中毒家兔体内呋喃丹水平检测及其体内分布规律研究

目的建立亲水性固相材料提取呋喃丹的方法,研究呋喃丹在中毒家兔死后体内的分布规律。方法选取雄性家兔12只,随机分为2LD_(50)和4LD_(50)组,经灌胃匀速注入2倍半数致死量(2LD_(50))(220mg/kg)和4倍半数致死量(4LD50)(440mg/kg)呋喃丹。观察给药到死亡时的生命体征的变化以及中毒症状,待呼吸和心跳全部消失时,迅速解剖动物,取心血、心、肝、脾、肺、肾、脑冷冻保存。采用亲水性固相材料萃取柱进行提取,二氯甲烷洗脱提取,气相色谱定量检测其中呋喃丹的水平。结果亲水性固相材料萃取柱提取回收率高,呋喃丹平均回收率99.3%,检出限为5ng/mL。各脏器组织呋喃丹水平由高到低分别为:(1)2LD_(50)组:肺(10.98±2.30)μg/mL、肝(7.92±2.39)μg/mL,心血(2.13±1.36)μg/mL,脑(2.01±1.57)μg/mL,心(1.23±0.38)μg/mL,肾(1.31±0.42)μg/mL,脾(0.59±0.21)μg/mL。(2)4LD_(50)组:肺(6.86±3.02)μg/mL,肝(6.12±2.38)μg/mL,肾(2.52±1.54)μg/mL,脑(1.91±0.72)μg/mL,心(1.48±1.75)μg/mL,心血(1.17±1.57)μg/mL,脾(1.15±0.66)μg/mL。结论亲水性固相材料提取呋喃丹的方法成本低、方便快捷、提取率高、杂质少,可应用于呋喃丹中毒案件法医学鉴定的实验研究。呋喃丹在死后家兔体内分布不均匀,在含血丰富的组织器官中水平高。     

生物检材中利血平的高效液相色谱／质谱和高效液相色谱的检测

目的：建立生物检材中利血平的高效液相色谱／质谱（HPLC／MS）和高效液相色谱（HPLC）的检测。方法：采用C18（200mm×4．6mm,5mL）色谱柱,乙腈：水（40：60）用盐酸调节pH值至3．00±0．05为流动相,检测波长258nm。结果：HPLc／Ms分析利血平的选择离子m／z为608,195。心血中利血平HPLC检测的回归方程、线性检测范围、相关系数、回收率、最低检出浓度,分别为Y=56．576X＋0．3168（μg／mL）,（0．5～48）μg／mL,0．995,（96．50土3．O）％,0．5μg／mL;肝组织中利血平HPI。C检测的回归方程、线性检测范围、相关系数、回收率、最低检出浓度,分别为Y=38．567X＋0．054（μg／g）,（o．5～48）μg／g,0．984,（97．5±2．5）％,0．5μg／g。染毒大鼠心血、心、肝、脾、肺、肾和脑中利血平的含量依次为：（316．39±6．43）,（284．96±3．03）,（353．82±7．73）,（185．74±4．07）,（221．64±1．38）,（215．94±2．87）,（170．15±7．05）μg／g。结论：生物检材中HPLC／MS检测方法选择性好,定性准确,HPLC检测简便,快速,灵敏,定量结果准确,可用于利血平中毒的临床快速检验诊断和利血平中毒死亡案件的法医学鉴定。     

乌司他丁联合血必净注射液治疗脓毒症的临床疗效及对炎性因子的影响

目的：探讨乌司他丁联合血必净对脓毒症的临床疗效及炎性因子的影响。方法选择2013年12月至2015年2月上海交通大学医学院附属新华医院收治的脓毒症患者76例,依据随机数字表法分为对照组和观察组,各38例。对照组在常规治疗基础上加用血必净30 mL ,加至0．9％ NaCl溶液10 mL ,静脉滴注,每日1次,连续使用1周;观察组在对照组治疗的基础上使用乌司他丁10万U ,加至0．9％ NaCl溶液50 mL ,静脉滴注,每日2次,连续使用1周。比较两组患者的治疗效果。结果治疗后,观察组总有效率显著高于对照组,差异有统计学意义[94．7％（36／38）比57．9％（22／38）,P＜0．05],且观察组患者的肿瘤坏死因子（TNF）-α（86±26） ng／L、白细胞介素（IL）-6（26±8） ng／L、IL-8（26±13） ng／L、C-反应蛋白（CRP）（26±3） mg／L、降钙素原（PCT ）（0．83±0．23）μg／L、内毒素（2．2±0．9）μg／L等指标以及急性生理学及慢性健康状况评分系统Ⅱ（APACH-Ⅱ评分）（15±5）均明显少于对照组患者[（119±24） ng ／L、（60±7） ng ／L、（50±12） ng ／L、（78±7） mg ／L、（2．84±1．18）μg／L、（3．9±1．0）μg／L、（20±5）分],差异有统计学意义（ P＜0．05）;机械通气时间（11±4）d、重症监护病房住院时间（13±3）d等也较对照组患者（14±3）d、（18±4）d有明显缩短,差异有统计学意义（ P＜0．05）。结论乌司他丁联合血必净可有效降低脓毒症患者的炎性因子水平,保护患者的脏器功能,降低患者的APACH-Ⅱ评分,缩短患者住院时间,提高患者的生活质量,值得临床应用。     

