Metabolomics in necrotizing enterocolitis: the state of the art

Introduction: Necrotizing enterocolitis (NEC) is a serious inflammation of the bowel that most often affects premature infants. The exact cause of NEC is still poorly understood but investigators believe that these different factors can play an important role: prematurity and immaturity of the intestine of the preterm newborns, differences in the intestinal bacterial colonization and an abnormal immune response. Since the early symptoms of NEC are often rather non-specific, the diagnosis of NEC can be difficult. Therefore, the search for diagnostic biomarkers for NEC remains warranted. This review focuses on the results of recent metabolomics investigations on NEC, providing important contributions to the understanding of the biochemical process characterizing this disease.

Areas covered: Only a few metabolomics applications of NEC are reported in the literature. The metabolic pathway principally altered in NEC patients is suggested to be linked to the carbohydrates metabolism and the role of gluconate as a predictive marker for this pathology is investigated.

Expert commentary: Biomarkers are crucial as a confident diagnostic tool to make NEC therapy more efficient. Although only a few metabolomics studies have been performed in NEC research, this technique proved to be helpful to understand the pathophysiological mechanisms of this disease.     

基于UPLC-Q-TOF-MS和GC-MS的小金丸化学成分识别

目的：采用超高效液相色谱与串联四极杆飞行时间质谱（UPLC-Q-TOF-MS）和气相色谱-质谱联用（GC-MS）技术快速鉴定小金丸中的化学成分,构建较为全面的小金丸化学成分谱,为小金丸药效物质基础研究奠定基础。方法：UPLC-Q-TOF-MS分析采用ACQUITY UPLC BEH C₁₈（Waters,2.1 mm×100 mm,1.7 μm）超高效液相柱,流动相为乙腈-0.1%甲酸水,梯度洗脱;飞行时间质谱采用正、负离子模式扫描,质谱数据以MSE模式采集。UPLC-Q-TOF-MS分析依据离子峰的保留时间、精确分子质量和多级质谱碎片,与文献报道数据和数据库对比,对小金丸的甲醇提取部分化学成分进行在线快速鉴定。GC-MS分析采用Agilent 19091S-433 HP-5MS 5% Phenyl Methyl Silox（Agilent,30 m×250 μm×0.25 μm）毛细管色谱柱,载气为氮气;质谱采用EMV模式：增益因子1.00。GC-MS分析通过增强型NIST4.L标准质谱库进行检索,对小金丸中挥发性成分进行了快速鉴定。结果：从小金丸中共鉴定化学成分118种,主要包括生物碱类24种、有机酸12种、甾体类9种、单萜类5种、倍半萜类16种、二萜类14种、三萜类7种和苯丙素类成分5种,另外还有个别酯类、糖类、氨基酸类和杂环类化合物等。结论：UPLC-Q-TOF-MS和GC-MS方法能快捷、准确、较全面地鉴定小金丸中的化学成分,为小金丸药效物质基础研究提供科学依据,并为中药复方的成分鉴定提供了一种简单快捷的分析鉴定方法。     

白及花与块茎石油醚部位成分分析及其生物活性的比较研究

目的 分析白及花和块茎石油醚部位成分及生物活性。方法 以石油醚（60~90℃）为提取溶剂,采用索氏提取法提取白及花和块茎脂溶性成分,采用GC-MS分析其化学成分,通过滤纸片扩散法、CCK8法和分光光度法分别对其进行抑菌、抗肿瘤和α-淀粉酶抑制作用研究,通过检测DPPH自由基、ABTS自由基和羟自由基的清除率考察其体外抗氧化活性。结果 白及花和块茎中共鉴定出20种石油醚部位成分,其中花11种,块茎19种,共有成分10种,α-乙酰基-γ-丁内酯（15.20%,21.84%）和苄醇（12.07%,24.10%）为主要成分。白及花中石油醚部位成分的抑菌活性高于块茎,对枯草芽孢杆菌效果最好,最小抑菌浓度和最小杀菌浓度分别为0.41,0.51 g·L^-1和0.53,0.66 g·L^-1。当质量浓度为5.0 mg·mL^-1时,白及块茎石油醚部位成分对A₅₄₉细胞的抑制率为29.451%,高于花中石油醚部位成分27.621%的抑制率,前者对α-淀粉酶的IC₅₀为1.819 mg·mL^-1,后者的IC₅₀为2.028 mg·mL^-1。两者对DPPH自由基、ABTS自由基和羟自由基的清除率均随质量浓度的增大而增大,当浓度达到1.0 mg·mL^-1时,前者对ABTS自由基和羟自由基的清除率分别为99.67%和92.22%,高于后者90.07%和38.11%的清除率,后者对DPPH自由基的清除率为31.23%,高于前者18.94%的清除率,两者均低于Vc的清除率。结论 白及花和块茎中石油醚部位成分主要为酯类和醇类,2种石油醚部位成分均具有一定的生物活性,具有潜在的药用价值,并可为白及花的资源开发提供参考。     

