高效液相色谱法同时测定人血浆中对乙酰氨基酚、伪麻黄碱和咖啡因

目的:建立一种同时测定人血浆中对乙酰氨基酚、伪麻黄碱和咖啡因的高效液相色谱(HPIC)法,并用于含上述组分的复方制荆的人体药动学研究.方法:以荼碱为内标,血样经醋酸乙酯提取后,采用高效液相色谱紫外(HPLC_UV)法进行测定.色谱柱为Diamonsil C<,18>柱(4.6 mm x 150mm,5μm);流动相为甲醇-0.05 mo·L-1磷酸二氢钾(23:77,pH 2.4);流速1 mL·min-1.检测波长210 nm.结果:人血浆中对乙酰氨基酚、盐酸伪麻黄碱和咖啡因质量浓度测定的线性范围分别为0.12～11.52 mg·L-1,0.008～0.432mg·L-1和0.03～2.16mg·L-1;最低可定量质量浓度分别为0.12,0.008,0.03 mg·L-1;各组分日内、日间RSD均小于15%,方法回收率均大于88%.结论:该方法能快速可靠地同时测定人血浆中对乙酰氨基酚、伪麻黄碱和咖啡因的浓度.可用于含上述组分的复方制剂的人体药动学或生物等效性研究.     

高效液相色谱法同时测定止痒酊中三组成分的含量

目的:建立高效液相色谱(HPLC)法同时测定止痒酊中间苯二酚、水杨酸、苯酚的含量.方法:采用高效液相色谱法,色谱柱为Lichrospher C18柱,流动相为甲醇-水(48∶52),流速1.0 mL·min-1, 检测波长为270 nm.结果:线性范围:间苯二酚40～200 mg·L-1,水杨酸20～100 mg·L-1,苯酚20～100 mg·L-1;日内、日间RSD在0.46%～2.58%,高、中、低浓度回收率在96.83%～104.0%.结论:本方法简便,快速,准确,可用于止痒酊的质量控制.     

HPLC法测定石柱参药材及其片剂中5种人参皂苷的含量

目的建立测定石柱参药材及其片剂中5种人参皂苷含量的方法,为石柱参的质量控制提供科学依据。方法色谱柱为Kromasil C18柱;流动相为乙腈-水,梯度洗脱;柱温为25℃;流速为1.0 mL.min-1;检测波长为203 nm。结果5种人参皂苷的色谱峰与相邻色谱峰分离良好。人参皂苷Rg1质量浓度在19.8~198 mg.L-1内、人参皂苷Re质量浓度在20.6~206 mg.L-1内、人参皂苷Rb1质量浓度在33.0~330 mg.L-1内、人参皂苷Rc质量浓度在18.0~180 mg.L-1内、人参皂苷Rb2质量浓度在13.0~130 mg.L-1内与峰面积呈良好的线性关系。石柱参药材中人参皂苷Rg1、Re、Rb1、Rc、Rb2的平均回收率分别为99.8%、98.3%、99.2%、95.4%、96.8%,RSD分别为3.0%、3.0%、2.6%、2.2%、2.8%(n=6);石柱参片剂中人参皂苷Rg1、Re、Rb1、Rc、Rb2的平均回收率分别为100.2%、99.0%、99.7%、96.4%、98.4%,RSD分别为2.2%、3.1%、3.6%、2.6%、2.8%(n=6)。结论本实验中所建立的方法可用于石柱参药材及其片剂的质量控制。     

SPE-HPLC法测定人血浆中头孢吡肟的浓度

目的:建立测定人血浆中头孢吡肟浓度的方法.方法:采用固相萃取(SPE)血浆中头孢吡肟.色谱柱为大连依利特Hypersil BDS C18,流动相为甲醇-磷酸盐缓冲液(20:80),柱温为30℃,流速为1 ml·min-1,检测波长为257 nm,进样量20 μl.结果:头孢吡肟检测浓度线性范围为1.0～200 mg·L-1(r＝0.999 9);最低检测限为1 mg·L-1;低、中、高3种浓度头孢吡肟的方法回收率分别为118.0%,97.0%,95.0%,日内RSD分别为1.7%,3.6%,2.1%,日间RSD分别为8.2%,4.9%,2.6%.结论:该方法简便、灵敏度高,适用于测定人血浆中头孢吡肟的含量.     

