RP-HPLC法测定大鼠血浆中多烯紫杉醇的含量

目的建立大鼠血浆样品中多烯紫杉醇的液-液萃取RP-HPLC测定法。方法大鼠血浆样品经乙腈-氯代正丁烷(体积比为1∶4)提取,以地西泮(diazepam)为内标物,流动相采用水-甲醇-四氢呋喃-氨水(甲酸调pH)系统进行梯度洗脱,流速为1.0 mL.min-1,色谱柱为Diamonsil C18柱(250 mm×4.6 mm,5μm),UV检测波长为232 nm。结果多烯紫杉醇在50~10 000μg.L-1内线性关系良好,定量下限为50μg.L-1,提取回收率为70.2%~79.0%。结论HPLC法适用于大鼠血浆中多烯紫杉醇浓度的测定及药物动力学研究。     

高效液相色谱法测定石杉碱甲微球在大鼠体内的血药浓度及药物残留量

目的:建立石杉碱甲微球大鼠体内血药浓度及注射部位药物残留量的测定方法.方法:采用碱化后有机溶剂提取的方法处理样本,运用高效液相色谱法进行测定.色谱柱为Kromasil-C18(4.6 mm×250 mm,5μm),血样测定的流动相为乙腈-0.2%三乙胺水溶液(20:100),肌肉匀浆液样品测定的流动相为乙腈-0.2%三乙胺水溶液(25:75),检测波长为310 nm.结果:血浆样品测定的线性范围为2.5～500 μg·L-1,最低定量浓度为2.5μg·L-1.肌肉匀浆液样本测定的线性范围为1.008～201.6 mg·L-1,最低定量浓度为1.008 mg·L-1.两种方法的相对回收率均在85%～115%范围内,RSD小于12%.结论:该法可用于石杉碱甲微球体内缓释效果的评价.     

高效液相色谱法测定人血浆中霉酚酸及其代谢产物的浓度

目的建立高效液相色谱法建立测定人血浆中霉酚酸(MPA)、霉酚酸葡萄糖苷(MPAG)、霉酚酸酰基葡萄糖苷(Ac MPAG)的浓度。方法血浆处理采用乙腈蛋白沉淀法。色谱柱为Gemini C18(250mm×4.6 mm,5μm),柱温为40℃。Ac MPAG、MPA采用荧光检测,流动相:甲醇-10 mmol·L-1KH2PO4(43:57,p H=8.0),荧光激发波长343 nm,发射波长425 nm。MPAG采用紫外检测,流动相:甲醇-10 mmol·L-1 KH2PO4(45:55,p H=3.0),检测波长254 nm。结果 MPA、MPAG、Ac MPAG血药浓度分别在0.2~20μg·m L-1、4.75~475μg·m L-1、0.02~2μg·m L-1内与峰面积线性关系良好。MPA、MPAG、Ac MPAG的相对回收率分别为88.1%~104.3%、95.7%~99.6%、94.2%~96.0%。日内及日间RSD均<10%。结论该方法测定人血浆中MPA、MPAG、Ac MPAG的浓度,操作简便,方法灵敏度高,适合用于MPA及其代谢产物的药动学研究及血药浓度监测。     

高效液相色谱法测定人血浆中格列本脲浓度

目的:建立测定人血浆中格列本脲浓度的高效液相色谱法.方法:格列本脲血浆样品在酸性条件下以二氯甲烷-正己烷(50:50)提取,以格列齐特为内标.Nova-pakC18柱(4.6 mm×250 mm,4μm),流动相为乙腈-0.03 mol·L-1磷酸二氢钾(pH 3.0)(44:56),流速1.2 mL·min-1,检测波长228 nm.结果:标准曲线线性范围25～600μg·L-1(r=0.9992),血浆中格列本脲最低检测限为15μg·L-1.平均提取回收率为(81.9±3.6)%,平均方法回收率为(101.1±6.8)%,日内RSD≤5.0%,日间RSD≤9.1%.结论:该方法具有良好的准确性、精密性和较高的灵敏度,适用于格列本脲的药动学研究及治疗药物浓度监测.     

