LC-MS/MS法测定Beagle犬血浆中美金刚的质量浓度

目的建立LC-MS/MS法测定Beagle犬血浆中美金刚的质量浓度,并将此方法应用于盐酸美金刚片在Beagle犬体内的药动学研究。方法采用蛋白沉淀法处理血浆样品,以金刚烷胺为内标物质(IS),采用Grace Altima HP C_(18)(50 mm×2.1 mm,5μm)色谱柱,流动相为体积分数0.1%甲酸水溶液-体积分数0.1%甲酸乙腈溶液,梯度洗脱,流速为0.35 m L·min~(-1)。离子源为电喷雾电离(ESI)源,采用正离子化方式测定,采用多反应监测(multiple reaction monitoring,MRM)模式检测,用于定量分析的离子反应为m/z 179.8→m/z 162.8(美金刚),m/z 152.6→m/z 135.7(金刚烷胺,内标物质)。4只健康雄性Beagle犬单次经口给予盐酸美金刚片25 mg,用Win Nonlin 6.1软件以非房室模型计算药动学参数。结果血浆中美金刚线性范围为0.001~0.2 mg·L~(-1),定量下限为1μg·L~(-1),日内和日间精密度在2.1%~10.8%内,准确度在-3.5%~2.9%内,提取回收率在94.9%~96.8%内。Beagle犬单次经口给予盐酸美金刚片后,主要的药动学参数t_(1/2)为(11.0±4.7)h,t_(max)为(9.0±2.0)h,ρ_(max)为(0.078±0.024)mg·L~(-1),AUC_(0-t)为(1.350±0.295)mg·h·L~(-1),AUC_(0-∞)为(1.396±0.306)mg·h·L~(-1)。结论所建立的定量分析方法符合生物样品分析方法指导原则要求,适用于盐酸美金刚片的药动学研究。     

LC-MS法快速测定尼群地平纳米结晶制剂在Beagle犬中的血药质量浓度

目的建立一种液相色谱质谱联用(LC-MS)方法,用于快速测定Beagle犬血浆中尼群地平的质量浓度。方法血浆样品采用液-液萃取法提取,色谱柱为Kinetex XB-C18柱(50 mm×2.1 mm,2.6μm),流动相为乙腈-体积分数0.2%甲酸溶液(体积比为70∶30)。质谱在ESI正离子模式下以多反应监测扫描方式检测。结果尼群地平的线性范围为1~500μg·L~(-1),日内和日间RSD均小于10%,准确度(RE)为-5.7%~0.1%,提取回收率大于90%,基质效应在86.11%~103.4%内。尼群地平在Beagle犬血浆中的主要药动学参数:tmax为(2.5±0.5)h;ρmax为(139.0±4.6)μg·L~(-1);t1/2为(7.1±0.9)h;AUC(0-48)为(891.4±157.1)μg·L~(-1)·h,AUC(0-∞)为(906.9±152.1)μg·L~(-1)·h。结论该测定方法适用于尼群地平纳米结晶制剂在Beagle犬体内的药物动力学研究。     

LC-MS/MS法测定可待因在Beagle犬血浆中的质量浓度

目的建立Beagle犬血浆中可待因质量浓度的液相色谱-串联质谱(LC-MS/MS)测定方法。方法色谱柱为Agela·C18柱(150 mm×4.6 mm,5μm),流动相为甲醇-水-甲酸(体积比为15∶85∶0.5),血浆样品经甲醇沉淀蛋白处理,以多反应监测(multiple reaction monitoring,MRM)扫描方式检测,测定Beagle犬经口给予洛芬待因缓释片后血浆中可待因的质量浓度。结果血浆中可待因质量浓度在0.2²0μg·L-1内线性关系良好,日内和日间精密度RSD≤11.5%,平均提取回收率为104.1%20μg·L-1内线性关系良好,日内和日间精密度RSD≤11.5%,平均提取回收率为104.1%¹09.2%,基质效应为119.2%109.2%,基质效应为119.2%¹28.3%。可待因在Beagle犬血浆中主要药动学参数t1/2为(2.9±0.5)h,ρmax为(8.7±3.4)μg·L-1,AUC0-∞为(28.5±7.3)μg·h·L-1。结论该方法适用于可待因在Beagle犬体内药动学的研究。     

