HPLC法测定健康人血清中吗咯替酯及其代谢产物浓度

增水法是利用岩样自身的毛细管压力，让岩样自然吸水，分三个阶段由低含水饱和度向高含水饱和度过渡，从而获取储层岩样不同的饱和度指数。利用增水法测定了陕北油气田的低孔低渗砂岩储层饱和度指数，其结果可靠。     

天然来源的新化合物4~"-异丁酰螺旋霉素Ⅲ(生技霉素A_(2β))

生技霉素是一组４＂－酰化螺旋霉素，继从生技霉素产生菌发酵液中分离到主要组分生技霉素Ａ和Ｂ之后，利用两相分配及柱层析的分离方法，又分离到分子量为９６８的化合物生技霉素Ａ２。根据其ＵＶ、ＩＲ、ＦＡＢ－ＭＳ、１Ｈ－ＮＭＲ、１３Ｃ－ＮＭＲ、ＤＥＰＴ谱、１Ｈ－１ＨＣＯＳＹ的分析及与４＂－异戊酰螺旋霉素Ⅲ（即生技霉素Ａ）光谱数据的比较表明生技霉素Ａ２含分子式皆为Ｃ５０Ｈ８４Ｎ２Ｏ１６的４＂－异丁酰螺旋霉素Ⅲ（生技霉素Ａ２β）和４＂－丁酰螺旋霉素Ⅲ（生技霉素Ａ２α）。生技霉素Ａ２碱水解产物中游离脂肪酸的气相色谱及ＧＣ－ＭＳ分析支持了以上结论。     

α－乙酰－γ－丁内酯合成工艺的改进

α－乙酰－γ－丁内酯合成工艺的改进周景尧，林国妹（复旦大学化学系，上海２００４３３）ＩＭＰＲＯＶＥＤＳＹＮＴＨＥＳＩＳＯＦα－ＡＣＥＴＹＬ－γ－ＢＵＴＹＲＯＬＡＣＴＯＮＥ￥ＺＨＯＵＪｉｎｇ－Ｙａｏ；ＬＩＮＧｕｏ－Ｍｅｉ（ＤｅｐａｒｔｍｅｎｔｏｆＣｈｅ...     

异丁茶碱的合成

异丁茶碱的合成ＰＲＥＰＡＲＡＴＩＯＮＯＦＩＳＢＵＦＹＬＬＩＮＥ骆宝基李遂一卢泽ａ刘伟ｂ（河南省医药工业研究所，开封４７５００３；ａ河南省医药学校，开封４７５００１；ｂ河南大学化学系，开封４７５００１）ＬＵＯＢａｏ－Ｊｉ，ＬＩＳｈｕｉ－Ｙｉ，ＬＵＺｅａ...     

天然来源的新化合物4″—异丁酰螺旋霉素Ⅲ（生技霉素A2β）

生技霉素是一组４″－酰化螺旋霉素，继从生技霉素产生菌发酵中分离到主要组分生技霉素Ａ和Ｂ之后，利用两相分配及柱层析的分离方法，又分离到分子量为９６８的化合物生技霉素Ａ２。根据其ＵＶ、ＩＲ、ＦＡＢ－ＭＳ、＾１Ｈ－ＮＭＲ、＾１３Ｃ－ＮＭＲ、ＤＥＰＴ谱、＾１Ｈ－＾１ＨＣＯＳＹ的分析及与４″－异戊酰螺旋霉系Ⅲ（即生技霉素Ａ）光谱数据的比较表明生技霉素Ａ２含分子式皆为Ｃ５０Ｈ８４Ｎ２Ｏ１６的４″－异丁酰螺旋霉     

滇钩藤化学成分研究

目的：对滇钩藤化学成分进行研究,寻找作用于心血管的活性化合物。方法采用分析分离技术进行化学成分的分离,运用光谱方法确定所分离化合物的结构,并进行心血管活必互理试验。结果：从中分离得到６个化合物,为ＳＵＢＩ（阿马利新）,ＳＵＢ２（四氢鸭木碱（,ＳＵＢ３（ｉｓｏｒｈｙｃｈｏｐｈｙｌｉｎｅ）,ＳＵ４（ｒｈｙｃｈｏｐｈｙｌｌｉｎｅ）,ＳＵＢ５（ｃｏｒｙｏｘｉｎｅ）,ＳＵＢ６（乌苏酸－３－Ｏ－β－Ｄ－葡萄醛     