Quantification of Silicone Oil and Its Degradation Products in Aqueous Pharmaceutical Formulations by 1H-NMR Spectroscopy

During the past years, there has been an increasing focus on the presence of silicone oil as a contaminant in pharmaceutical formulations kept in prefilled syringes (PFSs). As the PFSs are coated on the inner wall with silicone oil (polydimethylsiloxane), there is a potential risk that the oil can migrate from the inner surface of the primary packing material into the aqueous solution. Several studies have demonstrated that presence of silicone oil as droplets in a high-concentrated protein formulation can cause protein aggregation. Hence, because the use of silicone-coated primary packing material for protein formulations are increasing, the call for an easy and quantitative method for determination of silicone oil and its degradation products in pharmaceutical formulations is therefore needed. Several analytical techniques have in the past been developed with the aim of detecting the presence of silicone oil and degradation products hereof. Most of these methods require hydrolyzation, derivatization, and extraction steps followed by, for example, gas chromatography-mass spectrometry analysis. Applying these methods can cause a loss in detection or an overestimation of the hydrolytic degradation products of silicone oil, that is, trimethylsilanol and dimethylsilanediol. The 2 silanols are highly hydrophilic and prefers the aqueous environment. Analysis of an aqueous formulation obtained from a PFS by ¹H-NMR spectroscopy provides data about the content and levels of silicone oil and the 2 silanols even in levels below 10 ppm. The ¹H-NMR method offers an easy and direct, quantitative measurement of samples intended for clinical use and samples kept at elevated temperature for a prolonged time (i.e., stability studies). The result of the study presented here showed dimethylsilanediol to be the main silicone compound present in the aqueous formulation when kept in baked-on PFSs. The degradation product dimethylsilanediol, in full accordance with expected hydrolytic degradation of silicone oil, increased during storage and with elevated temperature. In addition, the method can be applied to aqueous samples where polydimethylsiloxane has been added as, for example, the major constituent of antifoam.     

饮用水中多种有机磷农药残留的气相色谱-三重四极杆质谱测定法

目的建立一种同时测定饮用水中7种有机磷农药残留量的气相色谱-三重四极杆质谱（GC-MS/MS）方法。方法采用液-液萃取方法,用二氯甲烷对水样中7种有机磷农药进行提取,无水硫酸钠脱水,滤液在40℃水浴中旋转蒸发近干,用二氯甲烷定容,GC-MS/MS测定,外标法定量。结果 7种有机磷农药在21 min内分离,在0.01-10 mg/L范围内相关系数r2均在0.99以上,在0.05、0.25和0.45 mg/L 3个水平加标回收率在78.9%-99.3%之间,相对标准偏差均小于10%,检出限为0.02-0.06μg/L。结论该方法操作简单,准确度和精密度高,消除了其他有机磷农药的干扰,避免假阳性的出现,适用于饮用水中7种有机磷农药残留量的检测和确证。     

Quality control tests for the crude drug of Conyza filaginoides

Context. Conyza filaginoides (D.C.) Hieron (Asteraceae) is a medicinal Mexican plant highly prized in contemporary Mexico for the treatment of upset stomach and diabetes.

Objective: This work was undertaken to develop a suitable high performance liquid chromatography (HPLC)-diode array detection (DAD) method for quantifying rutin (1), the main active principle from the aerial parts of C. filaginoides.

Materials and methods: The method was performed using a LiChrospher 100 RP-18 column. The mobile phase was water (containing 0.1% phosphoric acid)-methanol-acetonitrile (80:5:15, v/v) at a flow rate of 1.2?mL?min^?1.

Results: Limits of detection and quantification were 7.5 and 22.8?μg?mL^?1, respectively. The main recoveries measured at three concentrations were higher than 98%, with RSD <2%. Quantitative analysis of a few samples showed the presence of high concentrations of 1 (3.6?±?0.2?g/100?g of dry plant material). The volatile components were extracted by hydrodistillation or head space solid-phase microextraction (HS-SPME), and thereafter analyzed by gas chromatography coupled to mass spectrometry (GC-MS). Forty-three chemical constituents representing 90% of the total content of the oil were identified. The major light volatile compounds obtained by HS-SPME revealed a high content of monoterpene hydrocarbons.