RP-HPLC法同时测定Beagle犬血浆中曲马多、文拉法辛和罗通定浓度

目的:建立同时测定 Beagle 犬血浆中曲马多、文拉法辛和罗通定浓度的反相高效液相色谱法。方法:Beagle 犬血浆标本以维拉帕米为内标,经碱化和石油醚-乙醚(7:3)提取后,以0.025 mol·L~(-1)磷酸二氢钠-甲醇(70:30)为流动相,流速1min·mL~(-1),经碳十八烷基键合硅胶柱(250 mm×4.6 mm,5μm)分离,采用荧光法于λ_(ex)=295 nm,λ_(em)=302 nm 检测。结果:曲马多标准曲线范围3.125～3200μg·L~(-1),Y=0.0051X 0.0473(r=0.9996),最低定量限为3.125μg·L~(-1),高、中、低3种浓度的日内精密度均<10%,日间精密度均<10%,方法回收率79.3%～111.2%;文拉法辛标准曲线范围6.25～1600μg·L~(-1),Y=0.0036X-0.0070(r=0.9998),最低定量限为6.25μg·L~(-1),高、中、低3种浓度的日内精密度均<7%,日间精密度均<6%,方法回收率92.5%～108.8%;罗通定标准曲线范围4.6875～2400μg·L~(-1),Y=0.0064X-0.0004(r=0.9999),最低定量限为4.6875μg·L~(-1),高、中、低3种浓度的日内精密度均<9%,日间精密度均<5%,方法回收率89.3%～112.1%。结论:本法简便、快速、准确,用于 Beagle 犬同时服用曲马多、文拉法辛和罗通定的药动学研究,取得满意结果。     

RP-HPLC法测定Beagle犬血浆中的烟酸浓度及药动学研究

目的建立反相高效液相色谱法测定犬血浆中烟酸的浓度,并应用到烟酸缓释片在Beagle犬体内的药动学研究。方法采用高氯酸沉淀蛋白处理血浆样本;色谱条件:Megres C18柱(4.6mm×250mm,5μm),流动相:甲醇-10mmol.L-1磷酸二氢钾缓冲溶液(3∶97,V/V),柱温:12℃,流速:0.8mL.min-1,检测波长:263nm;比格犬灌胃给药500mg烟酸缓释片。结果线性范围为0.200~35.0μg.mL-1(r≥0.9996),检测限为50ng.mL-1,方法回收率为98.4%;烟酸缓释片表现出明显的缓释效果。结论方法操作简单、专属灵敏、重现性好,适用于Beagle犬体内烟酸的药动学研究。     

UPLC-MS法测定Beagle犬血浆中丁咯地尔和主要代谢物及其在药代动力学研究中的应用

目的建立快速灵敏的超高效液相色谱-质谱联用(UPLC-MS)法,研究Beagle犬分别以9.6、19.2、28.8 mg·kg-1的剂量经口给药盐酸丁咯地尔后,丁咯地尔及主要代谢物O-去甲基丁咯地尔在健康Beagle犬体内的药物代谢动力学行为。方法采用甲醇沉淀蛋白法进行血浆样品预处理,Acquity UPLC BEH C18(50 mm×2.1 mm,1.7μm)色谱柱,甲醇-5 mmol·L~(-1)醋酸铵(含体积分数为0.05%的甲酸)为流动相,梯度洗脱。采用ESI源正离子模式对血浆样品中丁咯地尔及其代谢物进行浓度检测,计算药代动力学参数。结果血浆中丁咯地尔和O-去甲基丁咯地尔的质量浓度分别在20.0~4 000.0μg·L~(-1)和2.0~400.0μg·L~(-1)内线性关系良好,日内、日间精密度RSD均不超过10.4%,提取回收率为90.4%~96.3%,基质效应为93.2%~97.0%。丁咯地尔及O-去甲基丁咯地尔血浆样品在所考察的储存条件下均可保持稳定。结论该方法适用于丁咯地尔及O-去甲基丁咯地尔在Beagle犬体内的药物代谢动力学研究。Beagle犬给药9.6~28.8 mg·kg-1剂量内,丁咯地尔药物代谢动力学行为呈线性关系,O-去甲基丁咯地尔药动学行为呈非线性关系。     

