RP-HPLC法同时测定人血浆中茶碱和多索茶碱的浓度

目的:建立以反相高效液相色谱法同时测定人血浆中茶碱和多索茶碱浓度的方法。方法:色谱柱为XterraRP18,流动相为甲醇-水(35:65),流速为1.0mL·min-1,检测波长为269.9nm,柱温为35℃,内标为非那西丁。结果:茶碱和多索茶碱浓度分别在1.563～50μg·mL-1(r=0.9995)、0.3125～10μg·mL-1(r=0.9999)范围内线性关系良好;茶碱、多索茶碱提取回收率分别为32.53%～35.08%、31.88%～34.55%,方法回收率分别为92.82%～109.41%、90.68%～102.72%,日内、日间RSD均<15%。结论:本方法快速、灵敏、准确,可用于同时测定人血浆中茶碱和多索茶碱的浓度。关键词茶碱;多索茶碱;反相高效液相色谱法;血药浓度监测     

HPLC法测定大鼠血浆中多索茶碱及代谢产物茶碱的浓度

目的:建立同时测定大鼠血浆中多索茶碱及其代谢产物茶碱的高效液相色谱法。方法:用18%高氯酸溶液处理血浆后,采用高效液相色谱法测定。色谱柱为Hypersil ODS C₁₈反相柱,流动相为甲醇-水（1:3）,紫外检测波长271 nm,柱温为室温,流速l ml·min^-1,进样量20μ1。结果:多索茶碱和茶碱的线性范围分别为0.5～50μg·ml^-1和0.2～20μg·ml^-1。日内和日间精密度RSD均小于9.6%。血浆样品稳定性符合要求。相对回收率97.02%～102.36%,绝对回收率大于85.0%。结论:本法专属性强,灵敏度高,重复性好,适用于多索茶碱的药物动力学研究。     

互为内标法测定人血清中茶碱与多索茶碱的浓度

目的建立人血清中茶碱与多索茶碱的HPLC测定法。方法血清经10%三氯乙酸沉淀蛋白,以Hypersil BDS-C18为色谱柱,甲醇-水（22：78,V/V）为流动相,流速1.0mL.min-1,柱温30℃,检测波长274nm;茶碱与多索茶碱互为内标。结果茶碱与多索茶碱均在1~40mg.L-1范围内线性关系良好,平均相对回收率均在95%~105%之间,日内、日间RSD均〈5%。结论本法操作简便、快速、灵敏度高,适用于临床茶碱与多索茶碱的药动学研究及常规血药浓度测定。     

HPLC法监测人血清中茶碱类药物浓度

目的建立同时测定茶碱类药物血药浓度的高效液相色谱法。方法筛选得到的最佳色谱条件为:采用色谱柱Ultimate XB-C18柱(4.6×200mm,5μm),以咖啡因为内标,甲醇-乙腈-水(20∶3∶77)为流动相,流速1.0m L·min-1,检测波长273nm,柱温30℃。采用先沉淀后提取的方法进样测定。结果多索茶碱和茶碱分别在1.5~24mg·L-1(r=1.0000)和2.5~40mg·L-1(r=0.9996)范围内线性关系良好。多索茶碱的低、中、高浓度平均回收率为100.07±5.30%,日间精密度和日内精密度RSD均小于6.5%。茶碱的低、中、高浓度回收率为101.72±4.92%,日间精密度和日内精密度RSD均小于5.5%。结论该方法灵敏度高、准确、重复性好,适用于多索茶碱和茶碱血药浓度的测定。     

气相色谱法测定多索茶碱注射液中多索茶碱的含量

目的用毛细管气相色谱法建立测定多索茶碱注射液中多索茶碱含量的方法。方法采用DM-17(30 m×0.32 mm×0.25μm)毛细管色谱柱,氮气为载体,FID检测器,柱温265℃,以盐酸罂粟碱为内标,按内标法计算多索茶碱的含量。结果多索茶碱在0.0597~1.194 g.L-1(r=0.9998)范围内有较好的线性关系(n=6),方法回收率(n=9)为98.0%,RSD为1.4%。结论该法操作简便,快速,准确,对于含多索茶碱药物的质量检测有一定参考价值。     

