纸色谱分离-荧光检测法测定人体尿液中的黄蝶呤

报道用丙醇-氨水(25％)-水(15：9：6．V／V)作展开剂,纸色谱分离人体尿液中的蝶呤类化合物,并利用黄蝶呤与其它蝶呤类化合物的荧光性质的差异来测定人体尿液中黄蝶呤的含量。黄蝶呤在0～0．6μg／10mL的范围内,其荧光强度与其荧光斑点的洗脱量呈良好的线性关系,从而建立了一种新的测定黄蝶呤的方法。该方法应用于人体尿液中黄蝶呤的测定,结果满意。     

高效液相色谱-荧光分析法同时测定人体尿液中黄蝶呤与异黄蝶呤

建立了高效液相色谱-荧光法同时测定癌症病人尿液中黄蝶呤及异黄蝶呤的新方法。选择荧光检测波长λex=345nm,λem=420nm。以磷酸盐缓冲溶液(pH=7.5)-甲醇(体积比为98∶2)为流动相,流速1.0mL/min,黄蝶呤与异黄蝶呤含量分别在0.0013～0.945μg/mL及0.00017～0.118μg/mL范围内与色谱峰面积呈良好的线性关系,线性相关系数分别为0.9999和0.9996,检出限分别为0.5ng/mL和0.05ng/mL,加标平均回收率在86.2%～107.5%之间。方法应用于癌症病人尿样分析,取得了较好的结果。     

高效液相色谱-荧光检测法测定人体尿液中的蝶呤-6-羧酸

利用蝶呤-6-羧酸具有天然荧光的特点,建立了人体尿液中蝶呤-6-羧酸的高效液相色谱-荧光分析方法。尿液经硫酸铵处理后,过0.45μm水相滤膜,直接进行液相色谱分析,色谱柱为NOVA-PAK C18柱,保护柱为RCSS Guard-PAKTMC18柱,流动相为V(5 mmol/L KH2P04-NaOH缓冲溶液,pH 7.3)∶V(甲醇)=99∶1),流速为0.6 mL/min,荧光激发波长为338 nm,荧光发射波长为420nm。蝶呤-6-羧酸在0.05~1.2μg/mL范围内呈线性关系,相关系数为0.9974,检出限为0.010μg/mL。分别添加0.625、2.50和4.50μg蝶呤-6-羧酸标准品,平均回收率(n=5)在99.7%~105%之间,相对标准偏差小于3.6%。此法可用于临床尿样中蝶呤-6-羧酸的分析。     

高效液相色谱-荧光检测法同时测定人体尿液中的多种蝶呤类化合物

建立了高效液相色谱-荧光检测法同时测定人体尿液中的蝶呤、新蝶呤、生物蝶呤和墨蝶呤.尿液在酸性条件下经碘-碘化钾溶液氧化30 min,滴加抗坏血酸还原液后,可进行液相色谱分析.色谱柱为Diamonsn C<,18>柱,用体积比9:10的水-甲醇为流动相,流速为1.0 mL/min,荧光检测波长为λ<'ex>=360 nm...     

高效液相色谱-紫外检测法测定人体尿液中的蝶呤-6-羧酸

建立了人体尿液中蝶呤-6-羧酸的高效液相色谱-紫外分析新方法. 运用Lichrospher C18柱(250×4.6 mm,5 μm),甲醇-水(70∶30,V/V)为流动相,流速为0.4 mL/min,可较好地将尿液中蝶呤-6-羧酸与其它共存干扰物质分离.在355 nm检测波长下,蝶呤-6-羧酸在0.19～4.8 μg/mL范围内与色谱峰面积呈良好线性关系,相关系数为0.9999,方法检出限为0.015 μg/mL.尿液经0.45 μm滤膜过滤后,可直接进样分析,方法简便,应用于癌症病人和健康人尿样中蝶呤-6-羧酸测定,结果较好.方法的加标回收率为94.6%～100.2%,相对标准偏差为0.81%～ 5.07% .     

