反相高效液相色谱法测定人血浆中氨溴索浓度

目的：建立测定血浆中氨溴索浓度的反相高效液相色谱法。方法：血浆样品用乙醚提取,醚层经００１ｍｏｌ·Ｌ－１盐酸提取后进样,色谱柱为ＲｅｓｏｌｖｅＣ１８４６ｍｍ×２５０ｍｍ（５μｍ）,流动相为乙腈－甲醇－００１ｍｏｌ·Ｌ－１ｐＨ７０磷酸盐缓冲液－四氢呋喃（３５０∶３５０∶２７５∶２５）,流速１５ｍＬ·ｍｉｎ－１,检测波长２４２ｎｍ,外标法峰高定量。结果：本法最低检测浓度５ｎｇ·ｍＬ－１,线性范围１０～３２０ｎｇ·ｍＬ－１,回归方程为Ｈ＝２４５＋１６４Ｃ,ｒ＝０９９９５,日内ＲＳＤ为２７％～５３％,日间ＲＳＤ为３２％～８２％。结论：该法适用于盐酸氨溴索片的药代动力学研究。     

高效液相色谱法测定盐酸氟桂利嗪血浆浓度及其药物动力学

以高效液相色谱法测定血浆中的盐酸氟桂利嗪浓度。色谱条件：紫外检测波长２１０ｎｍ；柱ＳｐｈｅｒｉｓｏｒｂＣ８１５０×４．６ｍｍ；流动相：甲醇（８０％）∶水（１５％）∶（０．０５ｍｏｌ·Ｌ－１ＮＨ４Ｈ２ＰＯ４＋０．０２５ｍｏｌ·Ｌ－１Ｈ３ＰＯ４）缓冲液（５％）。最低检测限１０ｎｇ·ｍｌ－１。药物动力学为一室模型，其参数（Ｔ１／２（Ｋ）２．３８～２．６４ｈ，Ｔｍａｘ２．３０～２．４３ｈ，Ｃｍａｘ１３５．１３～１４３．９６ｎｇ·ｍｌ－１。     

HPLC法测定复方福尔可定糖浆的含量

目的： 应用高效液相色谱法, 测定复方福尔可定糖浆的含量。方法： 采用ＯＤＳ 柱（２００ ｍ ｍ×４－６ ｍ ｍ） , 以０－０２ ｍｏｌ·Ｌ－ １ 庚烷磺酸钠甲醇溶液－ ０－０５ ｍｏｌ·Ｌ－ １ 磷酸二氢钾溶液－ 三乙胺（３０∶７０∶１ , 用磷酸调ｐＨ 为３－０） 为流动相, 检测波长为２１０ ｎｍ 。结果： 平均回收率（ ｎ ＝ ５） 福尔可定为９９－３ ％ , ＲＳＤ＝ ０－７ ％ ; 愈创木酚甘油醚为９９－８ ％ , ＲＳＤ＝ ０－５ ％ ; 盐酸麻黄碱为９９－５ ％ , ＲＳＤ＝ ０－４ ％ 。结论： 实验结果表明, 该方法简便, 准确, 灵敏度高, 重现性好。     

反相高效液相色谱法测定洛氟沙星的血药浓度

采用反相ＨＰＬＣ法测定洛氟沙星的血药浓度，血浆样品用二氯甲烷在ｐＨ７．０条件下提取后进样，流动相为１０ｍ ｍｏｌ／Ｌ磷酸二氢钾－１０ｍ ｍｏｌ／Ｌ溴化四丁铵－乙腈－三乙胺（４５：４４：１０：１，磷酸调节ｐＨ２．８），紫外检测器λ＝２９５ｎｍ，最低检测浓度１０μｇ／Ｌ，线性范围：０．１￣６．０ｍｇ／Ｌ，ｒ＝０．９９９９，日内ＲＳＤ为１．３８％￣３．４２％，日间ＲＳＤ为１．１１％￣２．８９％。     

