一种简便快速的液相色谱串联质谱法用于人体内血尿样品中奈诺沙星浓度的测定

目的建立一种简便快速高效液相色谱串联质谱(LC-MS/MS)的方法用于测定人血浆和尿液中奈诺沙星药物浓度。方法色谱柱为Waters Symmetry RP1 8柱(50mm×2.1 mmm,5μm),以加替沙星为内标,血浆和尿液样品测定采用0.1%甲酸乙腈混合溶液为流动相,比例分别为82:1 8和85:1 5,流速均为0.2 mL/min。采用蛋白沉淀法处理血浆样品,以液液萃取法清除尿液样品中杂质。ESI源正离子选择性反应临测(SRM),奈诺沙星检测通道:m/z 372.4→m/z354.1,内标检测通道:m/z376.1→m/z 332.2。结果本方法测定血浆和尿液中的奈诺沙星的线性范围均为5～100ng/mL,最低检测浓度均为5ng/mL血浆和尿液的奈诺沙星提取回收率分别为(98.49±4.69)%和(84.92±6.81)%,且血浆和尿液的RSD)分别小于或等于4.31%和9.76%。奈诺沙星血浆样品的方法日内、日间准确度分别为(99.27～103.75)%和(100.40～106.36)%;而尿液样品的方法日内、日间准确度依次为(103.52～11 5.94)%和(9 9.50～1 10.96)%,且基质效应、精密度、稳定性等均通过方法学验证。并已用于该药的人体药动学研究中药物浓度的测定。结论本方法操作简便、迅速,特异性强,灵敏度高,准确性好,符合奈诺沙星人体药物动力学研究中血尿样测定的方法学要求。     

HPLC法测定犬血浆中非索非那定的浓度

目的 建立HPLC-UV法测定犬血浆中非索非那定的浓度.方法 以乙腈:0.02 mol/L磷酸二氢钠(含0.5%三乙胺,pH值4.0)=35:65为流动相,非那西丁为内标,血浆样品经简单的乙腈沉淀蛋白萃取后,采用HPLE-UV法测定血浆中非索非那定的浓度.结果 非索非那定的线性范围是0.05～10.0 mg/ml,平均相对回收率为85.03%～105.28%,日内和日间精密度均<13%,检测限为25 ng/ml.结论 HPLC-UV法快速、准确、灵敏,可用于非索非那定的药动学研究.     

高效液相色谱法测量患儿血清甲氨蝶呤浓度

目的：建立测定患儿血清中甲氨蝶呤浓度的方法,用于临床患儿血药浓度监测。方法色谱柱采用岛津原装柱（250 mm×4．6 mm,5μm）,流动相为0．025 mol/L 磷酸二氢钠溶液（pH 为5．41）-甲醇（76∶24,V/V）,流速1．0 mL/min,检测波长313 nm,柱温35℃,样品经10％高氯酸沉淀蛋白后,取上清液130μL 加10μL 1 mol/L NaOH 后直接进样。结果血浆中甲氨蝶呤的浓度在0．00～4．84μmol/L 和4．84～10．00μmol/L 范围内与峰面积线性关系良好,r 1＝1,r2＝1,平均回收率分别为61．67％和71．83％,日内及日间精密度的 RSD 小于10％和12％（n＝3）。结论所用方法简便快速、准确可靠、灵敏度高、重现性好、线性范围宽和成本低,适于甲氨蝶呤的血药浓度监测。     