高效液相色谱荧光检测法测定人血浆中盐酸曲马多浓度

目的:建立测定人血浆中盐酸曲马多的高效液相色谱-荧光检测法。方法:血浆样品经液-液萃取,以氟康唑为内标,在Diamonsil C18柱(迪马公司,200mm×4.6mm,5μm),以磷酸二氢纳缓冲液(0.04mol.L-1,pH3.5)-乙腈(77∶23)为流动相,荧光检测,激发波长为275nm,发射波长为302nm。结果:曲马多的线性范围为5~1 000μg.L-1,最低检测限为1μg.L-1。低、中、高3种浓度的日内日间RSD均小于7.7%,提取回收率分别为(85.2±4.3)%,(80.6±3.5)%和(83.6±3.6)%。结论:该法重复性好、准确、简便、快速,适用于人血浆中盐酸曲马多浓度的测定及临床药动学研究。     

高效液相荧光色谱法测定人血浆中细辛脑的浓度

目的建立测定人血浆中细辛脑浓度的高效液相荧光色谱法.方法血浆样品经10%高氯酸沉淀后,以甲醇-0.5%醋酸液(7822)为流动相,流速为1 ml·min-1,色谱柱为BaiAn-C18(150mm×4.6 mm,5 μm),柱温为35℃,荧光检测波长激发波长(λex)265 nm,发射波长(hem)395 nm.结果血浆内源性杂质不干扰待测物测定,细辛脑的线性范围为1～300 μg·L-1,定量下限为1μg·L-1,日内、日间RSD均小于5%.样品3次冻融及在沉淀后,4℃下12 h内稳定性良好.结论该法灵敏、快速、准确,操作简便,线性范围宽,可用于细辛脑的临床药动学研究.     

高效液相色谱法测定血浆中雷贝拉唑的浓度

目的:建立高效液相色谱法测定血浆中雷贝拉唑的浓度.方法:血浆样品经处理后,选用Shim-pack VP-ODS柱(150mm×4.6 mm,5μm),0.05 mol·L-1磷酸二氢钾缓冲溶液(0.1 mol·L-1氢氧化钠溶液调pH 7.0)-乙腈(69:31)为流动相,检测波长为284 nm.结果:雷贝拉唑血药浓度的线性范围为5.46～2 184.61μg·L-1(r=0.999 9),提取回收率78.25%～89.60%,方法回收率94.56%～97.69%,日内、日间RSD分别为3.52%和4.60%.结论:该方法准确、灵敏,适于雷贝拉唑药动学研究.     

高效液相色谱法测定血浆中尼扎替丁浓度

目的:建立测定人血浆中尼扎替丁的高效液相色谱方法.方法:采用Diamonsil C18色谱柱(200 mm×4.6mm,5μm),流动相为0.1 mol·L-1醋酸铵缓冲液-甲醇(60:40),检测波长为320nm.血浆样品加盐析溶液碱化后以氯仿提取,雷尼替丁为内标.结果:尼扎替丁血药浓度线性范围为20～6 000l·L-1(r=0.999 9,n=6),最低检测浓度为10μg·L-1(S/N=3),方法回收率在96.84%～101.39%(n=5),日内和日间RSD均小于4%.结论:本法简便,快速,重现性好,适于尼扎替丁的药动学研究.     

高效液相色谱-荧光检测法测定血浆中五聚赖氨酸-β-羰基酞菁锌

目的:建立高效液相色谱-荧光检测法测定小鼠血浆中五聚赖氨酸-β-羰基酞菁锌[ZnPc-(Lys)5]的浓度。方法:血浆样品经预处理后采用CNW C18色谱柱(250 mm×4.6 mm,5μm),流动相为甲醇-水(分别含有0.05%三氟乙酸),流速为1.0 mL.min-1,荧光检测器激发波长为610 nm,发射波长为690 nm,柱温为25℃。结果:在2～500 nmol.L-1检测浓度范围内线性良好(r=0.999 0),检出限为0.5 nmol.L-1,提取回收率为86.99%～88.80%,相对回收率为96.20%～101.76%,日内和日间RSD均低于10%。结论:该方法简便、快速、准确、重复性好,可用于血浆中ZnPc-(Lys)5浓度的测定。     

高效液相色谱法测定人体血浆中甲氨蝶呤浓度方法的改进

目的:建立高效液相色谱法测定人体血浆中甲氨蝶呤浓度的方法。方法:血浆样品经处理后,色谱分离采用Zorbax-ODSC18(250mm×4.6mm,5μm)色谱柱,检测波长为302nm,流动相为0.01mol.L-1磷酸盐(用磷酸溶液调pH6.7)-甲醇(80∶20),流速为1.0mL.min-1,柱温为35℃。结果:甲氨蝶呤血药质量浓度在0.2～50mg.L-1(r=0.9997)范围内线性关系良好,最低检测质量浓度为5μg.L-1,提取回收率为96.5%～98.7%,方法回收率为98.2%～101.2%,日内、日间RSD分别为3.62%和4.57%。结论:该方法简单、快速、准确适用于临床甲氨蝶呤浓度监测及药动学研究。     

