气相色谱-离子阱二级质谱法测定猪肉中的氢化泼尼松和甲基氢化泼尼松残留

建立了固相萃取-气相色谱-离子阱二级质谱（GC-MS/MS）测定猪肉中氢化泼尼松和甲基氢化泼尼松残留量的分析方法。待测药物用乙腈提取,C18柱净化,K2Cr2O7氧化,GC-MS/MS测定。采用DB-5毛细管柱分离,电子轰击电离源二级质谱监测模式检测,外标法定量。方法的线性关系良好,线性相关系数均大于0.99,线性范围为2～50 μg/kg;氢化泼尼松和甲基氢化泼尼松的回收率为56%～84%。两种药物的检测限和定量限均分别为1 μg/kg和2 μg/kg。     

离子液体作为添加剂的反向微乳毛细管电动色谱分析化妆品中的糖皮质激素

建立了以离子液体为添加剂的反向微乳毛细管电泳(IL-MEEKC)法分离测定化妆品中氢化可的松、泼尼松和醋酸氢化可的松3种糖皮质激素的方法.微乳毛细管电泳的最佳缓冲体系组成为:2.4% SDS+6.6%正丁醇+0.5%正辛烷+35 mmol/L BMIM-BF4+20 mmol/L磷酸二氢钠缓冲液(pH 2.2);运行电...     

超高压液相色谱-串级质谱分析渔业水中7种激素药物

建立了同时测定渔业养殖水中氢化可的松、泼尼松、醋酸可的松、睾酮、甲睾酮、黄体酮及苯丙酸诺龙等7种激素残留药物的超高压液相色谱-电喷雾电离串联质谱(UPLC-ESI-MS/MS)分析方法.水样用HLB固相萃取柱净化、浓缩后,洗脱液经氮气吹干,残渣用乙腈-水(V∶V=1∶1)溶解,以乙腈和0.1％甲酸水溶液为流动相,经ZORBAXSB-C18色谱柱分离后进行LC-MS/MS多反应检测模式作定性定量分析.方法在0.5～40μggL或1～40μg/L范围具有良好的线性,相关系数大于0.997,检出限(S/N=3)为0.03～0.3μg/L,平均回收率为74.8％～113.0％,相对标准偏差为4.0％～16％.方法适用于渔业养殖水中7种激素的检测分析.     

超高效液相色谱法同时测定化妆品中的15种激素

建立了采用超高效液相色谱同时测定化妆品中糖皮质激素、雌激素、雄激素、孕激素等共15种激素(曲安西龙、氢化可的松、泼尼松、可的松、甲基泼尼松龙、倍他米松、地塞米松、醋酸泼尼松龙、醋酸氢化可的松、雌三醇、雌二醇、雌酮、己烯雌酚、睾酮、孕酮)的分析方法。不同形态的化妆品样品均以甲醇为提取溶剂进行超声提取,经Oasis HLB固相萃取柱富集净化,以Waters ACQUITY UPLC BEH C18色谱柱(1.7 μm,2.1 mm×50 mm)分离,乙腈和水为流动相梯度洗脱,6 min 内完成了15种激素的分离及检测。在1～25 ng进样范围内,15种激素的工作曲线的线性相关系数r均高于0.9995。在低、中、高（2,10,20 mg/kg）3个添加水平下15种激素的平均回收率为88.2%～102.4%,相对标准偏差为1.6%～7.4%。     

UPLC-MS/MS法分析婴幼儿配方乳粉中的12种雌性激素

建立了乳粉中12种雌性激素(孕酮、醋酸美伦孕酮、醋酸甲地孕酮、醋酸氯地孕酮、炔雌醇、雌三醇、17α-雌二醇、17β-雌二醇、雌酮、已烷雌酚、已烯雌酚、已二烯雌酚)的超高效液相色谱-串联质谱(UPLC-MS/MS)检测技术。样品用温水溶解、酶解后,经乙腈提取,HLB柱净化,使用ACQUITY UPLC BEH C18柱(2.1 mm×100 mm,1.7μm)分离,分别在电喷雾正、负离子模式下以多反应监测(MRM)方式检测孕激素和雌激素。结果表明,12种雌性激素的基质校准曲线在一定范围内线性关系良好,相关系数均大于0.99。3个浓度水平的平均回收率为80.2%~108.7%,相对标准偏差不高于7.9%。方法适用于婴幼儿配方乳粉中12种雌性激素的测定。     

