Excretion of malondialdehyde,formaldehyde, acetaldehyde,acetone and methyl ethyl ketone in the urine of rats given an acute dose of malondialdehyde

A high pressure liquid chromatographic system (HPLC) has recently been developed for the simultaneous detection of malondialdehyde (MDA), formaldehyde (FA), acetaldehyde (ACT) and acetone (ACON). We have examined the urinary excretion of these four lipid metabolites in the urine of rats following the acute oral administration of MDA (158 mg/kg body weight). During the first 12 h, increases in the urinary excretion of MDA and ACT of approximately 192- and 70-fold, respectively, were observed. The urinary excretion of both MDA and ACT decreased thereafter. An increase in FA excretion was observed only 12–24 h after MDA administration. A significant decrease in ACON relative to control values was observed 12–48 h after MDA treatment. Two new peaks were present in the HPLC chromatograms of urine samples 0–24 h after MDA administration. Both peaks were shown to be due to methyl ethyl ketone (MEK) which appears to be formed as a result of MDA metabolism. The results demonstrate that orally administered MDA is rapidly excreted in the urine, and alters the metabolism and excretion of other lipid metabolites.     

Sensory irritation to mixtures of formaldehyde,acrolein, and acetaldehyde in rats

 Sensory irritation of formaldehyde (FRM), acrolein (ACR) and acetaldehyde (ACE) as measured by the decrease in breathing frequency (DBF) was studied in male Wistar rats using nose-only exposure. Groups of four rats were exposed to each of the single compounds separately or to mixtures of FRM, ACR and/or ACE. Exposure concentrations of the mixtures were chosen in such a way that summation of the effects of each chemical would be expected not to exceed 80% reduction of the breathing frequency. FRM, ACR and ACE appeared to act as sensory irritants as defined by Alarie (1966, 1973). With FRM and ACR desensitization occurred, whereas with ACE the breathing frequency gradually decreased with increasing exposure time (up to 30 min). For mixtures, the observed DBF was more pronounced than the DBF for each compound separately, but was less than the sum of the DBFs for the single compounds. A model for three compounds competing for the same receptor was applied to predict the DBF of mixtures of FRM, ACE and ACR. The results also showed that with mixtures no desensitization occurred; in fact, the breathing frequency further decreased in the last 15 min of exposure. These observations were similar to those found for ACE alone, and might have been caused by effects on the upper respiratory tract. The results of the present study allow the conclusion that sensory irritation in rats exposed to mixtures of irritant aldehydes is more pronounced than that caused by each of the aldehydes separately, and that the DBF as a result of exposure to a mixture could well be predicted using a model for competitive agonism, thus providing evidence that the combined effect of these aldehydes is basically a result of competition for a common receptor (trigeminal nerve). Received: 24 April 1995/Accepted: 26 September 1995     

HPLC法测定大鼠血清中梓醇的浓度

目的:建立以高效液相色谱法测定大鼠血清中梓醇浓度的方法。方法:选用Hypersil-C18反相色谱柱,流动相为水-乙腈(99.5∶0.5,V/V),流速为1.0mL.min-1,检测波长为210nm。结果:梓醇在0.5～40μg.mL-1浓度范围内线性关系良好(r=0.9995);平均绝对回收率为84.32%～87.52%,平均方法回收率为95.60%～102.63%,日内RSD小于6.14%,日间RSD小于7.01%。结论:本法操作简便,准确度高,适于梓醇血药浓度测定。     

A Comparative Study on the Effects of Disulfiram,Cyanamide and 1-Aminocyclopropanol on the Acetaldehyde Metabolism in Rats

Abstract 1-aminocyclopropanol (ACP) is a potent inhibitor of aldehyde dehydrogenase (ALDH) in vivo and in vitro. Like cyanamide it has a rapid onset of action in vivo with the highest inhibition occurring after 2-24 hrs. and a long duration of action like disulfiram with measurable inhibition after 144 hrs. All the three inhibitors decreased the activity of the mitochondrial low-K_m ALDH strongly in vivo, however, in markedly different doses. Cyanamide inhibited the high-K_m ALDH only in vivo, whereas in vitro, the high-K_m ALDH was unaffected by cyanamide but significantly inhibited by disulfiram and ACP. The inhibition produced by the inhibitors appeared to be irreversible. Acetaldehyde protected the low-K_m enzyme at different extents depending on the inhibitor used. The inhibition of ALDH in intoxicated and control rats and its relation to acetaldehyde oxidation and the disulfiram-ethanol reaction are discussed.     