高效液相色谱法同时测定人血清中茶碱、二羟丙茶碱、多索茶碱的浓度

目的:建立人血清中茶碱、二羟丙茶碱和多索茶碱血药浓度同时测定的高效液相色谱法并应用于患者的个体化治疗。方法:采用蛋白沉淀法对血样进行预处理,以咖啡因为内标。色谱条件:Eclipse XDB-C₁₈(4.6 mm×150 mm,5 μm)色谱柱,流动相为甲醇-水梯度洗脱,流速为1.0 ml·min^-1,检测波长273 nm,柱温为30 ℃。结果:茶碱在1.47~47.00 μg·ml^-1范围内线性良好(r=0.998),二羟丙茶碱在1.28~40.80 μg·ml^-1范围内线性良好(r=0.999),多索茶碱在1.44~46.00 μg·ml^-1范围内线性良好(r=0.999)。茶碱、二羟丙茶碱和多索茶碱的平均绝对回收率均达到95%左右,平均相对回收率分别为100.34%~105.48%,97.09%~103.13%和97.61%~101.39%,日内和日间RSD均小于10%。结论:本方法简便、快速、灵敏,可用于茶碱、二羟丙茶碱和多索茶碱的临床血药浓度监测及药动学研究。     

RP-HPLC法测定人血清中茶碱与多索茶碱的浓度

目的:建立以反相高效液相色谱法同时测定人血清中茶碱、多索茶碱浓度的方法。方法:血清采用15%高氯酸进行沉淀后测定,其中色谱柱为InertsilODS-SP,流动相为甲醇-水(25∶75),检测波长为273nm,流速为1mL·min-1,柱温为30℃,内标为巴比妥钠。结果:茶碱、多索茶碱血清浓度分别在0.24～30.69mg·L-1、0.25～31.83mg·L-1范围内线性关系良好(r均为0.9999),最低定量限分别为0.24、0.25mg·L-1;平均相对回收率分别为99.62%～103.92%、96.26%～102.22%;日内RSD分别为0.44%～1.79%、0.54%～1.68%;日间RSD分别为1.68%～2.15%、0.83%～1.57%。结论:本方法简便、快速、准确、灵敏,可用于临床茶碱、多索茶碱血药浓度监测。     

茶碱类药物的血药浓度监测方法与应用

目的测定茶碱和多索茶碱的血清浓度,为临床血药浓度监测提供方法学依据。方法采用高效液相色谱法测定,以内标法定量。色谱柱为ShimpackCLCC18(150×6mm,5μm),流动相为0.02mol·L-1KH2PO4乙腈(85∶15),流速为1ml·min-1,检测波长为270nm。结果茶碱和多索茶碱在0.4～40μg·ml-1范围内呈良好的线性关系。茶碱的低、中、高浓度回收率分别为105.64%、104.23%和99.09%,日内RSD(相对标准偏差)分别为3.4%、1.1%和2.5%,日间RSD分别为5.6%、3.5%和3.8%,平均回收率为102.95%(n=36,RSD=5.0%)。多索茶碱的低、中、高浓度回收率分别为101.53%、103.78%和99.89%,日内RSD分别为4.1%、1.2%和1.6%,日间RSD分别为6.1%、4.4%和2.5%,平均回收率为101.76%(n=36,RSD=4.7%)。结论方法快速、简便、准确,适合于临床对茶碱和多索茶碱的血药浓度监测。     

多索茶碱与氨茶碱治疗支气管哮喘的临床分析

目的比较分析多索茶碱与氨茶碱治疗支气管哮喘的临床效果。方法我院2012年1月至2013年12月间收治的支气管哮喘患者66例,按照治疗方案分为多索茶碱(34例)和氨茶碱组(32例),比较两组治疗效果及不良反应发生情况。结果多索茶碱组患者的治疗有效率为94.11%,而氨茶碱组有效率为71.88%,氨茶碱有效率比多索茶碱低;多索茶碱组不良反应发生率达11.76%,明显低于氨茶碱组31.25%(P<0.05)。结论支气管哮喘中应用多索茶碱效果优于氨茶碱,且不良反应发生率更低,值得临床推广应用。     