LC-MS/MS法测定Beagle犬血浆中阿戈美拉汀的质量浓度

目的建立LC-MS/MS的方法测定Beagle犬血浆中阿戈美拉汀的质量浓度,并应用此方法研究阿戈美拉汀片在Beagle犬体内的药代动力学行为及其相对生物利用度。方法采用:Agela C18色谱柱(150 mm×4.6 mm,5μm I.D),以乙腈-浓度5 mmol·L-1醋酸铵水溶液-甲酸(体积比为58.0∶42.0∶0.1)为流动相,采用沉淀蛋白法,以多反应离子监测(multiple reaction monitoring,MRM)扫描方法进行检测,测定经口给予阿戈美拉汀片后Beagle犬血浆中阿戈美拉汀的质量浓度。结果 Beagle犬血浆中阿戈美拉汀在质量浓度0.50~100μg·L-1内与峰面积呈良好线性关系,相关系数r为0.999 0;日内和日间精密度RSD≤8.6%;阿戈美拉汀的平均提取回收率为90.2%~93.3%,基质效应为104.9%~106.8%;阿戈美拉汀在Beagle犬血浆中主要药动学参数t1/2为(4.2±1.5)h,ρmax为(149±85)μg·L-1,AUC0-24为(293±78)μg·h·L-1,AUC0-∞为(300±78)μg·h·L-1。结论该方法适用于阿戈美拉汀在Beagle犬体内的药代动力学及相对生物利用度的研究。     

P91024在Beagle犬体内的药物代谢动力学

目的:建立测定 Beagle 犬血浆中 P91024浓度的 HPLC 法并进行 Beagle 犬体内的药物动力学研究。方法:取犬血浆0.5mL,加甲醇0.5 mL,涡旋1 min,加2 mol·L~(-1)碳酸钠溶液50 μL,涡旋1 min,加环己烷提取血浆样品,氮气流吹干后用甲醇溶解残渣,进行 HPLC 分析。色谱柱为 Shim-pack VP-ODS 分析柱(150 mm×4.6 mm,5μm),Shim-pack GVP-ODS 预柱(10mm×4.6 mm);流动相为甲醇-水(87∶13);流速:1 mL·min~(-1);柱温:30℃;测定波长:231 nm,外标法定量。6条 Beagle 犬灌胃给予 P91024,计算主要药动学参数。结果:在10.0～4000.0 ng·mL~(-1)范围内,P91024峰面积与浓度呈良好线性关系(r=0.9998),最低定量浓度为10 ng·mL~(-1),该法的提取回收率为90.4%～93.6%(n=5),方法回收率为98.2%～104.2%(n=5),日内精密度和日间精密度分别为4.0%～6.6%和6.1%～8.4%。Beagle 犬灌服 P91024 20 mg·mL~(-1)后,其主要药物动力学参数为 C_(max)=1493.5 ng·mL~(-1),T_(max)=3.33 h,t_(1/2β)=11.68 h,MRT=8.58 h,AUC_(0-∞)=9305.1 ng·h·mL~(-1)。结论:本法灵敏、准确,适用于 P91024的血药浓度测定及药物动力学研究。     

米索前列醇片在健康人体内的药动学特征

目的建立人血浆中米索前列醇浓度的LC-MS/MS检测方法,研究米索前列醇片在健康志愿者体内的药动学特征。方法采用K-MS/MS C_(18)分析色谱柱(150 mm×4.6 mm,5μm);流动相为甲醇-水-1%氨水(55:45:0.075,V/V/V);流速为0.5 mL·min~(-1);柱温为室温。结果该测定方法的线性范围为10～3000 ng·L~(-1)(r=0.9964),最低检测浓度为10.0 ng·L~(-1)(按R_(S,N)≥3计),提取回收率为80%～90%,日内和日间RSD均<10%。药动学研究表明,米索前列醇片在体内代谢过程符合一室模型,主要药动学参数t_(1/2)、ρ_(max)、t_(max)、AUC_(0-360)分别为:(1.15±0.26)h,(1 209±973)ng·L~(-1),(32±13)min,(1 359±1 015)ng·h·L~(-1)。结论本法具有良好的准确性和较高的灵敏度,简便快速,适用于米索前列醇片血药浓度的测定和药动学研究。     

