应用液相色谱-质谱联用技术检测血浆样品中丹参酚酸类成分的浓度

目的：研究丹参酚酸中丹参素（tanshinol，TSN）、原儿茶醛（protocatechualdehyde，PCA）、丹酚酸A（salvianolic acid A，SAA）、丹酚酸B（salvianolic acid B，SAB）及丹酚酸D（salvianolic acid D，SAD）的电喷雾串联质谱（ESI-MS／MS）检测特性，为开展复方丹参滴丸药代研究，建立灵敏可靠的定量分析血浆样品中丹参酚酸浓度的方法。方法：研究丹参酚酸的电离模式及效率、二级质谱的碎片离子及打碎效率；建立有效可靠的从血浆样品中提取被测丹参酚酸物质的前处理方法；优化色谱条件、减少分析干扰、促进质谱检测；围绕准确性、精密度和提取回收率等进行方法学验证。结果：电喷雾（ESI）负离子模式能有效实现丹参酚酸被测化合物的电离。以下离子对m／z197→135、137→108、493→295、717→519、417→197分别为被测化合物偈N、PCA、SAA、SAB、SAD提供最佳的选择离子对监测（SRM）检测条件。为取得可靠的分析结果，建立了两个分析方法：其一用于测定血浆样品中的TSN；其二用于测定血浆样品中的PCA、SAA、SAB和SAD。两个分析方法均以HCI酸化血样后用乙酸乙酯提取的方法处理血浆样品，其中，TSN的提取回收率在57．7％～61．6％之间；PCA、SAA、SAB和SAD的提取回收率在38．0％～58．9％之间。偈N的线性范围为2．7～2000．0ng／mL，相关系数r为0．999，最低检测限（LLOQ）为2．7ng／mL；PCA、SAA、SAB及SAD的线性范围为1．4～1000．0ng／mL或4．1～1000．0ng/mL，相关系数r〉0．99，LLOQ分别为1．4ng／mL（PCA）和4．1ng/mL（SAA、SAB和SAD）。两个分析方法的日内准确性和精密度分别为90％～112％和2．5％～14．5％；日间准确性和精密度分别为91％～110％和0．7％～9．8％。结论：成功建立了能够灵敏可靠地测定血浆样品中5种丹参酚酸化合物浓度的分析方法，此方法可用于复方丹参滴丸的药代动力学研究。     

3种方法测定伪麻黄碱血药浓度的比较研究

目的 建立并比较测定伪麻黄碱血药浓度的HPLC法、柱前衍生-HPLC法和LC/MS/MS法,为体内低浓度伪麻黄碱的分析提供更为简便、准确的方法.方法 摸索并优化血浆前处理条件、衍生化条件、色谱条件、质谱条件等.结果 3种方法均具有较好的专属性、线性、精密度、回收率和稳定性,可准确、灵敏地检测生物样本中的伪麻黄碱.3种方...     

HPLC-MS/MS methods for the quantitative analysis of ophthalmic acid in rodent plasma and hepatic cell line culture medium

Ophthalmic acid (OA), an endogenous tripeptide analogue of glutathione, has been suggested as a potential biomarker for paracetamol/acetaminophen hepatotoxicity. Here HPLC-MS/MS methods have been developed for the precise, sensitive and specific detection and quantification of OA in in vitro cell culture medium and plasma. For the cell culture medium the LLOQ was found to be 1 ng/ml, with less than 1% between sample carry over at all concentrations and precision below 15% for within day and below 9% for between day analyses. For rat plasma the presence of endogenous OA resulted in the LLOQ being 25 ng/ml (defined as the lowest concentration on the calibration curve where the base peak was less than 20% of the LLOQ). For the plasma assay the percentage carry over was less than 1% for all concentrations and within and between batch precision was below 21%. The methods were linear for both sample types from the LLOQ up to 5 μg/ml. The method was successfully applied to the determination of OA in samples obtained following the chronic administration of the rat hepatotoxin methapyrilene, where plasma OA concentrations were observed to show a weak negative correlation with those of established liver injury biomarkers such as aspartate aminotransferase (AST).     