甲醛在红细胞和肝脏的代谢

本研究采用体外人红细胞和大鼠肝组织切片孵育的方法，观察甲醛的转化过程，结果表明，在红细胞和肝脏中甲醛较快地转化为甲酸，并与剂量和时间相关。     

Changes in the Nasal Epithelium of Rats Exposed by Inhalation to Mixtures of Formaldehyde, Acetaldehyde, and Acrolein

Formaldehyde, acetaldehyde, and acrolein are well-known upperrespiratory tract irritants and occur simultaneously as pollutantsin many indoor and outdoor environments. The upper respiratorytract, and especially the nose, is the prime target for inhaledaldehydes. To study possible additive or interactive effectson the nasal epithelium we carried out 1- and 3-day inhalationstudies (6 hr/day) with formaldehyde (1.0, 3.2, and 6.4 ppm),acetaldehyde (750 and 1500 ppm), acrolein (0.25, 0.67, and 1.40ppm), or mixtures of these aldehydes, using male Wistar ratsand exposure concentrations varying from clearly nontoxic totoxic. The (mixtures of) aldehydes were studied for histopathologicaland biochemical changes in the respiratory and olfactory epitheliumof the nose. In addition, cell proliferation was determinedby incorporation of bromodeoxyuridine and proliferating cellnuclear antigen expression. Effects were primarily observedafter 3 days of exposure. Histopathological changes and cellproliferation of the nasal epithelium induced by mixtures ofthe three aldehydes appeared to be more severe and more extensivein both the respiratory and the olfactory part of the nose thanthose observed after exposure to the individual aldehydes atcomparable exposure levels. As far as nasal histopathologicalchanges and cell proliferation are concerned neither dose additionnor potentiating interactions occurred at no-toxic-effect levels,except for a possible potentiating effect of acetaldehyde atnoneffect levels. The results did not indicate a major rolefor aldehyde dehydrogenases in the biotransformation of thealdehydes studied. Activities of glutathione S-transferase andglutathione reductase after 3 days of exposure to acrolein,alone or in combination with formaldehyde and acetaldehyde,were depressed whereas the glutathione peroxidase activity waselevated. No decrease of nonprotein sulphydryl levels were observed.These findings suggest that, for no-toxic-effect levels, combinedexposure to these aldehydes with the same target organ (nose)and exerting the same type of adverse effect (nasal cytotoxicity),but partly with different target sites (different regions ofthe nasal mucosa), is not associated with a greater hazard thanthat associated with exposure to the individual chemicals.     

补骨脂酚在大鼠血清、肝和肾中的含量测定

目的 建立一种快速检测大鼠灌胃给药后,补骨脂酚在血清、肝脏、肾脏中含量测定的方法。方法 Agilent HC-C₁₈柱(4.6 mm×250 mm,5 mm);流动相：0.1%甲酸水溶液(A)-乙腈(B),梯度洗脱(0~10 min,B 35%→45%,10~25 min,B 80%),流速：1 mL·min^?1,柱温：30 ℃,检测波长：0~10 min,295 nm(氯霉素),10~25 min,260 nm(补骨脂酚)。结果 补骨脂酚在血清、肝匀浆液、肾匀浆液中均能达到很好的分离,在0.1~10.0 mg·L^?1内有良好的线性关系。结论 本方法简便准确,可用于测定给药后血清、肝脏、肾脏中补骨脂酚的含量。     

P-HPLC法测定小鼠血清及脏器中拉米夫定的浓度

目的 :建立RP -HPLC法快速测定小鼠血清及部分脏器匀浆中拉米夫定 (LAM )的浓度。方法 :取0 1ml血清或组织匀浆 ,加入0 5ml甲醇 ,漩涡混合30s ,15000r/min离心10min ,上清液直接进样。选用KromasilC18(150mm×4 6mm ,5μm)为分析柱 ,pH6 8的磷酸盐缓冲液 -甲醇 (91∶9)为流动相 ,检测波长为270nm。结果 :各工作曲线线性范围为0 25～50μg/ml ,最低检测浓度为60ng/ml(S/N=3)。各样品平均绝对回收率大于88 % ,方法回收率在97 %～100 %之间 ,日内和日间精密度的RSD均小于10 %。结论 :此方法经济、可靠、简单、快速 ,可用于LAM血药浓度分析及动物体内分布研究     

