高效液相色谱法测定全血中加替沙星

提出了用环丙沙星为内标及用荧光检测的加替沙星(GXTC)的高效液相色谱测定方法.全血样品中GXTC用pH 7.0的磷酸盐缓冲溶液萃取并经固相萃取小柱净化,用甲酸及甲醇(4+96)混合溶液作淋洗液淋洗萃取小柱,所得淋出液用吹氮法蒸干,残渣溶于1.0 mL甲醇中,分取10.0μL溶液供高效液相色谱测定.测定中用甲醇、乙腈及甲酸(2+998)溶液三者以15比15比70的体积比混合后作为流动相,在Cloversil C18柱(150 mm×3.0 mm,5 μm)上进行分离.荧光检测激发波长为288 nm,发射波长为478 nm.加替沙星在15.0～240.0μg·L-1范围内呈线性,方法回收率在97.0%～102.0%之间,日内相对标准偏差(n=6)小于6.0%,方法的测定限(10S/N)为15.0μg·L-1.     

高效液相色谱法测定全血环孢霉素A的浓度

建立了高效液相色谱法测定全血环孢霉素Ａ 浓度的方法。全血样品经多步处理后以２０ μＬ注入ｒｅｏｓｏｌｖｅ Ｃ１８,５μ色谱柱,采用Ｖ（乙腈）∶Ｖ（水）＝ ８０∶２０ 的流动相,在７５℃柱温下,以１ ｍ Ｌ／ｍ ｉｎ 流速洗脱,检测波长为２０８ｎｍ 。环孢霉素Ａ 的回收率为９９％ ～１０４ ％ ,相对标准偏差＜ ３．４２％ （ｎ＝ ５） ,日间＜ ８．１９％ （ｎ＝ ４） 。最低检测浓度为２５ μｇ／Ｌ。     

高效液相色谱法测定全血中直链烷基苯磺酸钠

提出了高效液相色谱法测定全血中直链烷基苯磺酸钠同系物的含量。全血样品用酸性提取溶液稀释,过Bakerbond SPE C18固相萃取小柱,经甲醇-乙酸-水(49+2+49)混合溶液淋洗后用甲醇-氨水(98+2)混合溶液洗脱。经纯化后的洗脱液在Wakopak WS AS-Aqua(250 mm×4.6 mm,5μm)柱上进行分离,流动相为乙腈与水(60+40)混合溶液,检测波长为228 nm。5种n-ABS同系物的C10~C14的质量分数在10.0~100.0 ng.g-1范围内与其峰面积呈线性关系,全血中各同系物回收率为78%~107%;日内相对标准偏差(n=5)≤10%,日间相对标准偏差(n=5)≤13%。     

HPLC Determination and Pharmacokinetic Study of Indapamide in Human Whole Blood

A rapid and sensitive HPLC method for the quantification of indapamide in human whole blood is described. The procedure involves liquid–liquid extraction of human whole blood with methyl tertiarybutyl ether coupled with reversed-phase HPLC and UV detection. Separation was performed on a YMC^® ODS-A reverse column (5 µm particle size, 4.6×150 mm i.d.). The mobile phase was composed of acetonitrile - 2-propanol ?0.1 triethylamine in water (adjusting to pH 3.75 with 85% phosphoric acid) (35:5:60, v/v/v). The linear concentration range for indapamide was 5.0–500 ngmL^?1. The lower limit of quantification (LLOQ) was 5.0 ngmL^?1 for indapamide. Intra- and inter-assay RSD ranged from 2.36% to 4.53% and 1.68% to 8.01%, respectively. The sensitivity and precision were sufficient for determination of whole blood concentrations after therapeutic administration of both drugs and the method can be used for the estimation of pharmacokinetic parameters.     

石墨炉原子吸收光谱法测定全血中铅

用作为基体改进剂的含有氯化钯、Triton X-100及硝酸的混合溶液将全血样品稀释10倍,分取部分经稀释后的试液直接进样,按选定的仪器工作条件进行石墨炉原子吸收光谱法测定,此方法的检出限(S/N=3)为5.4 μg·L-1.应用此方法分析了一级国家标准物质(编号为GBW 09139e及GBW 09140e),测得结果与证书值相符,测定值的相对标准偏差(n=6)在1.2%～3.2%之间.     