异基因造血干细胞移植后免疫重建临床研究

为探讨异基因外周血造血干细胞移植（allo—PBSCT）后12个月内患者免疫重建特点及其与供受者年龄、供受者间HLA相容性、移植物抗宿主病（GVHD）及病毒感染的关系，随访37例异基因造血干细胞移植患者，用流式细胞术测定移植后1、3、6及12个月的T细胞亚群（CD3^＋、CD4^＋、CD8^＋）、B细胞（CD19^＋）及NK（CD16^＋CD56^＋）细胞，用散射比浊法检测免疫球蛋白IgG、IgA及IgM水平。结果显示，移植后1个月CD3^＋细胞的百分比为（47．5±23．2）％，3个月为（75．1±6．4）％，6个月为（69．7±12）％，12个月为（71．7±4．2）％。移植后1个月CD4^＋细胞百分比为（13．3±6．4）％，3个月为（20．2±11．4）％，6个月为（18．4±9．3）％，12个月为（29．1±8．7）％；移植后1个月CD8^＋细胞百分比为（43．1±17．4）％，3个月为（42．6±16．9）％，6个月为（46．9±10．3）％，12个月为（47±5．6）％；移植后1个月CD16^＋CD56^＋细胞百分比为（14．4±8．4）％，3个月为（15．9±7．6）％，6个月为（14．7±6．6）％，12个月为（13．6±3．4）％；移植后1个月CD19^＋细胞百分比为（6．4±5．6）％；3个月为（11．7±2．4）％，6个月为（13．3±7．3）％，12个月为（16．7±5．7）％。血清免疫球蛋白检测结果显示：移植后1个月IgA为（0．37±0．14）g／L，3个月为（0．28±0．21）g／L，6个月为（0．42±0．18）g／L，12个月为（0．53±0．34）g／L；移植后1个月IgG为（12．7±3．8）g/L，3个月为（16．3±5．2）g／L，6个月为（14．3±6．2）g/L，12个月为（15．4±6．9）g／L。移植后1个月IgM为（0．56±0．24）g／L，3个月为（0．64±0．16）g／L，6个月为（1．1±0．35）g／L，12个月为（1．2±0．28）g／L。大于等于45岁的患者T细胞亚群检测和血清免疫球蛋白与小于45岁患者比较无差异。发生慢性GVHD的患者移植后12?     

灯盏细辛注射液预防对比剂肾病的研究

【目的】观察中成药灯盏细辛注射液对防治冠状动脉介入诊疗后对比剂肾病（Contrast-induced Nephropathy,CIN）的有效性,为临床防治CIN提供新思路。【方法】随机将98例患者分成两组,从造影前6h至造影后6h,水化治疗组用0．9％氯化钠注射液（0．9％NS）以1．5mL／（kg·h）的滴速静脉滴注;灯盏细辛组用灯盏细辛注射液20mL（90mg）加入0．9％NS 500 mL,以1．5mL／（kg·h）的滴速静脉滴注。观测两组CIN发生率、血肌酐及内生肌酐清除率的变化。【结果】灯盏细辛组肌酐增加值[（31．2±16．6）vs（64．7±22．7）]和增加百分比[（10．70±5．3）％vs（26．8±10．3）％],内生肌酐下降值[（10．4±4．1）VS（15．0±4．2）]、下降比率[（9．9％±3．8）％VS（14．0％±4．7）％],CIN的发生率（6．1％vs22．4％）均明显低于对照组（P〈0．05）。【结论】灯盏细辛注射液对CIN的发生具有较好的保护作用。     

