与Beagel犬血浆蛋白结合率。色谱柱:Agilent C18柱(5μm,4.6 mm×150 mm);流动相:0.01 mol/L乙酸铵(pH 7.0):乙腈=20:80(V/V);紫外检测波长:215 nm。丙泊酚在5～600 ng/mL范围内线性关系良好。丙泊酚微乳注射液在质量浓度为50、400、1000 ng/mL时与血浆蛋白结合率分别为(96.97±0.52)%、(97.73±0.76)%、(97.74±0.58)%。HPLC法能较好的应用于丙泊酚微乳注射液血浆蛋白结合率的测定,丙泊酚微乳注射液具有较高强度的血浆蛋白结合率。     

HPLC法测定米格列奈钙及其有关物质

采用高效液相色谱法,室温下使用C18(250×4.6mm,5μm)色谱柱,以乙腈-0.01 moL/L KH2PO4溶液(pH 4)(体积比为43:57)为流动相,检测波长为210啪,流速为1.0 mL/min,测定米格列奈钙及其有关物质,米格列奈钙的浓度在5～450μg/mL范围内线性关系良好,相关系数为0.9999.还与非水溶液滴定法、元素分析方法对照验证,结果证明该方法适用于米格列奈钙及其有关物质的测定.     

反相高效液相色谱法测定Arthrobacter sp.黄嘌呤氧化酶活性

报道了采用反相高效液相色谱法(RP-HPLC)测定节杆菌(Arthrobacter sp.)黄嘌呤氧化酶活性的有效方法. 将酶初提液与含有黄嘌呤的反应体系在37 ℃下反应20 min, 反应终止后通过HPLC测定产物尿酸生成量的变化来分析酶的活性. 通过流动相组成、pH和柱温等分离条件的优化, 确定了最佳的色谱检测条件以NH4H2PO4 (50 mmol/L, pH 7.5)溶液为流动相, 流速1 mL/min, 柱温25 ℃, 检测波长290 nm. 为深入研究微生物细胞内黄嘌呤氧化酶提供了高效检测手段.     

反相高效液相色谱法测定化妆品中的苯甲酸和山梨酸

建立了反相高效液相色谱法测定化妆品中苯甲酸和山梨酸的分析方法.样品经甲醇超声提取后,直接过滤进样检测.采用Sinochrom C18色谱柱,以0.02 mol/L乙酸铵-甲醇(体积比为85:15)为流动相,流速为2.0 mL/min,检测波长为230 nm.结果表明:苯甲酸和山梨酸浓度在2.00～12.00 μg/mL...     

反相离子对液相色谱法同时测定11种氟喹诺酮类药物的研究

建立了高效液相色谱-荧光法同时测定11种氟喹诺酮类药物的分析方法.主要研究了流动相、流动相配比及流动相的pH对氟喹诺酮分离的影响.确定了液相色谱分析最佳条件.分离条件为:Xbridge Shield RP C18柱,以V(0.10%三氟乙酸)∶V(乙腈)∶V(甲醇)=89∶4∶7为流动相;检测波长为λex=280 nm和λem＝450 nm.方法检出限为:诺氟沙星、环丙沙星、培氟沙星和恩诺沙星0.007μg/mL,单诺沙星0.002 μg/mL,沙拉沙星和氧氟沙星为0.04 μg/mL,二氟沙星和奥比沙星为0.02 μg/mL,依诺沙星、麻保沙星为0.4 μg/mL,各组分回收率在97%～100.2%,相对标准偏差为0.2%～2.9%.     

反相高效液相色谱法测定兔血浆中的灯盏乙素

建立一种测定兔血浆中灯盏乙素浓度的高效液相色谱法.以芦丁为内标,采用甲醇沉淀蛋白法对血浆样品进行预处理.HPLC色谱柱为Kromasil C18柱(4.6 mm×250 mm,5 μn),流动相为水(用醋酸调pH 3.0)-乙腈(7822),检测波长335 nm,流速1.0 mL/min,柱温为室温;本法的线性范围为0.025～4μg/mL,最低定量浓度为0.025 μg/mL;方法回收率为95.9%～104.8%,日内RSD为1.9%～7.3%,日间RSD为1.9%～8.9%.方法学评价结果表明本方法灵敏、准确,操作简便,可用于深入研究灯盏乙素在动物体内的药代动力学过程.     

