用HPLC法同时测定比格犬血浆中地西泮、去甲西泮和氟马西尼的浓度

目的:建立测定比格犬血浆中地西泮、去甲西泮和氟马西尼浓度的HPLC法.方法:采用Kromasil 100-5 C18色谱柱(250 mm×4.6 mm,5μm);以氯硝西泮为内标,甲醇(A)-水(B)-四氢呋喃(C)为流动相(55∶40∶5),流速1.0 ml/min,检测波长254 nm;柱温25 C.结果:地西泮、去甲西泮和氟马西尼在浓度范围为0.025～1.000 μg/ml时线性良好(r=0.999 9,0.999 0,0.998 9),日内和日间RSD均＜6％(n=5),回收率接近100％.结论:本方法准确、快速、简便,可用于比格犬血浆中地西泮、去甲西泮和氟马西尼浓度的同时测定.     

《西北药学杂志》2005年第20卷总目次

*药物分析*甘草挥发性成分GC-MS分析1:(3)高效液相色谱法测定罗格列酮片剂含量1:(5)HPLC法测定双氯芬酸钠肠溶片的含量1:(6)高效液相色谱法测定咳喘三号糖浆中黄芩苷的含量1:(8)反相高效液相色谱法测定清开灵颗粒中栀子苷的含量1:(9)GC-ECD法检测血浆中地西泮及去甲西泮2:(51)盐酸去氢骆驼蓬碱肝动脉栓塞微球药物含量的研究2:(53)地西泮注射液含量的测定方法探讨2:(55)消癌平片中绿原酸的HPLC法测定2:(56)西宁地区维C银翘片质量分析2:(58)改变药典规定波长实现吡嗪酰胺片溶出度的在线过程分析3:(99)HPLC法测定乳糖酸红霉素的有关物…     

高效液相色谱法测定血浆地西泮及其代谢物

地西泮(diazepam)是临床常用的苯二氮(艹卓)类镇静药,其代谢物去甲地西泮(去甲安定,n-des-methyldiazepam)和奥沙西泮(去甲羟基安定,oxa-zepam)不仅已作为苯二氮(艹卓)类镇静药广泛用于临床.而且还是多种苯二氮(艹卓)类药物的活性代谢产物.据报道地西泮的代谢存在遗传多态性和东西方人种族间差异.因此,测定地西泮及其代谢物的血药浓度.对于研究其药物动力学和遗传药理学以及临床治疗监测均具有重要意义.有报道用HPLC法测定地西泮及其代谢物的血药浓度,但其方法或灵敏度尚不够高,或样品处理复杂.我们用RP-HPLC法测定血浆中地西洋及其代谢物浓度,具有灵敏度高和样品处理简单等特点,适于临床治疗药物监测和药理学研究.     

血浆中去甲地西泮及代谢物奥沙西泮的HPLC测定方法和大鼠口服药代动力学

采用高效液相色谱法测定去甲地西泮(去甲安定)及其代谢产物奥沙西泮。以RP-C₁₈为固定相,乙腈—0.01mol·L^-1醋酸钠(pH3.8,33.3:66.6)为流动相,地西泮为内标物,紫外波长240nm处定量测定。去甲地西泮、奥沙西泮和内标物的保留时间分别为2.8min,4.85min和8.5min;绝对回收率分别为74%,86%和86%。奥沙西泮在35.3～2260ng·ml^-1,去甲地西泮在20～2560ng·ml^-1血浆浓度范围内线性关系良好,r=0.9997和r=0.9998。二药的最低检测浓度分别为10ng·ml、和7ng·ml^-1;日内和日间相对标准偏差(RSD)均分别小于6%和10%(n=5)。多种常用药物对样品的色谱峰无干扰。并用此法研究了大鼠单次口服去甲地西泮的药代动力学。     