辛伐他汀和普罗布考对TNF-α诱导的内皮细胞凋亡的影响

【目的】探讨辛伐他汀和普罗布考对TNF-α诱导人脐静脉内皮细胞（HUVEC）凋亡的影响及其影响的量效关系和时效关系。【方法】体外培养HUVES,取对数生长期的细胞分为四个组：①TNF-α干预的浓度依赖性组（Ac组）[按浓度分为Acl（Ong／mL）,Ac2（1ng／mL）,Ac3（10ng／mL）,Ac4（25ng／mL）];②TNF-α干预的时间依赖性组（At组）[按时间分为Atl（Oh）,At2（12h）,At3（24h）,At4（48h）];③10ng／mLTNF_a＋不同浓度辛伐他汀干预组（B组）[浓度分为B1（O μmol／L）,B2（O．01 μmol／L。）,B3（O．1 μmol／L。）,B4（1．0t μmol／L）];④1O ng／mL TNF-a不同浓度普罗布考干预组（c组）[浓度分为C1 （0 μmol／L。）,C2（20 μmol／L）,c3（40 μmol／L）,C4（80 μmol／L）]。以上四组均干预12h后,用流式细胞术检测各组凋亡率。【结果】①Ac组凋亡率分别为：Acl（O．81±0．25％）,Ac2（2．35±0．04％）,Ac3（4．54±0．32％）,Ac4（6．17±0．28％）,各组之间存在统计学差异（P〈0．01）。②At组凋亡率分别为,Atl（0．87±o．35％）,At2（4．53±0．32％）,At3（7．45±1．97％）,At4（10．92±1．27％）,各组间存在统计学差异（P〈O．01）。③B组凋亡率分别为,B1（4．46±0．53％）,B2（1．38±0．12％）,B3（2．89±0．27％）,B4（3．65±0．08％）,各干预组与对照组之间存在统计学差异（P〈O．01）。④C组细胞凋亡率分别为,C1（4．60±0．64％）,C2（2．61±0．18％）,C3（2．21±0．22％）,c4（1．28±0．34％）,各干预组与对照组之间存在统计学差异（P〈0．01）。【结论】①TNF-α可以诱导内皮细胞凋亡,且存在浓度依赖性和时．：良赖性。②辛伐他汀可以抑制TNF-α口诱导的内皮细胞凋亡,但无浓度依赖性。③普罗布考可以抑制TNF-α诱导的内皮细胞凋亡,但无浓度依赖性。     

老年男性2型糖尿病患者骨密度与冠心病的相关性

目的探讨老年男性2型糖尿病患者骨密度与冠心病的相关性。方法102例老年男性2型糖尿病住院患者根据是否合并冠心病分两组,比较分析两组患者的腰1～腰4及右侧股骨颈的骨密度）,并采用多因素非条件Logistic回归分析骨质疏松与冠心病的相关性。结果合并冠心病组腰1～腰4和股骨颈的骨密度分别为（0．73±0．12）g／cm^2、（0．76±0．14）g／cm^2、（0．82±0．22）g／cm^2、（0．81±0．21）g／cm^2、（0．71±0．12）g／cm^2,未合并冠心病组骨密度分别为（0．86±0．17）g／cm^2、（0．92±0．22）g／cm^2、（0．96±0．23）g/cm^2、（0．98±0．25）g／cm^2、（0．80±0．13）g／cm^2,两组比较,差异均有统计学意义（t分别：-4．38、-4．32、-3．27-3．67、-4．30,P均〈0．05）。调整了年龄、体重指数、血脂、血钙、血磷后,经Logistic回归分析显示：骨质疏松和冠心病存在明显的相关性（OR=3．80,95％CI：1．50～9．61,P〈0．05）。结论老年男性2型糖尿病患者骨密度的降低可能跟冠心病的发生、发展有关。     

高效液相色谱法测定米非司酮的血药浓度

目的：建立测定米非司酮血药浓度的方法。方法：以乙睛-甲醇-二氯甲烷三乙胺一水为流动相，血清样品经乙醚萃取，液氮冷冻分离后，在波长302nm处检测米非司酮。结果：线行范围0．02～2．21／μg／ml，米非司酮的工作曲线为y（μg）=0．07x-0．25（r^2=0．9996）。回收率在89％～93％之间。日内与日间RSD均小于4％。结论：高效液相色谱法快速、简便、准确、灵敏，可用于米非司酮的药物动力学研究。     