Rapid,Sensitive and Reliable Ricin Identification in Serum Samples Using LC–MS/MS

Ricin, a protein derived from the seeds of the castor bean plant (Ricinus communis), is a highly lethal toxin that inhibits protein synthesis, resulting in cell death. The widespread availability of ricin, its ease of extraction and its extreme toxicity make it an ideal agent for bioterrorism and self-poisoning. Thus, a rapid, sensitive and reliable method for ricin identification in clinical samples is required for applying appropriate and timely medical intervention. However, this goal is challenging due to the low predicted toxin concentrations in bio-fluids, accompanied by significantly high matrix interferences. Here we report the applicability of a sensitive, selective, rapid, simple and antibody-independent assay for the identification of ricin in body fluids using mass spectrometry (MS). The assay involves lectin affinity capturing of ricin by easy-to-use commercial lactose–agarose (LA) beads, following by tryptic digestion and selected marker identification using targeted LC–MS/MS (Multiple Reaction Monitoring) analysis. This enables ricin identification down to 5 ng/mL in serum samples in 2.5 h. To validate the assay, twenty-four diverse naive- or ricin-spiked serum samples were evaluated, and both precision and accuracy were determined. A real-life test of the assay was successfully executed in a challenging clinical scenario, where the toxin was identified in an abdominal fluid sample taken 72 h post self-injection of castor beans extraction in an eventual suicide case. This demonstrates both the high sensitivity of this assay and the extended identification time window, compared to similar events that were previously documented. This method developed for ricin identification in clinical samples has the potential to be applied to the identification of other lectin toxins.     

丙戊酸治疗躁狂症及其血药浓度的临床对照研究

目的探讨丙戊酸治疗躁狂症的临床疗效、血药浓度及药物不良反应。方法60例躁狂症患者分别接受丙戊酸钠缓释剂及丙戊酸镁治疗6周,并于服药后1,2,4,6周末使用高效液相色谱法检测丙戊酸血药浓度,以Bech—Rafaelsen躁狂量表（BRMS）和不良反应量表（TESS）观察临床疗效和药物不良反应。结果观察组有效率为80．00％,对照组有效率为76．67％,两组疗效无明显差异,有效病例的血药浓度平均为（77．58±14．45）μg·mL^-1,治疗窗范围为50～100μg·mL^-1之间。结论丙戊酸能有效治疗躁狂症,血药浓度监测有助于临床安全有效用药。     

HPLC同时测定苯巴比妥、苯妥英、卡马西平和氯硝西泮血药浓度

目的 建立同时测定血清中苯巴比妥(PB)、苯妥英(PT)、卡马西平(CBZ)和氯硝西泮浓度的HPLC.方法 分析柱为Agilent C18柱(4.6 mm×250 mm,5 μm),流动相为水-甲醇-乙腈(45∶45∶10),流速为1.0 mL·min-1,检测波长为254 nm,柱温30 ℃,阿普唑仑为内标物.结果 PB,PT,CBZ和氯硝西泮的线性范围分别为2.5～80,1.25～40,0.75～24,0.02～0.64 μg·mL-1,最低检测浓度分别为0.2,0.2,0.1,0.005 μg·mL-1,方法回收率分别为99.1%,100.8%,100.1%,98.1%,日内、日间RSD均＜3.5%,绝对回收率分别为91.7%,90.2%,89.9%和89.3%,RSD均＜2%.结论 该方法快速、灵敏、实用,适用于治疗药物监测.     

姜黄胶囊中姜黄素在大鼠体内药动学和组织分布

摘 要 目的：研究姜黄胶囊中姜黄素在大鼠体内药物动力学和组织分布特点。方法: SD大鼠灌胃5 g·kg^-1姜黄胶囊后,采集血浆样品及肝脏、肾脏、胃、小肠等脏器,用HPLC测定各样品中姜黄素含量。结果:血浆中姜黄素在0.1~10μg·mL^-1范围内线性关系良好,姜黄素t1/2为（25.13±5.63） h,tmax为0.50 h,Cmax为（0.44±0.01）μg·mL^-1,AUC0-t为（2.11±0.04）μg·mL^-1·h;同时在胃肠道、肝脏等脏器中均能检测到姜黄素分布。结论:姜黄胶囊能使姜黄素快速吸收,并广泛分布于大鼠各个组织中,为该药临床用药提供依据。     