反相高效液相色谱法检测人血浆特比萘芬浓度

目的:建立人血浆特比萘芬检测的高效液相色谱方法.方法:以ZORBAX Eclipse XDB-C18为色谱柱;流动相为乙腈-0.2%三氟乙酸-水体系,流速为 0.8 ml·min-1;检测波长258 nm(0～5.2 min) 和283 nm(5.2～7.0 min).以伏立康唑为内标,血浆在酸性条件下经乙酸乙酯萃取后检测.结果:特比萘芬浓度在0.05～4.00 mg·L-1范围内线性关系良好(r=0.999 7);高中低三个浓度(0.10、1.00、3.00 mg·L-1)的日内精密度RSD分别为3.12%、2.85%和2.63%,日间精密度RSD分别为4.18%、3.29%和2.81%;相对回收率分别为(101.47±4.53)%、(99.50±3.63)%和(100.71±3.33)%,RSD分别为2.60%、2.41%和1.62%.健康志愿者单剂量口服250 mg 盐酸特比萘芬后,Cmax为(1.27±0.15)mg·L-1,Tmax为(1.56±0.32)h,t1/2α为(0.96±0.24)h,t1/2β为(16.73±1.87)h,AUC(0-48)为(13.44±1.95)mg·h·L-1,AUC(0-∞)为(15.32±2.16)mg·h·L-1.结论:该方法简便、快速、准确可靠,适用于人血浆特比萘芬浓度的测定及其药代动力学研究.     

高效液相色谱荧光检测法测定血浆加巴喷丁含量

目的:建立测定血浆加巴喷丁浓度的高效液相色谱法.方法:待测血浆中加入内标替米沙坦后,用乙腈沉淀蛋白,邻苯二甲醛衍生化,进行高效液相荧光检测.色谱柱为Symmetry C18柱,流动相为0.05mol·L-1磷酸二氢钾溶液-乙腈(57:43),pH 3.6.流速0.8 ml·min-1,激发波长330 nm,发射波长440 nm.结果:线性范围为0.027～16.35 mg·L-,r=0.997 3,日内、日间RSD均小于10%,低、中、高三种浓度的平均回收率分别为91.9%,99.2%,96.0%,最低检测限为0.027 mg·L-1.结论:本法精密、准确,适用于临床上血药浓度监测、人体药动学和生物利用度研究.     

高效液相色谱法测定盐酸平阳霉素软膏的血药浓度

目的:建立人血浆中盐酸平阳霉素的HPLC测定方法,从而研究患者使用盐酸平阳霉素软膏后的透皮吸收程度.方法:500μL人血浆用150μL 10%的高氯酸沉淀后高速离心,取上清液进样20μL进行分析.流动相为甲醇-乙腈-0.002 mol·L-1戊烷磺酸钠(15:10:75),用1 mol·L-1硫酸调pH 3.5;色谱柱:ODS-C18(150 mm×4.6 mm,5μm);检测波长:254 nm;流速:1.0 mL·min-1.10名志愿受试的寻常性银屑病患者使用盐酸平阳霉素软膏后,用HPLC法测定血浆中药物浓度.结果:本实验建立的血浆中盐酸平阳霉素HPLC测定方法,血浆中杂质不干扰样品的测定,药物在0.20～0.5 mg·L-1的范围内线性关系好;最低检测为0.20 mg·L-1;回收率高于80%;高、中浓度的日间和日内变异系数均小于10.0%;低浓度的日间和日内变异系数均小于15.0%,符合生物样品分析要求.10名患者用药后血浆中盐酸平阳霉素的药物浓度均低于最低检测浓度.结论:受试者对盐酸平阳霉素的吸收在0.20 mg·L-1以下,发生药物不良反应的可能性较小,本研究中10名志愿受试者亦未出现不良反应.     