噁唑烷酮类新药MRX-Ⅰ在人血浆、尿液浓度超高效液相色谱串联质谱测定方法的建立及验证

目的建立及验证超高效液相色谱串联质谱（UPLC-MS,/MS）的方法用于测定人血浆和尿液中恶唑烷酮类新药MRX-I药物浓度。方法 UPLC-MS/MS液相条件为色谱柱Waters ACQUITY UPLC BEH C8;流动相为乙腈：水（40：60,v/v）。质谱采用ESI源正离子多反应监测（MRM）。内标为利奈唑胺,以乙酸乙酯液-液萃取法清除血浆及尿液样本中杂质。方法学验证包括基质效应、绝对回收率、精密度和准确度及MRX-I在人血浆及尿液样本中放置稳定性。结果 UPLC-MS/MS法检测MRX-I在人血浆和尿液中的线性范围均为（0.005 00～1.00）mg/L,最低检测浓度均为0.005 00 mg/L。MRX-I与内标在血浆和尿液中的保留时间小于1.5 min。本方法学验证结果显示MRX-I在人血浆和尿液基质效应因子分别为90.4%±8.2%和82.7%±7.9%;血浆和尿液中MRX-I提取回收率分别为112.8%±13.4%和105.6%±13.4%。MRX-I血浆样本的测定方法日内、日间准确度分别为98.9%～105.0%和96.5%～102.6%;尿液样本的测定方法日内、日间准确度分别为92.7%～98.6%和95.1%～105.7%。MRX-I在人血浆和尿液样本室温放置24 h、预处理后自动进样器放置48 h、-40℃冰箱冻融3次、-40℃冰箱分别放置8个月和6个月仍然保持稳定。结论本研究建立的UPLC-MS/MS检测人血浆及尿液中MRX-I浓度方法的灵敏度高,专属性强。其方法学验证结果均符合生物样品分析的要求。     

HPLC 法测定间歇性高容量血液滤过患者血清、尿液及置换液中莫西沙星浓度

目的:建立高效液相色谱法(HPLC)测定间歇性高容量血液滤过(PHVHF)患者血清、尿液及置换液中莫西沙星浓度.方法:PHVHF联合莫西沙星治疗肾衰合并感染患者8例,测定肾功、电解质.采用依利特SinoChrom ODS-BP色谱柱(5 μm,4.6 mm × 150 mm)测定莫西沙星浓度,流动相为乙腈:(10 mmol/L磷酸二氢钾、1.5 mmol/L四丁基硫酸氢胺) ＝ 25:75(v/v),柱温 40℃,检测波长 290 nm,流速 1 mL/min.结果:血BUN 及SCr、血清K+ 水平显著下降(P ＜ 0.05).血清莫西沙星在0.1 ～ 10.0 mg/L范围的线性回归方程为:y ＝ 4.49x-0.257,r ＝ 0.999 86.准确度为98.7% ～ 101%,日内和日间RSD为1.12% ～ 5.7%.置换液在0.1 ～ 10.0 mg/L范围的线性回归方程为:y ＝ 3.877x-0.249,r ＝ 0.998 5.准确度为85.7% ～ 95.7%,日内和日间RSD 为1.34% ～ 8.9%.尿液在0.1 ～ 10.0 mg/L范围的线性回归方程为:y ＝ 3.565x-0.270,r ＝ 0.986 8.准确度为97.4% ～ 101.7%,日内和日间RSD为1.51% ～ 11.6%.结论:HPLC方法检测血滤患者血清、尿液及置换液中莫西沙星浓度,方法简便、快速、重复性好,可满足临床研究血滤时莫西沙星药代动力学变化的要求.     

荧光偏振免疫法测定尿中微量白蛋白

报道了用荧光偏振免疫法测定（FPIA）尿中微量白蛋白。偏振度倒数值与白蛋白浓度（0．04～0.2mg/L）问线性良好，相关系数（γ）0.9988，6个尿样（白蛋白1.21～46.5mg／L）的日内变异CV（％）为1．86～9.22（n=8），8个尿样（白蛋白0．71～8．43）的日间变异CV（％）为2．94～5．71（n=6），回收率为96．93±8.48％（均数±标准差）（n=26）。162例正常人24小时白蛋白排出量为5.01±3.95mg，216例糖尿病患者24小时白蛋白排出量55．99±89．26mg。与放射免疫法测定（RIA）相比相关方程为Y（RIA）=1．057X（FPIA） 0．086，γ=0。9981（n=25）。对24小时内尿样作分段收集及测定，结果表明晨尿的个体间差异及数值均接近于24小时的混合尿。     

