尸体脏器中混合碱性药物及其主要代谢物的毛细柱GC／MS检验

本文详细介绍了尸体脏器中吩噻嗪类、苯骈二氮杂类等混合碱性药物的毛细柱ＧＣ／ＭＳ检验方法，并对药物代谢物及其提取方法进行了讨论。     

Sorbent Extraction and Analysis of Volatile Metabolites Synthesized by Lactic Acid Bacteria in a Synthetic Medium

Volatile metabolites synthesized by strains of Luctobacillus brevis, L. fructivorans, L. hilgardii, L. plantarum, Pediococcus parvulus, and Leuconostoc oenos, originally isolated from commercial wines, were extracted from a synthetic medium using C₁₈ sorbent cartridges and analyzed using gas-liquid chromatography. All organisms synthesized isoamyl alcohol, 1-hexanol, 3-(methylthio)-1-propanol, and 2-phenyl ethanol in varying concentrations. Several other alcohols, acids, aldehydes, and esters were produced or utilized by some species or strains and enabled differentiation of the organisms. Recoveries of metabolites were dependent on the compounds, matrix, and presence of ethanol.     

粉葛愈伤组织不同极性提取物生物活性与功能成分分析

目的 为探究粉葛愈伤组织次生代谢产物的生物活性和功能成分。方法 采用系统溶剂法制备粉葛愈伤组织石油醚提取物(petroleum ether extract,PE)、乙酸乙酯提取物(ethyl acetate extract,EE)、甲醇提取物(methanol extract,ME)和水提取物(water extract,WE)等4种不同极性次生代谢产物,利用8株细菌、2个自由体系和1个细胞系测试四种提取物的抗菌、抗氧化和细胞毒活性,分析4种提取物中总黄酮、总酚、总多糖等主要功能成分含量与生物活性的相关性,并运用高效液相色谱法(high performance liquid chromatography,HPLC)检测其单体功能成分。结果 四种提取物的生物活性显著差异,其中EE对粪肠球菌、金黄色葡萄球菌、藤黄微球菌等革兰氏阳性菌作用突出,其最小抑菌浓度(minimum inhibitory concentration,MIC)为0.195~0.391 mg.mL_-1^,EE对1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)和2,2’-联氮双(3-乙基苯并噻唑啉-6-磺酸)二铵盐(2,2’-azinobis-(3-ethylbenzthiazoline-6-sulphonate),ABTS)自由基清除能力均最佳,其半抑制浓度(half maximal inhibitory concentration,IC₅₀)分别为0.2795和0.0640 mg.mL_-1^,EE对人肺癌细胞A549(human lung cancer cell)的抑制能力较佳,其半最大效应浓度(concentration for 50% of maximal effect,EC₅₀)为260.9 μg.mL_-1^{。总黄酮和总酚在}EE中含量均最高,分别为(273.89±7.48)μg.mg_-1^和(261.00±14.94)μg.mg_-1^,总多糖在WE中含量最高,达到(402.25±13.49)μg.mg_-1^,相关性分析显示,总黄酮为重要的抗菌和抗氧化活性成分。异黄酮单体功能成分在四种提取物中组成和含量差异均较大,且主要集中在EE和ME,其中EE以大豆苷(30.3428±1.4251)μg.mg_-1^{和大豆苷元}(45.7235±2.0230)μg.mg_-1^为主,ME以大豆苷(60.0125±3.9250)μg.mg_-1^{和染料木苷}(20.5655±1.3025)μg.mg_-1^为主。结论 粉葛愈伤组织次生代谢产物具有较强的抗菌、抗氧化活性以及微弱的细胞毒活性,主要含有大豆苷、染料木苷和大豆苷元等功能成分,该结果为粉葛愈伤组织次生代谢产物在食品与制药领域的开发利用提供了一定的参考。     

汾酒酿造过程中主要功能微生物的筛选和研究

以汾酒酿造过程中分离到的微生物为研究对象,模拟汾酒生产条件进行了纯种发酵试验,采用气相与液相色谱法检测了其主要代谢产物。初筛结果发现,主要代谢产物包括乙醛、乙醇、乙酸乙酯、乳酸和乙酸等。依代谢产物的合成情况,选出相应高产菌株进行了底物水平的代谢调控发酵试验,确定了30株与主要代谢产物相关的功能菌株,并对其进行了分类鉴定。它们分别属于毕赤氏酵母属、卵胞酵母属、酵母菌属、芽孢杆菌属、乳酸杆菌属和棒状杆菌属等。本文研究结果为探索汾酒酿造机理和优化酿造工艺提供了一定的理论依据和菌种资源。     