高效液相色谱-串联质谱联用测定脂肪中乙酰孕激素残留

建立了脂肪中5种乙酰孕激素,包括醋酸甲羟孕酮(MPA)、醋酸氯地孕酮(CMA)、醋酸甲地孕酮(MEGA)、甲烯雌醇醋酸酯(MLA)及17α-羟基孕酮醋酸酯(HPA)的液相色谱-串联质谱(LC-MS/MS)检测方法。考察了脂肪样品溶剂提取、固相萃取净化对样品分析的影响,确定样品各前处理过程的条件。采用电喷雾(ESI)阳离子,在选择反应监测模式(SRM)下,本方法对5种乙酰孕激素的检出限为0.2～0.3μg/kg,定量限为0.5μg/kg。在空白猪脂肪样品中以0.5、1.0和5.0μg/kg3个水平添加时,5种乙酰孕激素的回收率范围在60.5%～84.1%,RSD为8.9%～16.8%。     

氢化诺卜酸及其酰胺的合成

氢化诺卜醇(1)在醋酸溶液中被CrO3氧化制得氢化诺卜酸(2),由氢化诺卜酸与亚硫酰氯反应生成氢化诺卜酰氯(3),氢化诺卜酰氯分别与浓氨水、33%二甲胺水溶液、二乙胺和二异丙胺在冰水浴中反应制得相应的4种酰胺,各产物的得率均在70%以上,GC纯度95%以上。各产物均用IR,1H NMR,13C NMR与MS进行了结构表征。     

场放大进样-胶束毛细管电泳法测定化妆品中3种糖皮质激素

采用场放大进样-胶束毛细管电泳法对化妆品中氢化可的松、泼尼松和乙酸氢化可的松3种糖皮质激素进行了分离测定。电泳介质为0.20mmol.L-1硼砂缓冲溶液(pH 9.0),运行电压为-20kV,进样电压-20kV,进样时间45s,进水压力3kPa,进水时间20s,检测波长250nm。在优化试验条件下,氢化可的松、泼尼松和乙酸氢化可的松的检出限分别(3S/N)为0.015,0.017,0.017mg.L-1。应用此方法分析了化妆品样品,测得回收率在93.8%～107%之间,测定值的相对标准偏差(n=5)均小于5.1%。     

化学计量学-动力学分光光度法同时测定两种糖皮质类激素

在碱性介质中,高锰酸钾能将糖皮质类激素(醋酸泼尼松和地塞米松)氧化,而本身被还原成绿色的锰酸钾,基于这一反应,本文提出了同时测定醋酸泼尼松和地塞米松的化学计量学-动力学分光光度法.研究发现反应体系在610 nm处有一个吸收峰,实验以该波长为检测波长,优化实验条件.在该条件下,检测醋酸泼尼松和地塞米松的线性范围分别为0....     

大气压化学电离质谱法测定饲料中的醋酸甲羟孕酮

醋酸甲羟孕酮(medroxyprogesteroneacetate)是一种人工合成生长激素,一般被用来生产避孕药及补充人体激素,但是如果使用过量,将影响生育功能。欧盟已经明令禁止在食物和饲料中使用醋酸甲羟孕酮。2002年7月德国首次发现从荷兰进口的生猪猪肉中含有该激素,深入的调查后发现欧盟成员国的猪饲料和一些软饮料里发现了禁用的醋酸甲羟孕酮,可能在欧盟继疯牛病、口蹄疫和二(口恶)英之后引发新的食品安全危机。为保证我国动物源性食品安全,维护人民身体健康,必须建立一种适用、可行、规范的残留检测方法以适应当前加入WIO的形势需要。本实验建立了高效液相色谱-质谱联用法(LC-MS)测定饲料中醋酸甲羟孕酮,旨在为政府从源头上监控饲料中的醋酸甲羟孕酮提供准确可靠的检测方法。     

Stress degradation studies and development of stability-indicating TLC-densitometry method for determination of prednisolone acetate and chloramphenicol in their individual and combined pharmaceutical formulations

ABSTRACT: A rapid and reproducible stability indicating TLC method was developed for the determination of prednisolone acetate and chloramphenicol in presence of their degraded products. Uniform degradation conditions were maintained by refluxing sixteen reaction mixtures for two hours at 80°C using parallel synthesizer including acidic, alkaline and neutral hydrolysis, oxidation and wet heating degradation. Oxidation at room temperature, photochemical and dry heating degradation studies were also carried out. Separation was done on TLC glass plates, pre-coated with silica gel 60F-254 using chloroform: methanol (14:1 v/v). Spots at Rf 0.21 ± 0.02 and Rf 0.41 ± 0.03 were recognized as chloramphenicol and prednisolone acetate, respectively. Quantitative analysis was done through densitometric measurements at multiwavelength (243 nm, λmax of prednisolone acetate and 278 nm, λmax of chloramphenicol), simultaneously. The developed method was optimized and validated as per ICH guidelines. Method was found linear over the concentration range of 200-6000 ng/spot with the correlation coefficient (r2 ± S.D.) of 0.9976 ± 3.5 and 0.9920 ± 2.5 for prednisolone acetate and chloramphenicol, respectively. The developed TLC method can be applied for routine analysis of prednisolone acetate and chloramphenicol in presence of their degraded products in their individual and combined pharmaceutical formulations.     