高效液相色谱法同时测定血清色氨酸和犬尿氨酸浓度的方法建立及应用

目的建立一种高效液相色谱-紫外检测法同时测定血清中色氨酸(Trp)和犬尿氨酸(Kyn)浓度的方法。方法色谱柱采用Agilent Hypersil ODS(125.0mm×4.0mm,5μm),流动相为15mmol/L乙酸钠缓冲液(pH4.0)和乙腈95∶5(V/V),流速为0.8ml/min,柱温25℃,紫外检测λKyn=360nm,λTrp=278nm。结果Kyn和Trp的保留时间分别为3.3min和5.2min,线性范围分别为0.083~21.000μmol/L和2.650~678.000μmol/L,检测限分别为0.028μmol/L和0.053μmol/L,日内和日间相对标准偏差分别低于5%和10%,回收率分别为91.99%~113.89%和95.81%~118.82%。结论该方法简便、快速、特异,适合于临床检测。     

液相色谱-串联质谱法测定人血浆中二甲双胍的浓度

目的采用液相色谱-串联质谱法测定人血浆中二甲双胍的浓度。方法血浆样品用乙腈(含0.1%甲酸)沉淀蛋白后用二氯甲烷反洗后进行分析。使用Agilent C8(75 mm×4.6 mm,3.5μm)色谱柱。流动相:A泵:5 mmol/L醋酸铵(三乙胺调pH值至7.5),B泵:乙腈。线性梯度洗脱,流速0.4 mL/min。采用电喷雾离子源,多反应离子监测。用于定量分析的离子对二甲双胍为130.2/71.1,内标吗啉胍为172.2/60.2。结果线性范围为50～2 000 ng/mL,最低定量限为50 ng/mL,预处理回收率为81.7%～98.0%,二甲双胍的基质效应<9.97%,日内和日间相对标准偏差均<5.2%。结论液相色谱-串联质谱法快速、简便、灵敏度高,是一种适用于人血浆中药物浓度的测定及药物动力学和生物利用度研究的方法。     

单剂口服吉米沙星临床药动学研究

目的 研究单剂口服吉米沙星在中国健康志愿者中的药动学特性,为制订用于中国人的安全有效的给药方案提供依据.方法 12名受试者随机、开放、3交叉单剂空腹口服吉米沙星片剂160 mg、320 mg和480 mg.以高效液相色谱(HPLC)-荧光法测定血、尿样中吉米沙星药物浓度.应用Winnonlin分析软件计算药动学参数.结果 受试者单剂空腹口服吉米沙星160 mg、320 mg和480 mg后体内过程均符合非房室模型,平均血清Cmax分别为(0.70±0.19)mg/L、(1.40±0.32)mg/L和(1.84±0.35)mg/L;Tmax中位数分别为1(0.5,2)h、1(0.5,2)h和1.25(1,2)h;平均t1/2分别为(7.06±1.22) h、(7.02±0.94) h和(7.28±0.81) h;平均AUC0-24 h分别为(3.86±0.56) mg·h /L、(7.61±0.93) mg·h /L和(11.47±1.68) mg·h/L;平均AUC0-∞分别为(4.01±0.57) mg·h /L、(7.71±0.92) mg·h /L和(11.62±1.72) mg·h/L;平均Vd分别为(411.45±77.88)L、(423.47±59.48)L和(439.21±55.55)L;平均CLt分别为(40.71±6.22)L/h、(42.10±5.46)L/h和(42.26±7.14)L/h;给药后48 h内平均累积尿排出率分别为(38.95±6.14)%、(37.84±6.62)%和(35.57±5.07)%.结论 中国健康受试者单次空腹口服吉米沙星160～480 mg结果显示其符合线性药动学的特性,消除半衰期长.给药量的约40%以原形药物经肾排出.     