盐酸托烷司琼口腔崩解片在健康人体内的药动学及生物等效性

目的建立测定人血浆中托烷司琼浓度的LC/MS/MS法,并用该法研究托烷司琼在健康人体内的药动学特征及其相对生物等效性。方法采用LC/MS/MS法,测定20名健康男性受试者口服含盐酸托烷司琼10 mg的受试制剂和参比制剂后,不同时刻血浆中托烷司琼的浓度,绘制药动学曲线并计算主要药动学参数。结果受试制剂和参比制剂中托烷司琼的主要药动学参数如下:tmax分别为(2.1±0.8)和(2.1±0.8)h;ρmax分别为(15.7±6.2)和(16.1±6.2)μg.L-1,t1/2分别为(9.9±5.0)和(9.4±5.0)h;用梯形法计算,AUC0-t分别为(213.2±162.7)和(210.1±159.2)μg.h.L-1,AUC0-∞分别为(231.0±190.4)和(231.2±190.9)μg.h.L-1。以AUC0-t计算,盐酸托烷司琼口腔崩解片中托烷司琼的相对生物利用度为(102.2±25.7)%。结论盐酸托烷司琼的两种制剂生物等效。     

Pharmacokinetics and metabolism of the 5-hydroxytryptamine antagonist tropisetron after single oral doses in humans.

Tropiestron is a potent and selective antagonist of 5-hydroxytryptamine receptors. Tropisetron was developed for the indication of cancer chemotherapy-induced emesis. The pharmacokinetic and metabolic dispositions of tropisetron were studied in 12 healthy male volunteers receiving a single oral dose of 62 or 312 mumol (20 or 100 mg) of [14C]tropisetron. Serial plasma samples and complete urine and feces were collected for 120 hr postdose. Whereas the absorption of oral doses of 62-312 mumol tropisetron was rapid and complete, bioavailability was estimated to be only 66% for the 312 mumol dose and 52% for the 62 mumol dose, apparently because of saturable first-pass metabolism. Maximal concentrations of tropisetron averaged 87 and 608 nM after doses of 62 and 312 mumol, respectively, and the parent drug accounted for 21 and 36% of the radioactivity in AUC0-24 hr pools. Approximately 90% of the drug was metabolized before excretion, and approximately 70% of the dose was recovered in the urine. Following both the 62 and 312 mumol doses, the terminal half-life of tropisetron averaged 6-7 hr and that of total radioactivity was 10-11 hr. Tropisetron and its metabolites in plasma and urine were separated by gradient elution reversed-phase HPLC. Structures of eight metabolites were assigned on the bases of NMR and MS data. Tropisetron was metabolized by oxidative hydroxylation of the indole ring at positions 5, 6, and 7. The hydroxylated derivatives are further conjugated with glucuronic acid and sulfate. N-Oxygenation and oxidative N-demethylation at the tropinyl nitrogen also occur in trace amounts.(ABSTRACT TRUNCATED AT 250 WORDS)     

HPLC-MS-MS测定人血浆中的托烷司琼

目的建立HPLC-MS-MS方法测定人血浆中托烷司琼浓度。方法色谱柱为Hypurity C18柱（Thermo Hypersil-Keystone，2．1mm×150mm，5μm）；流动相为乙腈-0．1％甲酸水溶液-60：40；流速为0．2mL·min^-1，柱温为40℃，自动进样瓶温10℃。采用电喷雾离子源（ESI）正离子模式，用多反应监测模式（MRM）进行定量分析。用于定量分析的托烷司琼和内标的碎片离子分别为m／z 124和m／z 201。结果托烷司琼（TST）线性范围为0．22-57．25ng·mL^-1，决定系数（相关系数r的平方）为0．998，定量下限为0．22ng·mL^-1，提取回收率〉50％，日内和日间RSD均〈15％。结论此方法简便、高效、快速、和灵敏，可用于托烷司琼药动学及生物等效性研究。     

5-HT3 receptor antagonists protect against pressure overload-induced cardiac hypertrophy in murine