不同甲状腺机能状态对地西泮氧化代谢的影响

本文建立了大鼠的甲状腺机能低下（甲低）和甲状腺机能亢进（甲亢）的动物模型,用HPLC方法测定地西泮的代谢物,以研究甲低和甲亢对SD大鼠单次口服地西泮氧化代谢的影响,甲低组大鼠地西泮的N－去甲基化代谢物去甲地西泮的血药浓度（AUC）升高,消除减慢,说明甲低对地西泮的N－去甲基化影响不大,而主要影响去甲地西泮的C3－羟化代谢,不同程度甲亢对地西泮氧化代谢的影响有所不同,轻度和中度甲亢组去甲地西泮血药浓度（AUC）低于对照组,消除加快;而重度甲亢组去甲地西泮血药浓度（AUC）略高于对照组,消除减慢。轻、中、重度甲亢大鼠地西泮C3－羟化代谢物替马西泮血药浓度（AUC）都不同程度地低于对照组,表明仅在重度甲亢对去甲地西泮的C3－羟化代谢才有抑制性影响,但不同程度甲亢均抑制地西泮的C3－羟化代谢。     

HPLC法同时测定血浆地西泮及其代谢物浓度

目的 :建立同时测定血浆中地西泮及其代谢物浓度的方法。方法 :选用ZORBAXRP C18柱 (15 0mm× 4 6mm ,5 μm) ;甲醇 - 2 5mmol·L-1醋酸铵溶液 (6 0∶4 0 ,V/V)作流动相 ;流速 0 8mL·min-1;检测波长 2 30nm。取血浆样品 0 5mL ,在碱性条件下用二氯甲烷 -正己烷提取 ,HPLC检测。结果 :本法对替马西泮、去甲地西泮和地西泮 3种物质的最低检测限均为 2 μg·L-1,线性范围为 10～ 15 0 0 μg·L-1;奥沙西泮的最低检测限为 5 μg·L-1,线性范围为 2 0～ 15 0 0 μg·L-1。回收率均接近 10 0 % ,日内、日间RSD <5 %。结论 :本法能同时测定血浆中地西泮及其代谢物浓度 ,具有重现性好 ,灵敏、可靠 ,可用于地西泮中毒的监测     

不同甲状腺机能状态对地西泮氧化代谢的影响(英)

本文建立了大鼠的甲状腺机能低下（甲低）和甲状腺机能亢进（甲亢）的动物模型。用HPLC方法测定地西泮的代谢物，以研究甲低和甲亢对SD大鼠单次口服地西泮氧化代谢的影响。甲低组大鼠地西泮的N－去甲基化代谢物去甲地西泮的血药浓度（AUC）升高，消除减慢。说明甲低对地西泮的N－去甲基化影响不大，而主要影响去甲地西泮的C_3－羟化代谢。不同程度甲亢对地西泮氧化代谢的影响有所不同。轻度和中度甲亢组去甲地西泮血药浓度（AUC）低于对照组，消除加快；而重度甲亢组去甲地西泮血药浓度（AUC）略高于对照组，消除减慢。轻、中、重度甲亢大鼠地西泮C_3－羟化代谢物替马西泮血药浓度（AUC）都不同程度地低于对照组，表明仅在重度甲亢对去甲地西泮的C_3－羟化代谢才有抑制性影响，但不同程度甲亢均抑制地西泮的C_3－羟化代谢。     

HPLC法同时测定毛发样本中地西泮、去甲西泮和奥沙西泮

目的:建立测定毛发样本中地西泮、去甲西泮和奥沙西泮的HPLC分析方法。方法:采用超声法结合液液萃取法作为前处理方法。采用Athena C₁₈-WP(250 mm×4.6 mm, 5μm)色谱柱;以甲醇-0.02 mol·L^-1磷酸二氢钠溶液(磷酸调pH 3.4)-乙腈(30∶43∶27)为流动相;柱温50℃;流速0.7 mL·min^-1;检测波长254 nm。内标为氯硝西泮。结果:本方法在0.1～25 ng·mg^-1范围内具良好的线性关系,定量限为0.1 ng·mg^-1,检测限为0.05 ng·mg^-1,批内准确度(n=6)在88.2%～103.9%,批间准确度(n=24)在88.5%～106.2%,精密度小于或等于9.19%,提取回收率稳定,符合方法学要求。将该方法应用于实际病人的毛发样本,测得地西泮及其代谢物去甲西泮、奥沙西泮浓度分别是(0.307±0.016)ng·mg^-1、(0.244±0.012)ng·mg^-1     