组织因子凝血酶受体和血栓调节蛋白在重度脓毒症诱导ALI／ARDS大鼠肺组织细胞中的表达北大核心CSCD

目的探讨重度脓毒症诱导的ALI/ARDS大鼠肺组织细胞组织因子（TF）、凝血酶受体（TR）和血栓调节蛋白（TM）的表达。方法应用盲肠结扎穿孔（CLP）加生理盐水灌洗法制作重度脓毒症诱导的ALI／ARDS大鼠,以PaO2／FiO2≤300作为ALI/ARDS大鼠纳入标准,随机分为假手术组、ALI／ARDS组。制模成功后1、2、3h处死大鼠,取肺组织称质量制备组织匀浆,ELISA方法测定TF、TR和TM的表达。结果①TF表达：ALI/ARDS组灌洗后1、2、3h分别为（206363±8178）pg／mL、（148186±6383）pg／mL、（170889±7180）pg／mL,较假手术组[（55956±271）pg／mL、（53269±473）pg/mL、（54175±416）pg/mL]明显升高（P〈0．05）。②TR表达：ALI/ARDS组灌洗后1、2、3h分别为（1．657±0．063）ng／mL、（1．180±0．074）ng／mL、（1．497±0．087）ng／mL,较假手术组[（0．266±0．046）ng/mL、（0．273±0．048）ng／mL、（0．258±0．050）ng／mL]明显升高（P〈0．01）。③TM表达：ALI/ARDS组灌洗后1、2、3h分别为（16960±179）ng/mL、（10372±300）ng／mL、（12287±189）ng／mL,较假手术组[（567±21）ng／mL、（774±35）ng／mL、（695±43）ng/mL]明显升高（P〈0．05）。ALI／ARDS组组内各时点参数比较差异无统计学意义,但呈现灌洗后1h急性应激性升高、2h后下降、3h后再次上升的趋势。结论重度脓毒症诱导的ALI/ARDS大鼠肺组织在高表达TF和TR等促凝血因子的同时,也高表达具有辅助抗凝和增加纤溶功能的TM。     

血液灌流联合血液透析治疗尿毒症神经系统损害患者疗效

目的：观察血液灌流与血液透析联合治疗尿毒症神经系统损害患者临床疗效。方法将88例尿毒症神经系统损害患者随机分为两组各44例,观察组采用血液灌流与血液透析联合治疗,对照组采用单纯血液透析治疗,比较两组临床疗效,症状变化以及治疗前后中分子物质（MMS）、血尿氮素（BUN）、血肌酐（SCr）、β2-微球蛋白（β2-MG）、白蛋白（Alb）、C 反应蛋白（CRP）的水平变化。结果观察组治疗有效率90．9％（40／44）高于对照组75．0％（33／44）,治疗后麻木感发生率6．8％（3／44）低于对照组52．3％（23／44）,扑翼样震颤症状发生率6．8％（3／44）低于对照组38．6％（17／44）;观察组 MMS、BUN、SCr、CRP 水平及β2-MG 均显著低于对照组,分别为（1600±378）U／L vs （4479±607）U／L、（10．9±2．4）mmol／L vs （15．2±2．6）mmol／L、（328．6±127．2）μmol／L vs （542．6±131．8）μmol／L、（7．0±5．2）mg／L vs （9．4±6．0）mg／L 及（5．1±1．6）mg／L vs （8．8±2．5）mg／L,Alb 高于对照组（41．0±3．2）g／L vs （37．9±2．8）g／L（均 P ＜0．01或＜0．05）。结论血液灌流与血液透析联合治疗尿毒性神经系统损害临床疗效确切,能够全面改善临床症状,降低炎症水平及其它各项指标。     

自制大容量扣式负压抽提装置应用于RNA抽提的性能评价

目的对自制大容量扣式负压抽提装置进行扩增前性能评价。方法采用配套负压抽提装置(对照组装置)和自制大容量扣式负压抽提装置(实验组装置)对67份白细胞(WBC)大于2.0×109/L的新鲜全血进行核酸(RNA)提取并进行扩增前性能评价,验证实验组的工作效率、提取RNA的浓度、纯度及完整性。结果 67份RNA标本采用对照组装置69min完成抽提,处理效率平均为0.97份/分;实验组装置41min完成抽提,效率为1.63份/分。对照组抽提标本RNA浓度为(248.8±21.4)μg/mL,实验组为(260.3±21.8)μg/mL。对照组抽提标本OD260/OD280和OD260/OD230分别为(1.995±0.095)和(2.020±0.082),95%CI分别为1.964~2.025和2.001~2.040,实验组分别为(2.093±0.092)和(2.071±0.120),95%CI分别为2.075~2.113和2.044~2.103;2组比较差异有统计学意义(t=24.570,P0.001),线性回归方程Y=0.950 X+0.039,R2=0.903。2组装置提取RNA后完整性均较好,电泳条带整齐、清晰。结论自制大容量扣式负压抽提装置具有更高的抽提工作效率,且能保证RNA浓度、纯度及完整性,更符合实验室实际情况,具有更优的临床价值。     