Activation of cardiac sympathetic afferent reflex results in the increase of sympathetic activity. Serotonin (5-HT) activates cardiac sympathetic afferent through stimulating 5-HT₃ receptors, the aim of present study is to test whether 5-HT₃ receptor antagonists protect against cardiac hypertrophy. Cardiac hypertrophy induced by TAC for 4 weeks in mice was significantly inhibited by administration of 5-HT₃ receptor antagonists, ondansetron (2.5 mg/kg, ip.) or tropisetron (2.5 mg/kg, ip.). Histological analysis revealed that the increased cardiac fibrosis in hypertrophic heart was relieved by ondansetron or tropisetron treatment. Ondansetron or tropisetron reduced the elevated plasma level of noradrenalin in mice with cardiac hypertrophy. Ondansetron and tropisetron had no effect on cardiomyocte hypertrophy induced by phenylephrine treatment in vitro. Finally, we took tropisetron as the representative drug and examined the effects of tropisetron on the desensitization of cardiac β-adrenergic receptor in rat treated with abdominal aortic banding (AB). Results showed that tropisetron restored the desensitization of cardiac β-adrenergic receptor in AB-treated rats. In conclusion, 5-HT₃ receptor antagonists protected against cardiac hypertrophy and restored the desensitization of cardiac adrenergic responsiveness, the mechanism in which may be through reducing the sympathetic activity.     

Effects of metoclopramide and tropisetron on aldosterone secretion possibly due to agonism and antagonism at the 5-HT4 receptor

Objective: Part of the prokinetic activity of metoclopramide can possibly be ascribed to agonist activity at 5-HT₄ receptors. The 5-HT₃ antagonist tropisetron is thought to act as an antagonist at 5-HT₄ receptors. In the present study aldosterone secretion in response to the administration of these two drugs was explored to examine the role of the 5-HT₄ receptor in aldosterone secretion. Methods: Following a single-blind, random design, ten normal male volunteers received one of the following regimens on three occasions, with at least 2-week intervals: metoclopramide 10 mg i.v.; tropisetron 5 mg by slow i.v.i., or; tropisetron by slow i.v.i., followed by 10 mg metoclopramide i.v. Results: In response to metoclopramide alone the mean plasma aldosterone level rose significantly to 149% of basal level and remained significantly elevated for the next 20 min. With tropisetron alone, there was a significant 37.8% drop at 60 min and the aldosterone levels remained low for the duration of the experiment. Metoclopramide reversed the decline mediated by tropisetron significantly at 30 and 90 min. Aldosterone levels after the latter regimen also did not differ significantly from baseline at any time period. Conclusion: These results would suggest the existence of a tonic stimulatory influence of 5-HT via 5-HT₄ receptors on aldosterone secretion, which could be augmented by metoclopramide and blocked by tropisetron. However, the effect of tropisetron per se should be interpreted with caution given the lack of a saline group Received: 25 April 1995/Accepted in revised form: 4 December 1995     

Pharmacokinetics of therapeutic doses of tropisetron in healthy volunteers

AIMS: To establish the bioavailability of tropisetron (5 mg) administered orally as capsule compared with 2 mg given intravenously. METHODS: Using a randomized crossover design, 18 healthy volunteers received a single oral dose of tropisetron (5 mg) and an intravenous bolus of tropisetron (2 mg) separated by a wash-out period of 1 week. Plasma concentrations of tropisetron were determined by h.p.l.c. and the pharmacokinetic parameters were estimated. RESULTS: The mean pharmacokinetic parameters for 5 mg tropisetron given orally were Cmax 3.46 ng ml(-1), t(max) 2.6 h, t(1/2) 5.7 h and AUC(0,infinity) 32.9 ng ml(-1) h. After intravenous administration initial plasma concentration was 15.1 ng ml(-1), t(1/2) 5.6 h, AUC(0,infinity) 20.7 ng ml(-1) h, V 678 l and CL 1800 ml min(-1). An inverse correlation was demonstrated between CYP2D6 activity, measured by the sparteine metabolic ratio, and the bioavailability (mean 0.60, range 0.27-0.99) of oral tropisetron. CONCLUSIONS: Tropisetron exhibits a wide range of oral bioavailability at therapeutic doses, which is mainly determined by CYP2D6 activity.     

LC-MS 法测定血浆盐酸托烷司琼含量

目的盐酸托烷司琼片为Ⅳ类新药，为节约开发费用，更好的控制质量，建立了本品血浆含量检测方法。方法建立以LC-MS法测定盐酸托烷司琼血浆含量的方法，色谱柱为Hypersil Gold C18柱（150×2．1mm，5μm）；流动相为甲醇：水（用醋酸调整pH值为4．8）=52：48；流速为0．2mL·min^-1，选用ESI离子源，采用正离子方式、目标二级离子选择反应离子扫描（SRM），扫描时间为0．03s，测定盐酸托烷司琼的含量。结果线性回收方程：Y=0．0131231＋0．0328806X，采用1／X^2加权，相关系数为0．9924，线性范围：0．2-40ng·mL^-1。平均回收率98．55％，RSD〈10％。结论此种方法准确、稳定，适用于盐酸托烷司琼的血浆含量测定。     

Short-term treatment of primary fibromyalgia with the 5-HT3-receptor antagonist tropisetron. Results of a randomized, double-blind, placebo-controlled multicenter trial in 418 patients.

