高效液相色谱-蒸发光散射法测定牛黄中多种有效成分含量及其质量相关性研究

建立高效液相色谱-蒸发光散射法(HPLC-ELSD)同时快速测定牛黄中7种组分的含量的方法。色谱柱为UltimateAQ-C18柱,流动相为乙腈-0.2%甲酸水溶液,梯度洗脱,流速1mL/min;蒸发光散射检测器参数:漂移管温度100℃,载气(N2)流速1.9L/min。在选定色谱条件下,胆固醇、各种胆汁酸及其钠盐峰形良好且能达到很好地分离。各组分色谱峰峰面积的对数与浓度的对数呈良好线形关系(r0.9972);标准加入回收率为98.8%～112.7%;7种组分的最低检出限为0.001～0.007g/L。本方法可简便、快速地同时检测不同牛黄样品中7种组分的含量并可表征不同牛黄样品的质量,从而为该名贵药材的质量控制提供更全面的依据和参考。     

高效液相色谱-质谱联用法检测免疫抑制剂及其合并用药

建立高效液相色谱-质谱联用法同时测定人血浆中免疫抑制剂及合并用药12种药物浓度的方法.选用Kromasil-C18色谱柱(50 mm× 4.6 mm×5 μm),以甲醇-乙腈-1mmol/L乙酸铵溶液为流动相,采用梯度洗脱进行分离,样本用甲醇沉淀蛋白后进样,流速:1.1 mL/min;柱温:35℃;进样量:20μL.选用3200QTrap型液相色谱-串联质谱仪的多反应监测(MRM)扫描方式进行检测.12种药物的线性范围为0.2～1000μg/L;定量下限为0.2 μg/L.准确度与精密度结果显示方法日间、日内RSD均小于15％;相对偏差-13％～9.33％,稳定性较好.本方法快速、灵敏,专属性强、重现性好,可用于人体血浆中免疫抑制剂及其常用合并用药共12种药物浓度的测定.     

血浆辅酶Q10的高效液相色谱快速测定

建立了一种简单快速测定血浆辅酶Q10(CoQ10)的反相高效液相色谱方法.血浆经正丙醇萃取,上层清液直接进样分析.色谱柱为Hypersil ODS2 5μm,150 mm ×4.6 mm i.d,以异丙醇-甲醇(体积比19)作流动相,275 nm作检测波长,外标法定量.在0.05～20 mg/L范围内,峰高与质量浓度呈良好的线性关系(r=0.999 8),血浆中辅酶Q10的检出限为0.03 mg/L(S/N=3).该方法简单、快速、精密度高(RSD＜5%),适宜于血浆辅酶Q10含量的检测.     

高效液相色谱-串联质谱法同时测定血清中游离和结合型胆汁酸

建立了高效液相色谱-串联质谱同时测定血清中游离和结合型胆汁酸的方法。血清样品经固相萃取后,在C18色谱柱上梯度洗脱,用负离子多反应监测模式检测。15种胆汁酸在0.10~100.0μmol/L的浓度范围内与响应呈良好的线性,r为0.9990~0.9999;检出限为0.001~0.006μmol/L;本方法的相对标准偏差日内不超过3.0%,日间不超过6.0%;平均回收率为86.0%~110.0%,适合于临床检测血清胆汁酸。     

高效液相色谱法快速测定环境空气中13种醛酮类化合物

建立高效液相色谱法快速测定环境空气中13种醛酮类化合物的方法。选用Poroshell 120 SB–C_(18)色谱柱(250 mm×4.6 mm,4μm)为分离柱,以水–乙腈溶液为流动相,梯度洗脱,流量为1.5 mL/min,柱温为50℃,进样体积为10μL,采用紫外检测器检测,检测波长为360 nm。13种醛酮类化合物的质量浓度在0.05~2.0μg/mL范围内与色谱峰面积具有良好的线性关系,相关系数均不小于0.9998,方法检出限为0.16~0.47μg/m³。空白样品加标回收率为97.2%~101.1%,测定结果的相对标准偏差为0.5%~4.3%(n=6)。该方法分析速度快,准确度和灵敏度高,精密度好,适用于环境空气中醛酮类化合物的快速测定。     

硫辛酸片含量与降解物的高效液相色谱测定研究

建立了硫辛酸片中硫辛酸及其降解物快速测定的高效液相色谱方法。采用Waters XBridge色谱柱(5μm,150 mm×4.6 mm),流动相为0.05 mol/L磷酸二氢钾溶液(磷酸调至pH 2.5)-乙腈-甲醇(50∶45∶5),流速为1.0 mL/min,进样量为20μL,二极管阵列检测器,检测波长为220 nm,全波长扫描范围为200～400 nm,柱温为室温。结果表明,在上述色谱条件下,硫辛酸浓度在0.4～20.0 mg/L范围内线性关系良好(r=0.999 9);其检出限为0.2 mg/L;定量下限为0.6 mg/L;精密度RSD(n=12)为3.4%;加标回收率为100.9%,RSD为2.0%。该法简便、快速、准确、选择性好、灵敏度高,可用于硫辛酸片含量与降解物的测定。     

离子色谱法测定饮用水中痕量溴酸盐

建立了一种利用离子色谱法测定饮用水中痕量溴酸盐(BrO<,3><'->)的方法.色谱条件为:IonPac AS19(250mm x4 mm)色谱柱分离,KOH 梯度淋洗,流速1.0 mL/min,抑制电导检测,外标法定量.BrO<,3><'->浓度在2.0～100μg/L范围内有良好的线性,检出限为0.3 ng/mL,...     