Assay methodology for prednisolone, prednisolone acetate and prednisolone sodium phosphate in rabbit aqueous humor and ocular physiological solutions

Prednisolone, prednisolone acetate and prednisolone sodium phosphate are glucocorticoids used for ocular, anti-inflammatory therapy. A reversed-phase high-performance liquid chromatographic assay using ultraviolet detection has been developed that affords baseline resolution of the above analytes in balanced salt solutions and rabbit aqueous humor. The drugs can be quantified at 0.025-0.05 micrograms/ml in the above matrices; 6 alpha-methylprednisolone is used as the internal standard. Both esters of prednisolone are vulnerable to chemical and enzymatic hydrolysis giving prednisolone. Analysis of aqueous humor samples shows prednisolone acetate penetrating/metabolizing primarily to prednisolone; prednisolone sodium phosphate penetrates the cornea giving the ester and alcohol.     

Microstructure of microemulsion modified with ionic liquids in microemulsion electrokinetic chromatography and analysis of seven corticosteroids

In this work, the influences of ionic liquid (IL) as a modifier on microemulsion microstructure and separation performance in MEEKC were investigated. Experimental results showed that synergetic effect between IL 1‐butyl‐3‐methylimidazolium tetrafluoro‐borate (BmimBF₄) and surfactant SDS gave a decreased CMC. With increment of IL in microemulsion, negative ζ potential of the microdroplets reduced gradually. The influence of IL on the dimensions of microdroplet was complicated. At BmimBF₄ less than 8 mM, IL made microemulsion droplet smaller in size. While at BmimBF₄ more than 10 mM, the size increased and reached to a maximum value at 12 mM, where the microdroplets were larger than that without IL. After that, the micreodroplet size decreased again. Relative fluorescence intensity of the first vibration band of pyrene to the third one (I₁/I₃) enhanced as IL was added to microemulsion, which indicated that this addition increased environmental polarity in the inner core of microdroplets. Prednisone, hydrocortisone, prednisolone, hydrocortisone acetate, cortisone acetate, prednisolone acetate, and triamcinolone acetonide were analyzed with MEEKC modified with IL to evaluate the separation performance. Cortisone acetate and prednisolone acetate could not be separated at all in typical microemulsion. The seven analytes could be separated by the addition of 10 mM BmimBF₄ into the microemulsion system. The method has been used for analysis of corticosteroids in cosmetic samples with simple extraction; the recoveries for seven analytes were between 86 and 114%. This method provides accuracy, reproducibility, pretreatment simplicity, and could be applied to the quality control of cosmetics.     

加压毛细管电色谱-紫外检测法分析糖皮质激素及其在头发检测中的应用

采用反相加压毛细管电色谱与紫外检测联用技术,建立了一种高效、简便的糖皮质激素分析方法,适用于头发中糖皮质激素的检测。使用C18反相色谱柱,流动相为pH 8.0, 1.5 mmol/L的Tris-乙腈(65:35, v/v),检测波长为245 nm、分离电压为~10 kV、反压为10.5 MPa、泵流速为0.05 mL/min,进行等度洗脱,倍他米松、地塞米松、泼尼松、泼尼松龙、醋酸泼尼松龙、醋酸氢化可的松、醋酸可的松、皮质脂酮等8种激素在20 min内实现快速分离。各组分的质量浓度线性范围达到3个数量级,检出限(S/N=3)在μg/g水平,迁移时间和峰面积的相对标准偏差(RSD)分别小于4.8%和7.4%。将所建立方法应用于头发样品分析,检测前采用蛋白酶水解提取和净化处理样品,不同浓度糖皮质激素的回收率为71%～85%。该研究为糖皮质激素药物暴露监测以及压力检测提供了新手段,有望用于滥用药物的控制和临床诊断。     

Separation of corticosteroids by microemulsion EKC with diethyl L-tartrate as the oil phase

A novel microemulsion based on a mixture of diethyl L-tartrate (DET) and SDS was developed for the microemulsion EKC (MEEKC) determination of structurally related steroids. The system consisted of 0.5% w/w DET, 1.7% w/w SDS, 1.2% w/w 1-butanol, 89.6% w/w phosphate buffer (40 mM, pH 7.0), and 7% w/w ACN. With an applied voltage of +10 kV, a baseline separation of aldosterone (A), cortisone acetate (CA), dexamethasone (D), hydrocortisone (H), hydrocortisone acetate (HA), prednisolone (P), prednisolone acetate (PA), prednisone (Ps), triamcinolone (T), and triamcinolone acetonide (TA) could be achieved. Under the optimized conditions, the reproducibility of the retention time (n = 4) for most of the compounds was less than +/-0.8% with the exception of A, Ps, and T. The average number of theoretical plates was 18 800 plates/m. The results were compared with those achieved by the modified micellar EKC (MEKC). MEEKC showed obvious advantages over MEKC for the separation of highly hydrophobic substances. To further evaluate the system, we tested the MEEKC method by analyzing corticosteroids in a spiked urine sample.     

