气相色谱氮磷检测器法测定人血浆毒鼠强浓度

目的建立测定人体血浆中毒鼠强(TET)浓度的毛细管气相色谱/氮磷检测器(GC/NPD)分析法。方法血浆中TET用乙酸乙酯作液-液提取,以乙酸乙酯为溶媒,载气为氦气,色谱柱为HP-5弹性石英毛细管柱,GC/NPD测定毒鼠强的浓度。结果在100~500 ng.mL-1浓度范围内线性关系良好,r=0.997 2(n=5),最低检测浓度为100ng.mL-1。TET血浆样品的低、中、高3个浓度(100,200和400 ng.mL-1)日内、日间变异系数(RSD)均≤10%,方法回收率均≥80%。结论该法操作简便,结果准确,可用于TET中毒的实验室鉴别和临床救治过程中的体内毒物浓度监测。     

中空膜液相微萃取-HPLC法用于人血浆中水杨酸的测定

建立灵敏、准确地测定人血浆中水杨酸浓度的HPLC法。样品经过酸化处理后,以甲苯-正辛醇(4:6,V/V)为萃取溶剂、NaOH水溶液(pH=10)为接收相进行中空纤维膜微萃取。在该条件下,血浆中水杨酸浓度在2~250 ng.mL-1范围内,线性关系良好(r=0.9994);检测限为0.7ng.mL-1(S/N=3);精密度(RSD)<8%(n=5);回收率为87.6%~102.0%。该方法灵敏、有效、所需有机溶剂少、富集效率高,适于血浆中水杨酸浓度的测定。     

电感耦合等离子体质-谱法测定人血浆中铋的浓度

目的 建立电感耦合等离子体质谱(ICP-MS)法测定人血浆中铋的浓度.方法 血浆样品经硝酸硝化处理后,用ICP-MS进行检测,检测元素:Bi、Ti(内标),RF功率:1 550 W,等离子体气流速:15 L· min-1,反应气模式:关,样本深度:7.5 mm,雾化室温度:2℃.结果 血浆中铋在0.312～20 ng·mL-1线性关系良好(r=0.999 8),最低定量限为0.312 ng·mL-1,绝对回收率为92.7％～97.5％,日内及日间精密度RSD均＜6.2％.健康志愿者单次给予复方雷尼替丁胶囊2粒(含铋220mg)后铋吸收迅速,平均峰浓度均＜50 ng·mL-1.结论 硝酸硝化前处理及ICP-MS测定人血浆中铋的方法灵敏、准确、简便,能满足生物样品分析的要求,可用于铋体内过程研究.     

高效液相色谱法测定防己药材中粉防己碱与防己诺林碱的含量

目的用高效液相色谱法测定不同产地防己药材中粉防己碱与防己诺林碱的含量。方法色谱柱为Kromasil ODS-1（4．6mm×200mm，5μm），流动相为乙腈0．01％三乙胺水溶液，梯度洗脱，流速为1．0mL·min^-1，检测波长为280nm。结果粉防己碱在474-4740ng内线性关系良好，r=0．9999，平均回收率为99．3％，RSD=1．5％；防己诺林碱在238-2380ng线性关系良好，r=0．9998，平均回收率为99．5％，RSD=1．1％。结论建立了高效液相色谱法同时测定防己药材中粉防己碱与防己诺林碱的含量，比较了不同产地防己药材中粉防己碱与防己诺林碱的含量。     

HPLC法测定替米沙坦血药浓度的方法学考察

目的 建立HPLC法测定人血浆中替米沙坦浓度的分析方法.方法 色谱柱采用Agilent ZORBAX SB-C18(150mm×4.6mm,5μm),流动相为乙腈:0.03 mol· L-1 KH2PO4溶液(47∶55,v/v),流速为1.0mL·min-1,荧光检测激发波长为306nm,发射波长为374nm.结果 线性范围为5～1500ng·mL-1,回归方程为Y=0.01852X-0.01655,r=0.9997(n=5);最低定量下限(LLOD)为5ng·mL-1.结论 本方法准确、简便,重现性好,适用于替米沙坦的临床药动学研究.     

HPLC荧光法测定人血浆中的5-氨基水杨酸

目的 采用HPLC荧光法测定人血浆中的5-氨基水杨酸,研究单剂量口服柳氮磺吡啶肠溶片后5-氨基水杨酸在中国健康受试者体内的药动学特征.方法 24名健康受试者口服500 mg柳氮磺吡啶肠溶片,采用HPLC荧光法测定血浆中5-氨基水杨酸的浓度,利用DAS 2.1.1软件计算药动学参数.结果 5.055～808.8 ng·mL-1 5-氨基水杨酸与峰面积比值的线性关系良好(r =0.9985),最低检测浓度为5.055 ng· mL-1,日内、日间RSD均＜10％.单次口服给药后,药动学参数为:t1/2=11.19 _±2.48 h,Cmax=232.83±57.94 ng· mL-1,Tmax =17.58 ±5.33 h,AUC(0～t)=5.5907±1.7848 μg·h·mL-1.结论 所用方法准确、灵敏度高、重复性好,可用于口服柳氮磺吡啶肠溶片后5-氨基水杨酸血药浓度测定及人体药动学研究.     