粉防己碱，Fura 2－AM和Bay k 8644对脂多糖刺激大鼠腹腔巨噬细胞释放血小板活化因子的影响

目的：研究钙与脂多糖（ＬＰＳ）刺激大鼠腹腔巨噬细胞（ＰＭ）释放血小板活化因子（ＰＡＦ）的关系．方法：通过ＰＡＦ的生物测定法，观察了粉防己碱（Ｔｅｔ）、Ｆｕｒａ２ＡＭ和Ｂａｙｋ８６４４对ＬＰＳ刺激ＰＭ释放ＰＡＦ的影响．结果：ＬＰＳ刺激ＰＭ释放ＰＡＦ，但并不使其细胞内钙增高，Ｔｅｔ在０１，１０，１０，１００μｍｏｌ·Ｌ－１和Ｆｕｒａ２ＡＭ在００１，０１，１０，１０μｍｏｌ·Ｌ－１时降低ＬＰＳ刺激的ＰＭ释放ＰＡＦ（分别为９８±１１，６５±１６，４７±０８，３４±０４，９２±１７，５２±１３，３７±０４，３．２±０３μｇ·Ｌ－１，无药物时１１８±１２μｇ·Ｌ－１），Ｂａｙｋ８６４４在１０，５０，１０μｍｏｌ·Ｌ－１时能增加ＬＰＳ刺激的ＰＡＦ释放能增加ＬＰＳ刺激的ＰＡＦ释放（分别为１３２±１７，１６２±１４，１７６±１５μｇ·Ｌ－１），并且Ｂａｙｋ８６４４在５０μｍｏｌ·Ｌ－１时能全部或部分逆转Ｔｅｔ和Ｆｕｒａ２ＡＭ对ＰＡＦ释放的抑制作用．结论：尽管ＬＰＳ并不明显增加巨噬细胞内钙，但细胞内钙对ＬＰＳ刺激的ＰＡＦ释放是必要的．     

双波长分光光度法测定甲硝唑氧氟沙星注射液含量

目的：应用双波长分光光度法测定甲硝唑氧氟沙星注射液含量。方法：以０．１ｍｏｌ．Ｌ＾－１盐酸液为溶剂,在波长２７７,３０７．４,２９３,２６０ｎｍ处进行测定。结果：甲硝唑浓度在４～２０ｕｇ．ｍｌ＾－１之间,氧氟沙星浓芳在２～１０ｕｇ．ｍｌ＾－１之间符合比尔定律,浓度与吸收度差值呈良好线性关系,甲硝唑平均回收率为９９．５％,ＲＳＤ为０．６％（ｎ＝８）,氧氟沙星平均回收率为１００．７％,ＲＳＤ为０．４％     

Studies on Quantitative Determination of Active Principles in Chinese Herbal Medicine by Second Derivative Differential Pulse Polarography

本文建立了中草药黄芩中黄芩甙、青黛中靛蓝、牡丹皮中丹皮酚、荜茇中胡椒碱的二阶导数差示脉冲极谱的定量分析方法。黄芩、靛蓝、丹皮酚和胡椒碱的化学结构中都含有ＣＯ，在酸性条件下，于滴汞电极上均可发生还原反应生成ＣＯ，并分别于１５５０Ｖ，１３００Ｖ，１６３０Ｖ，０８６０Ｖ（ｖｓＡｇ／ＡｇＣｌ）处出现良好的二阶导数差示脉冲极谱峰。黄芩、靛蓝、丹皮酚和胡椒碱分别在４５×１０５～２７×１０４ｍｏｌ·Ｌ１，３８×１０５～２５×１０４ｍｏｌ·Ｌ１，１０～６０×１０４ｍｏｌ·Ｌ１和７０×１０５～２５×１０４ｍｏｌ·Ｌ１范围内，浓度与其二阶导数差示脉冲极谱峰幅值呈极为显著的线性关系（Ｐ〈００１），检测限分别为９０×１０８ｍｏｌ·Ｌ１，８４×１０９ｍｏｌ·Ｌ１，９２×１０９ｍｏｌ·Ｌ１和８７×１０９ｍｏｌ·Ｌ１。方法简便、快速、灵敏，结果准确。     