紫草中有效成分提取方法的研究

目的优选紫草中有效成分的提取方法。方法.用乙醇60℃以下提取和CO2超临界萃取,用高效液相色谱法,以左旋紫草素含量为考察指标,进行不同提取方法的比较研究。结果:CO2超临界萃取法左旋紫草素含量是乙醇提取法的3倍。结论紫草有效成分的提取以CO2超临界萃取法为优。萃取的条件是:萃取罐的温度为35℃,萃取压力为25Mpa,CO2流量为40L/h,萃取时间为3h。     

Development of a stacking-CZE method for the analysis of phenolic acids

Eight phenolic acids were analyzed by capillary zone electrophoresis. On-line analyte preconcentration was carried out by hydrodynamic injection of large volume of sample followed by removal of the bulk of the low conductivity sample matrix by polarity switching. The optimal electrolyte system consisted of 50mM sodium tetraborate of pH 9.0 (adjusted with 0.1 M phosphoric acid) containing 2% of alpha-cyclodextrin. The separations were carried out with a fused silica capillary (effective length 50 cm, i.d. 50 microm) and monitored at 200 nm. Under optimized preconcentration conditions (sample injection 99 s at 100 mbar and the polarity switching time 1.0 min) linear calibration ranges (0.1-2.0 microg/ml, R=0.9979-0.9995), favourable limits of detection (0.01-0.025 microg/ml) and good repeatability of the peak areas (R.S.D.: 2.76-5.69%, n=6) were achieved.     

衍生化技术在酚类化合物分析中的研究进展

结合不同仪器分析方法对酚类化合物分析方法中衍生化技术的最新应用进展进行综述。具体介绍了酚类化合物分析方法中所采用的衍生化方式和条件,并对各种衍生化方法的特点进行了论述。     

Qualitative and quantitative analysis of phenolic acids in Asclepias syriaca L

TLC and HPLC was applied to qualitative and quantitative determination of phenolic acids free and liberated by acid and alkaline hydrolysis in common milkweed. The presence of phenolic acids, namely p-hydroxybenzoic, p-coumaric, protocatechuic and caffeic acids, was confirmed in the leaves and flowers. Moreover, the flowers contained gallic acid, and the leaves contained alpha-resorcylic, vanilic and chlorogenic acids. Conjugated forms of phenolic acids predominate in the plant. They hydrolyse mainly to coffeic, p-coumaric, ferulic and p-hydroxybenzoic acids.     

Fully automated methods for the determination of hydrochlorothiazide in human plasma and urine

LC assays utilizing fully automated sample preparation procedures on Zymark PyTechnology™ Robot and BenchMate™ Workstation for the quantification of hydrochlorothiazide (HCTZ) in human plasma and urine have been developed. After aliquoting plasma and urine samples, and adding internal standard (IS) manually, the robot executed buffer and organic solvent addition, liquid—liquid extraction, solvent evaporation and on-line LC injection steps for plasma samples, whereas, BenchMate™ performed buffer and organic solvent addition, liquid—liquid and solid-phase extractions, and on-line LC injection steps for urine samples. Chromatographic separations were carried out on Beckman Octyl Ultrasphere column using the mobile phase composed of 12% (v/v) acetonitrile and 88% of either an ion-pairing reagent (plasma) or 0.1% trifluoroacetic acid (urine). The eluent from the column was monitored with UV detector (271 nm). Peak heights for HCTZ and IS were automatically processed using a PE-Nelson ACCESS*CHROM laboratory automation system. The assays have been validated in the concentration range of 2–100 ng ml⁻¹ in plasma and 0. 1–20 μg ml⁻¹ in urine. Both plasma and urine assays have the sensitivity and specificity necessary to determine plasma and urine concentrations of HCTZ from low dose (6.25/12.5 mg) administration of HCTZ to human subjects in the presence or absence of losartan.     

Comparison of fitting methods for the analysis of plasma concentration-time data resulting from constant rate intravenous infusion

Plasma concentration-time data resulting from constant rate intravenous infusion may be analysed in two ways: Samples may be collected both during and after infusion and fit to an infusion model. Samples may be collected after infusion is complete and the data may be fit as an i.v. bolus. The purpose of this study was to contrast these two fitting procedures in terms of the accuracy of the parameter values obtained. Concentration-time data were computer-generated with the introduction of random error to simulate the disposition profiles of two model drugs. The results of these simulations indicate that satisfactory values for area-dependent parameters may be obtained without fitting data during the infusion phase. The exception to this is the apparent steady-state volume whose values become less accurate with longer infusion times. The parameters most affected by ignoring data points in the infusion phase are the central volume of distribution, and the coefficient and disposition rate constant associated with the initial, rapid phase of disposition. The equation which describes the entire concentration-time profile provides the most accurate parameter estimates of the model equation. In addition, we also describe the influence of the fitting method on the intercompartmental transfer rate constants.     