Determination of Primaquine in Whole Blood and Finger-Pricked Capillary Blood Dried on Filter Paper Using HPLC and LCMS/MS

Primaquine (PQ) is the only 8-aminoquinoline antimalarial drug in clinical use because of its unique action on hypnozoites and gametocytes of Plasmodium species. We report here simple, sensitive and specific assay methods for the determination of PQ in human whole blood and dried blood spot (DBS) samples using high-performance liquid chromatography and liquid chromatography-mass spectrometry, respectively. Sample preparation was performed by a single or two-step liquid-liquid extraction with organic solvents. For whole blood analysis, separation was obtained on a reversed-phase C18 column with the mobile phase consisting of 0.25 % diethylamine and acetonitrile (7:3, v:v) running at a flow rate of 1.0 ml min^?1. UV detection was at the wavelength 263 nm. For DBS analysis, separation was obtained on a reversed-phase column with the mobile phase consisting of methanol and 0.1 formic acid (1:3, v:v) running at a flow rate of 0.5 ml min^?1. The selected ions generated by electrospray ionization were detected using mass spectrometer. Good precision and accuracy (both within-day and day-to-day assays) were obtained at the concentration ranges under investigation. Limits of quantification for PQ were accepted as 25 ng ml^?1 using 500 μl whole blood and 5 ng ml^?1 using 80 μl DBS samples. The mean recoveries for PQ and internal standard pyrimethamine (PYR) for both whole blood and DBS were over 70 %. The methods were successfully applied for a clinical pharmacology study of PQ in patients with Plasmodium vivax. Excellent correlation (r ²  = 0.997) was observed between the analysis of PQ in paired whole blood and DBS samples.     

高效液相色谱法测定血清中番茄红素

血清样品经正己烷萃取和高速离心除去蛋白质后在低温蒸发浓缩,最终用正己烷定容为10 mL,取10μL试液进样作高效液相色谱测定,所用固定相为Shimpack VP-ODS色谱柱,用乙腈与四氢呋喃混合液(体积比55:45)及单纯乙腈作流动相先后进行梯度淋洗.在471nm波长处作吸光光度检测,方法的线性范围为0.01～5.00 mg·L-1番茄红素.按此方法测定了632份健康人血清中蕃茄红素的含量,在0.06～2.94 mg·L-1范围内,中位值为o.45 mg·L-1.对方法的回收率及精密度做了试验,测得回收率在94.4%～99.7%之间,相对标准偏差值(n=6)为7.6%.     

HPLC Measurement of Cyclosporine in Blood Plasma and Urine and Simultaneous Measurement of its Four Metabolites in Blood

Abstract

A high performance liquid chromatographic assay has been developed for the estimation of cyclosporine and its four major metabolites in blood and for cyclosporine alone in plasma and urine samples. This assay employs a rapid and very reproducible solid-liquid extraction system. Isocratic chromatographic conditions allow the simultaneous measurement of cyclosporine and its four major metabolites in blood. The method is linear up to 2500 ng/ml and the minimum quantifiable limit for cyclosporine is 30 ng/ml, when 1 ml of sample is analyzed.     

HPLC及LC-MS法测定全血中青霉素

青霉素(penicilinG)属于青霉素类杀菌剂药物,极易溶于水,在甲醇、乙醇中溶解,在脂溶性溶剂中不溶解。易吸潮,遇酸、碱或氧化剂等迅速失效,水溶液也易分解。人体肌注后15～30min血浓度达到高峰值,有60％-90％原形物从肾脏中排出。青霉素毒性很小,人类即使使用大剂量也难以中毒,但是青霉素肌内注射、输液常引起过敏,并导致死亡。据1952～1966年的不完全统计,国内文献中关于发生青霉素过敏性休克的报告,至少400余例,其中50余例死亡。近2、3年间国内文献中仍有青霉素引起过敏反应的大量报告,尤其游医郎中的不合理用药,致死案例时有发生。这就要求我们对死者体内是否有青霉素进行分析测定。     

胶束电动毛细管色谱法测定全血环孢霉素A

建立了用于临床测定全血环孢霉素A的毛细管电泳分析方法。采用未涂渍熔融石英毛细管(50μm×58.5 cm,有效长度50.0 cm),以含20 mmol/L磷酸氢二钠、50 mmol/L十二烷基硫酸钠的乙腈-水(1∶1)体系为电泳介质体系,采用乙醚萃取的前处理方法,有效去除了全血中内源性物质的干扰。通过柱上富集浓缩技术,提高检测灵敏度,检出限达70μg/L,实现了环孢霉素A的低浓度检测。     