血浆头孢曲松HPLC测定法及其在胸外术后患者的应用

建立血浆中头孢曲松反相ＨＰＬＣ测定法,用于胸外术后患者头孢曲松血药浓度的测定。血浆样品经乙腈沉淀蛋白等预处理后,于ＯＤＳ柱上,以甲醇－０．１ｍｏｌ／Ｌ 到铵缓冲液（２０：８０）为流动相进行分离测定。线性范围０．２～８９ｍｇ／Ｌ,沉淀蛋白上率９１．６～９４．４％,加样回收率１０１．５％～１０３．２％,日蚋ＲＳＤ１．０９％～２．６５％。日间ＲＳＤ４．４２％～５．１３％。本法准确可靠,用于胸外术后患者头     

高效液相色谱法测定4－［4″－（2″，2″，6″，6″－四甲基哌啶氮氧自由基）氨基］－4＇－去甲表鬼臼毒素大鼠血浆浓度及药代动力学参数

建立了大鼠血浆中４－［４″－（２″，２″，６″，６″－四甲基哌啶氮氧自由基）氨基］－４＇－去甲表鬼臼毒素（ＧＰ－７）浓度的ＨＰＬＣ测定方法。用ＯＤＳ柱分离，甲醇－水－冰醋酸（５９：４１：０．６）为流动相，检测波长２８５ｎｍ。血浆甲乙醚－二氯甲烷混合液萃取，４５℃水浴蒸干，残渣用流动相溶解进样，内标法定量；血药浓度在２～２００μｇ·ｍｌ－１范围内呈线性关系，ｒ＝０．９９９７，血浆最低检测浓度０．２μｇ·ｍｌ－１，方法回收率９４％～１００％，日内、日间ＲＳＤ２．２９％～７．７０％。大鼠ｉｖＧＰ－７１０，２０，３０ｍｇ·ｋｇ－１，药代动力学过程符合二室开放模型，消除半衰期为３９．８±１０．８ｍｉｎ。     

反相高效液相色谱法测定阿霉素的血药浓度

本文采用反相ＨＰＬＣ法测定阿霉素的血药浓度，血浆样品用甲醇－氯仿（１：４）在ｐＨ９．０的条件下提取后进样，５ｍｍｏｌ／Ｌ磷酸－异丙醇－甲醇－乙腈（４５：３５：１０：１０，ｐＨ２．９）为流动相，荧光检测器λｅｘ＝４５０ｎｍ，λｅｍ＝５３０ｎｍ。最低检测浓度１０ｎｇ／ｍｌ，线性范围在３０－１５００ｎｇ／ｍｌ内ｒ＝０。９９８７，日内ＲＳＤ为１．５％－２．４％，日间ＲＳＤ为１．８％－３．７％。并对７例肺癌     

高效液相色谱—间接光度检测法同时测定血清和尿中5种氨基苷类抗生素

目的：建立一种直接分离测定体液中庆大霉素、丁胺卡那霉素、妥布霉素、西梭霉素和乙基西梭霉素等５种氨基苷类抗生素的高效液相色谱－间接光度检测（ＨＰＬＣ－ＩＰＤ）法。方法：在流动相中加入具有紫外检测响应的检测剂烟酰胺,用紫外检测器直接测定紫外吸收很差的上述５种药物。Ｃ１８固定相,流动相为含烟酰胺０５ｍｍｏｌ·Ｌ－１、庚烷磺酸钠５ｍｍｏｌ·Ｌ－１和磷酸００５ｍｏｌ·Ｌ－１的甲醇－乙腈－水（２７∶１８∶５５）混合溶液。结果：血清和尿样平均回收率均大于９６％,日内和日间ＲＳＤ均小于６％。并测定了肌注此类药物病人的血清和尿样品。结论：该法适于体液中氨基苷类药物检测。     

Decreased Lithium Disposition to Cerebrospinal Fluid in Rats with Glycerol-induced Acute Renal Failure