Separation of exogenous and endogenous steroid hormones by micellar high-performance thin-layer chromatography

The conditions for the separation of prednisolone, dexamethasone, cortisol, cortisone, and cortisone acetate are selected using high-performance thin-layer chromatography on PTSKh-AF-V-UF plates with a 12 mM aqueous solution of sodium dodecyl sulfate as a micellar mobile phase. The hormones are identified under UV radiation (254 nm).     

Determination of prednisolone and prednisone in plasma by liquid chromatography with fluorescence detection

After a single-step extraction from plasma (250 μl) with dichloromethane, the drugs and dexamethasone (internal standard) are oxidized by copper(II) acetate to the corresponding glyoxal and converted into the fluorescent quinoxalines by reaction with 1,2-diamino-4,5-methylenedioxybenzene. The quinoxalines are separated within 55 min by reversed-phase liquid chromatography with isocratic elution. The detection limits for prednisolone and prednisone added to plasma are 3 ng ml^?1 in plasma (signal-to-noise ratio=3).     

Quantitation of cortico-steroids as common adulterants in local drugs by HPLC

Summary A sensitive and specific method for quantitation of the steroids betamethasone, prednisolone and cortisone acetate commonly used as adulterants in locally produced herb extracts and in certain homeopathic drugs is described. Reverse-phase liquid chromatography with UV detection has been used.     

Microemulsion electrokinetic chromatography of corticosteroids. Effect of surfactants and cyclodextrins on the separation selectivity

The separation of neutral hydrophobic corticosteroids (cortisone, cortisone acetate, hydrocortisone, hydrocortisone acetate, prednisolone and prednisolone acetate) by microemulsion electrokinetic chromatography (MEEKC) was studied. In the preparation of microemulsion, heptane was the solvent, n-butanol the co-surfactant and, as anionic surfactants, sodium dodecyl sulfate (SDS) or taurodeoxycholic acid sodium salt (STDC) were employed. Using an acidic running buffer, (phosphate pH 2.5) a strong suppression of the electroosmotic flow (EOF) was observed; this resulted in a fast anodic migration of the analytes partitioned into the negatively charged microemulsion droplets. Under these conditions, STDC showed better separation of corticosteroids than the conventional SDS; however, the use of a single anionic surfactant did not provide the required selectivity. The addition of the neutral surfactant polyoxyethylene glycol octadecyl ether (Brij 76) significantly altered the migration of each analytes allowing a better tuning of separation; however, in order to obtain adequate resolution between couples of adjacent critical peaks, the addition of neutral cyclodextrins (CDs) was found to be essential. This apparently complex system (CD-MEEKC), was optimized by studying the effect of the most important parameters affecting separation: STDC concentration, Brij 76 concentration, nature and concentration of cyclodextrins. Following a rational step-by-step approach, the optimised conditions providing the complete separation of the analytes were found to be: 4.0% STDC, 2.5% Brij 76, 6.6% n-butanol, 1.36% heptane and 85.54% of a solution 5 mM beta-CD in 50 mM phosphate buffer (pH 2.5). The optimized system was preliminary applied to the detection of corticosteroids related substances at impurity level and it could be considered a useful orthogonal alternative to HPLC methods.     

Comparison of the analysis of corticosteroids using different techniques

In this work we have optimized the analysis of 18 human corticosteroids, some endogenous (tetrahydrocortisol, tetrahydrocortisone, cortisol, and cortisone) and others synthetic (betamethasone, budesonide, cortisone acetate, desonide, dexamethasone, dexamethasone acetate, flunisolide, fluocinolone acetonide, halcinonide, methylprednisolone, prednisolone, prednisone, triamcinolone, and triamcinolone acetonide). Three analytical techniques were developed: ELISA, gas chromatography coupled with mass spectrometry (GC–MS), and liquid chromatography coupled with mass spectrometry (LC–MS). Several sample-preparation methods were optimized for each technique and enabled compounds of interest to be extracted from small urine samples (several mL). The results enabled us to assess the possibilities and the sensitivity of each technique for application to doping tests.