葛根素注射液对多索茶碱在大鼠体内药动学的影响

目的研究葛根素对多索茶碱在大鼠体内药动学的影响。方法采用高效液相色谱（HPLC）法测定12只大鼠单独给药和联合葛根素给药后多索茶碱及其代谢产物茶碱的血浆浓度,采用DAS药动学软件对多索茶碱血浆浓度 时间数据进行非房室模型分析,求多索茶碱在大鼠体内的主要药动学参数,并进行统计分析。结果两组大鼠多索茶碱t1/2差异有统计学意义（P＜0.05）,合并用药组较单独用药组增加了6.65%;CL/F差异有统计学意义（P＜0.05）,合并用药组较单独用药组减少38.00%。单独给药组和合并给药组茶碱AUC（0 8）、Cmax、tmax差异无统计学意义（P＞0.05）,t1/2差异有统计学意义（P＜0.05）,合并用药组较单独用药组增加了35.29%;CL/F差异有统计学意义（P＜0.05）,合并用药组较单独用药组减少了35.29%。结论葛根素能抑制多索茶碱在大鼠体内代谢,且能延长多索茶碱代谢产物茶碱在体内的消除,两药同时使用应注意监测血茶碱浓度。     

高效液相色谱法测定芹菜素在不同血浆中的蛋白结合率

目的:建立测定芹菜素在大鼠血浆、人血浆和牛血清白蛋白(BSA)中蛋白结合率的方法,并计算不同介质中的的相关参数。方法:采用高效液相色谱(HPLC)测定不同介质中芹菜素的浓度,并结合平衡透析法测定蛋白结合率。结果:高、中、低浓度下,芹菜素的血浆蛋白结合率分别为:大鼠血浆:(99.4±3.8)%、(99.6±5.6)%、(99.5±4.5)%;人血浆:(96.3±7.2)%、(97.9±10.5)%、(97.8±9.7)%;牛血清白蛋白:(98.7±8.6)%、(99.6±9.4)%、(99.1±8.0)%。结论:本方法快速、简便、可靠,芹菜素与大鼠血浆、人血浆和牛血清白蛋白结合率很高,且与血药浓度无显著相关性。     

A third, "deep," compartment for phenothiazine drug disposition: a new look at an old problem

Volunteers were given oral promazine doses, and blood and urine were collected for promazine assay for as long as was necessary for the drug to cease to be detectable. Postabsorption data were subjected to two- and three-compartment modeling. Renal clearance was calculated. Renal excretion data were analyzed by use of the sigma-minus method, which permits the calculation of the half-life in plasma from excretion data. The mean alpha-phase rate constant for plasma decay (plasma assays) was .0155 min-1. The mean beta-phase rate constant for plasma decay (plasma assays) was .0046 min-1; the mean beta-phase rate constant for plasma decay (urine assays) was .0056 min-1. The mean gamma-phase rate constant for plasma decay (urine assays) was .00075 min-1. These data demonstrate the existence of a third, deep, distribution compartment from which promazine is released slowly and which is only detectable by means of excretion studies.     

Factors affecting the free plasma fraction of phenytoin in patients with epilepsy

The variability and possible factors modifying the free plasma fraction of phenytoin were investigated in 134 patients with epilepsy undergoing long-term treatment. The total and free plasma concentrations of phenytoin were determined using fluorescent polarisation immunoassay. Concentrations of albumin, bilirubin and creatinine were also obtained. The free plasma concentration was separated by ultrafiltration, at 25 degrees C, using Centrifree((R)) filters. Factors related to the free plasma fraction of phenytoin (free plasma concentration/total plasma concentration) were gender, age, dose, therapeutic regimen, total plasma concentration and the biochemical parameters mentioned. The mean of free plasma fraction was 9.1% with a very high variability (between 3.3 and 37%). No significant relationship was found between the free plasma fraction and dose, age, gender, total plasma concentration or the biochemical data. The only variable with a significant effect (p < 0.05) on the free plasma fraction was polytherapy with valproic acid. The variability in the free plasma fraction of phenytoin was high in epileptic patients, and was poorly related to the clinical or analytical variables studied. In the absence of pathologies that modify phenytoin binding (uraemia, hypoalbuminaemia), the only factor predictive of a possible alteration in the binding of phenytoin to plasma proteins was polytherapy with valproic acid.     