HPLC测定血浆中的游离肉碱

目的 建立血浆中游离肉碱的测定方法.方法 用高效液相色谱(HPLC)测定各患者的血浆游离肉碱,并进行比较.结果 方法线性范围为2 ～ 32μg·mL-1;最低检测浓度为2μg·mL-1,总分析时间为9.248 min.肉碱的平均保留时间(8.467±0.042) min.高、中、低3种浓度的平均回收率为98.7％.批内批间RSD分别为4.8％和10.9％(n=5).结论 HPLC法测定血浆游离肉碱浓度灵敏、可靠、简便,适用于快速检测肉碱缺乏症病人的临床筛选工作.     

灵敏专属的HPLC法测定比格犬中非索非那定的浓度

目的建立简便的测定血浆中非索非那定血药浓度的高效液相色谱法.方法流动相为乙腈∶0.02磷酸缓冲液(35∶65,V∶V),流速为1.0 mL·min-1.柱温40 ± 1 ℃,检测波长210 nm.结果标准曲线的线性范围在10～1000 ng·mL-1,r = 0.999 4.最低检测限为3 ng·mL-1.高(800 ng·mL-1)、中(400 ng·mL-1)、低(10 ng·mL-1) 浓度提取回收率分别为85.2%±4.2%, 88.0%±3.9%和89.7%±3.5%;日间和日内RSD分别低于6.1%和4.2%.结论所建立的方法灵敏、快速、重现性好、选择性强.     

高效液相色谱法测定大鼠血浆内厄洛替尼的浓度及其药动学研究

目的 建立以高效液相色谱法测定大鼠血浆中厄洛替尼浓度的方法,并研究其在大鼠体内的药物动力学.方法 色谱柱为Dikma Diamonsil C18,流动相为乙腈-水=60∶40(pH=2.0),流速为1.2 mL· min-1,柱温为35℃;检测波长331nm.结果 厄洛替尼血药浓度在80～4 000 ng·mL-1与峰面积线性关系良好(r=0.997 7),最低检测限为80 ng·mL-1;日内、日间RSD均≤10％,提取回收率在83.77％～85.68％.6只SD大鼠单剂量静脉给予厄洛替尼(10 mg·kg-1)后药动学参数分别为:Cmax=(1.203×103士118.2)ng·mL-1; t1/2=(23.24±2.33)h;AUC0-r=(4.713×103士213.6) ng· h· mL-1;AUC0-∞=(5.363×103±323.6) ng· h· mL-1;MRT=(25.91±2.34)h;Vd=(28.44±1.58)L.结论 本方法简便、准确、灵敏、专属性强,同样适用于人血浆中厄洛替尼浓度的测定及其药动学研究,对于评价厄洛替尼疗效和安全性有重要意义.     

高效液相色谱法测定人血浆中阿托伐他汀浓度

目的 建立高效液相色谱法（HPLC）测定人血浆中阿托伐他汀的浓度.方法 采用Nova-pak C18色谱柱（3.9×150mm,4μm）;流动相为甲醇-0.02 mol·L-1,醋酸铵缓冲液（63：27,V/V）;流速为0.9mL·min-1;检测波长245nm;柱温为30℃;血浆样品用乙酸乙酯提取浓缩后分析,黄体酮为内标.结果 阿托伐他汀的血药浓度在1.0～535.0ng·mL-1范围内回归方程为y=0.1366x＋0.0484（r=0.9992）,检测限为0.5ng·mL-1 （S/N≥3）.阿托伐他汀在2.0、201.1、535.0ng·mL-1三种浓度水平的平均回收率分别为100.8%、101.1%、99.5%,日内和日间RSD均小于2.80%.结论 本法准确、快速、稳定,适用于阿托伐他汀的血药浓度检测.     

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。     

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.     

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.     

Hyperkalaemia in patients in hospital

A survey of all laboratory blood specimens with a plasma potassium concentration greater than or equal to 5.5 mmol/L was conducted over a three month period. Of 331 specimens with hyperkalaemia, 71 were excluded because the specimens was haemolysed, old or contaminated. The laboratory served a population of 348,561 and during this time measured the plasma potassium on 25,016 occasions. Sixty-six outpatients and 20 neonates were not evaluated. The survey was undertaken on 86 of 102 inpatients (46 males), 48 of whom were over 66 years of age. Fifty-seven patients were admitted under a medical service and 29 under a surgical service. Fifty-nine had a single episode of hyperkalaemia. Thirty-two underwent a surgical procedure. The commonest contributing factor was impaired renal function which was present in 71 (83%) patients. Although a definitive causative role for drugs could be identified in only five patients, in 52 (60%) patients drugs were a contributing factor (potassium supplements 24, ACE inhibitors 16, nonsteroidal antiinflammatory drugs 12). Thirty-five of the 86 (41%) patients died during their hospital admission. Nineteen of the 35 deaths occurred within three days of the hyperkalaemia being recorded. A normal plasma potassium was eventually documented in 50 of the 86 patients. Of the remaining 36 patients, 25 (69%) subsequently died. In general the treatment of patients with hyperkalaemia focused on identifying and treating the underlying cause. Hyperkalaemia must always be considered seriously and regard given to the overall clinical status of the patient, with particular attention to drug therapy, renal and cardiac function, acid base status and the possibility of sepsis.