高效液相色谱法测定人血浆中阿托伐他汀的浓度

目的建立高效液相色谱法(HPLC)测定人血浆中阿托伐他汀浓度。方法色谱柱:Aglient Eclipse HPLC色谱柱(150×4. 6 mm,5μm);流动相:乙腈:0. 02 mol/L醋酸铵缓冲溶液(冰醋酸调至p H=5. 2)=40:60;流速:1 ml/min。结果按上述色谱条件,阿托伐他汀保留时间为7. 5 min,专一性好,且基线无干扰。阿托伐他汀在0. 41～14. 45μg/ml范围内线性关系良好(R2=0. 9993)。以0. 64、3. 03、12. 89μg/ml为阿托伐他汀低、中、高浓度的血浆质控样品,日内和日间RSD均小于8%,相对提取回收率大于89%,且稳定性良好。结论该方法专一性强,简便快捷,适用于阿托伐他汀在特殊人群体内药代动力学及其相关性研究。     

Determination of creatine kinase isoenzyme MB activity in serum using immunological inhibition of creatine kinase M subunit activity. Activity kinetics and diagnostic significance in myocardial infarction.

This is a new method for the determination of creatine kinase isoenzyme MB activity in serum. The method uses direct activity measurement of creatine kinase B subunit activity after blocking of CK-M subunit activity by inhibiting antibodies. The test takes no longer than 15 min. The method yields an intra-serial C.V. of 2.0-12.9%, and a C.V. from day to day of 5.5%. The detection limit is 3.4 U/l creatine kinase MB. In the 95 cases with proven myocardial infarction several types of creatine kinase MB activity kinetics could be determined. The percentage of creatine kinase MB of peak CK-total is 6-25%, with a mean of 11.1%. The amount of creatine kinase MB with respect to total CK activity after reinfarction is higher than the amount after initial infarction.     

俯卧位通气对高海拔地区肺复张治疗无效急性呼吸窘迫综合征患者氧合的影响

目的 探讨俯卧位通气对高海拔地区肺复张术(RM)治疗无效急性呼吸窘迫综合征(ARDS)患者的治疗作用.方法 从海拔2260m的地区医院筛选RM治疗无效的41例ARDS患者[平均氧合指数( PaO2/FiO2)较RM前升高＜20％视为RM无效],依不同病因分为肺内源性ARDS组(ARDSp组)和肺外源性ARDS组(ARDSexp组),每组再按信封法随机分为俯卧位组和仰卧位组,即ARDSp俯卧位组(11例)、ARDSp仰卧位组(9例)、ARDSexp俯卧位组(10例)、ARDSexp仰卧位组(11例).在通气前及通气1、2、3、4h监测动脉血氧分压( PaO2)、PaO2/FiO2、静态顺应性(Cst)、气道阻力(Raw)的变化.结果 通气lh时,ARDSexp俯卧位组PaO2/FiO2( mm Hg,l mm Hg=0.133 kPa)即较通气前显著升高(157.4±40.6比129.3±48.7,P＜0.05),并随通气时间延长呈持续增高趋势,4h达峰值(219.1 ±41.1);且ARDSexp俯卧位组通气3h内PaO2/FiO2较其他3组显著增高,另3组间则差异无统计学意义.ARDSp俯卧位组、ARDSexp俯卧位组通气4h时PaO2/FiO2均较相应仰卧位组显著增高(208.8±39.7比127.4±47.1,219.1±41.1比124.9±50.8,均P＜0.05).4组通气前后Cst无显著改变,各组间差异也无统计学意义.ARDSp俯卧位组通气4h时Raw(cmH2O·L-1·s-1)较通气前显著降低(6.8±1.7比10.7±1.8,P＜0.05),且明显低于其他3组;其他3组各时间点Raw组内及组间比较差异均无统计学意义.结论 俯卧位通气作为ARDS机械通气重要策略之一,可以改善RM无效高原ARDS患者的氧合,为抢救患者赢得宝贵的时间.     