地西泮代谢物的结合物水解条件研究

目的:考察β 葡萄糖醛酸苷酶水解尿液中地西泮代谢物的结合物的酶解条件.方法: 实验设计采用正交表L9(34).取尿样2 mL,加内标,再加不同量的β 葡萄糖醛酸苷酶,在不同温度、不同酶解时间下水解,然后用正己烷 二氯甲烷(5∶3)5 mL提取,挥干提取液后用50 μL甲醇溶解,10 μL进样,采用高效液相色谱法检测.结果:对水解产物峰面积与内标物峰面积的比值进行方差分析,发现水解尿液中去甲地西泮、替马西泮、奥沙西泮的葡萄糖醛酸苷结合物的最佳酶解条件分别为55℃,5 h,5 000 U酶量;55℃,2 h,1 000 U酶量;55℃,2 h,5 000 U酶量.结论:水解2 mL尿液中地西泮代谢物的结合物的最佳条件为55℃,5 h,5 000 U酶量.     

奥沙西泮与地西泮治疗广泛性焦虑的对照研究

目的比较奥沙西泮与地西泮治疗广泛性焦虑的疗效和不良反应。方法 87例符合CCMD-3诊断标准的广泛性焦虑患者随机分为2组,分别给予奥沙西泮和地西泮治疗6周。采用汉密尔顿焦虑量表（HAMA）评定临床疗效,副作用量表（TESS）评定不良反应。结果治疗后2组HAMA评分差异无统计学意义（P〉0.05）;奥沙西泮组肝功能异常出现率小于地西泮组,差异有统计学意义（P〈0.05）。结论奥沙西泮和地西泮治疗广泛性焦虑的疗效相似,氮奥沙西泮对肝功能的影响小,更为安全。     

Diazepam and N-desmethyldiazepam concentrations in saliva, plasma and CSF.

1 Salivary and plasma diazepam and nordiazepam concentrations were measured in 51 paired samples from four experimental situations. In seven of the patients CSF samples were estimated. 2 Correlation of 0.89 (P less than 0.001) was observed between salivary and plasma diazepam and 0.81 (P less than 0.001) between salivary and plasma nordiazepam. 3 Mean salivary diazepam was 1.6% (+/- 0.3%) of the plasma diazepam. It was found to vary markedly in an acute dosage study. Mean salivary nordiazepam was 2.9% (+/- 1%) of the plasma measure and was dependent on salivary flow rate. 4 CSF diazepam was in equilibrium with unbound plasma diazepam and salivary diazepam. 5 Mean protein binding of diazepam in vitro was 99.3% with no variations as a function of concentration. 6 The results suggest salivary diazepam and nordiazepam measures to be of value in epidemiological studies. However, they do not predict accurately the plasma total or unbound drug concentration from a salivary sample in an individual.     

Effect of orally administered misoprostol and cimetidine on the steady state pharmacokinetics of diazepam and nordiazepam in human volunteers.

