HPLC-MS法测定人血浆中克拉霉素含量的方法学研究

目的:建立人血浆中克拉霉素测定的 HPLC-MS 法。方法:血浆经1mol·L~(-1)氢氧化钠溶液碱化后,用乙醚提取。采用 Zorbax SB-C_(18)(150mm×2.1 mm,5μm)分析柱,以乙腈-水-冰醋酸(60:40:0.05)为流动相,流速为0.4 mL·min~(-1);用HPLC-ESI~+-MS 法,选择性离子检测方法。质谱检测参数如下:干燥气流速13 L·min~(-1),干燥气温度350℃,雾化气压207kPa,毛细管电压3000 V,碎片电压150 V。选择检测的离子为 m/z 748.5(克拉霉素),m/z 837.5(罗红霉素)。结果:血浆中克拉霉素检测方法的线性范围为0.01～10μg·mL~(-1),最低检测限可达1.1 ng·mL~(-1)。血浆中克拉霉素的方法回收率为92.7%～99.4%,日内 RSD<5.1%,日间 RSD<3.6%。结论:本法灵敏,准确,可用于克拉霉素的药代动力学研究。     

柱前衍生化高效液相色谱-质谱联用法测定人体内血浆中的硫普罗宁

目的：建立高效液相色谱-质谱联用法测定人血浆中硫普罗宁的浓度。方法：色谱条件——色谱柱为Waters SunFire~(TM)C_(18)(2.1 mm×20 mm,5μm),流动相为甲醇-0.05%冰乙酸水溶液(50∶50,V/V),流速为0.3 mL·min~(-1),柱温为室温；质谱条件——电喷雾离子化法(ESI)采集负离子,单离子方式检测硫普罗宁衍生化产物m/z 327.95和内标葛根黄豆苷元m/z 253.00。结果：方法的定量下限为0.1 mg·L~(-1),线性范围为0.1～4.0 mg·L~(-1)(r=0.996 2),提取回收率均大于(66±s 5)%,日内、日间精密度(RSD)均小于9.4%。结论：该法灵敏、专属,适用于硫普罗宁人体血浆药物浓度的测定。     

HPLC-MS/MS法测定人血浆中阿莫西林的质量浓度

目的建立测定人血浆中阿莫西林质量浓度的高效液相色谱-质谱(HPLC-MS/MS)联用检测方法,并用于生物等效性研究。方法以阿莫西林-d4为内标,血浆样品经甲醇沉淀后,经HPLC-MS/MS分析。色谱柱为ZORBAX SB-Aq (150 mm×4.6 mm,3.5μm);流动相为甲醇-2 mL·L~(-1)甲酸溶液(40∶60);流速为0.6 mL·min~(-1)。采用电喷雾离子源(ESI),多重反应选择离子监测(MRM)负离子模式,阿莫西林和阿莫西林-d4的定量离子对分别为m/z 364→223和m/z 368→227。结果阿莫西林血药质量浓度在0.088 57～39.370 00μg·mL~(-1)范围内线性关系良好,选择性、准确度、精密度、提取回收率、基质效应和稳定性等均符合要求。结论该方法简单、快速、灵敏、准确,可用于阿莫西林血药质量浓度的检测及生物等效性研究。     

液相色谱-质谱联用法测定大鼠血浆中盐酸金刚烷胺浓度及体内药动学研究

目的:建立测定大鼠血浆中盐酸金刚烷胺的液相色谱-质谱联用法。方法:色谱柱为 Agilent Zorbax SB-C_(18)(2.1 mm×150 mm,3.5μm);乙腈-0.05%甲酸溶液(28∶72)为流动相,流速0.2 mL·min~(-1);柱温25℃;内标为盐酸美金刚。质谱条件为电喷雾电离源(ESI),选择正离子抽提方式检测,金刚烷胺和美金刚的选择检测离子分别为 m/z 152.20([M H]~ )、m/z180.20([M H]~ )。选用 SD 大鼠6只,单剂量尾静脉注射给予1 mg·kg~(-1),进行了血药浓度的测定。结果:盐酸金刚烷胺血浆线性范围为0.9824～491.2 ng·mL~(-1),检测限为0.025 ng·mL~(-1),方法回收率均大于95%,日内、日间精密度试验的 RSD<5.0%。结论:本方法专属性强,灵敏度高,线性关系良好,操作简便,适用于药代动力学研究。     