LC-MS法测定人血浆中单硝酸异山梨酯

目的:建立高效液相色谱-质谱法测定人血浆中单硝酸异山梨酯浓度的方法.方法:以茶碱为内标,血浆样品经乙酸乙酯提取后,用Zorbax XDB-C18分析柱(150 mm×2.1 mm,5 μm),流动相为A:0.01%冰乙酸,流速为0.15 ml·min-1;B:甲醇,流速为0.05 ml·min-1.采用四极杆质谱检测器大气压电喷雾离子源,负离子检测,检测离子:单硝酸异山梨酯为m/z 250,内标为m/z 179.结果:血浆中单硝酸异山梨酯浓度在0.0125～1.2 mg·L-1范围内线性关系良好(r=0.999 6),最低检测浓度为0.0125 mg·L-1,平均提取回收率为86.0%,日内RSD＜8.9%,日间RSD＜4.8%.结论:该法简便、快速,灵敏度高,可用于单硝酸异山梨酯制剂的药动学研究.     

多波长UPLC法同时测定冠心丹参胶囊中三七皂苷R_1等5种有效成分的含量

目的利用多波长超高效液相色谱法,建立冠心丹参胶囊中三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1、丹酚酸B和丹参酮ⅡA5种有效成分的含量测定方法。方法采用ACQUITY UPLC HSS T3(100 mm×2.1 mm,1.8μm)色谱柱,柱温为30℃,以乙腈和体积分数为0.1%磷酸溶液为流动相,采用梯度洗脱,流速为0.4 m L·min-1,检测波长为203 nm(三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1)、286 nm(丹酚酸B)和270 nm(检测丹参酮ⅡA)。结果三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1、丹酚酸B和丹参酮ⅡA分别在质量浓度10~200 mg·L-1(r=0.999 7)、20~800 mg·L-1(r=0.999 3)、20~800 mg·L-1(r=1.000 0)、20~800 mg·L-1(r=1.000 0)和1~50 mg·L-1(r=0.999 5)内与峰面积呈良好线性关系;加样回收率(n=6):三七皂苷R1为95.0%(RSD=1.2%)、人参皂苷Rg1为105.4%(RSD=1.9%)、人参皂苷Rb1为98.8%(RSD=0.9%)、丹酚酸B为97.4%(RSD=2.6%)、丹参酮ⅡA为104.9%(RSD=4.8%)。结论本方法简便可行,为冠心丹参胶囊质量控制提供了一种可靠的检测方法。     

HPLC法检测大鼠血浆中吡非尼酮的浓度及其药动学研究

目的:建立大鼠血浆吡非尼酮检测的高效液相色谱方法,研究吡非尼酮的药代动力学。方法:血浆经乙酸乙酯,采用ZORBAX SB-C18色谱柱;流动相为乙腈-水-0.1%TFA(三氟乙酸),流速为1.0 mL·min-1;检测波长为318(0～8 min)、246nm(8～10 min)。6只雄性大鼠单剂量灌胃给予15 mg.kg-1吡非尼酮,分别在给药后多点尾静脉采血;用该方法检测血浆中吡非尼酮的浓度。用DAS(数据采集系统)计算药代动力学参数。结果:吡非尼酮浓度在0.025～4.00 mg·L-1范围内线性关系良好(r=0.999 9);定量下限为0.025 mg·L-1;低、中、高3个浓度的相对回收率分别为(100.40±4.66)%、(102.17±4.02)%和(101.12±2.89)%;日内RSD分别为4.6%,3.9%,2.9%,日间RSD分别为5.5%,4.8%,3.7%。大鼠吡非尼酮灌胃后,血浆吡非尼酮在0.39 h达到1.69 mg·L-1的峰值浓度,血浆半衰期为2.21 h。吡非尼酮在大鼠体内符合一级吸收的二室模型。结论:本方法准确可靠、简便快速,适用于大鼠血浆吡非尼酮浓度的测定及其药代动力学研究。     