PURPOSE: The lithium disposition to cerebrospinal fluid (CSF) was evaluated in rats with acute renal failure (ARF) to examine whether electrolyte homeostasis of the CSF is perturbed by kidney dysfunction. In addition, the effects of renal failure on choroid plexial expressions of the Na(+)-K(+)-2Cl(-) co-transporter (NKCC1) and Na(+)/H(+) exchanger (NHE1) were also studied. METHODS: After lithium was intravenously administered at a dose of 4 mmol/kg, its concentration profile in plasma was evaluated by collecting plasma specimens, while that in CSF was monitored with a microdialysis probe in the lateral ventricles. NKCC1 and NHE1 expressions were measured via the Western immunoblot method using membrane specimens prepared from the choroid plexus in normal and ARF rats. RESULTS: The lithium concentration in CSF of ARF rats was 30% lower than that of normal rats, while their plasma lithium profiles were almost the same, indicating that the lithium disposition to CSF was decreased in ARF rats. It was revealed that the choroid plexial expression of NKCC1 was increased by 40% in ARF rats, but that of NHE1 was unchanged. CONCLUSION: ARF decreases the lithium disposition to CSF, possibly by promoting lithium efflux from CSF due to increased NKCC1 expression in the choroid plexus.     

Free and bound propofol concentrations in human cerebrospinal fluid

AIMS: The aim of this study was to define the relationship between unbound propofol concentrations in plasma and total drug concentrations in human cerebrospinal fluid (CSF), and to determine whether propofol exists in the CSF in bound form. METHODS: Forty-three patients (divided into three groups) scheduled for elective intracranial procedures and anaesthetized by propofol target control infusion (TCI) were studied. Blood and CSF samples (taken from the radial artery, and the intraventricular drainage, respectively) from group I (17 patients) were used to investigate the relationship between unbound propofol concentration in plasma and total concentration of the drug in CSF. CSF samples taken from group II (18 patients) were used to confirm the presence of the bound form of propofol in this fluid. The CSF and blood samples taken from group III (eight patients) were used to monitor the course of free and bound CSF propofol concentrations during anaesthesia. RESULTS: For group I patients the mean (and 95% confidence interval) total plasma propofol concentration was 6113 (4971, 7255) ng ml(-1), the mean free propofol concentration in plasma was 63 (42, 84) ng ml(-1), and the mean total propofol concentration in CSF was 96 (76, 116) ng ml(-1) (P < 0.05 for the difference between the last two values). For group II patients the fraction of free propofol in CSF was 31 (26, 37)%. For group III patients the fraction of free propofol in CSF during TCI was almost constant (about 36%). CONCLUSIONS: The unbound propofol concentration in plasma was not equal to its total concentration in CSF and cannot be directly related to the drug concentration in the brain. Binding of propofol to components of the CSF may be an additional mechanism regulating the transport of the drug from blood into CSF.     

Liquid chromatography tandem mass spectrometry assay for topiramate analysis in plasma and cerebrospinal fluid: validation and comparison with fluorescence-polarization immunoassay

SUMMARY: The authors report the development and validation of a liquid chromatography tandem mass spectrometry assay (LC/MS/MS assay) for the analysis of topiramate (2,3:4,5-bis-o-(-1-methyl)-beta-D-fructopyranose sulfamate) in plasma and cerebrospinal fluid (CSF). Comparison is made with the commercially available fluorescence-polarization immunoassay (FPIA). LC/MS/MS ASSAY: Using the internal standard, 1,2:3,4-bis-o-(1-methylethylidene-alpha-D-galactopyranose sulfamate), a structural isomer, the calibration curve in plasma was linear in the concentration range of 0.02-20.0 mg/L (r(2) = 0.9998). The coefficients of variation in plasma were < or = 3%, and the accuracy ranged from 100% to 101% in the therapeutically relevant concentration range of 0.4-16.0 mg/L. In CSF, the mean recovery was 98%, and there was linearity between the nominal and the estimated concentration in the range of 1.5-20.0 mg/L (r(2)= 0.9996). FPIA: The calibration curve was linear in the concentration interval of 1.6-24.3 mg/L (r(2) = 0.9994), and the mean recovery was 96%. Accuracy in plasma was 99- 104%, and precision was 3.2-6.0%. In CSF, there was linearity between the nominal concentration and the estimated concentration in the range of 1.5-20.0 mg/L (r(2) = 0.9995), and the mean recovery was 100%. COMPARISON BETWEEN FPIA AND LC/MS/MS: There was a high correlation between the FPIA and the LC/MS/MS assay (r(2) = 0.9965 in plasma and r(2) = 0.9996 in CSF, P < 0.001 for both). In plasma and CSF, the two methods showed equal results, evaluated as the ratio between the two methods (plasma: median ratio = 1.00; 95% confidence interval [CI], 0.98-1.02, paired-sample test, P = 0.79; and CSF: median ratio = 1.00, 95% CI, 0.99-1.02, paired-sample test, P = 0.75). The coefficient of variation on the ratios between the two methods had similar levels: 5% in plasma and 3% in CSF. CONCLUSION: The new LC/MS/MS assay has favorable characteristics, being highly precise and accurate. FPIA also proved precise and accurate, and there was a high agreement with the LC/MS/MS assay in plasma and CSF. Either method displayed sufficient precision and accuracy and may thus be implemented in daily routine.     

