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In order to observe the effects of the ground and intact Zhi Zi (Fructus Gardeniae) and different combinations of the ingredients and refined single Chinese drug granules in Yin Chen Hao Decoction compound prescription on the contents of gardenoside (an effective component of the prescription), the contents of gardenoside were determined with reversed phase high performance liquid chromatography (HPLC), with acetonitrile-water (15:85) as mobile phase, at wave length 238nm. The results indicated that the gardenoside-decocted-out rates in the decoctions prepared by different combinations of the ingredients with the ground Zhi Zi (Fructus Gardeniae) all were higher significantly than those in the decoction with intact Zhi Zi (Fructus Gardeniae), and generally, different combinations of the ingredients in the decoction had only little effect on the gardenoside-decocted-out rate.
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制备型高效液相色谱在生物医药制品领域的应用
制备型高效液相色谱(preparative high performance liquid chromatography,Prep-HPLC)是一种使用高压、大流量液体输送系统在高分辨率、大内径、高载量分离柱上进行样品高纯度分离的液相色谱制备方法.应用该方法分离的产品在纯度、回收率、分离效率等方面远远优于传统的制备方法,因此在生物制品和药物研究、生产领域得到广泛应用[1-3].本文就近年来Prep-HPLC方法的研究进展及其在生物医药制品领域的应用做一综述.
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韧带样型纤维瘤病中APC基因和β-catenin基因突变的研究
韧带样型纤维瘤病中Wnt 信号通路异常在肿瘤的发生、发展过程中起着重要作用,我们采用聚合酶链反应(PCR)-变性高效液相色谱技术(denaturing high performance liquid chromatography, DHPLC)-直接测序的方法分析韧带样型纤维瘤病中APC基因、β-catenin基因的突变,探讨肿瘤发生发展的原因和机制.
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变性高效液相色谱结合混和样品池法检测ACE基因多态性
血管紧张素转换酶(angiotensin converting enzyme,ACE)基因多态性与冠心病、高血压等常见心血管疾病的关系是目前研究的热点所在.变性高效液相色谱(denaturing high performance liquid chromatography,DHPLC)能够在使DNA部分变性的温度下,利用空间构向的不同区分不完全匹配的杂合DNA双链.DHPLC被广泛地应用于未知单核苷酸多态性(single nucleotide polymorphism,SNP)的检测工作.
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高效液相色谱法测定面粉中过氧化苯甲酰
本文采用乙醇提取过氧化苯甲酰,用盐酸羟胺还原成苯甲酸后注入高效液相色谱系统进行分离测定,以保留时间定性,峰面积定量.经过实验,RSD为3.1%,回收率为85.0%~92.0%,r=0.999,用于样品中过氧化苯甲酰含量测定,结果满意.
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高效液相色谱法测定健胃消食片的有关物质
健胃消食片收载于卫生部药品标准[1].其组成分为太子参、陈皮、山药、麦芽(炒)、山楂,具有健胃消食的功能,用于脾胃虚弱,消化不良等症.然而其含量测定无标准,为了有效控制其质量,用高效液相色谱法测定该制剂的橙皮苷含量,操作方法简便、准确,重复性好,其回收率98.7%(RSD=1.3%),重复性的RSD为1.3%.
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高效液相色谱法测定复合维生素胶囊中泛酸含量
婴幼儿配方食品和乳粉中泛酸(或泛酸钙)的测定已有国家标准[4],但是测定含大量粘稠性明胶的复合维生素类胶囊中的泛酸含量时,国标样品处理方法对泛酸提取比例非常低,而且采用该流动相体系不能很好地分离被测组分.为此对方法的流动相和样品提取溶剂进行改进,以期取得良好的萃取和分离效果.
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用高效液相色谱法测定化妆品中性激素的探讨
化妆品中添加激素具有促进毛发生长、消除皱纹、增加皮肤弹性、防止皮肤老化等作用,但长期使用含激素的化妆品易导致人体代谢紊乱及癌症的发生.卫生部颁布的<化妆品卫生规范>(以下简称"规范")规定在化妆品中禁止使用性激素,并规定用高效液相色谱法检测雌二醇、雌三醇、雌酮、已烯雌酚、睾丸酮、甲基睾丸酮、孕酮7种激素.但在实际应用中按"规范"的色谱条件无法分离雌二醇和已烯雌酚这两个组分.本文针对"规范"中方法的不足,对方法的色谱条件进行了改进,使7种激素组分得到有效的分离,满足检测工作的需要.现将结果报道如下.