高效液相色谱-电喷雾质谱同时测定人体尿液中黄蝶呤与异黄蝶呤

建立了高效液相色谱-电喷雾质谱(HPLC-ESI-MS)法同时测定人体尿液中黄蝶呤与异黄蝶呤的分析方法.尿样经离心、过膜后直接进样,采用电喷雾质谱进行定性和定量.结果表明,黄蝶呤与异黄蝶呤含量分别在0.0204～2.04μg/mL和0.0202～ 2.02 μg/mL范围内与色谱峰面积呈良好的线性关系,线性相关系数r2...     

高效液相色谱-荧光检测法测定环境样品中的过氧化物

对高效液相色谱-荧光检测法测定过氧化氢和有机过氧化物的方法进行了改进,从而提高了方法的检测灵敏度。以氯化血红素(hemin)作催化剂进行柱后衍生反应,过氧化物将对羟基苯乙酸氧化生成能吸收荧光的二聚物,然后用荧光检测器检测。实验确定了最佳反应管温度和荧光检测波长。应用该法测定了大气和雨水样品中过氧化物的浓度。     

高效液相色谱荧光检测法测定瓜子中的4-氨基丁酸

在众多的氨基酸中，4-氨基丁酸是一种非蛋白质类氨基酸，它在医学上是一种抑制性神经递质，由于其自身的特点，一些常规的直接测定法灵敏度较低。尽管对其测定有较多报道，但采用柱前衍生的分离测定的报道相对较少，且测定结果不很理想。本实验采用新型荧光试剂咔唑-9-乙基氯甲酸酯(CEOC)作为衍生试剂在Hyperil C18柱上分离测定了4-氨基丁酸(GABA)。结果表明，所建立的方法简单、快速，灵敏度高。     

离子色谱荧光检测法测定肾上腺素和多巴胺

１引 言 多巴、多巴胺、肾上腺素与去甲肾上腺素等统称为儿茶酚胺类物质,它们都是由酪氨酸衍生而来的系列物。儿茶酚胺作为神经递质和激素,与人们的健康与疾病有密切关系,它们不仅直接参与行为活动以及参与血压、心率、呼吸和睡眠等植物神经功能的调控,而且还与一些功能性疾病如神经分裂症、忧郁症以及器质性病变的出现有关。儿茶酚胺的分析研究始于２０世纪４０年代,早期的测定方法主要是三羟基吲哚法和乙二胺缩合法。随着人们对儿茶酚胺研究的深入,又报道了许多测定方法,如放射酶学法、气相色谱法、色质联用法、高效液相色谱法以及…     

同步荧光法测定人尿中异黄蝶呤

研究了一种测定人尿中异黄蝶呤的同步荧光分析方法.在KH2PO4-NaOH缓冲溶液中(pH 7.8),波长差(Δλ)为65 nm的条件下进行同步荧光扫描,可有效排除尿液中其它物质对异黄蝶呤测定的干扰.异黄蝶呤的质量浓度在0～1.4 μg/mL范围内与荧光强度呈良好的线性关系.线性方程为Ⅰ=4105.3ρ 59.696, 相关系数r=0.9997.异黄蝶呤添加到健康人和胃癌病人尿样中的平均回收率分别在102.9%～108.4%及85.6%～95.6%之间,其相对标准偏差(RSD)分别为0.37%～1.2%及1.0%～2.2%.方法已应用于人尿中异黄蝶呤的测定.     

Determination of metabolites relating to the tryptophan-NAD pathway in rat urine by cellulose high performance thin-layer chromatography

High performance thin-layer chromatography (HPTLC) was used to determine 12 metabolites of the tryptophan-NAD pathway in rat urine. The method of separation described in the previous report was improved. The metabolites were separated on the cellulose plate by using five multiple developments with four solvent systems. The plate was scanned by the TLC scanner at 254 nm and the amounts of the metabolites were calculated by comparing peak areas of the scanning curves obtained for the urine sample and for the authentic standards. This procedure can be used as a semiquantitative determination method for the metabolites in biological fluids.     