Pharmacokinetics of cytosine arabinoside, methotrexate, nimustine and valproic acid in cerebrospinal fluid during cerebrospinal fluid perfusion chemotherapy

This report investigates the pharmacokinetics of cytosine arabinoside (Ara-C), methotrexate (MTX), nimustine (ACNU) and valproic acid (VPA) in cerebrospinal fluid (CSF) during CSF perfusion chemotherapy. A 28-year-old Japanese woman with disseminated glioblastoma was, on admission, on a stable oral regimen of prolonged-release VPA tablets (Depakene-R), 400 mg twice a day, for seizure control. Twelve courses of CSF perfusion chemotherapy with Ara-C, MTX, and ACNU were administered. Plasma samples and CSF samples via Ommaya reservoirs were obtained during the eleventh course of treatment. The Ara-C and ACNU concentrations were measured by HPLC. The MTX and VPA concentrations were measured by fluorescence polarization immunoassay. During CSF perfusion chemotherapy, the highest CSF concentrations of Ara-C, MTX, and ACNU were observed at the end of the perfusion and decreased in a monoexponential pattern. The half-lives of Ara-C, MTX, and ACNU were 2.65, 3.52, and 0.71 h, respectively. No anticancer drugs were detectable in plasma during CSF perfusion chemotherapy. Before CSF perfusion chemotherapy, the free VPA concentration in plasma was 14.4% of the total VPA concentration. The mean total and free VPA concentrations in plasma were 78.0+/-0.8 and 10.9-0.3 microg/ml, respectively. The free VPA concentrations in plasma and in CSF were of similar values. At the end of perfusion, the lowest free VPA concentration in CSF was 30.3% of that at the initiation of perfusion. The free VPA concentrations in CSF at 3, 7, 23, and 47 h after the end of perfusion were 79.8, 94.5, 100.9, and 100.9% respectively of that at the initiation of perfusion. During CSF perfusion chemotherapy, the ratio of free VPA concentrations to the total VPA in CSF was 86.3+/-6.9%. The VPA concentrations in CSF rapidly decreased during the CSF perfusion but recovered to pre-treatment levels within 7 h.     

Plasma protein-binding and CSF concentrations of valproic acid in man following acute oral dosing

Simultaneous cerebrospinal fluid (CSF), total and free plasma valproic acid (VPA) concentrations were measured in 17 patients receiving two weight-adjusted VPA doses as seizure prophylaxis prior to diagnostic myelography or cisternography. Free drug concentrations were similar when measured by equilibrium dialysis (ED) at 37 degrees C for 24 h (Dianorm) or by a novel ultrafiltration (UF) method (EMIT freelevel system 1, SYVA) (ED:2.3-35.5 mg-1; UF:1.3-33.6 mg-1; r = 0.78, P less than 0.002). There was wide variation in total VPA concentration (39-154 mg-1) and in free fraction (ED: 3.3-25.6%; UF: 5.9-24%). Concentration dependent protein binding was not demonstrated. CSF VPA varied between 4.2 and 25.6 mg-1 and was accurately reflected by free plasma VPA concentrations (ED: r = 0.75, P less than 0.005: UF: r = 0.93, P less than 0.001). CSF concentration also correlated with the total plasma VPA (r = 0.76, P less than 0.005). The Emit freelevel system 1 provides a rapid measure of unbound VPA in the plasma which may be suitable for routine clinical use.     