The Measurement of Warfarin Enantiomers in Serum Using Coupled Achiral/Chiral,High-Performance Liquid Chromatography (HPLC)

An assay for the serum concentration of the enantiomers of warfarin, R-warfarin and S-warfarin, has been developed using a bovine serum albumin chiral stationary phase (BSA-CSP) coupled to a Pinkerton internal-surface reverse-phase (ISRP) achiral column. The ISRP column is used to separate R,S-warfarin from the serum components and warfarin metabolites and to quantitate the total R,S-warfarin concentration. The eluent containing R,S-warfarin is then selectively transferred to the BSA-CSP, where the enantiomers are stereochemically resolved ( = 1.19) and the enantiomeric composition is determined. This system is sensitive and accurate, does not require extensive precolumn manipulations, and can be automated for use in large-scale clinical studies.     

HPLC法快速测定5—氟尿嘧啶血药浓度

目的 :建立反相高效液相色谱法测定人血清中 5 -氟尿嘧啶浓度。方法 :血清样品用乙酸乙酯提取 ,水浴氮气吹干 ,残留物用水溶解后进样。色谱柱为C1 8柱 (2 5 0mm× 4 6mm) ,水为流动相 ,流速 1 0mL·min- 1 ,紫外检测波长 2 73nm ,桂皮酸为内标。结果 :本法最低检测浓度为 0 0 5 μg·mL- 1 ,线性范围为 1 0～ 5 0 0 μg·mL- 1 ,日内RSD为 2 7%～ 4 1% ,日间RSD为 3 8%～ 4 7(n =4)。结论 :该法适用于 5 -氟尿嘧啶的药代动力学研究及临床血药浓度检测。试验结果表明 ,该方法经济、简便、快速、灵敏度高、重现性好。     

黄芪注射液对重症急性胰腺炎大鼠外周血清氧化相关物质的影响

目的 探讨黄芪注射液对重症急性胰腺炎(severe acute panereatitis,SAP)大鼠外周血清氧化相关物质的影响.方法 雄性SD大鼠100只,随机分为A、B、C、D、E组,每组20只,其中A、B、C、D组通过逆行胰胆管注射5％牛磺胆酸钠建立SAP大鼠模型,B、C、D组同时给予不同浓度的黄芪注射液,E组给予相同体积的生理盐水.检测5组血清谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-PX)、超氧化物歧化酶(superioxide dismutase,SOD)、活性氧类物质(reactive oxygen species,ROS)活性和丙二醛(malondialdehyde,MDA)含量.结果 A组血清GSH-PX、SOD活性明显低于E组,ROS活性和MOD浓度明显高于E组(P＜0.05);B、C、D组血清GSH-PX、SOD活性明显高于A组,ROS活性和MDA浓度低于A组(P＜0.05).结论 黄芪注射液能提高SAP大鼠外周血GSH-PX、SOD活性,降低ROS活性和MDA浓度.     

高效液相色谱法测定大鼠纤维化肝组织中羟脯氨酸含量

目的 :测定肝纤维化大鼠肝脏中羟脯氨酸含量。方法 :采用2 ,4—二硝基氯苯柱前衍生化反相高效液相色谱法 ,以乙晴 -醋酸盐缓冲系统梯度洗脱 ,于360nm波长处测定羟脯氨酸含量。结果 :羟脯氨酸衍生物与其它氨基酸较好分离 ,并可准确定量。结论 :本方法快速、灵敏 ,分辨率高 ,尤其适用于临床肝纤维化患者肝穿刺后样本量极小的情况。     

Development of a High-Performance Liquid Chromatographic Method for Determination of Letrozole in Wistar Rat Serum and its Application in Pharmacokinetic Studies

A fast, sensitive, and specific reversed-phase high-performance liquid chromatographic (RP–HPLC) method for the determination of letrozole in Wistar rat serum was developed. In this method, liquid–liquid extraction of letrozole was achieved using diethyl ether as the extracting solvent. The analysis was carried out on a reversed-phase C18 (250 mm × 4.6 mm, 5 μm) column with an isocratic mobile phase of methanol–water (70:30,v/v), at a flow rate of 1.0 mL min⁻¹. Detection was carried out at 239 nm with a UV–visible spectrophoto-metric detector. The method was shown to be selective and linear over the concentration range of 0.15–100 μg mL⁻¹. The intra-day and inter-day precision studies showed good reproducibility with coefficients of variation less than 11% for the analyte. The relative errors of intra– and inter–day accuracy were within −11.52 to −2.26%. The limit of quantification was evaluated to be 0.15 μg mL⁻¹. The method was successfully applied for the pharmacokinetic study of letrozole after oral administration of 10 mg kg⁻¹ of letrozole in six healthy Wistar rats.     

