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。     

UHPLC法检测血清中万古霉素的浓度

目的 建立通过超高效液相色谱（UHPLC）检测血清中万古霉素浓度的方法,指导临床安全合理用药.方法 使用Agilent 1290超高效液相色谱平台,采用Angilent Zorbax SB-Aq色谱柱（2.1 mm×50 mm,1.8 μm）,以乙腈∶水（体积比为10∶90）为流动相进行梯度洗脱,检测波长为230 nm,柱温30 ℃,流速0.4 mL/min,进样量1 μL.结果 万古霉素在5~100 μg/mL质量浓度范围内线性良好（r=0.998 7）,检测下限为1 μg/mL.检测低浓度（5 μg/mL）、中浓度（60 μg/mL）、高浓度（90 μg/mL）三个浓度值的平均方法回收率均大于90%,各浓度值得日内、日内相对标准偏差（RSD）均小于8%.结论 利用UHPLC可以快速准确的检测血清中万古霉素的浓度.     

人血浆毒鼠强气相色谱-氮磷检测器检测及其临床应用

目的:利用气相色谱-氮磷检测器(GC/NPD)建立一种快速检测人血浆中毒鼠强(TET)的方法.方法:血浆中TET用乙酸乙酯作液液提取,毛细管气相色谱柱,GC/NPD法测定血浆中毒鼠强的浓度,以甲基1605为内标,内标法定量.结果:方法学显示毒鼠强与内标分离良好,不受样品中内源性物质干扰,在50～700 ng/mL范围内具有良好的线性关系(r＞0.99),最低检出浓度为5 ng/mL,最低检测限为10 ng/mL.血浆样本回收率的在70%以上.日内、日间精密度分别为RSD 3.5%～12.8%和6.5%～12.0%.结论:该本方法灵敏、稳定,能满足毒鼠强药物代谢动力学研究和临床检测的需要.     

奥美拉唑胶囊剂有关物质再评价

[摘要] 目的：建立合适的高效液相色谱法检测奥美拉唑肠溶胶囊中的有关物质的含量。方法：phenomenex luna C8色谱柱（150mm×4.6mm,5μm）,流动相为乙腈：0.01M磷酸氢二钠溶液（用磷酸调节pH至7.6）（25:75）,检测波长为280nm,流速为1.0mL/min,柱温为30℃。结果：该色谱条件能有效检出奥美拉唑肠溶胶囊中的降解产物,检测限为1ng（0.025%）。结论：该方法简便、可靠、快速,能够准确测量奥美拉唑肠溶胶囊中有关物质的含量。     

高效液相色谱法测定人血浆中头孢地尼的浓度

目的：建立测定人血浆中头孢地尼含量的高效液相色谱（HPLC）方法。方法：血浆样品用高氯酸沉淀蛋白,采用Ulti-mate C18色谱柱（5μm,2 504.6mm）,以乙腈：0.025mol.L-1磷酸二氢钠缓冲盐溶液（pH3.24）=15∶85作为流动相进行分离,检测波长为286nm,流速为1.0mL.min-1。结果：头孢地尼血药浓度30~2 500ng.mL-1范围内线性关系良好,最低检测浓度为30ng.mL-1。方法回收率（n=5）分别为76.41%,63.78%,66.44%,日内和日间精密度均小于15%。结论：该法操作简单、灵敏、准确、重现性好,适用于头孢地尼临床药代动力学研究。     

LC-MS/MS法快速定量人血浆中克拉霉素的浓度

目的建立液相色谱串联质谱法测定人血浆中克拉霉素的浓度。方法选用ACQUITY UPLC BEH C18柱(2.1×50mm,1.7μm)的色谱柱,以含0.1%的甲酸纯水,含0.1%的甲酸95%乙腈为流动相,采用梯度洗脱进行分离,样本经蛋白沉淀及30%乙腈复溶后进样,选用API4000型质谱仪的多重反应监测(MRM)扫描方式进行检测。结果克拉霉素线性范围为4.00～2 000.00ng/mL,定量下限4.00ng/mL。准确度与精密度结果显相对偏差为1.20%～2.90%,低、中、高3个浓度提取回收率平均值均大于100%,基质效应小,稳定性好。结论该方法快速、灵敏、专属性强、重现性好,可用于人体克拉霉素血药浓度的监测及人体药代动力学研究。     