The effects of misoprostol and cimetidine on diazepam pharmacokinetics were evaluated in order to determine whether the kinetic variables for diazepam and nordiazepam alone differ with the repeated oral administration of misoprostol and cimetidine to healthy adult volunteers. The trial was conducted as an open crossover study in 12 normal subjects, divided into two groups with all subjects receiving both regimens. Total study duration was 5 weeks. An initial clinical assessment, including blood biochemistry and assessment of subject oxidation status was carried out on study day 1. On this day, subjects began taking diazepam (10 mg) orally for one week, with pharmacokinetic studies performed at day 8, when steady state levels of diazepam were reached. This was followed by one week with active drug, misoprostol to Group I and cimetidine to Group II, with pharmacokinetic studies performed at the end of a 1-week treatment. After a 2-week wash-out period, both groups took for one week, the alternate drug, i.e. cimetidine plus diazepam to Group I and misoprostol plus diazepam to Group II. On days 8, 15 and 36, subjects were admitted to the hospital for 12 h, during which time a clinical examination was carried out and blood samples were taken at time zero and at 4, 8, 12, 24, and 36 h post-dosing for the measurement of serum diazepam and nordiazepam. The main parameters measured and evaluated were diazepam and nordiazepam pharmacokinetics at steady state (days 8, 15 and 36). These were areas under the curve in the dose intervals (AUC0-24h), maximum plasma concentrations (Cmax), time to peak concentrations (Tmax), elimination half-life (t1/2), elimination constant (Kel), distribution volume (Vd), total body clearance (ClB) and clearance after oral administration (Cloral). The results demonstrated that plasma diazepam and nordiazepam concentrations had a significant increase after steady states have been reached with the simultaneous administration of 800 mg of cimetidine daily for one week. The simultaneous administration of 800 micrograms of misoprostol did not cause any significant change in diazepam and nordiazepam plasma levels after steady states had been reached. Comparing the pharmacokinetic parameters of Groups A and B as well as within groups on days 8, 15 and 36, a significant increase in plasma diazepam and nordiazepam levels was detected. This was due to a cimetidine-induced impairment in microsomal oxidation of diazepam and nordiazepam, which caused a decrease in total metabolic clearance and increased mean steady state plasma concentrations. A more prolonged half-life was observed for both groups taking cimetidine as well as an increase of mean maximum plasma concentrations.(ABSTRACT TRUNCATED AT 400 WORDS)     

Behavioural and pharmacokinetic studies in the monkey (Macaca mulatta) with diazepam, nordiazepam and related 1,4-benzodiazepines.

1. Behavioural activity (delayed differentiation and spatial delayed alternation) and pharmacokinetics of diazepam and its metabolites, N-desmethyldiazepam (nordiazepam), 3-hydroxydiazepam (temazepam) and 3-hydroxy-N-desmethyldiazepam (oxazepam), and of dipotassium clorazepate (clorazepate), were studied in the monkey (Macaca mulatta). Diazepam and its metabolites (1.8 and 3.0 mg/kg) and clorazepate (2.6 and 4.3 mg/kg) were given by intraperitoneal injection. 2. Hydroxylation of diazepam (temazepam and oxazepam) led to a loss of, or a considerable reduction in, behavioural activity, whereas activity was preserved, though modified, by demethylation (nordiazepam). It was not possible to establish change in behaviour at specific time intervals after clorazepate, but combined performance data revealed an effect. 3. The maximum mean plasma concentrations of diazepam, temazepam, oxazepam and clorazepate were observed at 0.5 h, and the maximum mean plasma concentration of nordiazepam was observed at 1 hour. Plasma concentrations of nordiazepam were the highest and decreased monoexponentially. Plasma concenqrations of the other drugs declined rapidly at first but more slowly later, and these data were analysed as biexponential models. In the analysis for metabolites, nordiazepam reached measurable levels after the injection of diazepam and clorazepate. 4. It is suggested that differences in the effects of closely related benzodiazepines may not be due solely to their plasma pharmacokinetic properties, but may arise from differences in their intrinsic activity.     