LC／MS／MS测定大鼠血浆中普萘洛尔及其代谢物4-羟普萘洛尔、N-去异丙基普萘洛尔的浓度

目的:用高效液相色谱-质谱联用方法同时测定大鼠血浆中普萘洛尔及其代谢物4-羟普萘洛尔、N-去异丙基普萘洛尔的浓度。方法:大鼠血浆用乙醚萃取法处理后,采用 LC/MS/MS 方法,测定大鼠血浆中普萘洛尔及其代谢物4-羟普萘洛尔、N-去异丙基普萘洛尔的浓度。HPLC 条件:大连依利特 Hypersil BDS C_(18)柱(2.1 mm×50 mm,3μm),流动相:水(含0.1%甲酸)-乙腈(39:61,v/v),柱温:12℃,流速:200 μL·min~(-1),进样量:10 μL;质谱检测参数为电喷雾电离源(ESI);喷雾电压:3500 V;碰撞压力:0.1333 Pa;源内碰撞诱导电压:普萘洛尔,10 V;4-羟普萘洛尔,15 V;N-去异丙基普萘洛尔,15 V;扫描时间:0.3 s;检测离子:普萘洛尔 m/z 260[M H]~ ;4-羟普萘洛尔 m/z 276[M H]~ ;N-去异丙基普萘洛尔218[M H]~ 。结果:普萘洛尔线性范围2—1000 ng·mL~(-1),最低定量限为2 ng·mL~(-1)。4-羟普萘洛尔和 N-去异丙基普萘洛尔的线性范围1～200 ng·mL~(-1),最低定量限为1 ng·mL~(-1)。普萘洛尔的萃取回收率在90%以上,4-羟普萘洛尔和 N-去异丙基普萘洛尔的萃取回收率均为50%以上,日内、日间 RSD 皆小于15%。结论:适用于测定大鼠血浆中普萘洛尔及其代谢物4-羟普萘洛尔、N-去异丙基普萘洛尔的浓度及药动学的研究。     

液相色谱-电喷雾串联质谱法测定人血浆中辛伐他汀浓度

目的:建立液相色谱-电喷雾串联质谱联用测定人血浆中辛伐他汀浓度的方法。方法:色谱条件为 Discovery ODS C_(18)(2.1 mm×100 mm,3μm)色谱柱;流动相为乙腈-0.1%甲酸溶液(65:35,v/v);柱温30℃;流速0.2 mL·min~(-1);进样量5μL。质谱条件为电喷雾电离源(ESI),选择性检测定量离子为 m/z 441.3/325.0(辛伐他汀)和 m/z 427.2/325.0(洛伐他汀,内标)带正电荷的离子峰。样品用乙酸乙酯提取。结果:血浆中辛伐他汀浓度在0.20～20.0 ng·mL~(-1)内呈良好线性关系,r=0.9996。提取回收率为90.0%～92.4%(RSD 6.0%～14.9%),方法回收率在85%～115%之内,日内和日间精密度试验的RSD 均<15%,最低检测浓度为0.2 ng·mL~(-1)。结论:该测定方法经全面考察符合血浆样品的测定要求,可以应用于辛伐他汀的人体药动学研究和生物等效性评价。     

HPLC—MS（TOF）法测定大鼠血浆中环维黄杨星D的浓度

目的:本文建立了HPLC-MS(TOF)法测定大鼠ig给药后体内环维黄杨星D的浓度,为进一步研究环维黄杨星D的药代动力学提供了可靠的方法。方法:大鼠血浆中加入内标多奈哌齐后碱化,以液-液萑取法提取血浆后采用LC-ESI-MS(TOF)联用技术,以电喷雾(ESI)作为接口技术,选择环维黄杨星D的准分子离子([M H]~ ,m/z 403)和内标多奈哌齐的准分子离子([M H]~ ,m/z 380)作为测定离子,测定大鼠血浆中环维黄杨星D的浓度。HPLC条件:Lichrospher CN柱(4.6 mm×150mm,10μm),甲醇-醋酸盐缓冲液(80:20)为流动相,流速:0.8mL·min~(-1);质谱检测参数:喷嘴电压力5500V;喷管电压为180 V;检测电压为2 225 V;喷管温度为140℃。结果:环维黄杨星D的线性范围为0.5～200 ng·mL~(-1),最低检测限为0.2 ng·mL~(-1),日内精密度RSD为6.1％,日间精密度的RSD为7.6％。结论:该测定方法具有灵敏、专一和快速的优点,应用本法测定了大鼠ig给药环维黄杨星D的血浆浓度。     