湿法消解-原子荧光法测定全血中硒

以全血样品为原料,探讨湿法消解-原子荧光法测定全血中的硒含量。血样经硝酸-高氯酸消解后,用硼氢化钠将硒还原成硒化氢,由氩气载入原子化器,产生的原子荧光强度与试液中硒元素含量在一定范围内呈正比,外标法定量。以消解效率为指标,优化样品的消解条件,测定血中硒在0-10μg/L范围内线性情况良好,相关系数为0.9992,最低检出限为0.143μg/L,相对标准偏差为1.51%-1.58%,平均加标回收率为90.86%-104.62%。血中硒的原子荧光测定法灵敏度高,精密度和稳定性好,可应用于血中硒的生物监测。     

Sonoelectroanalysis: Anodic Stripping Voltammetric Determination of Cadmium in Whole Human Blood

The quantitative detection of cadmium in human blood is shown to be possible by anodic stripping voltammetry under conditions of insonation. An immersion horn probe is introduced into a thermostatted conventional three‐electrode cell opposite a Nafion‐coated mercury plated glassy carbon electrode. The enhanced mass transport associated with power ultrasound yields efficient preconcentration of the cadmium before it is detected using anodic differential‐pulse stripping voltammetry. Insonation further offers the crucial benefits of first surface activation and cleaning, helping to prevent electrode fouling by the organic components in blood and second fully equilibrating “free” and “bound” Cd²⁺ ions in the complex matrix. Acoustic streaming and cavitation promote the mass transport of cadmium to the surface of electrode, facilitating measurements in solutions of low cadmium concentration, where “silent” measurements fail to yield an analytical signal. The system is calibrated using standard micro additions of cadmium to give the total amount of cadmium in blood. The calibration plot, showing the dependence of the cadmium stripping peak height on cadmium concentration in 1 : 50 diluted blood samples, was linear in the range 1×10^?10 M to 4×10^?9 M Cd²⁺. The values of cadmium concentration obtained using sonoelectroanalytical methodology were compared with the results obtained by independent atomic absorption spectroscopy (AAS) measurements and good agreement was found.     

微波萃取气相色谱法分析全血中的甲基苯丙胺

苯丙胺(又称安非他明,amphetamine)类毒品是一类非法人工合成的有机胺类兴奋剂,它主要作用于中枢神经和交感神经,属于中枢神经兴奋类毒品,其中甲基苯丙胺(俗称冰毒)最常被滥用.由于苯丙胺类毒品的滥用现象日趋严重,对社会危害极大.因此,对该类毒品的分析研究不仅意义重大,而且十分迫切.目前,国内外报道的生物样品中苯丙胺类毒品的前处理方法主要有液液萃取、固相萃取及固相微萃取等方法.近年来,微波辅助萃取(microwave-assisted extraction,MAE)因其萃取时间短、回收率高、试剂用量少、污染小和可自动控制制样条件而得到了分析工作者的认同,并已应用于土壤、沉积物中有机芳烃[1]以及生物样品中农药残留[2]的萃取研究,而在法庭科学上的应用极少报道.本文对全血中的甲基苯丙胺进行了初步的微波萃取技术研究,建立了微波萃取气相色谱法分析生物样品中甲基苯丙胺的方法.     

632例全血微量元素锌检测及分析

为分析不同年龄组人群的锌缺乏情况,给补锌提供参考依据,用EXCEL 2002软件处理9个年龄组632例微量元素锌含量。结果表明:(1)各年龄组均有缺锌,各组均值(除成人组)只能达到该组正常参考值的的低值水平;(2)各年龄组男女性之间的锌值均有明显差异(P<0.05);(3)0~1岁婴儿缺锌率最高,成人组缺锌率最低,与其他各年龄组有极显著差异(P<0.01),5~6岁年龄组缺锌率比相邻两年龄组高;(4)1~2岁组与2~3岁组、4~5岁组与5~6岁组、6~10岁组与11~16岁组锌值比较,无显著性差异。提示各年龄组均要注意合理膳食,提高富锌食物的摄入,0~1岁婴幼儿还要适当补锌。     