Pharmacokinetics of diazepam and nordiazepam in the cat

The cat has been used extensively as an experimental model for studying the pharmacology of compounds that exhibit CNS activity including diazepam and nordiazepam. However, since little is known about the distribution and elimination of diazepam in this species, the pharmacokinetics of diazepam and nordiazepam were studied in the cat following intravenous doses of 5, 10, and 20 mg/kg of diazepam and 5 and 10 mg/kg of nordiazepam. The disappearance of diazepam and nordiazepam from blood was fitted with classical equations. Theoretical and trapezoidal areas under the curve (AUCth and AUCtr) were calculated. The volumes of distribution (Vd beta) were calculated as model-independent parameters for diazepam and nordiazepam. Intrinsic hepatic clearance, extraction ratio, and tissue binding parameters were also calculated for diazepam. From the observed data, it is apparent that the blood concentrations and the resulting areas under the curves are proportional to the dose of diazepam administered and that the pharmacokinetics of diazepam were linear over the dose range studied. In addition, nordiazepam formed after diazepam administration appeared to be proportional to the dose of diazepam administered. The terminal elimination rate constant of nordiazepam remained constant over the dose range studied. It appears that both diazepam and nordiazepam are highly bound to tissue. The total body clearance of diazepam (4.72 +/- 2.45 mL/min/kg) is approximately six times that of nordiazepam (0.85 +/- 0.25 mL/min/kg). Approximately 50% of an administered dose of diazepam was biotransformed to nordiazepam in the cat.     

甲状腺功能对地西泮及其代谢产物在大鼠体内代谢的影响

目的：研究甲状腺功能对地西泮及其代谢物的药代动力学影响。方法：采用HPLC技术，测定不同甲状腺功能状态时，大鼠血液中地西泮及其体内主要代谢产物去甲基地西泮的浓度。结果：甲亢组大鼠地西泮在体内消除加速，峰浓度下降，AUC减少，消除T1/2缩短。甲减组大鼠则消除减慢，峰浓度增高，AUC增大，消除孔。延长。而其主要代谢产物去甲地西泮的药动学参数则相反。结论：甲状腺功能提高时，大鼠对地西泮的代谢能力明显增加，消除加速；而甲腺功能降低则相反。     

The urinary elimination profiles of diazepam and its metabolites, nordiazepam, temazepam, and oxazepam, in the equine after a 10-mg intramuscular dose

A method for the extraction of diazepam and its metabolites (nordiazepam, temazepam, and oxazepam) from equine urine and serum and their quantitation and confirmation by liquid chromatography-tandem mass spectrometry is presented. Valium, a formulation of diazepam, was administered at a dose of 10 mg intramuscularly to four standard-bred mares. Diazepam is extensively metabolized in the horse to nordiazepam, temazepam, and oxazepam. Diazepam urinary concentrations were found to be less than 6 ng/mL. Nordiazepam was found to be mainly in its glucuronide-conjugated form and was measured out to a collection time of 53-55 h. Oxazepam and temazepam were entirely conjugated, and their urinary concentrations were measured out to collection times of 121 h and 77-79 h, respectively. Diazepam and nordiazepam were measured in equine postadministration serum out to collection times of 6 and 54 h, respectively. Oxazepam and temazepam were not detected in postadministration serum.     

Plasma concentrations of benzodiazepines.

1. Twenty anxious patients were treated with medazepam, diazepam, chlordiazepoxide, amylobarbitone and placebo, each given in flexible dosage for 2-4 weeks. 2. At the end of each treatment, a series of clinical, physiological and behavioural variables were measured and plasma samples were taken for estimation of the appropriate drug and metabolite concentrations. 3. Nordiazepam was shown to be an important metabolite of both medazepam and diazepam: the ratio of medazepam to noradiazepam was 0.14 and the ratio of diazepam to nordiazepam following diazepam administration was 0.72. 4. No significant correlations were found between the plasma concentrations of any of the treatments and the clinical ratings or behavioural measures. 5. Some relationship was shown between levels of medazepam and its physiological effects.     