Nonlinear Distribution of Atenolol Between Plasma and Cerebrospinal Fluid

Long Evans rats were given atenolol doses ranging from 0.27 to 5.4 mg/kg by intraperitoneal injection. Animals were dosed once every 2 hr for a total of five doses. Atenolol concentrations 1 hr after the last dose were measured from simultaneously obtained plasma and cerebrospinal fluid (CSF) samples. CSF concentrations of atenolol were not proportional to plasma concentrations. The ratio of CSF/plasma concentrations was higher (0.33) at lower plasma atenolol levels (<100 ng/ml) than at the higher atenolol plasma levels (0.05) (P < 0.001). The relationship between plasma and cerebrospinal fluid atenolol concentrations was best described by the sum of a Michaelis–Menten and linear function. Animals were also given atenolol doses and then subjected to global cerebral ischemia. The relationship of atenolol concentrations from plasma and CSF in these animals was linear, with a constant partition ratio of 0.02. Together these data show that atenolol partitioning between plasma and CSF is nonlinear and possibly an energy-dependent process.     

病毒灭活血浆与普通冰冻血浆的临床应用价值分析

目的对病毒灭活血浆与普通冰冻血浆的临床应用价值进行对比研究与分析,为临床提供参考依据。方法选取进行病毒灭活血浆输注的72例患者作为观察组,选取同期输注普通冰冻血浆的72例患者作为对照组。比较两组患者血浆蛋白质检测结果、部分凝血因子含量、输血不良反应发生情况。结果观察组患者血浆总蛋白、纤维蛋白原及白蛋白含量分别为(48.21±6.62)、(0.89±0.16)、(29.78±6.91)g/L,均明显低于对照组的(59.71±4.52)、(2.41±0.27)、(34.82±7.17)g/L,差异均具有统计学意义(P<0.05)。观察组患者凝血因子Ⅴ为(6.22±1.31)、Ⅷ为(5.85±1.54)、Ⅹ为(38.44±3.42),对照组患者凝血因子Ⅴ为(8.19±1.52)、Ⅷ为(8.11±2.05)、Ⅹ为(56.81±6.24);观察组患者凝血因子Ⅴ、Ⅷ、Ⅹ均低于对照组,差异具有统计学意义(P<0.05)。观察组患者的输血不良反应发生率0显著低于对照组的6.94%,差异具有统计学意义(P<0.05)。结论虽病毒灭活血浆的血浆蛋白与部分凝血因子低于普通冰冻血浆,但输血治疗时,其风险很低;而普通冰冻血浆能够补充全部的稳定凝血因子,因此二者应该合理选择,充分有效的利用血液资源,提高临床疗效。     

肠易激综合征血浆胃肠激素含量与直肠肛管压力的相关性

目的 研究直肠肛管压力及胃肠激素的改变在肠易激综合征（IBS）发病中的作用,以及探讨胃肠激素与直肠肛管压力之间的关系。方法 采用五通道环状固态压力传感器测压导管测定43例IBS患者直肠肛管的压力,以及采用放射免疫分析法检测血浆胃泌素、胃动素、胰高糖素的水平,并与20例正常人作对照。结果 腹泻型IBS患者直肠静息压、肛管静息压较对照组高,便秘型IBS患者最大缩榨压较对照组低;腹泻型IBS患者的初始感觉量、排便感觉量、紧迫感觉量、最大耐受量均较正常对照组为低,而便秘型IBS患者均较对照组高;IBS患者各组血浆胃泌素及胃动素含量均高于对照组,血浆胰高糖素含量腹泻型IBS患者低于对照组,而便秘型IBS患者高于对照组;IBS患者的肛管括约肌静息压、最大缩榨压与血浆中胃动素的浓度呈正相关,IBS患者的初始感觉量、排便感觉量及最大耐受量与胰高糖素呈负相关。结论IBS患者的直肠肛管压力以及胃肠激素均异常。直肠肛管压力、直肠容量感觉能力与血浆胃肠激素含量呈正相关,直肠容量感知能力与血浆胃肠激素含量呈负相关。     

A subnanogram API LC/MS/MS quantitation method for depsipeptide FR901228 and its preclinical pharmacokinetics