Quantitative in vivo and in vitro sex differences in artemisinin metabolism in rat

1. The pharmacokinetics of the antimalarial compound artemisinin were compared in the male and female Sprague-Dawley rat after single dose i.v. (20 mg.kg) or i.p. (50 mg.kg) administration of an emulsion formulation. 2. Plasma clearance of artemisinin was 12.0 (95% confidence interval: 10.4, 13.0) l.h. kg in the male rat and 10.6 (95% CI: 7.5, 15.0) l.h. kg in the female rat suggesting high hepatic extraction in combination with erythrocyte uptake or clearance. Artemisinin half-life was 0.5 h after both routes of administration in both sexes. Values for plasma clearance and half-lives did not statistically differ between the sexes. 3. After i.p. administration artemisinin AUCs were 2-fold higher in the female compared with male rat (p 0.001). Artemisinin disappearance was 3.9-fold greater in microsomes from male compared with female livers and it was inhibited in male microsomes by goat or rabbit serum containing antibodies against CYP2C11 and CYP3A2 but not CYP2B1 or CYP2E1. 4. The unbound fraction of artemisinin in plasma was lower (p 0.001) in plasma obtained from the male (8.8 2.0%) compared with the female rat (11.7 2.2%). 5. The possibility of a marked sex difference, dependent on the route of administration, has to be taken into account in the design and interpretation of toxicological studies of artemisinin in this species.     

Quantitative in vivo and in vitro sex differences in artemisinin metabolism in rat

1. The pharmacokinetics of the antimalarial compound artemisinin were compared in the male and female Sprague-Dawley rat after single dose i.v. (20 mg x kg(-1)) or i.p. (50 mg x kg(-1)) administration of an emulsion formulation. 2. Plasma clearance of artemisinin was 12.0 (95% confidence interval: 10.4, 13.0) 1 x h(-1) x kg(-1) in the male rat and 10.6 (95% CI: 7.5, 15.0) 1 x h(-1) x kg(-1) in the female rat suggesting high hepatic extraction in combination with erythrocyte uptake or clearance. Artemisinin half-life was approximately 0.5 h after both routes of administration in both sexes. Values for plasma clearance and half-lives did not statistically differ between the sexes. 3. After i.p. administration artemisinin AUCs were 2-fold higher in the female compared with male rat (p < 0.001). Artemisinin disappearance was 3.9-fold greater in microsomes from male compared with female livers and it was inhibited in male microsomes by goat or rabbit serum containing antibodies against CYP2C11 and CYP3A2 but not CYP2B1 or CYP2E1. 4. The unbound fraction of artemisinin in plasma was lower (p < 0.001) in plasma obtained from the male (8.8 +/- 2.0%) compared with the female rat (11.7 +/- 2.2%). 5. The possibility of a marked sex difference, dependent on the route of administration, has to be taken into account in the design and interpretation of toxicological studies of artemisinin in this species.     

Interindividual variability in metabolic activation in humans in vivo

In assessing interindividual variability in metabolic activation, the toxic metabolite is often too unstable for conventional analysis. Possible alternatives include a stable product of the reactive metabolite e.g. cysteinyl derivatives of N-acetyl-4-benzoquinoneimine, the toxic metabolite of paracetamol, adducts with DNA or protein, and indirect measurement of the activity of the enzyme(s) producing the active metabolite. An example of the last approach is the use of furafylline, a highly specific inhibitor of human CYP1A2, to determine the extent of the metabolic activation of the cooked food mutagens PhIP and MeIQx. The extent of inhibition, determined from levels of unchanged amine in urine, is an indirect measure of the activity of the activation pathway. Further refinement of this approach, allied to improved measures of the biological process of interest should prove of value in evaluating interindividual variability and its role in the risk assessment process.     