Fetal trunk and head volume measured by three-dimensional ultrasound at 11 + 0 to 13 + 6 weeks of gestation in chromosomally normal pregnancies.

OBJECTIVE: To establish the relationship between fetal trunk and head volume measured by three-dimensional (3D) ultrasound and gestational age at 11 + 0 to 13 + 6 weeks of gestation. METHODS: The fetal trunk and head volume were measured using 3D ultrasound in 417 chromosomally normal fetuses from singleton pregnancies at 11 + 0 to 13 + 6 (median, 12 + 0) weeks of gestation. Regression analysis was used to determine the significance of the association between fetal volume and gestational age. The Bland-Altman analysis was used to compare the measurement agreement and bias for a single examiner and between different examiners. RESULTS: The fetal trunk and head volume increased linearly with gestation from a mean of 5.8 mL at 11 + 0 weeks to 33.3 mL at 13 + 6 weeks and 1 SD was 4.4 mL. There was also a significant linear association between fetal volume and crown-rump length (CRL), from a mean of 5.1 mL at a CRL of 45 mm to 37.5 mL at a CRL of 84 mm and 1 SD was 2.7 mL. However, within this gestational range, a doubling in CRL, from a mean of 48 mm at 11 + 0 weeks to 79 mm at 13 + 6 weeks, was associated with a 5-6-fold increase in fetal volume. The mean difference in fetal volume between paired measurements by the same sonographer was -0.87 mL (95% limits of agreement, -2.31 to 4.05 mL) and the mean difference between paired measurements by two sonographers was -1.09 mL (-5.49 to 3.32 mL). CONCLUSIONS: 3D ultrasound can provide a reproducible measurement of the fetal trunk and head volume in early pregnancy. At between 11 + 0 and 13 + 6 weeks there is a 5-6-fold increase in fetal volume but only a doubling in CRL.     

A simple solid‐phase extraction method for the analysis of red cell phospholipids by liquid chromatography‐ tandem mass spectrometry

Background

There has been increasing interest in the analysis of phospholipids in red blood cells as potential long‐term biomarkers of different disease states. Here, we describe a simple method for the analysis of two phospholipids: 1‐Palmitoyl‐2‐oleoyl‐sn‐glycero‐3‐phosphoethanol (PE 16:0/18:1) and 1‐Palmitoyl‐2‐linoleoyl‐sn‐glycero‐3‐phosphoethanol (PE 16:/0/18:2) in erythrocytes by liquid chromatography‐tandem mass spectrometry (LC‐MS/MS).

Methods

Whole blood samples were removed free of plasma and washed in isotonic saline. Red cells were lysed with ultrapure water. Lysate samples were processed using a hybrid solid‐phase extraction (SPE) phospholipid cartridge (1 mL, 30 mg). Both PE 16:0/18:1 and PE 16:0/18:2 and their deuterated internal standards were separated on an ACE C4 (150 mm × 2.1 mm, 2.7 μm particle size) by gradient elution at a flow rate of 0.5 mL per minute using mobile phases consisting of 0.01 mol/L ammonium acetate in: water (A), methanol (B), and isopropanol (C). The phospholipid species were quantified by the following transitions: PE 16:0/18:1: 701.5→281.3 and PE 16:0/18:2: 699.5→279.3.

Results

Both PE species displayed linearity ranging from 10 to 500 μg/L. The coefficient of variation (CV%) of PE 16:0/18:1 concerning intraday and interday precision was between 1.9%‐2.6% and 3.0%‐4.3%, respectively. For PE 16:0/18:2, this was between 1.8%‐3.4% and 3.7%‐4.1%, respectively. Both phospholipid species had accuracy (PE 16:0/18:1: 91%‐98% and PE 16:0/18:2: 94%‐103%) and extraction recovery (PE 16:0/18:1: 95%‐106% and PE 16:0/18:2: 92%‐102%) exceeding 90% over the analytical range. The limit of detection was 5 μg/L.