高效液相色谱法同时测定反应液中的巯基乙酸和巯基乙酸异辛酯

建立了同时测定反应液中的巯基乙酸(TGA)和巯基乙酸异辛酯(TGB)的高效液相色谱分析方法.实验采用Shimpack C18色谱柱,紫外检测器,检测波长210 nm,以V(乙腈)∶V(水)=70∶30作为流动相(用H3PO4调节流动相pH为3),流速1.0 mL/min,柱温30 ℃.结果表明,TGA和TGB在上述色谱条件下可实现较好分离,测定结果的最大相对标准偏差分别为0.53%和0.46%,检出限分别为2.03×10-3 g/L和6.11×10-3 g/L,加标回收率分别在99.0%～100.8%和99.1%～100.7%.     

高效液相色谱-质谱甄别七种化学战剂相关化合物

建立了高效液相色谱-质谱(HPLC-MS)甄别7种化学战剂相关化合物的分析方法,采用C18柱分离了包括酸性、中性和碱性在内的7种化学战剂相关化合物,选择离子监测法(SIM)测定这7种化合物的检出限在0．01—1mg/L之间。利用HPLC～APCI^ MS测定了4种土壤中的双(2-羟乙基)亚砜,此方法回收率大于93％,相对标准偏差(RSD)小于4％.     

HPLC法测定丙酮酸钙的含量

建立一种用高效液相色谱法(HPLC法)测定丙酮酸钙含量的方法。色谱条件为C18柱,乙腈—0.1%二环已胺和0.1%甲酸的水溶液(5:95)为流动相, 检测波长为230nm。线性范围0.08~0.8mg/mL(r = 0.9999)。     

Aqueous Extract of Crataegus azarolus Protects Against DNA Damage in Human Lymphoblast Cell K562 and Enhances Antioxidant Activity

The present study was carried out to characterize the cellular antioxidant effect of the aqueous extract of Crataegus azarolus and its antigenotoxic potential using human myelogenous cells, K562. The antioxidant capacity of this extract was evaluated by determining its cellular antioxidant activity (CAA) in K562 cells. Also, preceding antigenotoxicity assessment, its eventual genotoxicity property was investigated by evaluating its capacity to induce the DNA degradation of treated cell nuclei. As no genotoxicity was detected at different exposure times, its ability to protect cell DNA against H₂O₂ oxidative effect was investigated, using the “comet assay.” It appears that 800 μg/mL of extract inhibited the genotoxicity induced by H₂O₂ with a rate of 41.30 %, after 4 h of incubation. In addition, this extract revealed a significant cellular antioxidant capacity against the reactive oxygen species in K562 cells.     

Antigenotoxic Effect Against Ultraviolet Radiation‐induced DNA Damage of the Essential Oils from Lippia Species

The antigenotoxicity against ultraviolet radiation (UV)‐induced DNA damage of essential oils (EO) from Lippia species was studied using SOS Chromotest. Based on the minimum concentration that significantly inhibits genotoxicity, the genoprotective potential of EO from highest to lowest was Lippia graveolens, thymol‐RC ≈ Lippia origanoides, carvacrol‐RC ≈ L. origanoides, thymol‐RC > Lippia alba, citral‐RC ≈ Lippia citriodora, citral‐RC ≈ Lippia micromera, thymol‐RC > L. alba, myrcenone‐RC. EO from L. alba, carvone/limonene‐RC, L. origanoides, α‐phellandrene‐RC and L. dulcis, trans‐β‐caryophyllene‐RC did not reduce the UV genotoxicity at any of the doses tested. A gas chromatography with flame ionization detection analysis (GC‐FID) was conducted to evaluate the solubility of the major EO constituents under our experimental conditions. GC‐FID analysis showed that, at least partially, major EO constituents were water‐soluble and therefore, they were related with the antigenotoxicity detected for EO. Constituents such as p‐cymene, geraniol, carvacrol, thymol, citral and 1,8‐cineole showed antigenotoxicity. The antioxidant activity of EO constituents was also determined using the oxygen radical antioxidant capacity (ORAC) assay. The results showed that the antigenotoxicity of the EO constituents was unconnected with their antioxidant activity. The antigenotoxicity to different constituent binary mixtures suggests that synergistic effects can occur in some of the studied EO.     