A comparison of the physical dependence inducing properties of flunitrazepam and diazepam

Dogs dosed chronically (4-7 weeks) with oral flunitrazepam (7.6 mg/kg/day) or diazepam (24-36 mg/kg/day) administered in 4 equally divided doses had dose-related flumazenil precipitated benzodiazepine abstinence scale scores (BPAS) of comparable intensities despite the fact that plasma levels of flunitrazepam and its metabolites were much lower than nordiazepam levels in the diazepam-dependent dog. Both groups of dependent dogs had clonic and tonic-clonic seizures after oral and IV flumazenil. Precipitated abstinence signs persisted longer in the diazepam than in the flunitrazepam-dependent dogs. Differences in the pharmacokinetics of the drugs of dependence, their metabolites, and their interactions at receptor sites offer a partial explanation for the high level of dependence seen in the flunitrazepam dog. The finding that the estimated plasma free concentration of flunitrazepam and its metabolites is equal to or greater than that of diazepam and its metabolites together with the fact that flunitrazepam has a higher affinity for the benzodiazepine receptor than either diazepam, nordiazepam or oxazepam can explain why the intensity of the precipitated abstinence syndrome is comparable in flunitrazepam- and diazepam-dependent dogs. Although the flumazenil-induced precipitated abstinence syndromes observed in flunitrazepam- and diazepam-dependent dogs differed qualitatively they did not differ quantitatively. It is therefore concluded from these data that the doses of flunitrazepam and diazepam, chosen for producing comparable degrees of weight loss during dose escalation, did not differ in the degree of physical dependence that they produced in the dog.     

Chronic administration of and dependence on halazepam, diazepam, and nordiazepam in the dog.

Halazepam administered chronically to dogs in oral doses of 180 and 450 mg/kg/day produced physical dependence which was revealed by a flumazenil precipitated abstinence syndrome and measured by the Nordiazepam Precipitated Abstinence Scale score (NPAS) (McNicholas et al., 1988; Sloan et al., 1990). This abstinence as measured by the NPAS score was more severe in diazepam- and halazepam-dependent than in nordiazepam-dependent dogs whereas the incidence of precipitated clonic seizures was greater in the diazepam- and nordiazepam-dependent than in the halazepam-dependent dogs. Pharmacokinetic studies showed that in the dog the major conversion of halazepam, like diazepam, was to nordiazepam and an oxazepam conjugate. Appreciable quantities of oxazepam, 3-OH halazepam and its conjugated metabolite were also identified in plasma. The NPAS score obtained in the halazepam-dependent dogs, however, was greater than the NPAS score obtained in nordiazepam-dependent dogs who had nordiazepam plasma levels over three times higher than those obtained in the halazepam-dependent dogs. Further, the precipitated abstinence observed in the halazepam-, diazepam- and nordiazepam-dependent dogs differed in qualitative as well as in quantitative aspects including marked differences in the time course of abstinence signs. These data argue that the different dependencies produced by halazepam, diazepam and nordiazepam are not due solely to either the parent compound or to a single metabolite but most likely to their combined effects.     

A rapid HPLC procedure for the simultaneous determination of chloroquine, monodesethylchloroquine, diazepam, and nordiazepam in blood

Chloroquine, monodesethylchloroquine, diazepam, and nordiazepam levels are simultaneously determined in whole blood or plasma by HPLC. Papaverine is used as internal standard, and the analysis is performed after protein-binding hydrolysis, absorption on Extrelut, and elution with diethyl ether/methylene chloride (70:30 v/v). UV detection is used at 343 nm for 12 min, then changed to 242 nm. There are two mobile phases with two flow rates. The procedure requires 30 min, is reproducible, sensitive (8-10 ng/mL for chloroquine and its metabolite, 4 ng/mL for diazepam and nordiazepam), and selective, especially towards other antimalarial agents and drugs like adrenaline or barbiturates, which may be used in chloroquine poisoning therapy. It can be used for pharmacokinetic studies, therapeutic control, to establish the diagnosis and prognosis of a chloroquine poisoning, and to follow and optimize treatment.