高效液相色谱-荧光法同时测定人体尿液中的酪氨酸和色氨酸

建立了高效液相色谱-荧光检测法同时测定人体尿液中的酪氨酸(Tyr)和色氨酸(Typ)。采用Diamonsil C18柱(250×4.6mm,5μm),以水-乙腈(80∶20,V/V)为流动相,流速1.0mL/min,荧光检测波长为λex=260nm、λ_(em)=340nm。Tyr和Typ的含量分别在0.10~6.0μg/mL及0.050~6.0μg/mL范围内与色谱峰面积呈良好的线性关系(r=0.9993),检测限分别为0.05μg/mL和0.02μg/mL。在三个添加水平的回收中,两种氨基酸的平均回收率在86.3%~122.3%之间,相对标准偏差(RSD)小于8.51%。将所建立的方法应用于健康人、普通病人和癌症患者晨尿检测,结果发现:癌症患者尿样中色氨酸的水平与健康人和普通病人相比偏低。     

荧光法测定尿中的富马酸比索洛尔

建立了直接测定尿中富马酸比索洛尔的荧光分光光度法。荧光强度与富马酸比索洛尔含量在100－1000ng/mL范围内呈线性关系。检测限20ng/mL。对浓度100ng/mL与500ng/mL的标准溶液进行测定,其回收率分别为95．6％和99．6％,相对标准偏差为5．6％和2．3％,可用于 尿中富马酸比索洛尔含量的测定。     

高效液相色谱-荧光法测定多囊卵巢综合征患者血浆中吲哚类物质

建立了一种同时测定血浆吲哚类物质（吲哚丙酸、吲哚乙酸、吲哚与3-甲基吲哚）的高效液相色谱-荧光法（HPLC-FLD）。以液-液萃取法进行样本预处理,采用Shim-pack VP-ODS色谱柱（150 mm×4.6 mm,4.6 μm）分离,流动相为15 mmol/L磷酸二氢钠溶液-甲醇（42：58,v/v）,柱温为30℃,流速为0.8 mL/min。吲哚丙酸、吲哚乙酸、吲哚与3-甲基吲哚在各自范围内线性关系良好。该方法的日内与日间相对标准偏差均小于6.31%,平均回收率为97.5%~107.0%。利用该方法对多囊卵巢综合征（PCOS）患者（n=61）与作为对照的正常育龄期女性（n=25）的血浆样本进行测定,结果发现PCOS患者血浆吲哚类物质显著高于对照组,其中吲哚是PCOS的危险因子和潜在的诊断标志物。该方法操作简便,灵敏度高,适用于临床检测和实验室研究。     

丹磺酰肼衍生-高效液相色谱-荧光检测法测定包装纸中的甲醛和乙醛

建立了丹磺酰肼(DNSH)衍生-高效液相色谱-荧光检测测定包装纸中甲醛和乙醛的分析方法,并与2,4-二硝基苯肼(DNPH)衍生法进行了比较。纸张样品经衍生化试剂振荡萃取30 min,衍生化反应24 h,萃取液经PSA/C18净化管净化处理后,以Diamonsil~ C18(2)色谱柱(150 mm×4.6 mm,5μm)为固定相,用醋酸水溶液(pH2.55)-乙腈为流动相进行梯度洗脱。采用荧光检测器检测,激发波长为330 nm,发射波长为484 nm。结果表明,衍生剂、甲醛-DNSH和乙醛-DNSH在20 min内可完全分离,方法的加标回收率为81.64%~106.78%,相对标准偏差(RSD)为2.02%~5.53%(n=5),甲醛和乙醛的检出限分别为19.2μg/kg和20.7μg/kg,定量限分别为63.9μg/kg和69.1μg/kg。该法操作简单,灵敏度高,比常规方法具有更低的检出限,能很好地应用到实际样品检测中,为低含量醛类化合物的检测提供了一种新思路。     

Determination of cortisol in guinea pig urine by high performance thin-layer chromatography, high performance liquid chromatography, and thin-layer chromatography/radioimmunoassay