高效液相色谱法测定食品中的姜黄素

食品中的姜黄素经甲醇提取、净化后,以液相色谱-紫外检测器外标法定量检测.色谱条件:反相C<,18>色谱柱(250 mm x4.6 mm,5μm),以乙腈-0.05%乙酸氨水溶液为流动相等度洗脱,流速为1.0 mL/min.姜黄素质量浓度在0.5-50.0μg/mL范围内与峰面积呈线性关系,回归方程y=7440x-295...     

Determination of puerarin in rat cortex by high-performance liquid chromatography after intravenous administration of Puerariae flavonoids

In the present study, a rapid and simple high-performance liquid chromatographic (HPLC) assay for determination of puerarin in rat cortex was developed. The analysis was carried out on a Zorbax SB-C18 column with mobile phase acetonitrile-0.5% aqueous phosphoric acid (11:89, v/v). The detection was by UV at 252 nm. The calibration curve for puerarin was linear (r=0.9999) over the concentration range 0.516-206.250 microg/mL. The limit of detection was 0.206 microg/mL (signal-to-noise ratio 3) and the limit of quantification (signal-to-noise ratio 10) was 0.516 microg/mL. Stability studies showed that puerarin was stable at temperatures of 4 degrees C in methanol for at least 30 days. The intra- and inter-day assays of puerarin from rat cortex were less than 2.5% at concentration range 0.516-206.250 microg/mL and good overall recoveries (97.4-101.7%) were found at same concentrations. The method was applied to determine the pharmacokinetic parameters and the time course of puerarin in rat cortex, following a single dosage of intravenous administration of flavonoids from Puerariae radix at 32 mg/kg of puerarin to male Wistar rats.     

Photoprotective and Antigenotoxic Effects of the Flavonoids Apigenin,Naringenin and Pinocembrin

This work evaluated the photoprotective and antigenotoxic effects against ultraviolet B (UVB) radiation of flavonoid compounds apigenin, naringenin and pinocembrin. The photoprotective efficacy of these compounds was estimated using in vitro photoprotection indices, and the antigenotoxicity against UVB radiation was evaluated using the SOS chromotest and an enzymatic (proteinase K/T4 endonuclease V enzyme) comet assay in UV‐treated Escherichia coli and human (HEK‐293) cells, respectively. Naringenin and pinocembrin showed maximum UV‐absorption peak in UVC and UVB zones, while apigenin showed UV‐absorption capability from UVC to UVA range. These compounds acted as UV filters reducing UV‐induced genotoxicity, both in bacteria and in human cells. The enzymatic comet assay resulted highly sensitive for detection of UVB‐induced DNA damage in HEK‐293 cells. In this work, the photoprotective potential of these flavonoids was widely discussed.     

高效液相色谱法测定人体红细胞中儿茶酚氧位甲基转移酶的活性

 采用高效液相色谱紫外检测法测定人体红细胞中儿茶酚氧位甲基转移酶 (COMT)的活性。以 3,4 二羟基苯甲酸 (DBA)作为酶反应底物 ,S 腺苷甲硫氨酸 (SAM)作为甲基供体 ,在镁离子的存在下 ,将SAM上的甲基转移到DBA 3位的氧上。色谱法测定反应产物 4 羟基 3 甲氧基苯甲酸 (4 OH 3 MBA)的生成量。人体红细胞中COMT活性的线性范围在 1U/mL～ 6 0U/mL ,最低检测限为 0 5U/mL(S/N≥ 5 ) ,方法的精密度良好 (平均RSD <10 % )。     

Development of a CE method for the determination of mycophenolic acid in human plasma: a comparison with HPLC