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。     

氯吡格雷及其羧酸代谢物高效液相色谱-串联质谱法快速检测

目的建立高效液相色谱-串联质谱法快速定量检测人血浆中氯吡格雷及其羧酸代谢物的方法。方法氯吡格雷样本用乙腈沉淀蛋白后,选用Eclipse XDB-C18柱(150.0 mm×4.6 mm×5.0μm),以乙腈-1mmol/L乙酸铵溶液为流动相进行梯度洗脱。氯吡格雷羧酸代谢物样本用乙腈沉淀蛋白后,选用Kromasil-C18色谱柱(50.0mm×4.6mm×5.0μm),以乙腈-1 mmol/L乙酸铵溶液(60∶40v/v)为流动相等度洗脱。选用3200QTrap型液相色谱-串联质谱仪的多重反应监测(MRM)扫描方式进行检测。结果氯吡格雷线性范围为:0.01~5.00ng/mL,定量下限为0.01ng/mL。氯吡格雷羧酸代谢物线性范围为:10.0~5 000ng/mL,定量下限为10.0ng/mL。准确度与精密度结果显示方法日间、日内变异系数均小于15%,相对偏差-2.00%~2.65%,稳定性较好。结论本研究建立了快速、灵敏、专属性强、重复性好的方法,适用于临床药代动力学和生物等效性研究。     

HPLC-MS/MS检测急性百草枯中毒患者血浆的方法及意义

目的探讨高效液相色谱串联质谱(HPLC-MS/MS)检测急诊百草枯中毒患者血浆中百草枯水平的可靠性和优越性。方法采用Phenomenex Kinetex mHILIC色谱柱(100.0mm×2.1mm,2.6μm),以1%甲酸铵水溶液(含甲酸铵250mmol/L)和乙腈分别为流动相A、B,建立HPLC-MS/MS(流速0.35mL/min、检测波长258nm、柱温40℃),通过受试者工作特征(ROC)曲线评价血浆百草枯水平。结果 54.28～13 190.00ng/mL百草枯的线性关系良好(R~2=0.999 9),批内、批间RSD均10%。63例急性百草枯中毒患者血浆百草枯水平平均为2 820ng/mL,死亡组水平显著高于存活组(P0.05)。结论 HPLC-MS/MS能快速、准确、可靠检测急诊百草枯中毒患者血浆的百草枯水平,对患者的治疗具有指导意义。     

人血浆百草枯高效液相色谱-串联质谱法的建立与评价

目的建立测定人血浆百草枯浓度的高效液相色谱-串联质谱法并评价。方法用甲醇蛋白质沉淀法对血浆样品预处理后,以乙腈-水(含200 mmol/L甲酸铵和0.1%甲酸)为流动相梯度进样,流速为0.4 mL/min。通过色谱柱(Waters XBridge BEH HILIC,2.5μm,2.1 mm×100 mm)分离样品。以ESI正离子模式、多反应离子监测(MRM)模式监测百草枯(m/z 186.1→171.1作为定量离子对)。用该方法检测临床患者血浆百草枯浓度,用ROC曲线评价血浆百草枯浓度和百草枯中毒严重指数(SIPP)对临床结局的判断价值。结果血浆百草枯浓度在50～10 000 ng/mL时,线性关系良好(R~2=0.997),最低定量下限为50 ng/mL。低(100 ng/mL)、中(2 000 ng/mL)、高(8 000 ng/mL)3个浓度水平质控品的不精密度均符合要求,且无基质效应。处理后样品室温放置6 h内稳定,样本反复冻融3次对结果无影响。31例中毒患者SIPP为17.76(0.30～90.91)h·mg/L,死亡组SIPP高于存活组(P0.05)。SIPP的ROC曲线下面积为0.889,最佳临床临界值为11.679 h·mg/L。结论本法灵敏、准确、快速、特异性好,适用于临床检测百草枯中毒患者。     

Measurement of L-carnitine and acylcarnitines in body fluids and tissues in children and in adults

We have developed a reliable and validated radio-enzymatic method for the assay of L-carnitine and acylcarnitines, using a modification of existing methods. The sensitivity of the assay is 10 mumol/l using 10 microliters of plasma or urine. It is also suitable for measurements of carnitine in a 10 mg sample of liver or muscle obtained by percutaneous biopsy. The use of N-ethylmaleimide in the reaction mixture together with an excess of [1-14C]acetyl CoA ensures that the reaction proceeds to completion and a linear response is obtained. Using this method control ranges have been established for plasma and urine carnitine concentrations in healthy children and adults, and for the carnitine content of liver and muscle in adults. No significant difference was found between fasting and post-prandial plasma carnitine levels. An age-related increase was found in urinary total carnitine and acylcarnitine concentration throughout childhood. These data provide a reliable basis for studies of patients with abnormal carnitine and acylcarnitine metabolism, distribution and excretion.     