We investigated the efficacy and tolerability of short-term treatment with tropisetron, a selective, competitive 5-HT3-receptor antagonist in fibromyalgia. The trial was designed as a prospective, multicenter, double-blind, parallel-group, dose-finding study. We randomly assigned 418 patients suffering from primary fibromyalgia to receive either placebo, 5 mg, 10 mg or 15 mg tropisetron once daily for 10 days. Clinical response was measured by changes in pain score, visual analog scale, tender point count and ancillary symptoms. Responders were prospectively defined as patients showing a 35% or higher reduction in pain score. Treatment with 5 mg tropisetron resulted in a significantly higher response rate (39.2%) than placebo (26.2%) (p < 0.05). In the visual analog scale, the group administered 5 mg tropisetron showed a significant improvement (p < 0.05) and the group administered 10 mg tropisetron showed a nonsignificant clinical benefit. The number of painful tender points was significantly reduced (p = 0.002) in the 5 mg tropisetron group. Regarding ancillary symptoms, the 5 mg tropisetron group showed a significant improvement (p < 0.05) in sleep and dizziness. The patients' overall assessment of efficacy was significantly higher for 5 mg (p = 0.016) and 10 mg (p = 0.002) tropisetron than for placebo. The safety and tolerability of tropisetron was good; gastrointestinal tract symptoms were the most frequently reported adverse events. Short-term treatment of fibromyalgia patients with 5 mg tropisetron for 10 days proved to be efficacious and well tolerated. In this study a bell-shaped dose-response curve was seen.     

阿瑞匹坦联合托烷司琼预防顺铂化疗引起呕吐的临床观察

目的:观察阿瑞匹坦联合托烷司琼方案预防顺铂化疗引起呕吐的疗效及不良反应。方法:采用随机、自身交叉对照的方法,将60例接受两周期含顺铂联合化疗的患者,随机分为AB、BA组。AB组第1周期应用阿瑞匹坦联合托烷司琼,第2周期应用托烷司琼;BA组第1周期应用托烷司琼,第2周期应用阿瑞匹坦联合托烷司琼。结果:可评价疗效的59例患者中,阿瑞匹坦联合托烷司琼方案和托烷司琼方案对急性呕吐的完全缓解率分别为74.6%和57.6%,有效控制率分别为91.5%和81.4%(Z=-2.017,P=0.044);对延迟性呕吐的完全缓解率分别为69.5%和42.4%,有效控制率分别为86.4%和71.2%(Z=-3.112,P=0.002)。两种方案的主要不良反应为呃逆、便秘、头痛、头晕、口干等,不良反应发生率比较差异无统计学意义(P>0.05)。结论:阿瑞匹坦联合托烷司琼方案对顺铂化疗引起急性呕吐与延迟性呕吐均有很好的疗效,不良反应可以耐受。     

地塞米松复合托烷司琼对乳腺术后呕吐的影响

目的 探讨围手术期预防性使用不同的药物干预方式对乳腺区段切除术后恶心呕吐(PONV)发生情况的影响,为短小手术预防PONV药物干预方案提供一定理论依据.方法 选取复合纳入标准的患者90例.将患者随机分为Ⅰ组(地塞米松组)、Ⅱ组(托烷司琼组)、Ⅲ组(地塞米松和托烷司琼联合用药组)3组,每组30例.在麻醉诱导后,Ⅰ组静脉注射地塞米松,Ⅱ组静脉注射托烷司琼,Ⅲ组静脉联合注射地塞米松和托烷司琼,观察和记录术后24 h患者发生PONV的患者数量,对实验数据统计分析,得出结果.结果 3组患者中,Ⅲ组PONV等级(WHO标准)和术后PONV视觉模拟评分(VAS)均低于Ⅰ组和Ⅱ组,差异具有统计学意义(P<0.05).结论 与单独静脉注射地塞米松或托烷司琼相比较,静脉联合注射地塞米松和托烷司琼对预防乳腺区段切除术后患者恶心呕吐效果更优.