高效液相色谱-质谱法测定大鼠血浆中硫酸酯型脱氢表雄酮和孕烯醇酮

建立同时测定 SD 大鼠血浆中脱氢表雄酮硫酸酯(DHEAS)和孕烯醇酮硫酸酯(PREGS)的高效液相色谱-质谱法。方法:选择雌酮硫酸酯(ES)作为内标。雄性 SD 大鼠的血浆使用乙腈沉淀除去蛋白后,经固相萃取、水解及衍生化后,以高效液相色谱质谱检测器测定脱氢表雄酮硫酸酯和孕烯醇酮硫酸酯的浓度。色谱柱为 Zorbax SB C_(18)柱温40℃,流动相为乙腈水,使用梯度洗脱;采用大气压化学离子源(HPLC/APCI-MS),负离子检测方式,用于选择离子检测(SM)定量分析的离子为 m/z 490.0(DHEAS)和 m/z 518.0(PREGS)及 m/z 472.1(ES)[M-H]~-。结果:血浆中脱氢表雄酮硫酸酯和孕烯醇酮硫酸酯线性范围分别为:0.05～2.0ng·mL~(-1),0.03～2.0 ng·mL~(-1),r 分别为0.9990和0.9979。低、中、高浓度血样的提取回收率在66.8%～82.3%之间,相对回收率为:98.4%～111.6%,日内、日间变异均小于15%。正常雄性 SD 大鼠的血浆脱氢表雄酮硫酸酯和孕烯醇酮硫酸酯分别为0.070±0.020 ng·mL~(-1)、0.13±0.031 ng·mL~(-1)。结论:本实验方法具有灵敏度高、准确度好及变异较小的特点,适合测定雄性 SD 大鼠血浆中脱氢表雄酮硫酸酯和和孕烯醇酮硫酸酯的含量。     

高效液相色谱-质谱法测定人血浆紫杉醇浓度

目的 建立高效液相色谱-质谱(HPLC-MS)法测定人血浆中紫杉醇的浓度。方法 以克拉霉素为内标,血浆样品经乙腈沉淀蛋白后,进行HPLC-MS法测定。色谱柱为Agilent Zorbax Eclipse SB C18柱(2.1 mm×150 mm,5 μm)；流动相为甲醇-0.05%甲酸水溶液(70：30),流速0.2 mL·min^-1；柱温25 ℃；质谱条件为电喷雾电离源（ESI）,以选择性正离子方式检测,紫杉醇和内标克拉霉素的选择性检测离子分别为m/z 876.5［M+Na］+,m/z 748.6［M+H］+。结果 紫杉醇的线性范围为7.50～6 000.00 ng·mL^-1,r=0.999 9,定量限为7.50 ng·mL^-1,方法回收率均>90%,日内、日间精密度（RSD）均<10%。结论 该法专属性强,灵敏度高,线性关系良好,操作简便,适用于临床血浆紫杉醇浓度的测定.     

液相色谱-质谱／质谱联用法测定人血浆中加兰他敏

目的:建立测定人血浆中加兰他敏浓度的液相色谱-质谱/质谱联用法,用于其人体血药浓度测定。方法:血浆样品经液-液萃取后,以甲醇-水-甲酸(65:35:1)为流动相,Zorbax SB-C_8柱(150mm×4.6mm,5μm)分离,采用大气压化学电离源,以选择反应监测方式进行正离子检测。内标为双氢吗啡酮,用于定量分析的离子反应分别为 m/z 288→m/z 213(加兰他敏)和 m/z 286→m/z 185(内标)。结果:血浆中加兰他敏最低定量限为0.5ng·mL~(-1),其线性范围为0.5～100ng·mL~(-1)。其高、中、低3个浓度的平均提取回收率为80.9%,方法回收率为100.5%,日内及日间 RSD 均<8%。结论:方法选择性强,灵敏度高,快速准确,可作为加兰他敏人体内药动学研究的手段。     

Efficacy of gut lavage,hemodialysis, and hemoperfusion in the therapy of paraquat or diquat intoxication