Digital imaging among medical facilities

The Department of Veterans Affairs (VA) in the USA operates a network of 172 medical centres which all utilize a hospital information system (HIS) which has been developed and is currently maintained by the VA. During the past several years, an image management and communication module has been developed, installed and clinically utilized at the Washington DC and Maryland VA Medical Centres. This image management and communication system, referred to as the decentralized hospital computer program (DHCP) imaging system, is fully integrated with a commercial picture archiving and communication system (PACS). The system is utilized to capture, archive, and display all images generated within the hospital including radiology, nuclear medicine, pathology, endoscopy, bronchoscopy, and dermatology, intraoperative photographs, ECG data, and a limited number of paper documents. The ultimate goal of the project is to have all patient text and image data available at any clinical workstation to any authorized user anywhere within the network of medical centres. Clinical requirements for an imaging workstation include ease of use, rapid and reliable access to the complete set of patient information, and images which are of acceptable quality to meet the requirements of the user and the subspecialty. Patient confidentiality and data security must be safeguarded at all times. Integration of the images with the remainder of the patient's database was found to be critical to the success of the project. The experience at the Washington and Maryland facilities suggests that an imaging system that is successfully integrated with a hospital information system can provide substantial clinical and economic benefits both within and among medical centres. Clinical acceptance and utilization of the system has been excellent, particularly in diagnostic radiology where DHCP Imaging has been interfaced to a commercial PAC system. Based upon this initial experience, the VA has begun to deploy the system throughout its large network of medical centres.     

Myocardial elastography at both high temporal and spatial resolution for the detection of infarcts

Myocardial elastography is a novel method for noninvasively assessing regional myocardial function, with the advantages of high spatial and temporal resolution and high signal-to-noise ratio (SNR). In this paper, in-vivo experiments were performed in anesthetized normal and infarcted mice (one day after left anterior descending coronary artery [LAD] ligation) using a high-resolution (30 MHz) ultrasound system (Vevo 770, VisualSonics Inc., Toronto, ON, Canada). Radiofrequency (RF) signals of the left ventricle (LV) in longitudinal (long-axis) view and the associated electrocardiogram (ECG) were simultaneously acquired. Using a retrospective ECG gating technique, 2-D full field-of-view RF frames were acquired at an extremely high frame rate (8 kHz) that resulted in high-quality incremental displacement and strain estimation of the myocardium. The incremental results were further accumulated to obtain the cumulative displacements and strains. Two-dimensional and M-mode displacement images and strain images (elastograms), as well as displacement and strain profiles as a function of time, were compared between normal and infarcted mice. Incremental results clearly depicted cardiac events including LV contraction, LV relaxation and isovolumetric phases in both normal and infarcted mice, and also evidently indicated reduced motion and deformation in the infarcted myocardium. The elastograms indicated that the infarcted regions underwent thinning during systole rather than thickening, as in the normal case. The cumulative elastograms were found to have higher elastographic SNR (SNR(e)) than the incremental elastograms (e.g., 10.6 vs. 4.7 in a normal myocardium, and 6.0 vs. 2.4 in an infarcted myocardium). Finally, preliminary statistical results from nine normal (m = 9) and seven infarcted (n = 7) mice indicated the capability of the cumulative strain in differentiating infracted from normal myocardia. In conclusion, myocardial elastography could provide regional strain information at simultaneously high temporal (>/=0.125 ms) and spatial ( approximately 55 microm) resolution as well as high precision ( approximately 0.05 microm displacement). This technique was thus capable of accurately characterizing normal myocardial function throughout an entire cardiac cycle, at the same high resolution, and detecting and localizing myocardial infarction in vivo.     

Clinical Pharmacokinetics of Morphine

Morphine, the most widely used mu-opioid analgesic for acute and chronic pain, is the standard against which new analgesics are measured. A thorough understanding of the pharmacokinetics of morphine is required in order to safely and effectively use this analgesic in a wide variety of patients with different levels of organ function. A MEDLINE search was conducted to identify literature published between 1966 and January 2002 relevant to the pharmacokinetics of morphine. These publications were reviewed and the literature summarized regarding unique and clinically important elements of morphine disposition relative to its parenteral administration (including intravenous, intramuscular, subcutaneous, epidural and intrathecal administration), absorption profile (immediate release, controlled release, and sublingual/buccal, and rectal administration), distribution, and its metabolism/ excretion. Special populations, including infants, elderly, and those with renal/liver failure, have a unique morphine pharmacokinetic profile that must be taken into account in order to maximize analgesic efficacy and reduce the risk of adverse events.     