LC-MS/MS法测定Beagle犬血浆中妥洛特罗质量浓度及其在药动学研究中的应用

目的建立LC-MS/MS法测定Beagle犬血浆中妥洛特罗的质量浓度,研究Beagle犬给予妥洛特罗贴剂后的药动学特征。方法血浆经乙酸乙酯-二氯甲烷(体积比4∶1)提取。色谱柱为Agilent TC-C18柱,流动相为甲醇-体积分数1%甲酸溶液(体积比67∶33),流速为0.5 m L·min-1,进样量为30μL。质谱采用多反应监测模式(multiple reaction monitoring,MRM),电喷雾离子源,分析时间4.5 min。采用DAS 2.0软件以非房室模型计算药动学参数。结果妥洛特罗质量浓度在0.1~10μg·L-1内与峰面积呈良好的线性关系,定量下限为0.1μg·L-1。日内、日间精密度(RSD)均小于15%,准确度(RE)为-4.27%~-0.60%,方法回收率大于(82.0±6.35)%。健康Beagle犬给予妥洛特罗贴剂(规格为每贴2 mg)后主要药动学参数:tmax为(5.25±1.49)h,ρmax为(7.82±1.98)μg·L-1,t1/2为(5.59±2.37)h;采用梯形法计算,AUC0-t为(99.26±15.05)μg·h·L-1,AUC0-∞为(101.94±14.99)μg·h·L-1。结论该方法可用于妥洛特罗临床前药动学研究。     

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.     

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.     

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。     

Polymorphisms in genes involved in neurotransmission in relation to smoking

Smoking behavior is influenced by both genetic and environmental factors. The genetic contribution to smoking behavior is at least as great as its contribution to alcoholism. Much progress has been achieved in genomic research related to cigarette-smoking within recent years. Linkage studies indicate that there are several loci linked to smoking, and candidate genes that are related to neurotransmission have been examined. Possible associated genes include cytochrome P450 subfamily polypeptide 6 (CYP2A6), dopamine D₁, D₂, and D₄ receptors, dopamine transporter, and serotonin transporter genes. There are other important candidate genes but studies evaluating the link with smoking have not been reported. These include genes encoding the dopamine D₃ and D₅ receptors, serotonin receptors, tyrosine hydroxylase, trytophan 2,3-dioxygenase, opioid receptors, and cannabinoid receptors. Since smoking-related factors are extremely complex, studies of diverse populations and of many aspects of smoking behavior including initiation, maintenance, cessation, relapse, and influence of environmental factors are needed to identify smoking-associated genes. We now review genetic polymorphisms reported to be involved in neurotransmission in relation to smoking.     

Tramadol concentrations in blood and in cerebrospinal fluid in a neonate

Based on blood and cerebrospinal fluid samples collected in a full-term neonate, the penetration of tramadol in the central nervous system is described. Following intravenous administration of tramadol, a lag time of about 4 h was observed until full blood–brain equilibration was achieved. This pharmacokinetic observation is in line with a recent pharmacodynamic evaluation of the central opioid effects of tramadol in adults.     

Disease control in asthmatic children seen in private practice in Switzerland

ABSTRACT

Background: Asthma is the most common chronic childhood disease in Switzerland with a prevalence of 10%. Asthma has a high economic burden accounting for high medical costs. Assessment of disease control is likely to be of help in the implementation of strategies to improve asthma. Therefore, we aimed to evaluate asthma control and therapy regimens among children in private practice.

Methods: We assessed asthma control as well as therapy regimens in 575 asthmatic children in an experience programme in Switzerland by using an abbreviated questionnaire based on the asthma control questionnaire and the child health questionnaire on Visit 1 and Visit 2.

Results: Good asthma control at Visit 1 was only present in 25.7% of asthmatic children. Occasional asthma symptoms, limitation of physical activity, nocturnal awakening and anxiety of the parent was present in 80.5%, 41.2%, 46.8% and 57% of the children, respectively. After adjustment of therapy regimens at Visit 1, mainly by adding a leukotriene receptor antagonist, asthma control was reported to be much better in 53.4% of the children at Visit 2.

Conclusions: As asthma control is inadequately achieved within a major portion of asthmatic children, it is imperative to find measures to improve asthma control and hence, to reduce the burden of disease.