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高效液相色谱法测定人发中的锌和铜
液相色谱因其具有很高的选择性,若与光度法结合,便可建立痕量金属离子的高选择性和高灵敏度的分离分析法,可同时分离离子型、分子型及其混合物[1].用二甲酚橙(XO)光度法单独测定锌离子、铜离子已有报道[2],由于它们的络合物吸收峰相近,测定时会造成干扰;用甲醇-水作流动相,离子交换色谱法分离镍离子、锌离子也已有报道[3].本法以乙腈-水(10:90)作流动相,六次甲基四胺(HMTA)作为离子对试剂,缓冲溶液pH值控制为5.6,XO作柱前衍生剂,建立了反相高效液相色谱快速分离分析人发中痕量锌和铜的方法,加标回收率铜平均为98.9%,锌平均为97.5%.
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变性高效液相色谱基因扫描技术在人类疾病基因诊断中的应用
自1992年变性高效液相色谱(denaturing high performance liquid chromatography, DHPLC)基因扫描技术建立以来,因其在筛查基因突变或单核苷酸多态(single nucleotide polymorphism, SNP)位点方面具有准确、敏感、高通量、自动化的特点,巳经有800余篇研究论文或应用实例发表,而其中的700多篇是在近5年内问世,涉及370多个基因,由此发现或确认了许多致病基因突变或疾病相关基因SNP位点.DHPLC是一种准确、敏感、经济、实用的遗传变异筛查技术,可用于单碱基替换、小片段缺失和插入等多种基因序列突变的检测.DHPLC基因扫描技术可在生殖细胞和体细胞中筛查遗传变异.而像特定位点DNA甲基化状态异常等遗传功能改变,甚至多个特异基因突变的同步微序列测定,也可用在DHPLC基础上建立的方法来检测.我们对DHPLC基因扫描技术检测遗传变异的原理、流程、特点进行介绍,对DHPLC检测基因突变的准确性、敏感性、有效性、实用性,及其在临床基因诊断领域的验证和应用进行讨论.
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A rapid and simple method for the determination of polyhexamethylene biguanide (polyhexanide, PHMB) in liquid and gel-like pharmaceutical formulations by means of high performance liquid chromatography coupled to diode-array detection (HPLC-DAD) was developed. Best separation was achieved using a cyanopropyl bonded phase (Agilent Zorbax Eclipse XDB-CN column 4.6 mm75 mm with particle size of 3.5 mm) as well as gradient elution consisting of acetonitrile/deionized water at a flow rate of 1.0 mL/min. The optimized and applied chromatographic conditions permitted separation of polyhexanide from interacting matrix with subsequent detection at a wavelength of 235 nm with good sensitivity. The method validation was carried out with regard to the guidelines for analytical procedures demanded by the International Conference on Harmonisation (ICH). Mean recoveries of 102% and 101% for gel-like as well as liquid preparations were obtained. Suitable repeatability as well as intermediate precision could be achieved with limits of detection r0.004 mg/mL for both formulations, equivalent to r0.004% PHMB concerning sample preparation. Determination of PHMB was accomplished without tedious sample preparation. Interacting matrix could be eliminated by the chromatographic procedure with excellent performance of system suitability. All analytical requirements were fulfilled permitting a reliable and precise determination of PHMB in pharmaceuticals. Furthermore, the developed method was applied to stability testing of pharmaceutical preparations containing PHMB.
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A novel method for analysis of three active components curcumin, demethoxycurcumin and bisdemethoxycurcumin in Curcuma longa L. was developed by HPLC coupled with electrochemical detection. Three curcuminoids were well separated on a C18 column and detected with high sensitivity. A mobile phase containing acetonitrile and 10 mM Na2HPO4-H3PO4 (pH 5.0) (50:50, v/v) was used. Good linearity was obtained in the range of 0.208-41.6, 0.197-39.4, and 0.227-114μM for curcumin, demethoxycurcumin and bisdemethoxycurcumin respectively. The limit of detection reached up to 10 ? 8 M, which was lower than that by UV detection. The relative standard deviations (RSDs) ranged from 1.06%to 1.88%for intra-day precision and from 4.30%to 5.79%for inter-day precision, respectively. The proposed method has been applied in real herb sample and recoveries ranging from 86.3%to 111%were obtained.