Conclusion

Here we propose a simple SPE LC‐MS/MS method for analyzing phospholipids in erythrocytes, which can be easily adopted.

     

Bioequivalence evaluation of two brands of cefaclor 500 mg capsules: quantification of cefaclor using solid phase extraction technique

OBJECTIVE: To assess the bioequivalence of two cefaclor 500 mg capsule formulations, and to develop a new high performance liquid chromatographic (HPLC) method using solid phase extraction technique for the quantification of cefaclor in human plasma. METHOD: An open, randomized, two-way, crossover trial with a one-week washout period in 25 healthy volunteers. The two commercial brands used were Recocef(Julphar, United Arab Emirates) as test and Ceclor(Eli Lilly, UK) as reference product. The drug was administered with 240 mL of water after a 10-h overnight fast. After dosing, serial blood samples were collected for a period of 8 h. Plasma harvested from blood was analysed for cefaclor by a new HPLC method using a solid phase extraction technique. The limit of detection of cefaclor was 17.6 ng/mL; average recovery was 96.5%; the intraday CV was less than 8% and interday CV was less than 13%. Various pharmacokinetic parameters, including AUC0-t, AUC0-infinity, Cmax, Tmax, T1/2, and Kel, were determined from plasma concentrations for both formulations. Statistical analysis (ANOVA and 90% confidence intervals) were applied to AUC0-t, AUC0-infinity and Cmax for bioequivalence evaluation of two brands. The new HPLC method with solid phase extraction circumvented the problem of mixed polarity of cefaclor and facilitated its extraction from the complex plasma matrix while keeping the background free from interference due to endogenous plasma compounds. RESULTS: No significant difference was observed between the two brands of cefaclor capsules. CONCLUSION: Recocef was judged bioequivalent to Ceclor and the two products can therefore be considered to be interchangeable in medical practice.     

Efficacy of preoperative donation of blood for autologous use in radical prostatectomy

To determine the amount of blood lost, the number of transfusions, and the effectiveness of preoperative autologous blood donation in radical prostatectomy, 163 patients' records from 1987 to 1991 were reviewed at four university hospitals and three community hospitals. Calculated red cell volume lost was 1003 +/− 535 mL (mean +/− SD), which corresponds to 44 +/− 18 percent (mean +/− SD) of total red cell volume. Preoperative donation of blood for autologous use reduced the rate of transfusion of allogeneic blood from 66 to 20 percent (p < 0.001). Of the patients who donated 1 to 2 units, 32 percent received allogeneic blood; 14 percent of those who donated 3 units received allogeneic blood. Donation of 4 units reduced the allogeneic transfusion rate to 11 percent. However, as the number of units donated increased (1-3 units), the units not transfused also increased (0-21%). Ninety-one (56%) of 163 patients donated fewer than 3 units. Autologous blood donation is effective in minimizing the transfusion of allogeneic blood to radical prostatectomy patients, but many patients do not donate enough blood (< 3 units). The donation of 3 units of blood for autologous use is recommended for patients who undergo radical prostatectomy.     

Liquid chromatography as candidate reference method for the determination of vitamins A and E in human serum

BackgroundOwing to the increasing interest in public health research of antioxidant micronutrients and the inaccuracy of routine serum concentrations of the fat‐soluble vitamins A (retinol) and E (DL‐α‐tocopherol) measurements, we developed a reliable, highly sensitive, robust and rapid method for the quantification of two clinically important lipophilic antioxidants in serum using a reverse‐phase HPLC/DAD method.MethodSample preparation and analytical conditions that would affect extraction efficiency and quantitative results of vitamins A and E were investigated and optimized. Vitamins A and E were extracted from serum via liquid‐liquid extraction (LLE). After adequate sample preparation, the samples were injected directly into the HPLC system with diode‐array detector (DAD). Chromatographic separation was completed in 7 minutes for vitamins A and E. With vitamin A acetate and vitamin E acetate as internal standards, the method was applied to the measurement of vitamins A and E in human serum.ResultsWe evaluated method linearity, accuracy (recovery rate and trueness), precision, carryover, limit of quantitation and limit of detection, and measurement uncertainty. The method was evaluated for trueness using NIST Standard Reference Material SRM 968f. The serum concentration of the studied compounds had a good linear relationship in the range of 0.05 ~ 3.0 μg/mL concentration (r ＝ 0.9998), with 0.0077 μg/mL detection limit and 0.025 μg/mL quantitative limit for vitamin A, respectively, and 1.0 ~ 60.0 μg/mL concentration (r ＝ 0.9999), with 0.40 μg/mL detection limit and 0.50 μg/mL quantitative limit for vitamin E, respectively. The intra‐ and inter‐assay coefficients of variation were calculated by using three concentrations (1, 2, and 3) of the studied compounds in human serum samples. Intra‐assay and inter‐assay precision were 1.23%‐4.97% and 0.97%‐3.79% for vitamin A, respectively, and 0.64%‐4.07% and 0.81%‐5.96% for vitamin E, respectively. The average recovery rates were 100.98% for vitamin A, and 99.21% for vitamin E, respectively. The carryover rate of vitamins A and E was below 1%. As for the evaluation of accuracy, the biases were <± 5% by comparing with NIST standard reference material SRM 968f.ConclusionThe method is a simple sample treatment procedure for the determination of fat‐soluble vitamins A and E in human serum with high sensitivity and specificity. The proposed method could be recommended as a candidate reference method for the determination of serum concentrations of the fat‐soluble vitamins A and E in human serum.     