Pharmacokinetic modeling of plasma and cerebrospinal fluid methotrexate after high-dose intravenous infusion in children.

A pharmacokinetic study was performed in plasma and cerebrospinal fluid (CSF) of patients suffering from brain tumors to describe the disposition of methotrexate. An open three-compartment model was developed to fit together the data obtained in plasma and CSF. The pharmacokinetic parameters obtained by the model agreed with those obtained with classical analysis and the fitting correctly depicted the plasma and CSF concentration decays. According to the results, such a model could be applied to other anticancer drugs.     

Pharmacokinetics of nimustine, methotrexate, and cytosine arabinoside during cerebrospinal fluid perfusion chemotherapy in patients with disseminated brain tumors

Objective: This study was conducted to evaluate the pharmacokinetics of anticancer drugs in cerebrospinal fluid (CSF) perfusion chemotherapy. Methods: We administered CSF perfusion chemotherapy with nimustine (ACNU), methotrexate (MTX), and cytosine arabinoside (Ara-C) to three patients with disseminated malignant brain disease. The drugs were infused via Ommaya's reservoirs to the lateral ventricle and removed by drainage from the temporal lobe or lumbar spine. CSF and plasma concentrations of the anticancer drugs were determined by high-performance liquid chromatography and fluorescence polarization immunoassay. Results: The concentrations of anticancer drugs in the discharged CSF peaked about 40 min after the start of a 1-h CSF perfusion. After the perfusion, the drug level in CSF decreased exponentially in a monophasic manner. ACNU and Ara-C were not detectable in the discharged CSF in the temporal lobe at 6 h and 48 h after perfusion, respectively, but MTX was detectable at 48 h. The maximum concentration ratio of anticancer drugs and the duration of perfusion were inversely correlated. The plasma concentrations of anticancer drugs were much lower than those in CSF. The half-life of ACNU was very short (0.2–1.1 h), whereas the half-lives of MTX and Ara-C were relatively long (2.81–13.5 h and 1.84–6.25 h, respectively). The half-lives of the anticancer drugs in CSF tended to decrease with repeated CSF perfusion chemotherapy. Conclusion: Results suggest that CSF perfusion chemotherapy enables a high concentration of anticancer drug to be administered for dissemination in the spinal cord within a short period of time, with minimal adverse effects. Received: 17 September 1996 / Accepted in revised form: 21 April 1998     

Methotrexate concentration in cerebrospinal fluid of the space created by tumor removal

This paper investigates the post-surgical pharmacokinetics of methotrexate (MTX) in the plasma, the cerebrospinal fluids (CSF), and the spaces created by tumor removal (STR) in patients with glioblastoma, during hyperosmotic disruption of the blood brain barrier (HODBBB) and intra-arterial chemotherapy with MTX. Eight Japanese patients with glioblastoma, three with open STRs and five with closed STRs, received a total of thirteen courses of HODBBB and intra-arterial combination chemotherapy with MTX. The patients were initially administered mannitol, then the anticancer drugs were infused into the carotid artery. Samples of blood and CSF from the STRs were obtained. MTX concentrations were measured by fluorescence polarization immunoassay and the pharmacokinetic parameters of MTX in plasma and CSF were estimated. The plasma concentrations of MTX peaked at the end of drug infusion, and then decayed bi-exponentially during the remainder of the treatment period. The CSF concentration of MTX in the STR peaked 2 h after drug administration, then mono-exponentially decreased. The area under the concentration-time curve (AUC) for plasma and CSF MTX concentrations increased in parallel with the MTX dose. In patients with open STRs, the mean AUC of MTX in CSF was 4.44% of that found in plasma, while in patients with closed STRs, the mean was 61.2% of that found in plasma. In the latter group, the MTX administered using HODBBB and intra-arterial chemotherapy was maintained in the STRs for long periods.