Clinical and in vitro investigations were carried out to test the efficacy of gut lavage, hemodialysis, and hemoperfusion in the treatment of poisoning with paraquat or diquat. In a patient suffering from diquat intoxication 130 times more diquat was removed by gut lavage 30 h after ingestion than was removed by complete aspiration of the gastric contents.Determination of in vitro clearances for paraquat and diquat by hemodialysis showed that, at serum concentrations of 1–2 ppm, such as are frequently encountered in poisoning in man, toxicologically relevant quantities of herbicide cannot be removed from the body. At a concentration of 20 ppm, on the other hand, hemodialysis proved to be effective, the clearance being 70 ml/min at a blood flow rate of 100 ml/min. The efficacy of hemoperfusion with coated activated charcoal was on the whole better. Especially at concentrations around 1–2 ppm, the clearance values for hemoperfusion were some 5–7 times higher than those for hemodialysis.In a patient suffering from paraquat poisoning, both hemodialysis as well as hemoperfusion were carried out. The in vitro results could be confirmed: At serum concentrations of paraquat less than 1 ppm no clearance could be obtained by hemodialysis while by hemoperfusion with activated charcoal quite high clearance values were measured and the serum level dropped down to zero.

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Zusammenfassung Klinische Untersuchungen und Laboratoriumsversuche wurden durchgeführt, um die Wirksamkeit von Darmspülung, Hämodialyse und Hämoperfusion bei Paraquat- und Deiquat-Vergiftungen zu prüfen.Bei einem Patienten wurde 30 Std nach Deiquat-Aufnahme durch Darmspülung 130mal mehr Deiquat entfernt als durch vollständige Aspiration des Mageninhaltes. In vitro-Versuche ergaben, daß bei Blutserumkonzentrationen von 1–2 ppm, die bei Vergiftungen oft gemessen werden, durch Hämodialyse keine toxikologisch relevanten Paraquat- oder Deiquat-Mengen entfernt werden können. Dagegen erwies sich die Hämodialyse bei 20 ppm und einer Blutumlaufgeschwindigkeit von 100 ml/min mit einer Clearance von 70 ml/min als wirksam. Die Hämoperfusion mit beschicheter Aktivkohle war in diesen Versuchen aber eindeutig überlegen, denn insbesondere bei Konzentrationen um 1–2 ppm waren die Clearance-Werte 5–7mal höher als bei der Hämodialyse.Die in vitro-Ergebnisse wurden bei einem Patienten mit einer Paraquat-Vergiftung bestätigt: Bei Konzentrationen unter 1 ppm war die Hämodialyse wirkungslos, während durch Hämoperfusion relativ hohe Clearance-Werte erreicht wurden, so daß der Serumspiegel rasch unter die Nachweisgrenze abfiel.

     

Aquatic Insects and Trace Metals: Bioavailability,Bioaccumulation, and Toxicity

Abstract

The uptake of metals from food and water sources by insects is thought to be additive. For a given metal, the proportions taken up from water and food will depend both on the bioavailable concentration of the metal associated with each source and the mechanism and rate by which the metal enters the insect. Attempts to correlate insect trace metal concentrations with the trophic level of insects should be made with a knowledge of the feeding relationships of the individual taxa concerned. Pathways for the uptake of essential metals, such as copper and zinc, exist at the cellular level, and other nonessential metals, such as cadmium, also appear to enter via these routes. Within cells, trace metals can be bound to proteins or stored in granules. The internal distribution of metals among body tissues is very heterogeneous, and distribution patterns tend to be both metal and taxon specific. Trace metals associated with insects can be both bound on the surface of their chitinous exoskeleton and incorporated into body tissues. The quantities of trace meals accumulated by an individual reflect the net balance between the rate of metal influx from both dissolved and particulate sources and the rate of metal efflux from the organism. The toxicity of metals has been demonstrated at all levels of biological organization: cell, tissue, individual, population, and community. Much of the literature pertaining to the toxic effects of metals on aquatic insects is based on laboratory observations and, as such, it is difficult to extrapolate the data to insects in nature. The few experimental studies in nature suggest that trace metal contaminants can affect both the distribution and the abundance of aquatic insects. Insects have a largely unexploited potential as biomonitors of metal contamination in nature. A better understanding of the physico-chemical and biological mechanisms mediating trace metal bioavailability and exchange will facilitate the development of general predictive models relating trace metal concentrations in insects to those in their environment. Such models will facilitate the use of insects as contaminant biomonitors.     