Metabolite profiling of doenjang,fermented soybean paste,during fermentation

BACKGROUND: A fermented soybean paste known as doenjang is a traditional fermented food that is widely consumed in Korea. The quality of doenjang varies considerably by its basic ingredients, species of microflora, and fermentation process. The classification of predefined metabolites (e.g. amino acids, organic acids, sugars and sugar derivatives, and fatty acids) in doenjang samples according to fermentation was performed by using GC‐FID and GC‐MS data sets with the application of a multivariate statistical method. RESULTS: The predominantly produced amino acids included alanine, valine, leucine, isoleucine, proline, glutamine, phenylalanine and lysine, showing remarkable increases in amounts during the later stages of fermentation. Carbonic acid, citric acid, lactic acid and pyrogultamic acid were identified as the major organic acids. Significant amounts of erythrose, xylitol, inositol and mannitol were detected during fermentation. Regarding fatty acids, relatively higher amounts of palmitic acid, stearic acid, oleic acid, linoleic acid and linolenic acid were found in the doenjang at each fermentation time point. Principal component analysis (PCA) successfully demonstrated changes in composition patterns as well as differences in non‐volatile metabolites according to fermentation period. CONCLUSION: A set of metabolites could be determined representing the quality of doenjang during fermentation, and which might also be correlated with taste ingredients, flavour, nutrition, and physiology activities that are claimed to be dependent on the quality control of commercial doenjang. Copyright © 2010 Society of Chemical Industry     

Pharmacological Perspectives on the Detoxification of Plant Secondary Metabolites: Implications for Ingestive Behavior of Herbivores

Plant secondary metabolites (PSMs) are a major constraint to the ingestion of food by folivorous and browsing herbivores. Understanding the way in which mammalian detoxification pathways are adapted to deal with PSMs is crucial to understanding how PSMs influence ingestive behavior of herbivores and hence their fitness and the impact that they have on vegetation. Pharmacological concepts can provide insights into the relationship between the absorption and metabolic fate of PSMs and ingestive behavior. Lipophilic PSMs will be absorbed into the bloodstream and must be removed fast enough to prevent their accumulation to toxic levels. Elimination depends on their metabolism, usually by cytochrome P450 enzymes, to more polar metabolites that can be excreted by the kidney. The concentration of PSM in blood (C) is a better measure of exposure to a toxin compared to the amount ingested because there can be great variability in the rate and degree of absorption from the gut. C rises and falls depending on the relative rates of absorption and elimination. These rates depend in part on metabolic and transport processes that are saturable and liable to inhibition and induction by PSMs, indicating that complex interactions are likely. Herbivores can use diet choice and the rate and amount of PSM consumption to prevent C from reaching a critical level that produces significant adverse effects.     

Application of Pharmacological Approaches to Plant–Mammal Interactions

The dominant theory in the field of mammalian herbivore–plant interactions is that intake, and therefore tolerance, of plant secondary metabolites (PSMs) is regulated by mechanisms that reduce absorption and increase detoxification of PSMs. Methods designed by pharmacologists to measure detoxification enzyme activity, metabolite excretion, and most recently, drug absorption, have been successfully applied by ecologists to study PSM intake in a variety of mammalian study systems. Here, we describe several pharmacological and molecular techniques used to investigate the fate of drugs in human that have potential to further advance knowledge of mammalian herbivore–plant interactions.     

Quantitative GC-ECD Analysis of Halogenated Metabolites: Determination of Surface and Within-Thallus Elatol of Laurencia obtusa

Information on natural concentrations or variability of secondary metabolites in marine organisms may be important both to ecological/evolutionary and applied approaches. A gas chromatographic procedure with an electron capture detector (GC-ECD) was developed to quantify the sesquiterpenoid elatol at the surface and within-thalli of 70 specimens of the red seaweed Laurencia obtusa. The concentration of elatol was highest within-thalli [9.89 mg g⁻¹ of L. obtusa, dry weight (d.w.)], compared to lower values found at the surface [0.006 mg g⁻¹ of L.obtusa (d.w.), or 0.5–10.0 ng cm⁻²]. This method provides a rapid and inexpensive quantification of small quantities of elatol, and probably may also be used to quantify other halogenated compounds usually found in red seaweeds.     

Unraveling the Secrets of Protein–Metabolite Interactions

Saghatelian and colleagues recently introduced a global metabolite‐profiling approach that allows protein–metabolite interactions (PMI) to be identified. This approach represents an excellent strategy and valuable tool for unraveling the many secrets of the metabolome. The key features of the methodology will be summarized here.