反相高效液相色谱法直接测定水蛭中14种未衍生氨基酸的含量

建立了一种反相高效液相色谱/蒸发光散射检测器方法,同时测定中药水蛭中14种未衍生氨基酸含量。以异亮氨酸等氨基酸为标准品、PrevailTMC18(250mm×4.6mmi.d.,5μm)为色谱柱、梯度洗脱、漂移管温度115℃、气体流量2.5L/min,在25min内即可将水蛭中14种氨基酸分离测定。氨基酸质量浓度0.072~2.29g/L时,其对数值与峰面积对数值线性关系良好;14种氨基酸的加样回收率为94.8%~104.4%;信噪比为3时,异亮氨酸检出限为20mg/L。该法快速、简便、准确,可作为水蛭中氨基酸直接测定方法,亦可为其它中药中氨基酸分析提供参考。     

反相高效液相色谱法同时测定丁酸氯维地平中10种相关杂质

建立反相高效液相色谱辅-光电二极管阵列检测器(RP-HPLC)法同时测定丁酸氯维地平原料药中的10种杂质。色谱柱为Symmetry C18柱(250 mm×4.6 mm,5μm),流动相为0.05 mol/L Na H2PO4溶液(pH 2.5)-乙腈/甲醇(3∶2,V/V),梯度洗脱,柱温35℃,流速为1.5 m L/min,检测波长220 nm。丁酸氯维地平及其10个已知杂质能够达到良好的分离,且各组分在各自测定浓度范围内与峰面积的线性关系良好(r≥0.9970);丁酸氯维地平及杂质1~10的检出限(S/N=3)在0.15~0.90 mg/L之间。本方法快速、简便、有效,可用于丁酸氯维地平原料药的质量控制管理。     

HPLC法测定盐酸美他环素片的含量

建立用外标法测定盐酸美他环素片含量的简单高效液相色谱方法.采用PurospherStar C18色谱柱(250mm×4.6mm,5μgm);流动相为:V(甲醇):V(水)=50:50混合后用H3PO4调节pH至3.0±0.02;检测波长:280nm;进样量:20μL,流量:0.7 mL/min;温度35℃.美他环素在20.02～200.2μg/mL浓度范围内线性关系良好(r=0.99999);平均回收率为98.6%,回收率的RSD为0.6%.本方法可用于盐酸美他环素片的含量测定.     

Thin-Layer Chromatography of Bile Alcohols,Bile Acids and Conjugated Bile Acids

Abstract

Thin-layer chromatography of bile alcohols, bile acids and bile acid conjugates has been reviewed. Particular emphasis has been placed on the separation of the glycine and taurine conjugated bile acids as a class and as individual compounds, and on the isolation of bile alcohols and C₂₇ bile acids diastereo-isomeric at C-25.     

反相高效液相色谱法分析人体胆汁中结合型胆汁酸

根据反相液相色谱的保留值方程ＩｇＫ＇＝ａ＋ｃ．ｃｂ,结合型胆２汁酸的选择性变化规律ｌｇａ＝△ａ＋△ｃｖ．ｃｂ及结合型胆汁酸的标准样品色谱保留值,对人体胆汁中６种结合型胆汁酸进行了定性定量分析。６种结合型胆汁酸的浓度在０～１０．１５ｇ／Ｌ范围２内与相应的色谱峰高成良好的线性关系（相关系数在０．９９９６～０．９９７９之间）。     

Separation and Determination of Bile Acids in Human Bile by High-Performance Liquid Chromatography

Abstract

A method for simultaneous determination of major bile acids in human bile is described. The unconjugated, glycine- and taurine-conjugated bile acids are extracted with Sep-pak C¹⁸ and separated into groups by ion-exchange chromatography on a lipophilic gel. Subsequently, resolution of each group into ursodeoxycholate, cholate, chenodeoxycholate, deoxycholate and lithocholate is attained into two stages by high-performance liquid chromatography on a Radial-PAK A column. First, 0.3% ammonium phosphate (pH 7.7)/acetonitrile (19:8, v/v) is used for separation of the latter three as a mobile phase. Ursodeoxycholate and cholate are efficiently separated in 0.3% ammonium phosphate (pH 7.7)/acetonitrile (23:8, v/v). The present method is applicable to quantitation of bile acids in human bile with satisfactory accuracy and precision.     