Support for selection of a methamphetamine cleanup standard in Colorado

Methamphetamine production for illicit use occurs in makeshift labs and is associated with the release of numerous chemicals, including methamphetamine residues. These methamphetamine residues may pose a health risk to residents who reoccupy these structures after property seizures. Several states have established technology-based cleanup standards for methamphetamine, but none have examined the health-protectiveness of these standards. In response to Colorado House Bill 04-1182, exposure intakes correlated with three technology-based standards were calculated for various groups of individuals. Intakes were assessed for a 1-year-old infant, 6-year-old child, and a female of childbearing age. Exposure intakes were compared to toxicity reference values developed from developmental endpoints following methamphetamine exposure from the available literature. Uncertainty factors were applied to the lowest adverse effect levels observed in these studies to arrive at the toxicity reference values. These reference values were greater than the calculated intakes from each proposed technology standard, suggesting that all of the proposed standards would be protective of human health exposure. The cost and practicality of attaining each of the proposed standards was also factored into the decision making process. In their final regulation (6 CCR 1014-3), the CDPHE selected 0.5 microg/100 cm(2) as the final cleanup standard for methamphetamine residues.     

甲醛的检测方法及相关进展

甲醛作为一种生活中广泛存在的有毒污染剂,对人体健康造成很大危害,甲醛的污染已引起人们的高度重视,检测甲醛污染至关重要。近几年随着科学技术的飞速发展,在不同的研究领域甲醛测定技术也有了一定程度的发展。本文综述了近年来甲醛检测方法的进展,涉及到的检测方法主要有:分光光度法、液相色谱法、气相色谱法、示波极谱法、电位法、荧光法、化学发光法。本文分析了相关检测方法的优点和缺点,展望了快速检测分析方法的前景。     

药学经济学评价方法分类及成本效果分析法的应用例析

面对有限的卫生资源,临床治疗费用的居高不下,需要对临床治疗方案进行选择,此时药学经济学评价就发挥了巨大的作用,它能够提高卫生资源的应用效率,以使得社会获得最大收益。本文描述了药学经济学评价的四种方法,并阐述了药学经济学评价在临床治疗方案筛选决策中的应用现状,最后,对药学经济学评价方法中的成本效果分析法在临床治疗方案筛选决策中的应用进行了探讨。     

Integration of data and methods for genome analysis

The development of genomic and post-genomic technologies has created an explosion in the quantity, diversity and availability of both biological data and methods of analysis. Biologists are currently facing the problem of using all these resources to convert raw data into new valuable knowledge. This review presents software platforms designed to handle data and/or methods in the context of genome analysis.     

UPLC与HPLC测定人血浆中多索茶碱的比较

目的 比较UPLC与HPLC分析多索茶碱血药浓度时灵敏度的差异.方法 UPLC法的色谱柱为Kinetex C18柱(100mm×2.1 mm,1.7 μm),HPLC法的为Diamonsil C18柱(150 mm ×4.6 mm,5μm),流动相均为甲醇-10 mmol·L-1乙酸铵(25∶75),流速分别为0.3、1.0 mL·min-1,检测波长均为274 nm,进样量分别为10、20 μL.结果 与HPLC比较,UPLC法中多索茶碱的灵敏度提高了300倍,同时分析时间缩短1/3;UPLC法能够定量测定临床接受多索茶碱治疗40名患者的血浆药物浓度为0.163 ～ 12.8 μg·mL-1,而其中8名患者的血药浓度低于1.0 μg·mL-1,HPLC法无法准确测定.结论 与HPLC比较,UPLC不仅能缩短分析时间,而且能大幅度提高分析方法的检测灵敏度,可满足临床治疗药物监测对分析方法的灵敏度需求.     