Theophylline kinetics in peripheral tissues in vivo in humans

Several biochemical and cellular effects have been described for methylxanthines under in vitro conditions. However, it is unknown, whether threshold concentrations required to exert these effects are attained in target tissues in vivo. We therefore employed the microdialysis technique for measuring theophylline concentrations in peripheral tissues under in vivo conditions.Following in vitro and in vivo calibration, microdialysis probes were inserted into the medial vastus muscle and into the periumbilical subcutaneous adipose layer of healthy volunteers. Following single oral dose administration of 300 mg or i.v. infusion of 240 mg theophylline, in vivo time courses of theophylline concentrations were monitored in tissues and plasma. Major pharmacokinetic parameters (c_max, t_max, AUC) were calculated for plasma and tissue time courses. The mean AUC_tissue /AUC_plasma-ratio was 0.56 (p.o.) and 0.55 (i.v.) for muscle and 0.55 (p.o.) and 0.72 (i.v.) for subcutaneous adipose tissue.We conclude that microdialysis provides important information on the distribution and the tissue pharmacokinetics of theophylline.Abbreviations FPIA Fluorescence polarisation immuno assay - AUC Area under the curve - t_max Time to peak concentration - c_max Peak concentration     

休克时血中氧自由基的变化

本实验测定10名休克患者血浆和红细胞的丙二醛(MDA)、血浆总抗的氧化活性(AOA)的含量。结果表明:休克病人红细胞膜和血浆 MDA 含量(4.298±0.722;5.348±0.834)与对照组(3.235±0.682;4.356±1.081)比较明显增高(P<0.05);血浆 AOA(39.65±7.858)与对照组(48.21±10.81)比较明显降低(P<0.01)。提示:休克时,患者机体内自由基反应增强是引起组织细胞损伤的原因之一。     

Changes in angiopoietin expression in glomeruli involved in glomerulosclerosis in rats with daunorubicin-induced nephrosis

AIM: To study the potential pathological role of endogenous angiopoietins in daunorubicin-induced progressive glomerulosclerosis in rats. METHODS: Seventy male Wistar rats were allocated randomly into a daunorubicin group (DRB; n=40) or a control group (n=30). The rats in the DRB group were injected with DRB (15 mg/kg), in their tails. Subsequently, at intervals of 1, 2, 4, 6, 8, and 12 weeks, 5 male Wistar rats in each group were chosen randomly for 24 h urinary protein quantitative measurements (24 h UPQM), and determination of plasma tumor necrosis factor alpha (TNF-alpha), angiopoietin-1 (Ang1), and angiopoietin-2 (Ang2) levels. Kidney sections were examined by electron microscopy, Periodic Acid Schiff (PAS) staining, immunohistochemical staining and in situ hybridization histochemistry. RESULTS: As glomerulosclerosis progressed in the DRB group, expression of Ang1 mRNA and protein in glomeruli decreased and expression of TNF-alpha protein, Ang2 mRNA and protein in glomeruli increased. Expression of Ang1 mRNA and protein in glomeruli were negatively correlated with 24 h UPQM, Fn protein expression, and mean area of extracellular matrix (MAECM). In comparison, expression of Ang2 mRNA and protein in glomeruli were positively correlated with 24 h UPQM, Fn protein expression and MAECM; furthermore, there was a positive correlation between plasma Ang2 and 24 h UPQM. Plasma TNF-alpha and expression of TNF-alpha in glomeruli were positively correlated with expression of Ang2 mRNA and protein in glomeruli. There was a negative correlation between Ang1 protein expression and Ang2 protein expression in glomeruli. CONCLUSION: During DRB-induced glomerulosclerosis, podocyte injury led to a shift in the balance of Ang1 and Ang2 in glomeruli. Increased TNF-alpha in plasma and glomeruli may upregulate Ang2 expression in glomeruli. Elevated Ang2 in both plasma and glomeruli may mediate protein permeability through the glomerular filtration barrier. Moreover, local expression of Ang2 may facilitate the progress of glomerulosclerosis by upregulating a component expression of extracellular matrix.     

Trichinellosis in immigrants in Switzerland

We describe a case of trichinellosis diagnosed at the Division of Infectious Diseases, Hospital of Lugano, in January 2009. This case was associated with a cluster of cases and was traced to the consumption of contaminated meat after a wild boar hunt in Bosnia.