Rosamicin in Urethral and Vaginal Secretions and Tissues in Dogs and Rats

In animal studies we investigated the distribution of rosamicin in plasma and urethral and vaginal tissues in rats as well as in urethral and vaginal secretions in dogs. We found concentration ratios between urethral secretion and plasma of 1.9 and between vaginal secretion and plasma of 2.4. The rosamicin concentrations in urethral and vaginal tissue significantly exceeded the levels of all other tissues investigated. Because rosamicin could be valuable for the treatment of bacterial urethritis and the colonization of the vaginal introitus with fecal bacteria in women, it should be investigated clinically in this respect.     

Similarities and differences in clients treated and in medications prescribed by APRNs and psychiatrists in a CMHC

This study is part of a larger study comparing prescribing practices of psychiatrists and advanced practice psychiatric nurses (APRNs) using the following three groups of patients: patients treated by psychiatrists, those treated by APRNs, and those treated by both APRNs and psychiatrists at different times in 1 year. Demographics for 5507 patients were examined. A subsample of APRNs and psychiatrists prescribed similar total numbers of medications. Psychiatrists prescribed more types of antidepressant medications other than the SSRI antidepressants, and they prescribed more than twice the number of benzodiazepines. APRNs prescribed more SSRIs and spent more time with clients during medication visits.     

Racial differences in oxidative stress and inflammation: in vitro and in vivo

African American race is an independent risk factor for enhanced oxidative stress and inflammation. We sought to examine whether oxidative-stress and inflammatory markers that are typically measured in humans also differ by race in cell culture. We compared levels between African American and Caucasian young adults and then separately in human umbilical vein endothelial cells (HUVECs) from both races. We found heightened oxidative stress and inflammation in the African Americans both in vitro and in vivo. African American HUVECs showed higher nitric oxide (NO) levels (10.8 ± 0.4 vs. 8.8 ± 0.7 μmol/L/mg, p = 0.03), Interleukin-6 (IL-6) levels (61.7 ± 4.2 vs. 23.9 ± 9.0 pg/mg, p = 0.02), and lower superoxide dismutase activity (15.6 ± 3.3 vs. 25.4 ± 2.8 U/mg, p = 0.04), and also higher protein expression (p < 0.05) of NADPH oxidase subunit p47phox, isoforms NOX2 and NOX4, endothelial nitric oxide synthase (NOS), inducible NOS, as well as IL-6. African American adults had higher plasma protein carbonyls (1.1 ± 0.1 vs. 0.8 ± 0.1 nmol/mg, p = 0.01) and antioxidant capacity (2.3 ± 0.2 vs. 1.1 ± 0.3 mM, p = 0.01). These preliminary translational data demonstrate a racial difference in HUVECs much like that in humans, but should be interpreted with caution given its preliminary nature. It is known that racial differences exist in how humans respond to development and progression of disease, therefore these data suggest that ethnicity of cell model may be important to consider with in vitro clinical research.     

Postural strategies and falls in elderly and in parkinsonism]

OBJECTIVE: To use a posture analysis to show the evolution of postural pattern connected with falls.MATERIAL AND METHOD: It is a prospective study on two groups of 16 persons of more than 60 years. A group concerns 16 small disability off drug parkinsonian patients, a group concerns 16 healthy witnesses. All the persons benefited from a posture recording by means of a force platform and were followed during 1 year. RESULTS: Data analysis underlines three groups of persons corresponding to three postural patterns, independently of the presence of Parkinson disease. A group (n = 18) did not contain fallers, the second (n = 10 ) contained 20% of fallers, the third (n = 4) contained 100% of fallers. Differences between the groups were identified on 16 posturographic parameters. DISCUSSION: A group has a good functional value and one does not record any fall. Its characteristics, which correspond to a category of persons who compensate well for the phenomena of ageing, are found in the literature. A group has an intermediate functional value and regrets 20% of fallers. Kinetic profile reveals a tendency to the stiffness of the posture. This group is going to operate rather ankle strategies. A group has an inferior functional value and regrets 100% of fallers. Kinetic profile seems disrupted and not to be able to adapt itself in a satisfactory way to the situation otherwise than by stereotypical reactions. This group is going to operate systematically much less stabilizing hip strategies. CONCLUSION: A close determinism between physiological neuromotor ageing and Parkinson disease does exist. We showed with a prospective follow-up, the arisen of fall and showed the evolution of postural patterns related to fall. It appears as well that evolution mainly follows three stages leading from a small risk of fall gait pattern to a major risk of fall gait pattern.