兔血清中琥乙红霉素的HPLC测定及药代动力学研究

目的： 用高效液相色谱法测定兔血清中琥乙红霉素的浓度。方法： 采用ＹＷＧ－ Ｃ１８ 色谱分析柱（４－６ ｍ ｍ×１５０ ｍ ｍ ,１０ μｍ）, 岛津ＳＰＤ－ ６ＡＶ 紫外检测器, 甲醇－ ０－０２ ｍｏｌ·Ｌ－ １ － 醋酸铵水溶液（７０∶３０ , 用氨水调ｐＨ７－３） 为流动相, 红霉素作内标。血液样品在碱性条件下用二氯甲烷提取纯化后进样, 在２１０ ｎｍ 波长处检测。结果： 琥乙红霉素线性范围为０－２５～１２ ｍｇ·Ｌ－ １ （ｒ＝ ０－９９８ ９）, 最低检测量０－０２ μｇ, 最低检出浓度０－０５ ｍｇ·Ｌ－１ , 平均方法回收率９６－２６％ 。精密度考察琥乙红霉素日内、日间相对标准偏差均小于６％ 。结论： 本方法快速、准确、重复性好。     

罗格列酮对动脉粥样硬化大鼠血脂及血清炎症因子水平的影响

目的探讨罗格列酮对动脉粥样硬化(AS)大鼠血脂及血清炎症因子水平的影响。方法12周龄雄性W istar大鼠55只,随机分为3组:①正常对照组(N,n=20),②模型组(M,n=20),③治疗组(罗格列酮组,RSG,n=15)。其中N组大鼠喂食基础饲料12周;M组和RSG组大鼠喂食高脂饲料6周后,分别给予生理盐水(10 mL.kg-1.d-1)和罗格列酮(3 mg.kg-1.d-1)灌胃治疗6周,且在喂食开始时一次性腹腔注射维生素D340万IU.kg-1。造模6周后,取N组和M组大鼠各5只处死,检测血清总胆固醇(TC)、三酰甘油(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C),ELISA法检测血清C反应蛋白(CRP)、单核细胞趋化蛋白-1(MCP-1)、血管内细胞黏附分子-1(ICAM-1)表达;提取大鼠心脏,HE染色观察冠状动脉粥样硬化斑块的病理形态改变。灌胃治疗6周后,各组大鼠处死后按前述方法再次检测各指标变化及冠状动脉病理形态变化。结果造模6周后,M组大鼠血清TC,TG,LDL-C,CRP,MCP-1,ICAM-1水平均升高(P<0.05),HDL-C水平的变化没有统计学意义(P>0.05),光镜下观察可见冠状动脉粥样硬化斑块形成。灌胃治疗6周后,RSG组大鼠血清TC,TG,LDL-C,CRP,MCP-1水平较M组降低(P<0.05或P<0.01),但仍略高于N组(P<0.05),HDL-C,ICAM-1水平未见明显变化(P>0.05)。光镜下观察见RSG组大鼠AS病变较M组明显减轻。结论罗格列酮治疗6周能降低大鼠TC、TG、LDL-C水平,抑制血清炎症因子CRP、MCP-1的表达,这可能是其抗AS的机制之一。     

高效液相色谱法测定人血清中卡马西平及环氧化卡马西平浓度

目的建立测定人血清中卡马西平及其代谢物10,11-环氧化卡马西平浓度的方法.方法采用高效液相色谱法,血样用乙醚提取,以艾司唑仑为内标,色谱柱为Kromasil C18柱(150 mm ×4.6 mm,5μm),流动相为乙腈-水(1:2),检测波长215 nm,流速1.2ml·min-1.结果血清中卡马西平、环氧化卡马西平线性范围分别为1.0～20.0 mg·L-1(r=0.999 9)、0.1～5.0 mg·L-1(r=0.999 4),平均回收率接近100%,日内、日间RSD均小于5%.结论本法快速、简便、准确,适用于临床常规监测需要.