A highly sensitive and specific atmospheric pressure ionization (API) liquid chromatographic-tandem mass spectrometric (LC/MS/MS) method for the quantitation of depsipeptide FR901228 (NSC-630176, FR), a naturally occurring antitumor agent, was developed and validated. FR was extracted from human or rat plasma along with the internal standard, t-Boc-Met-Leu-Phe (BMLP) with ethyl acetate. Components in the extract were separated on a 5-microm C8 Spherisorb 50 x 4.6 mm i.d. column by isocratic elution with methanol/acetonitrile/12 mM ammonium acetate (60:10:30, v/v/v). The liquid flow was passed through a presource splitter and 5% of the eluate was introduced into the API source. The components were analyzed in the multiple-reaction monitoring (MRM) mode to enhance specificity. Linear calibration curves were obtained in the range of 0.1-100.0 ng/ml with 0.5 ml human plasma and 0.5-100.0 ng/ml with 0.1 ml rat plasma. The limit of quantitation (LOQ) was 0.1 ng/ml using 0.5 ml human plasma and 0.5 ng/ml using 0.1 ml rat plasma. The overall within-day precision was below 12% in human plasma and below 7% in rat plasma; and the between-day precision was below 10.2% in human plasma and 7.2% in rat plasma. The accuracy at low, medium and high levels ranged from 99.3 to 111.7% in human plasma and 96.2-107.3% in rat plasma. The high sensitivity permitted pharmacokinetic study of FR in the rat at a single i.v. dose as low as 1 mg/kg. At this dose, plasma FR levels declined biexponentially with a mean terminal t(1/2) of 187.7 min (n = 6) and were detectable up to 24 h. After an oral dose at 5 mg/kg, plasma FR levels were highly erratic and yielded a mean bioavailability of 1.6% (n = 6). At a higher oral dose of 50 mg/kg, a mean bioavailability of 10.6% was obtained, both being estimated by a non-crossover method.     

Unbound valproate fraction in plasma and subcutaneous microdialysate in steady state and after a single dose in humans

The aim of the current study was to characterize the observed discrepancy between unbound plasma valproate (VPA) in single dose and steady state in humans. Unbound and total plasma VPA and subcutaneous microdialysate VPA concentrations were estimated in single dose (6 subjects, n = 33) and steady state (11 subjects, n = 110). Trough plasma samples from 14 patients with total VPA concentrations of 300 micromol/L and 14 patients with VPA concentrations ranging from 600 to 700 micromol/L were analyzed for the unbound VPA fraction and compared with the unbound VPA fraction in spiked plasma samples from healthy subjects containing similar total VPA concentrations. The unbound plasma VPA fraction was significantly higher (P < 0.001) in the steady-state group compared with the single-dose group. The unbound VPA fraction was significantly higher in steady state compared with spiked plasma samples at high and low total VPA concentrations (P < 0.001). The difference between microdialysate and unbound plasma VPA concentrations was significant in the steady-state group (P < 0.001), while no difference was observed in the single-dose group. The mean (+/- SD) subcutaneous microdialysate-to-unbound plasma ratio in the single-dose and steady-state groups was 1.08 (+/- 0.401) and 0.74 (+/- 0.123), respectively. The ratio difference between the groups was significant (P < 0.001). The results of the current study show that unbound plasma fractions of VPA are consistently higher in steady state compared with single dose. Together with the finding of higher unbound VPA fraction in steady state compared with spiked plasma samples, these results provide indirect evidence of displacement of VPA from plasma proteins by product(s) of VPA biotransformation. In addition, subcutaneous microdialysate VPA levels were consistently lower than unbound plasma levels in steady state but not after single dose. The mechanisms underlying this observation need to be studied further.     

LC-MS/MS测定大鼠血浆中虎杖苷浓度

目的 建立测定大鼠血浆中虎杖苷浓度的LC-MS/MS方法。 方法 采用LC-MS/MS测定大鼠血浆中虎杖苷的浓度,以二苯乙烯苷为内标,血浆样品用乙腈沉淀蛋白,色谱柱为Agilent Zorbax SB-C₁₈ (100 mm×2.1 mm, 3.5 μm),流动相为甲醇-乙腈-0.1%甲酸水溶液(18∶15∶67),流速0.3 ml/min,柱温30 ℃。 结果 虎杖苷的线性范围为1.0~5 000.0 ng/ml (r=0.998 4),最低定量检测浓度为1.0     

9-硝基喜树碱在大鼠组织中的分布及内酯稳定性

考察9-硝基喜树碱(9-NC)静脉注射后在人鼠组织中的分布及内酯稳定性.建立了HPLC法间时测定组织和血浆中9-NC内酯浓度和总浓度.大鼠静脉注射9-NC溶液后测定各时间点组织中内酯浓度、总浓度和内酯比例.大多数组织中的9-NC内酯比例明显高于血浆;肝中的内酯比例最低,甚至低于血浆;血浆、肾和小肠中的内酯比例随时问延长而下降.9-NC在肝以外的组织中内酯稳定性显著优于血浆.