高血压性左室肥厚患者全血镁的测定

通过对比观察了50例高血压性左室肥厚患者和60例高血压性无左室肥厚患者全血镁含量的变化。结果表明,左室肥厚组全血镁明显低于无左室肥大组（P＜0．05）。提示高血压性左室肥厚患者镁的缺乏,可能是形成左室肥厚的因素之一。建议高血压性左室肥厚患者,在降压治疗过程中,同时给予适量补充镁剂,有利于左室肥厚的逆转。     

强直性脊柱炎患者全血微量元素测定分析

选取人类白细胞抗原（HLA－B27）呈阳性的强直性脊柱炎患者54例，采晨空腹血，用原子吸收光谱法测定了全血中锌，铜，铁，钙及锶五种必需元素的含量。与50名无关节炎者组成的正常对照组进行比较。结果表明，患者组全血锌，铁，钙含量显著低于对照组（P＜0．01），锶低于对照组（P＜0．05），铜无统计学意义。提示患者体内微量元素代谢失常，很可能与该病的发生与发展有一定的关系。     

一体化平台石墨炉原子吸收光谱法直接测定全血中顺铂

用ψ（HNO3）=0．5％的硝酸稀释样品，一体化平台石墨炉原子吸收光谱法直接测定全血中顺铂，无须进行基体匹配，Pt的特征质量m=59pg，检出限为32pg(3σ)，相对标准偏差(RSD，n=7)小于5％，样品加标回收率在95％～105％之间。     

GC-NCI-MS衍生化法同时测定全血中的亚硝酸根和硝酸根

建立了全血中亚硝酸根（NO?22-）和硝酸根（NO?33-）的衍生化气相色谱－负化学电离源－质谱（GC－NCI－MS）定量分析方法。以同位素标记的15NO?22-和15NO?33-为内标,将全血中的NO?22-和NO?33-提取衍生后,对目标衍生物进行GC－NCI－MS分析。优化了目标物提取溶剂、衍生物提取溶剂、相转移试剂用量、衍生化方式、衍生化反应时间和温度,确定了最佳前处理条件。结果表明,全血中NO?22-在0.05～5 μg/mL范围内线性关系良好（r2 = 0.995）,检出限为0.01 μg/mL,准确度为89.6%～98.0%,日内和日间相对标准偏差（RSD）分别为1.8%～9.2%和2.1%～6.2%,提取回收率为99.8%～118%;NO?33-在1～300 μg/mL范围内线性关系良好（r2 = 0.997）,检出限为0.2 μg/mL,准确度为92.8%～112%,日内和日间RSD分别为0.60%～14%和0.60%～3.7%,提取回收率为80.3%～88.4%。应用该方法测得16份健康人体全血中内源性NO?22-和NO?33-的质量浓度分别为0.05～0.10 μg/mL和1.70～7.70 μg/mL。对真实亚硝酸盐中毒人体的全血进行测定,2种目标物的质量浓度远高于人体全血中内源性的浓度水平,可判定为亚硝酸盐中毒。该方法特异性强、灵敏度高,结果准确、可靠,能够满足亚硝酸盐中毒案件的检验鉴定需求。     

280例患病新生儿全血中14种元素变化的研究

于1997年6月～1998年10月间对280例患4种常见病新生儿全血中14种元素进行了检测,并与正常足月、顺产新生儿100例对比.结果表明:(1)G-6PD缺陷引起的高胆红素血症组与正常组对照,男、女Fe,Zn均低于正常组;男、女Cu,均高于正常组.(2)缺氧缺血性脑病组与正常组对照,男、女Zn、Fe、Ca均低于正常组;男、女Cu、Mg、Co均高于正常组.(3)硬肿症组与正常组对照,男、女Zn、Fe均低于正常组;男、女Mg、Cu均高于正常组.(4)足月小样儿组与正常组对照,男、女Fe、Zn、Co均低于正常组,余无异常.     

280例患病生儿全血中14种元素变化的研究

于１９９７年６月－１９９８年１０月间对２８０例患４种常见病新生儿全血中１４种元素进行了检测,并与正常足月、顺产新生儿１００例对比。结果表明：（１）Ｇ－６ＰＤ缺陷引起的高胆红素血症组与正常组对照,男、女Ｆｅ、Ｚｎ均低于正常组;