用杆状病毒载体在家蚕细胞中表达HBeAg基因及其表达产物纯化的 …

以ＰＣＲ技术扩增含有Ｐｒｅ－Ｃ信号肽序列及完整的ＨＢｅＡｇ基因序列，克隆至家蚕核多角体病毒转移载体ｐＢｍ０３０，与野生型ＢｍＮＰＶ ＤＮＡ共转染家蚕ＢｍＮ细胞，空斑纯化后得重组病毒，研究结果表明ＢｍＮ细胞能正确识别与切割ＨＢｅＡｇ信号肽序列，所表达的ＨＢｅＡｇ效价高，纯度好，ＥＬＩＳＡ法测得培养上清中ＨＢｅＡｇ效价达１：３２０００。上清经过Ｓｅｐｈａｃｒｙ １ Ｓ－２００和ＤＥＡＥ－Ｓｅｐｈａｒｏ     

α—常春藤皂甙和无患子皂甙B对小鼠肝微粒体细胞色素P—450的作用

目的：研究α－常春藤皂甙（Ｈｅｄ）和无患子皂甙Ｂ（ＳａｐＢ）对小鼠肝细胞色素Ｐ－４５０的影响与保肝作用的关系，方法：给小鼠ｓｃＨｅｄ，ＳａｐＢ和Ｈｅｄ＋ＳａｐＢ（１：１．５）检测小鼠肝微粒体细胞色素Ｐ－４５０的含量。结果：Ｈｅｄ２０ｍｇ．ｋｇ＾－１，ＳａｐＢ２０ｍｇ．ｋｇ＾－１和Ｈｅｄ＋ＳａｐＢ２０ｍｇ．ｋｇ＾－１使肝细胞色素Ｐ－４５０分别降低了４０％，５５％和５０％，这种抑制作用在３ｄ后基本恢复     

病毒合剂中黄芩甙的高效液相色谱法定量分析

对病毒合剂中有效成分之一－－黄芩甙的提取分离方法、薄层层检鉴别及高效液相含量分析方法进行研究，方法简便，重现性好，加样回收率９８．７％，ＲＳＤ％＝１．２４。     

1α,25-二羟维生素D_3对患者PBMC分泌的IFN-α和IL-10水平的影响

目 的 探 讨 １α，２５－二羟 维生 素 Ｄ_３（１α，２５－（ＯＨ ）２Ｄ ）对 系 统性 红斑 狼 疮（ＳＬＥ）患 者 外周 血 单个 核 ３细 胞（ＰＢＭ Ｃ）分 泌的 白细 胞 介素 （ＩＬ）－１０ 和 干扰 素 （ＩＦＮ ）－α水平 的 影响 。方法 ２０ 例 ＳＬＥ 患 者和 １０ 名健 康 志愿者 ＰＢＭ Ｃ 的提 取 采 用 Ｆｉｃｏｌ密 度 梯 度 离 心 法 ，ＳＬＥ 组 和 对 照 组 在 不 加 或 加 入 １α，２５－（Ｏ Ｈ ）２Ｄ３ 的 情 况 下 孵 育 ７２ ｈ收 集培 养 上清 ，上 清液 ＩＬ－１０ 和 ＩＦＮ－α水平 检 测采 用酶 联 免疫 吸附 试 验（ＥＬＩＳＡ ）。结 果 与 正常 对 照组 相比 ，ＳＬＥ组 ＰＢＭ Ｃ 中 ＩＬ－１０ 和 ＩＦＮ－α水 平 明 显 增高 （Ｐ＜０．０１）。０．０１ ｍ ｏｌ／Ｌ １α，２５－（Ｏ Ｈ ）２Ｄ３ 的 加 入 明 显 抑 制 了 ＳＬＥ 组ＰＢＭ Ｃ 中 ＩＦＮ －α的 产 生 （Ｐ＜０．０１），增 加 了 ＳＬＥ 组 ＰＢＭ Ｃ 中 ＩＬ－１０ 的 产 生 （Ｐ＜０．０１），但 对 正 常 对 照 组 ＩＦＮ －α和ＩＬ－１０ 水平 无明 显 影响 。结 论 １α，２５－（ＯＨ ）２Ｄ 可 能 抑制 体外 培 养的 ＳＬＥ 患 者 ＰＢＭ Ｃ ＩＦＮ －α的产 生     

Glycine‐modified growth hormone secretagogues identified in seized doping material

A number of unknown pharmaceutical preparations seized by Danish customs authorities were submitted for liquid chromatography–high resolution mass spectrometry (LC–HRMS) analysis. Comparison with reference standards unequivocally identified the content of the powders as analogs of the growth hormone secretagogues GHRP‐2 (Pralmorelin), GHRP‐6, Ipamorelin, and modified growth hormone releasing factor (modified GRF 1–29), which can be used as performance‐enhancing substances in sports. In all cases, the detected modification involved the addition of an extra glycine amino acid at the N‐terminus, and analytical methods targeting growth hormone secretagogues should hence be updated accordingly.     