Mycophenolate mofetil (MMF) is a widely used drug for the maintenance of immunosuppressive therapy in renal-transplant recipients. MMF is rapidly metabolized in vivo to mycophenolic acid (MPA), a reversible, noncompetitive inhibitor of inosine monophosphate dehydrogenase, which represents a limiting enzyme in lymphocyte proliferation. MPA shows large interindividual pharmacokinetic variability: its monitoring is therefore of primary importance to achieve adequate immunosuppression with minimized risk of graft rejection or toxicity. We developed a CE method for the determination of total MPA (tMPA) in plasma, based on easy sample preparation; CE evaluation of tMPA was performed in 30 mmol/L sodium-borate with 10 mmol/L SDS (pH 10.00) at 25 degrees C using a 60 cm (54.5 to window) uncoated capillary with UV detection at 254 nm wavelength. MPA was readily detectable in plasma; the CE method was linear in the range of 0.7-120 microg/mL (r >0.992). Intra- and interassay imprecision was <7% except for the lowest spiked MPA concentration, which had an intra-assay RSD% of 14.7 compared to 18.3 interassay. Data by CE were compared with results obtained by a validated HPLC method. CE assay of tMPA exhibited a very good correlation (r(2) >0.988) with respect to HPLC; Bland-Altman difference versus average showed a mean of -0.18 microg/mL +/- 1.14 SD. CE determination of tMPA is a robust, sensitive and reproducible method with the advantage over HPLC of being fast, simple and unexpensive, also enabling quick assessment of MPA for pharmacokinetic studies.     

HPLC法测定头孢呋辛酯中高分子聚合物含量

目的考察高效液相测定法用于头孢呋辛酯中高分子聚合物测定的可行性。方法色谱柱:聚苯乙烯-二乙烯苯共聚物的填充色谱柱,检测波长:280 nm,进样量:20μL;流动相:0.032 mol/L Li Br溶液;流量为0.30 m L·min-1。结果头孢呋辛酯色谱条件经方法学考察均符合要求,在0.005 4~1.532 2 mg·m L-1范围内,溶液质量浓度与聚合物峰面积呈线性相关,回归方程为:y=64 217 x-41 214(r=0.999 6)。结论头孢呋辛酯中高分子聚合物浓度可用高效液相色谱的方法测定,所采用的方法准确可靠,可考虑用于头孢呋辛酯及其类似物的高分子聚合物测定。     

An HPLC method for the determination of ethacridine lactate in human urine

An HPLC method was developed and validated for the determination of ethacridine lactate in human urine. Solid-phase extraction cartridges were used to extract urine samples. Separation was carried out on a C(18) column maintained at 30 degrees C with methanol-0.05% sodium dodecylsulfonate (70:30, v/v, pH 3) as mobile phase at a flow rate of 1.0 mL/min. Detection was at UV 272 nm. The calibration curve was linear in the concentration range of 4-4000 ng/mL, with linear correlation coefficient r equal to 0.9998. The limit of detection for the assay was 1.1 ng/mL. The within-day accuracy ranged from 94.8 to 101.6% and precision from 2.3 to 5.4%. The between-day accuracy ranged from 96.8 to 102.6% and precision from 4.0 to 5.3%. The absolute recovery was 95.4-101.2%. Urine samples were stable for at least 15 days if stored in the dark at -20 degrees C. This simple and accurate method allows the sensitive determination of ethacridine lactate in human urine. It was successfully applied to assess the urine level of ethacridine lactate in women received intra-amniotic injection.     

Enantioselective determination of arotinolol in human plasma by HPLC using teicoplanin chiral stationary phase

A sensitive enantioselective high-performance liquid chromatography (HPLC) method was developed and validated to determine S-(+)- and R-(-)-arotinolol in human plasma. Baseline resolution was achieved by using teicoplanin macrocyclic antibiotic chiral stationary phase (CSP) known as Chirobiotic T with a polar organic mobile phase consisting of methanol:glacial acetic acid:triethylamine, 100:0.1:0.1, (v/v/v) at a fl ow rate of 0.8 mL/min and UV detection set at 317 nm. Human plasma was spiked with stock solution of arotinolol enantiomers and labetalol as the internal standard. The assay involved the use of liquid-liquid extraction procedure with ethyl ether under alkaline condition for human plasma sample prior to HPLC analysis. Recoveries for S-(+)- and R-(-)-arotinolol enantiomers were in the range 93-103% at 200-1400 ng/mL level. Intra-day and inter-day precision calculated as %RSD was in the ranges 1.3-3.4 and 1.9-4.5% for both enantiomers, respectively. Intra-day and inter-day accuracies calculated as percentage error were in the ranges 1.2-3.5 and 1.5-6.2% for both enantiomers, respectively. Linear calibration curves in the concentration range 100-1500 ng/mL for each enantiomer showed a correlation coefficient (r) of 0.9998. The limit of quantitation (LOQ) and limit of detection (LOD) for each enantiomer in human plasma were 100 and 50 ng/mL (S/N = 3), respectively.