A single-dose,three-period,six-sequence crossover study comparing the bioavailability of solution,suspension, and enteric-coated tablets of magnesium valproate in healthy Mexican volunteers under fasting conditions

Background: Valproic acid has been associated with a highly variable intersubject absorptive phase; therefore, magnesium salt (magnesium valproate [MgV]) was developed to diminish variation during enteric absorption.Objectives: The aims of this study were to assess the pharmacokinetics of single oral doses of MgV 500-mg solution, suspension, and enteric-coated tablets in a healthy Mexican population, and to compare formulation-related differences.Methods: This was a randomized, single-dose, 3-period, 6-sequence crossover study in healthy Mexican volunteers aged 18 to 45 years. In each period, subjects received single oral doses of 500-mg MgV solution, suspension, and enteric-coated tablet formulations, with a 7-day washout period between each dosing period. Serial blood samples were collected at 0 hour (prior to MgV administration) and at 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 6, 9, 12, 24, 48, and 72 hours after dosing. Valproate was measured by a new method of ultraperformance liquid chromatography coupled with mass spectrometry. Pharmacokinetic parameters of interest were C_max, T_max, AUC_0–72, AUC_0?∞, t½, V_d/F, CL/F, and mean residence time (MRT). Formulation-related differences were assayed in accordance with the Mexican regulatory bioequivalence criteria. Log-transformed values of C_max and AUC were used to construct a classic 90% CI. Bioequivalence was established if the 90% CI for the mean test:reference ratio of log-transformed C_max and AUC were within the range of 0.80 to 1.25. Tolerability was assessed based on subject interview, vital sign monitoring, and clinical assessment.Results: A total of 24 healthy volunteers (12 women and 12 men; mean [SD] age, 28.79 [6.5] years; height, 164 [9.8] cm; weight, 65.42 [8.95] kg; and body mass index, 24.28 [2.11] kg/m²) were included. For the MgV solution, the mean (SD) pharmacokinetic parameters of C_max, T_max, AUC_0–72, AUC_0–∞, t½, V_d/F, CL/F, and MRT were 59.75 (8.24) μg/mL, 0.542 (0.14) hours, 1099.67 (241.70) μg · h/mL, 1156.30 (264.01) μg · h/mL, 16.19 (2.36) hours, 9633.68 (1892.70) mL, 418.35 (92.01) mL/h, and 18.36 (1.44) hours, respectively. For the MgV suspension, the mean (SD) pharmacokinetic parameters of C_max, T_max, AUC_0–72, AUC_0?∞, t½, V_d/F, CL/F, and MRT were 55.04 (7.72) μg/mL, 0.773 (0.51) hour, 1057.76 (223.37) μg · h/mL, 1111.09 (245.07) μg · h/mL, 16.32 (2.20) hours, 1069.05 (1775.64) mL, 435.43 (99.59) mL/h,\ and 18.41 (1.43) hours, respectively. For the MgV entericcoated tablets, the mean (SD) pharmacokinetic parameters of C_max, T_max, AUC_0–72, AUC_0?∞, t½, V_d/F, CL/F, and MRT were 54.88 (6.73) μg/mL, 2.79 (0.89) hours, 1100.79 (216.70) μg · h/mL, 1163.61 (238.36) μg · h/mL, 16.48 (2.10) hours, 9675.15 (1659.36) mL, 412.36 (85.24) mL/h, and 19.95 (1.53) hours, respectively. The 90% CIs for the tablets:solution ratio were 82.15 to 95.44, 94.60 to 105.39, and 95.43 to 105.95 for C_max, AUC_0–72, and AUC_0?∞, respectively. The 90% CIs for the suspension:solution ratio were 84.79 to 98.50, 88.89 to 99.02, and 89.15 to 98.97, respectively. The 90% CIs for the tablets:suspension ratio were 89.90 to 104.43, 100.84 to 112.34, and 101.60 to 112.80, respectively.Conclusion: This single-dose study found that the 3 formulations (solution, suspension, and enteric-coated tablets) of MgV met the regulatory criteria for bioequivalence in these healthy, fasting, Mexican volunteers.     