Degradation pathways,analytical characterization and formulation strategies of a peptide and a protein calcitonine and human growth hormone in comparison

Peptides and proteins differ from conventional chemical entities in their sensitivity to numerous environmental factors and their susceptibility to different degradation pathways. Therefore, complex analytical methodologies are necessary to characterize their molecular entity as well as to detect and quantify the possible degradation products. The formulation of these molecules for a pharmaceutical product requires stabilization by various excipients. Most of the products are brought to market as solutions or lyophilisates. In the first part, this article presents a comparison between the degradation profile of a peptide (calcitonine) and a protein (human growth hormone), in solution and as a freeze-dried product. The various analytical methods used to characterize and identify the degradation products are reviewed and discussed. The second part contains an overview of the different formulation strategies for calcitonine and human growth hormone. Finally, the different stress conditions used to obtain stability data are discussed critically. This leads on to general comments on the design of stability studies for peptide and protein drugs as pharmaceuticals taking into consideration the official guidelines.     

The use of LC/MS, GC/MS, and LC/NMR hyphenated techniques to identify a drug degradation product in pharmaceutical development

Understanding drug degradation in the formulated product is critical in pharmaceutical development as it has significant impacts on drug efficacy, safety profile and storage conditions. As a result, identification of degradation compounds has taken an important role in the drug development process. In this study, various hyphenated analytical techniques, such as liquid chromatography mass spectrometry (LC/MS), gas chromatography mass spectrometry (GC/MS), and liquid chromatography nuclear magnetic resonance with a solid phase extraction interface (LC/SPE/NMR), have been applied to the identification of a drug degradation product which grew over time in the stability study of the drug product. The target unknown is less polar and more unsaturated than the drug substance based upon reverse phase HPLC relative retention time and UV spectra. It is not ionizable by electrospray ionization (ESI) or atmospheric pressure chemical ionization (APCI) in either a positive or a negative mode. The unknown was isolated by an HPLC fraction collector and enriched by solid phase extraction. GC/MS with chemical ionization (CI) was employed to determine the molecular weight of this compound. Its fragmentation pattern was determined by CI-MS/MS using an ion trap mass spectrometer. The isolated material was also analyzed by LC/SPE/NMR, from which the structure of this compound was further characterized. The study utilizes a combination of various hyphenated analytical techniques to obtain complimentary information for structure elucidation of the unknown. The combination approach is critical for unambiguous impurity structure elucidation in drug degradation studies of pharmaceutical drug products.     

Somatropin and its variants: structural characterization and methods of analysis

Human Growth hormone (hGH, somatotrophin) is a 22 kDa, 191 amino-acid single chain protein produced by somatroph cells of the anterior pituitary gland. It is the major physiological regulator of growth, and deficiencies in growth hormone levels have long been recognized as the underlying cause of growth disorders (dwarfism). The ability of exogenous hGH to restore normal rates of growth in both human and animal models of growth retardation has long been recognized and the use of hGH in therapy goes back several decades. Initial preparations were prepared by extraction and purification from cadaveric pituitary tissue, and since 1984, hGH has been prepared by recombinant Deoxyribosenucleic acid (rDNA) technology. As is usually the case with "biologicals", characterization of the drug substance depended on a combination of physico-chemical and biological methods, and the hGH molecule became well characterized fairly early in its life as a drug. Indeed, by 1980 the major degradation forms and structural variants of the hGH molecule had been described and reviewed. Little satisfactory progress had been made in refining biological assays for hGH, and, although in vitro assays were described, potency-defining assays remained dependant on the whole body growth response in rats, and were both invasive and imprecise. In the early 1990's a series of collaborative studies on analysis of recombinant hGH (somatropin) established that available bioassays were much less selective that physico-chemical methods in detecting and quantifying structural degradation, and 1994 saw an international consensus to replace the bioassays with physico-chemical analytical methods for the routine batch release of somatropin. During the last decade in most markets somatropin has, unusually for a protein, been subject to batch release and control dependent entirely on physico-chemical analysis, without the routine use of any form of bioassay. During that time there has been a continuous development and refinement of methods, and the identification of a range of structural variants and degradation products of the molecule. The present review sets out to summarise the current knowledge on physico-chemical analytical methods for somatropin, and the structural variants that have been identified and characterized. A survey of available biological analytical methods is beyond the scope of this review, as is consideration of the earlier pituitary preparations and the recombinant 192 amino-acid methionyl form of the molecule (somatrem), although it is likely that many of the methods and variants described would be equally applicable to somatrem.     