Conclusions: A precise, reliable, and accurate analytical HPLC method to detect and quantify 1 in the crude drug and some preparations were developed and fully validated. The volatile components of the plant are described for the first time. The proposed method would be useful for quality control assurance of this important Mexican plant.     

The in vivo evaluation of anti-angiogenic effects of Hypericum essential oils using the chorioallantoic membrane assay

Context. Hypericum species including Hypericum confertum Choisy, H. hircinum L., H. hyssopifolium Chaix. subsp. elongatum (Ledeb.) Woron var. microcalycinum (Boiss. &; Heldr.) Boiss. and H. perforatum L. (Clusiaceae) are used as medicinal plants in Turkey.

Objective: The anti-angiogenic evaluation of Hypericum essential oils using the chick embryo chorioallantoic membrane (CAM) assay are performed with this study for the first time.

Materials and methods: The anti-angiogenic activity of Hypericum essential oils (0.5–5.0?mg/ml) was evaluated in vivo using the CAM assay, compared to standard anti-angiogenic substances at the same concentrations, in trice replicated independent assays. GC and GC-MS analyses were carried out simultaneously to identify the chemical compositions of the Hypericum essential oils.

Results: The CAM treated with H. perforatum essential oil showed anti-angiogenic effect (score 0.6?±?0.3) at 50?µg/pellet concentration, whereas other tested Hypericum essential oils showed no effect compared to the standards (e.g. suramin score 0.5?±?0.2). Furthermore, the tested oils showed neither membrane toxicity nor irritation at the tested concentrations. The major compound of the essential oil of H. confertum was identified as germacrene D (30.2%). The major compound of the essential oils of the H. hircinum. H. hyssopifolium subsp. elongatum var. microcalycinum and H. perforatum was identified as α-pinene (88.3, 57.8, 33.3%), respectively.

Discussion and conclusion. Hypericum species and in particular H. perforatum essential oil may have important effect toward wound healing and various inflammations. The data obtained in this experiment suggest further investigations on various cancers due to its anti-angiogenic effects observed.     

顶空GC-MS法测定去甲斑蝥素中的有机溶剂残留

目的：建立去甲斑蝥素中有机溶剂残留量的测定方法。方法：采用顶空毛细管气相色谱法,HP-5MS柱（30m×0．25mm×0．25μm）,火焰离子化检测器（FID）,以水为溶剂,进行气相色谱分析,并以电子轰击（EI）为离子源,进行质谱（MS）分析。结果：证实残留溶媒为丙酮。结论：本方法简单、准确、灵敏,重复性好,适用于去甲斑蝥素中有机溶剂残留量的测定。关键词去甲斑蝥素;顶空气相色谱一质谱法;有机溶剂;残留量     

阔叶五层龙的化学成分研究

目的对阔叶五层龙所含化学成分进行研究。方法应用液-液萃取及多种柱层析方法进行化学成分的分离、纯化,并利用多种光谱技术进行结构的分析鉴定。结果从阔叶五层龙植物中分离得到9个化合物：2-hydroxyfriedelan-3-one (1),软木三萜酮 (2), lup-20(29)-en-3, 21-dione (3),D-friedoolean-14-en-3-one (4),3-(3″,4″-dihydroxy-trans-cinnamoyloxy)-D- friedoolean-14-en-28-oic acid (5), 3, 22-dioxo-29-normoretane (6), 羽扇豆醇 (7), β-谷甾醇 (8), β-胡萝卜苷(9)。结论该9个化合物均为首次从阔叶五层龙中分离得到。     

野生和栽培鲜肿节风挥发油成分分析

目的鉴定野生和栽培鲜肿节风挥发油中的成分。方法应用气相色谱一质谱联用技术和NIST-98、WILEY275检索库对两种挥发油的化学成分进行分析和鉴定。结果从分离出的115种物质中野生和栽培鲜肿节风挥发油分别鉴定了65和64种化学成分,其相对含量占挥发油总量分别为76．3％和77．8％,其中两者共有化学成分为48个,其相对含量总和占各自总检出成分分别为74．38％和79．32％。结论野生品与栽培品的挥发油成分、共有成分的相对百分含量有差异,有些相差很多倍;共有成分所占比例较大,且共有成分总含量接近。该研究为肿节风的开发利用提供了理论依据。