手转胎头术失败的原因与分娩结局临床分析

目的 探讨手转胎头术失败的原因与分娩结局.方法 选择2008年1月至2010年12月于我院住院分娩的持续性枕横位、枕后位产妇198例,根据行手转胎头术后结果分为成功组126例、失败组72例.比较两组分娩结局,对比分析失败原因.结果 失败组胎儿体质量≥3500 g的发生率[76.4％(55/72)]明显高于成功组[31.7％(40/126)],差异有统计学意义(x2=30.177,P=0.001)、失败组宫缩乏力发生率[58.3％(42/72)]高于成功组[38.1％ (48/126)],差异有统计学意义(x2=7.569,P=0.006)、失败组骨盆临界或轻度狭窄发生率[38.9％ (28/72)]高于成功组[23.8％(30/126)],差异有统计学意义(x2 =5.030,P=0.002)、失败组手转胎头时机不当(宫口开大＜6 cm、胎头位于坐骨棘上及宫口开大8～10 cm、胎头位于坐骨棘下≥2 cm)发生率[61.1％(44/72)]高于成功组[38.9％(49/126)],差异有统计学意义(x2=9.084,P=0.003).失败组母儿并发症(产后出血、产褥病率、胎儿窘迫、新生儿窒息)发生率高于成功组(x2 =9.586,P=0.002、x2=9.334,P=0.002、x2=5.910,P=0.015、x2=5.240,P=0.022)、失败组剖宫产发生率[72.2％(52/72)]明显高于成功组[34.1 ％(43/126),x2=26.641,P=0.001)].结论 手转胎头术能使难产变顺产,降低剖宫产率,减少母儿并发症,但须积极预防、处理导致手转胎头术失败的原因,对矫正失败后继续矫正及试产应慎重.     

Evidence-Based Pain Management and Palliative Care in Issue Four for 2009 of The Cochrane Library

ABSTRACT

The Cochrane Library of Systematic Reviews is published quarterly. Issue 4 for 2009 contains 4027 complete reviews, 1906 protocols for reviews in production, and 11447 one-page summaries of systematic reviews published in the general medical literature. In addition, there are citations of 600,000 randomized controlled trials, and 12,200 cited papers in the Cochrane methodology register. The health technology assessment database contains over 7500 citations. This edition of the Library contains 90 new reviews, of which 19 have potential relevance for practitioners in pain and palliative medicine.     

Wie zuverlässig sind Angaben zu nichtentzündlichen ausgedehnten Schmerzen?

ZusammenfassungFragestellung Es wurde geprüft, wie sich der Differenziertheitsgrad zweier Schmerzmessmethoden auf Angaben zur Ausgedehntheit klinischer Schmerzen auswirkt. Zugleich wurde der Referenzzeitraum variiert, über den die Patienten berichten sollten.Methode Erfasst wurde der Einfluss zu Lasten der Befragungsdifferenziertheit durch den Vergleich zweier Körperschema-Bildvorlagen. Drei Referenzzeiträume (Schmerz aktuell, letzte Woche, letztes halbes Jahr) wurden vorgegeben.Ergebnisse Patienten mit ausgedehnten Schmerzen gaben bei differenzierter Befragung um so mehr Schmerzen an, je weiter die Schmerzen zurück lagen und je größer der Berichtszeitraum war. Patienten mit gelenknahen Schmerzen gaben bei hoch differenzierter Befragung weniger ausgedehnte Schmerzen in der Vergangenheit an als bei globaler Einschätzung. Patienten mit Rückenschmerzen berichteten bei differenzierter Befragung zum aktuellen Schmerz über weniger ausgedehnte Schmerzen als bei globaler Befragung.Schlussfolgerung Die Angaben zur Schmerzausdehnung variieren vor allem bei Patienten mit ausgedehnten Schmerzen in Abhängigkeit von der Differenziertheit der Befragung. In diesen Fällen ist die Wahrscheinlichkeit erhöht, dass sich die Beschwerdesymptomatik zumindest teilweise erst in der Reaktion auf die situativen Befragungsbedingungen konstituiert und daher nicht auf andere Befragungsbedingungen generalisiert werden kann.