Specificity and Cross Reactivity in Bile Acid Radioimmunoassays Serum Bile Acids,Radioimmunoassay

Abstract

In man there are four main bile acid fractions and within each fraction there is the possibility of at least three bile acid moities - two conjugated and one unconjugated. Bile acids thus present a considerable challenge to radioimmunoassay techniques. Few of the antisera described to date are satisfactory in that they do not show equal reactivity to each of the moities to be assayed, and many have unacceptable cross reactivity properties. Clearly there is need for caution and development in this field. The first application of radioimmunoassay (RIA) techniques to the measurement of serum bile acids was made by Simmonds, Korman, Go and Hofmann in 1973¹. Because of its unique sensitivity and ease of application, RIA has been used not only to determine the increased serum bile acid levels found in liver disease, but also to monitor the clearance of bile acids from the peripheral circulation of normal subjects, to provide 24 hour serum bile acid profiles in normal subjects and even to assay the low serum bile acid levels found in patients in whom the bile acid enterohepatic circulation has been interrupted. Some 20 preparations of bile acid antisera have been described to date and commercial kits are now available, for RIA of each of the 4 major bile acid fractions found in human sera. Considerable differences however, in the specificity of these antisera are indicated by their reported cross reactivities and the analytical validity of their use is often questionable.     

反相高效液相色谱法测定鼠胆汁中游离与结合型胆汁酸

建立了一种柱前衍生反相高效液相色谱荧光检测大鼠胆汁中游离与结合型胆汁酸的方法.以4-溴甲基-7-甲氧基香豆素为衍生剂,月桂酸为內标,Waters Nova C18色谱柱分离,采用甲醇/乙腈和水梯度洗脱,荧光激发波长(λex)330 nm,发射波长(λem)400 nm,內标标准曲线法定量.各胆汁酸在5-2 400μmol/L范围内线性良好,r为0.998 9-0.999 7.最低检出限为1.2-2.5μmol/L.精密度日内在1.05%-3.67%之间,日间在3.16%-6.83%之间.平均回收率为87.8%-106.7%.该方法灵敏、重复性好,分析时间短,线性范围宽,适合于科研工作中对鼠胆汁中各胆汁酸进行定量检测.     

Bile salt-phospholipid aggregation at submicellar concentrations

The aggregation behavior of the bile salts taurodeoxycholate (NaTDC) and sodium cholate (NaC), are followed at concentrations below critical micelle concentrations (CMCs) using the environment sensitive, fluorescent-labeled phospholipid, 2-(6-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)hexanoyl-1-hexadecanoyl-sn-glycero-3-phosphocholine (NBD-C₆-HPC). A buffer solution containing NBD-C₆-HPC is titrated with increasing NaC or NaTDC and the fluorescence changes followed. Both bile salts induced fluorescence changes below their critical micelle concentration indicating the presence of a bile salt–phospholipid aggregate. A critical control experiment using 6-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino) hexanoic acid (NBD-X) shows that the bile salts are interacting with the longer, C16 hydrocarbon tail, not the NBD probe. The fluorescence curves were fitted to the Hill equation as a model for cooperative aggregation. The cooperativity model provides a minimum estimate for the number of bile salts to give maximal fluorescence. This number was calculated for NaC and NaTDC to have a minimum value of 2. A small aggregation number supports the existence of primary micellar aggregates at submicellar concentrations for bile salt–phospholipid aqueous solutions.     

基于胆汁酸的刺激响应聚合物

刺激响应聚合物是当今材料科学研究的热点之一。其在外界刺激下,自身的物理或者化学性质发生变化产生响应,在药物可控释放、生物传感器、催化、吸附分离等方面有广阔的应用。胆汁酸是天然的甾类生物分子,其分子结构中含有羟基和羧基等官能团,容易进行化学修饰,且具有双亲性和一定的刚性。在聚合物中引入胆汁酸分子,能够从结构和功能两个方面丰富刺激响应高分子的研究。一方面,胆汁酸及修饰的胆汁酸既可以作为单体直接聚合,也可以被连接到含有一定官能团的聚合物上,由此可以得到以胆汁酸作为主链、侧基、端基,以及胆汁酸作为星形聚合物的核等多种聚合物结构。另一方面,胆汁酸的引入不仅可以提高聚合物的生物相容性,有效地赋予聚合物胆汁酸结构的独特性质,而且可以用于构建具有一定刺激响应功能的组装结构,进而应用于形状记忆、手性分离、药物载体等材料中。本文综述了基于胆汁酸的刺激响应聚合物近年来的相关工作,从基于胆汁酸的刺激响应聚合物的分子设计和结构构筑出发,结合其性质和应用进行论述,也对该领域的改进和发展提出展望。     

Bile Acids-Selective Chemosensors Based on NBD-amine-Modified Cyclodextrins

A new type of fluorescent chemosensor, based on modified cyclodextrins bearing the fluorophore unit NBD-amine, was prepared. One of these new chemosensors, NC0γCD is very sensitive to bile acids, but is not sensitive to other guests (e.g. adamantane and borneol derivatives). The response of the new type of chemosensor to a guest was an increase in the fluorescence intensity and the sensitivity parameter (ΔI/I ₀ ) dose not correlate to the binding affinity of NC0γCD.     

胆酸衍生物的合成及其应用

综述了胆酸衍生物,包括应用于高分子材料、分子识别及医药领域的胆酸衍生物的合成.