LHPC—MS／MS法测定人体血浆中丙泊酚浓度的评价

【目的】建立人血浆中丙泊酚浓度的高效液相色谱串联质谱法（HPLC—MS／MS）测定方法,以评价其效果。【方法】采用LC—MS／MS法测定丙泊酚的浓度,色谱柱为Zorbax Eclipse XDB-C18（50mm×4．6mm,5μm）,流动相为甲醇及0．1％氨水溶液;流速0．4mL／min;电喷雾离子化电离源（ESI）,质谱采用MRM模式,以负离子检测,检测离子为丙泊酚m／z177．2→m／z161．4、麝香草酚m／Z147．2→m／z106．7。20名健康志愿者静脉注射丙泊酚2mg／kg后,按预定时间点抽取动脉血进行血浆药物浓度的检测。【结果】丙泊酚血药浓度在线性范围分别为0．012～12．06μg／mI。范围内线性关系良好（r=0．9993）,定量下限为0．012μg／mL;低、中、高3个浓度的日内及日间相对精密度（RSD）〈15％;平均提取回收率86．7％,RSDl．87％。【结论】本方法特异性强、灵敏、高效、精密度及准确度好,适用于人体血浆中丙泊酚的检测。     

Comparison of the pharmacokinetics of lansoprazole 15- and 30-mg sachets for suspension versus intact capsules

OBJECTIVE: The pharmacokinetic profiles of single doses of lansoprazole 15- and 30-mg sachets for suspension were compared with those of corresponding doses of lansoprazole oral capsules. METHODS: Healthy adult male and female subjects were randomized (1:1 ratio) into 2 Phase 1, open-label, single-dose, 2-sequence, 2-period complete crossover studies. In the first study, each subject received 1 lansoprazole 15-mg sachet mixed with water and 1 lansoprazole 15-mg oral capsule; in the second study, each subject received 1 lansoprazole 30-mg sachet mixed with water and 1 lansoprazole 30-mg oral capsule. Administration of the 2 formulations was separated by a washout period of > or =7 days. Blood samples were collected before and after each administration to assess the pharmacokinetic parameters of lansoprazole and bioequivalence between suspension and capsule. RESULTS: Thirty-six subjects (19 males, 17 females) with a mean (SD) age of 32.0 (9.6) years and mean (SD) body weight of 68.6 (10.5) kg received lansoprazole 15 mg. Thirty-six subjects (22 males, 14 females) with a mean (SD) age of 38.0 (8.3) years and mean (SD) body weight of 75.1 (9.7) kg received lansoprazole 30 mg. The pharmacokinetic parameters of the 15- and 30-mg lansoprazole sachets for suspension were similar to those of the corresponding doses of the oral capsules. The mean (SD) values for C(max) and AUC from time 0 to infinity (AUC(0-infinity) for the lansoprazole 15-mg sachet (591.9 [242.3] ng/mL and 1614 [2065] ng.h/mL, respectively) did not differ significantly from those for the lansoprazole 15-mg capsules (578.6 [275.2] ng/mL and 1620 [2290] ng.h/mL, respectively). These parameters also did not differ significantly between the lansoprazole 30-mg sachet and 30-mg capsule: mean (SD) C(max), 1103 (428.3) and 1077 (465.6) ng/mL, respectively; mean (SD) AUC(0-infinity), 2655 (1338) and 2669 (1311) ng.h/mL, respectively. The 90% Cls for C(max) and AUC(0-infinity) ratios were contained within the 0.80 to 1.25 equivalence range, supporting bioequivalence. CONCLUSIONS: These findings suggest that the 15- and 30-mg lansoprazole sachets for suspension are bioequivalent to the corresponding doses of oral capsules. The sachet for suspension may provide an alternative route of administration to patients who have difficulty swallowing solid oral formulations.