Summary In order to collect urinary samples from unrestrained guinea pigs, animals were kept in their familiar home cages with wood shavings for bedding. Cortisol was removed from shavings by a simple washing step, and an attempt was made to measure its concentrations by high performance thin-layer chromatography (HPTLC), high performance liquid chromatography (HPLC), or thin layer chromatography/radioimmunoassay (TLC-RIA). After intramuscular administration of 25 mg cortisol, cortisol excretion increased from about 20–30 μg/day to 400–500 μg/day (HPTLC: 531 μg/day, HPLC: 493 μg/day; TLC-RIA: 394 μg/day). Similarly, the treatment of the animals with 20 IU ACTH resulted in an augmented cortisol excretion, with mean values of 294 μg/day (HPTLC), 256 μg/day (HPLC) and 143 μg/day (TLC-RIA), respectively. The present study shows, for the first time, that cortisol excretion in unrestrained laboratory animals can be determined. Whilst the cortisol values measured by HPTLC and HPLC agree, the amounts measured by TLC-RIA were significantly lower. These differences are probably due to the presence of substances in urine or shavings which interfere with the radioimmunological determination. Hence, cortisol should be determined either by HPTLC or HPLC. Beside having a desirable specificity, these methods are more suited than TLC/RIA for steroid analysis since they confer the possibility of measuring additional steroids (e.g. precursors and/or metabolites of cortisol) in a single urine extract. This is especially the case for the HPTLC method since substances can be transformed into fluorescent derivatives.     

在线富集与高效液相色谱-荧光检测法联用同时检测儿童水杯中9种双酚类物质

基于在线富集-高效液相色谱-荧光法检测法,以C₁₈柱分别作为富集柱和分析柱,经优化荧光波长、色谱分离和富集条件,建立了同时快速检测塑料儿童水杯浸出液中9种双酚类物质的方法。方法学考察结果显示,所建方法符合含量测定要求。9种双酚类物质在各自范围内线性关系良好,相关系数（r²）均大于0.998,检出限为0.13~66.7 ng/L。9种双酚类物质的回收率为90.7%~112.4%（RSD<11.3%,n=6）。结果显示,除双酚芴外,其他双酚类物质均有检出,浸出量随盛放时间增加而增大,高温多次浸泡后部分双酚类浸出量降低。该方法灵敏度高,操作简便,环境友好,可实现多种新型双酚类物质的同时快速检测。     

流动注射化学发光法快速测定尿液中左亚叶酸钙

在碱性介质中, 左亚叶酸钙对鲁米诺-K3Fe(CN)6体系有显著的增敏作用, 据此, 建立了一种简单、快速测定左亚叶酸钙的流动注射化学发光新方法. 在优化的实验条件下, 该法测定左亚叶酸钙的线性范围为5.0×10-8～1.0×10-5 g/mL; 检出限(3σ)为2.0×10-8 g/mL; 对浓度为1.0×10-6 g/mL的样品进行11次平行测定, 相对标准偏差为1.3%. 将此法用于尿液中左亚叶酸钙的测定, 同时进行回收率实验.     

Determination of diastereoisomeric alkaloids in urine by capillary electrophoresis with electrochemiluminescence detection

A new and simple capillary electrophoresis with electrochemiluminescence detection was developed for the separation and the quantification of a pair of diastereoisomenc alkaloids（ephedrine and pseudoephedrine）.The limits of detection（S/N = 3） were 4.5×10^-8 mol/L for ephedrine and 5.2×10^-8 mol/L for pseudoephedrine,respectively.The RSDs of migration time and peak area were less than 1.3 and 2.5%（n = 5）,respectively.The applicability of the propose method was illustrated in the determination of ephedrine and pseudoephedrine in human urine,ephedrine in nasal drops,and the monitoring of pharmacokinetics for pseudoephedrine.     

Determination of methimazole in urine by liquid chromatography

A liquid chromatography methodology is developed and validated for detection and quantification of methimazole in urine. The approach is based on derivatization with 2-chloro-1-methylquinolinium tetrafluoroborate, reversed-phase high-performance liquid chromatography (HPLC) separation of so formed methimazole 2-S-quinolinium derivative from other urine matrix components, followed by detection and quantification with the use of ultraviolet–visible detector. Neither extraction, nor preconcentration of the sample are necessary. The methimazole standards added to normal urine before derivatization step show that the response of the detector, set at 345 nm, is linear within the concentration range studied, that is, from 0.25 to 50 mg/l urine. The relative standard deviation values for precision and recovery within the calibration range were from 1.8 to 5.0% and from 95.7 to 103.3%, respectively. Lower limits of detection and quantitation were 0.15 and 0.25 mg/l urine, respectively.