Alzheimer’s disease – current trends in basic and clinical research. Highlights from the 16th ECNP

Advances in the molecular biological knowledge of neuronal nicotinic acetylcholine receptors (nAChRs) have led to a growing interest by the pharmaceutical industry in the development of novel compounds that selectively modulate nAChR function. The ability of (-)-nicotine, an activator of nAChRs, to enhance attentional aspects of cognition in animals and humans, to exert neuroprotective and anxiolytic-like effects, and presumably to mediate the negative correlation between smoking and Alzheimer's (and Parkinson's) Disease, has focused interest on the potential therapeutic utility of modulators of nAChR function for treatment of some of the deficits associated with these progressive, neurodegenerative conditions. Numerous compounds are known which activate nAChRs and which might serve as lead compounds toward the development of such agents. The pharmacologic diversity of neuronal nAChR subtypes suggests the possibility of developing selective compounds which would have more favourable side-effect profiles than existing agents. This broader class of agents, collectively called cholinergic channel modulators (ChCMs), is anticipated to encompass compounds which would have more favourable side-effect profiles than existing agents, which generally exhibit low selectivity. This selectivity may be achieved by preferentially activating some subtypes of nAChRs (i.e., Cholinergic Channel Activators, ChCAs) or inhibiting the function of other subtypes (Cholinergic Channel Inhibitors, ChCIs). An overview of the biology of nAChRs and the rationale for the use of ChCMs for the treatment of dementia related to neurodegenerative diseases are presented, followed by a discussion of lead compounds and compounds under consideration for clinical evaluation.     

2-(Acetoxyphenyl)-(Z)-styryl sulfides as cyclooxygenase inhibitors

2-(Acetoxyphenyl)-(Z)-styryl sulfides are described as selective cyclooxygenase-2 (COX-2) inhibitors, useful for treating inflammation and COX-2-mediated disorders including neoplasia. 2-(Acetoxyphenyl)-(Z)-styryl sulfide is claimed to be the most potent COX inhibitor in the series with a COX-2 selectivity ratio of 33. This compound is also claimed to be superior to celecoxib (Celebrex^®, Pfizer) in inhibiting cell growth of colorectal carcinoma cells. In this evaluation, the COX inhibitory activity of this compound is compared to that previously disclosed for diarylheterocycles and 2-(acetoxyphenyl)alkyl sulfides. The validity of the DLD-1 cell line in the growth inhibition studies is questioned based on recent literature reports indicating the lack of COX-2 expression in this cell line.     

Sleep-disordered breathing with chronic opioid use

Chronic opioid use for pain relief or as substitution therapy for illicit drug abuse is prevalent in our societies. In the US, retail distribution of methadone and oxycodone has increased by 824 and 660%, respectively, between 1997 and 2003. μ-Opioids depress respiration and deaths related to illicit and non illicit chronic opioid use are not uncommon. Since 2001 there has been an emerging literature that suggests that chronic opioid use is related to central sleep apnoea of both periodic and non-periodic breathing types, and occurs in ～ 30% of these subjects. The clinical significance of these sleep-related abnormalities are unknown. This review addresses the present knowledge of control of ventilation mechanisms during wakefulness and sleep, the effects of opioids on ventilatory control mechanisms, the sleep-disordered breathing found with chronic opioid use and a discussion regarding the future research directions in this area.     

Drug targets for cognitive enhancement in neuropsychiatric disorders

The investigation of novel drug targets for treating cognitive impairments associated with neurological and psychiatric disorders remains a primary focus of study in central nervous system (CNS) research. Many promising new therapies are progressing through preclinical and clinical development, and offer the potential of improved treatment options for neurodegenerative diseases such as Alzheimer's disease (AD) as well as other disorders that have not been particularly well treated to date like the cognitive impairments associated with schizophrenia (CIAS). Among targets under investigation, cholinergic receptors have received much attention with several nicotinic agonists (α7 and α4β2) actively in clinical trials for the treatment of AD, CIAS and attention deficit hyperactivity disorder (ADHD). Both glutamatergic and serotonergic (5-HT) agonists and antagonists have profound effects on neurotransmission and improve cognitive function in preclinical experiments with animals; some of these compounds are now in proof-of-concept studies in humans. Several histamine H3 receptor antagonists are in clinical development not only for cognitive enhancement, but also for the treatment of narcolepsy and cognitive deficits due to sleep deprivation because of their expression in brain sleep centers. Compounds that dampen inhibitory tone (e.g., GABA_A α5 inverse agonists) or elevate excitatory tone (e.g., glycine transporter inhibitors) offer novel approaches for treating diseases such as schizophrenia, AD and Down syndrome. In addition to cell surface receptors, intracellular drug targets such as the phosphodiesterases (PDEs) are known to impact signaling pathways that affect long-term memory formation and working memory. Overall, there is a genuine need to treat cognitive deficits associated with many neuropsychiatric conditions as well as an increasingly aging population.