利用LC-MS和二维色谱相关光谱技术识别HPLC色谱图中杂质峰

采用二维色谱相关光谱技术,实现质控HPLC方法与LC-MS色谱系统色谱峰的相互识别。本文采用HPLC-DAD(中国药典2010版方法)得到头孢唑肟和头孢地尼混合降解杂质的色谱图,建立降解杂质标准色谱光谱数据;采用LC-MS的方法结合MassWorks软件的应用鉴别出头孢唑肟和头孢地尼的主要降解产物;通过计算光谱相关性对杂质色谱峰进行归属。色谱二维光谱相关法在不同色谱系统下能够准确定性,在没有杂质对照品的情况下,可为质控HPLC方法中的杂质峰的归属提供新的途径和新的思路。     

Profiling impurities and degradants of butorphanol tartrate using liquid chromatography/mass spectrometry and liquid chromatography/tandem mass spectrometry substructural techniques

A rapid and systemic strategy based on liquid chromatography/mass spectrometry (LC/MS) profiling and liquid chromatography/tandem mass spectrometry (LC/MS/MS) substructural techniques was utilized to elucidate the degradation products of butorphanol, the active ingredient in stadol® NS. This strategy integrates, in a single instrumental approach, analytical HPLC, UV detection, full-scan electrospray mass spectrometry, and tandem mass spectrometry to rapidly and accurately elucidate structues of impurities and degradants. In these studies, several low-level degradation products were observed in long-term storage stability samples of bulk butorphanol. The resulting analytical profile includes information on five degradants including molecular structures, chromatographic behavior, molecular weight, UV data, and MS/MS substructural information. The degradation products formed during long-term storage of butorphanol tartrate included oxidative products proposed as 9-hydroxy- and 9-keto-butorphanol, norbutorphanol, a ring-contraction degradant, and Δ1, 10-butorphanol. These methodologies are applicable at any stage of the drug product cycle from discovery through to development. This library of butorphanol degradants provides a foundation for future development work regarding product monitoring, as well as a useful diagnosite tool for new degradation products.     

Analysis and determination of biological activity of short-chain peptides from porcine brain hydrolysate

Gel permeation chromatography fractions of short-chain peptides from a hydrolysate product, which in turn was from the purified porcine brain through enzyme hydrolysis, were tested for their biological activities. The results showed that the fractions A4 and A5 had significant biological activities. The two fractions were analyzed with analytical techniques such as high-performance liquid chromatography (HPLC), capillary zone electrophoresis (CZE), isoelectric focusing (IEF), discontinuous sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Preliminary results showed that the main components of these two fractions were short-chain acidic peptides with a relative molecular mass (M(r)) of less than 2400.     

Primary structure of fox pituitary growth hormone

Tryptic digests of fox growth hormone (fGH) were separated by reverse phase high performance liquid chromatography (HPLC) and by paper electrophoresis. From the amino acid composition of these tryptic peptides and from their alignment with the expected tryptic peptides from bovine growth hormone (bGH), the primary structure of fGH is proposed. There are only 17 amino acid residues which are different in these two growth hormone molecules.     

Induction of growth hormone by the roots of<Emphasis Type="Italic">Astragalus membranaceus</Emphasis> in pituitary cell culture

The traditional Asian medicinal herb, roots of Astragalus (A.) membranaceus (Leguminosae), is used for many purposes, some of which are purported to stimulate the release of growth hormone in vivo. Extracts of A. membranaceus were tested to determine whether they stimulate the release of growth hormone in rat pituitary cell culture. A. membranaceus was extracted sequentially with 80% ethanol (fraction A), n-hexane (fraction B); the test compound from the herbal extraction was isolated using silica gel column chromatography and was identified with spectral data. Test compound was also extracted by traditional boiling water methods. Induction of growth hormone in pituitary cell culture was conducted with isolated compounds and extracted fractions of A. Radix (dried roots of A. membranaceus). The fraction A was not active in the rat pituitary cell culture, but the fraction B derived from the ethanol fraction stimulated the release of growth hormone in culture. Six compounds from fraction B (1-6) were isolated and identified previously. The compounds 1,2-benzendicarboxylic acid diisononylester (1), beta-sitosterol (2), and 3-O-beta-D-galactopyranosyl-beta-sitosterol (5) did not induce growth hormone release in the culture. Formononetin (3), 9Z,12Z-octadecadienoic acid (4), stigmast-4-en-6beta-ol-3-one (6) and 98-E, a mixture of 1'-9,12-octadecadienoic acid (Z,Z)-2',3'-dihydroxy-propylester (7) and 1'-hexadecanoic acid-2',3'-dihydroxy-propylester (8) stimulated the release of growth hormone in the rat pituitary cell culture significantly compared to the control. In conclusions, four compounds isolated from extracts of A. Radix induced growth hormone release in the rat pituitary cell culture. The 98-E isolate was the most active inducer of growth hormone release.     

指纹图谱在中药质量控制中的应用

随着现代分析技术和信息科学的发展．中药指纹图谱技术在中药质量控制中发挥着重大作用。本文概述了近年来中药指纹图谱的构建方法、相似度评价方法、化学模式识别、存在问题。