固相萃取用于血浆中抗癫痫药物的提取

目的：探讨固相萃取法用于血浆中抗癫痫药物的提取。方法：以活化的ＯＤＳ柱（４０ｕｍ）为固相萃取柱,二氯甲冠为洗脱剂,洗脱液浓缩后以ＨＰＬＣ法测定药物的浓度。结果：苯巴比妥、苯妥英在５．００～６０．００ｕｇ／ｍｌ范围内,卡马西平在２．５０～２０．００ｕｇ／ｍｌ范围内线性关系良好,ｒ分别为０．９９８０,０．９９９９,０．９９９０;绝对回收率分别为５９．６０％～６８．７％,５８．９％～６８．０％,５３．８     

固相萃取用于血浆中抗癫痫药物的提取

目的：探讨固相萃取法用于血浆中抗癫痫药物的提取。方法：以活化的ＯＤＳ 柱（４０μｍ） 为固相萃取柱,二氯甲烷为洗脱剂,洗脱液浓缩后以ＨＰＬＣ 法测定药物的浓度。结果：苯巴比妥、苯妥英在５ ．００ ～６０ ．００μｇｍｌ 范围内,卡马西平在２ ．５０ ～２０ ．００μｇｍｌ 范围内线性关系良好,ｒ 分别为０ ．９９８０ ,０ ．９９９９ ,０ ．９９９０ ;绝对回收率分别为５９ ．６０ ％～６８ ．７ ％ ,５８ ．９ ％ ～６８ ．０ ％ ,５３ ．８ ％ ～５７ ．３ ％ ;相对回收率分别为９２ ．２ ％ ～９８ ．６ ％ ,９７ ．１ ％ ～１０２ ．６ ％ ,９７ ．９ ％ ～１０２ ．０ ％ ,ＲＳＤ＜ ６ ％ ;血浓监测结果与经传统的液液萃取后所得结果非常接近。结论：固相萃取法操作简单、省时、提取干净,可用于血浆中抗癫阅药物的提取。     

Comparing ultrasonic- and microwave-assisted methods for extraction of phenolic compounds from Kabkab date seed (Phoenix dactylifera L.) and stepwise regression analysis of extracts antioxidant activity

This study aimed to extract bioactive compounds (phenolics and flavonoids and condensed tannins) from roasted date seed (Phoenix dactylifera L. cv Kabkab) using various solvent systems (W: water, AE: aqueous-ethanol, AA: aqueous-acetone) and extraction method (ultrasonic-assisted (UAE), microwave-assisted (MAE) and the combination of these two methods (UMAE) maceration (ME) and Decoction-Infusion (DIE) Extraction). Moreover, the feasibility of antioxidant activity prediction was investigated based on stepwise regression analysis and phytochemical properties. Extraction yield, Total phenolic content (TPC), total flavonoid content (TFD) and antioxidant activity (DPPH*, ABTS*⁺, FRAP and averaged antioxidant activity: AAA) of the extracts were evaluated. The main effect of solvent systems and extraction methods on phytochemical compounds and antioxidant activity were significant (P < 0.01). Water and aqueous-ethanol solvents extracted higher phytochemical compounds than aqueous-acetone (P < 0.05). Although the highest extraction performance was observed for the ME method, the novel methods show an acceptable result in a much shorter time. Among novel methods, the highest and lowest performances were recorded for UAE and MAE. There was no significant difference between novel-green methods (e.g., UAE and UMAE). Although the lowest phytochemical yield was obtained for MAE, this performance was obtained in less than 5 min. The highest and lowest correlation between the phytochemical compositions and antioxidant activity parameters were for DPPH* and AAA, respectively. Stepwise-regression analysis showed the weakest and strongest prediction models for AAA (10-fold S = 0.098 and 10-fold R² = 87.07) and ABTS*⁺ (10-fold S = 0.129 and 10-fold R² = 78.25), respectively.     

川芎DNA提取方法的研究

目的 以伞形科蒿本属植物川芎Ligusticum chuanxiong Hort.叶为材料,研究川芎DNA的提取方法 .方法 分别采取CTAB法、高盐低pH法、木本植物DNA提取法、改良高盐低pH法提取基因组DNA,并对4种方法进行了改进.通过0.9%琼脂糖凝胶电泳和RAPD两种方法检测所提取的DNA样品.结果 比较DNA产量、质量等,确定了木本植物DNA提取法最佳.结论 木本植物DNA提取法为最佳方法.