基于UHPLC-Q-TOF/MS的不同产地普洱生茶化学成分差异研究

为了更加全面地了解产地对普洱生茶品质与化学成分的影响,本研究选取来自临沧市、普洱市、西双版纳州三大产区12个茶山(自然村)的普洱生茶样品,采用超高效液相色谱-四级杆-飞行时间质谱(UHPLC-Q-TOF/MS)对不同产地普洱生茶的非挥发性代谢物表型进行了研究。结果表明:不同产地普洱生茶的化学成分在含量上具有较大差异,且具有明显的产地特征。采用主成分分析,可以对来自西双版纳自治州(包括勐腊县、勐海县、景洪市)、普洱市、临沧市3个地级行政区的普洱生茶进行有效区分,也可以对来自普洱茶产区的东南、西南、西北3个区域的普洱生茶进行有效区分。进一步鉴定了普洱生茶中79种主要成分,并对其在12个茶山(自然村)的普洱生茶中的含量分布,以及与不同产地普洱生茶滋味品质的关系进行了分析。本研究表明,基于UHPLC-Q-TOF/MS的茶叶非挥发性化学成分轮廓可以作为普洱生茶产地判别的依据。     

用液相色谱串联质谱法测定渔业水样中6种农药的含量

本文建立了一种快速检测渔业水质样本中酰胺类除草剂乙草胺、丙草胺、丁草胺、三嗪类除草剂莠去津、烟碱类杀虫剂噻虫嗪、三唑类杀菌剂苯醚甲环唑6种农药含量的液相色谱串联质谱法。采用乙腈-水-甲酸梯度洗脱、电喷雾电离方式、多反应监测模式测定考察检出限、线性范围、日内精密度及日间精密度。在三种加标浓度下,该方法的回收率为78.2%~104.7%。用液相色谱-四级杆-飞行时间质谱对疑似目标物质进行定性,用精确质量数、同位素丰度比和二级碎片解析方式等解析定性结构。该方法能够满足检测这六种农药的要求,能准确定性,自建的谱库可为渔业污染事故鉴定和水产品质量安全风险评估做支撑。     

Characterizing the pigmented traditional rice cultivars grown in temperate regions of Kashmir (India) for free and bound phenolics compounds and in vitro antioxidant properties

The purpose of the research was to identify the phenolic and flavonoid compounds of seven different traditional pigmented whole rice cultivars grown in the temperate regions of Kashmir so as to study their relationship with in vitro antioxidant capacities. The completely pigmented rice cultivars were found to have higher phenolic, flavonoid, anthocyanin contents and exhibited higher antioxidant capacities than the light colored and sparely colored rice cultivars. A total of 40 compounds had been identified in the analyzed rice cultivars that were found to be distributed in 6 major categories with 6-phenolics, 6-flavonoids, 11-hydroxycinnamic acid derivatives, 7-hydroxybenzoic acid derivatives, 3-anthocyanins and 7-flavonoid glucosides of different flavonoid compounds. Among the free and bound fractions for each cultivars the light and sparsely colored depicted higher content of phenolics and in vitro antioxidant properties in bound faction, while the completely pigmented cultivars showed higher antioxidant properties in free fractions. The anthocyanins quercetin-3-O-galactoside, cyanidin-3-O-rutinoside and pelargonidin-3-O-diglucoside had been identified by LC-MS existing in the free fractions of the analyzed rice cultivars whereas, the free fraction of acetone + H₂O possessed higher percentage of phenolic compounds as compared to methanolic extracts and bound fractions. The black colored cultivars possessed higher DPPH scavenging activity and lipid peroxidation inhibition.     

Analysis of Mycosporine-Like Amino Acids in Selected Algae and Cyanobacteria by Hydrophilic Interaction Liquid Chromatography and a Novel MAA from the Red Alga Catenella repens

Mycosporine-like amino acids (MAAs), a group of small secondary metabolites found in algae, cyanobacteria, lichens and fungi, have become ecologically and pharmacologically relevant because of their pronounced UV-absorbing and photo-protective potential. Their analytical characterization is generally achieved by reversed phase HPLC and the compounds are often quantified based on molar extinction coefficients. As an alternative approach, in our study a fully validated hydrophilic interaction liquid chromatography (HILIC) method is presented. It enables the precise quantification of several analytes with adequate retention times in a single run, and can be coupled directly to MS. Excellent linear correlation coefficients (R² > 0.9991) were obtained, with limit of detection (LOD) values ranging from 0.16 to 0.43 µg/mL. Furthermore, the assay was found to be accurate (recovery rates from 89.8% to 104.1%) and precise (intra-day precision: 5.6%, inter-day precision ≤6.6%). Several algae were assayed for their content of known MAAs like porphyra-334, shinorine, and palythine. Liquid chromatography-mass spectrometry (LC-MS) data indicated a novel compound in some of them, which could be isolated from the marine species Catenella repens and structurally elucidated by nuclear magnetic resonance spectroscopy (NMR) as (E)-3-hydroxy-2-((5-hydroxy-5-(hydroxymethyl)-2-methoxy-3-((2-sulfoethyl)amino)cyclohex-2-en-1-ylidene)amino) propanoic acid, a novel MAA called catenelline.     

几种经济红藻中茉莉酸合成途径的关键物质

为研究几种经济红藻中茉莉酸合成途径的关键物质,实验采用液相色谱—质谱联用技术(LC-MS)对茉莉酸生物合成途径相关物质建立检测方法,并对龙须菜、坛紫菜受到机械损伤时各物质的变化情况进行检测。以100%甲醇提取茉莉酸(JA)、茉莉酸甲酯(MeJA)、12-氧-植物二烯酸(12-OPDA)和13-氢过氧化亚麻酸(13-HpOTE),利用XBridge TM C18色谱柱,以甲醇/水为流动相分离4种物质。优化条件下4种物质得到良好分离,加标回收率为81.23%～90.25%,检测限为0.04～0.56 ng/mL,灵敏度高。坛紫菜、龙须菜和真江蓠3种红藻中,坛紫菜中未检测到JA和13-HpOTE,4种物质均在另外2种藻中检测到。龙须菜受机械损伤胁迫后,4种物质在短时间内得到积累,其中13-HpOTE响应迅速。研究表明,红藻中可能存在类似于植物的茉莉酸合成途径,并参与对损伤的胁迫响应。     

液相色谱-串联质谱法同时检测鸡肉中61种抗菌药物

为建立固相萃取结合液相色谱-串联质谱同时测定鸡肉中磺胺类、喹诺酮类、大环内酯类药物残留的分析方法,通过准确称取2g鸡肉样品,经PBS缓冲液提取,以Agilent Pleax PCX固相萃取柱净化,最后以液相色谱-串联质谱进行检测。结果显示:在最优试验条件下测得磺胺类、喹诺酮类、大环内酯类抗菌药物线性良好,相关系数(R²)均大于0.99;方法定量限为1.0~10.0μg/kg;在3个浓度添加水平下,平均回收率为62.9%~118.2%,批间变异系数为4.3%~18.8%。结果表明,本试验建立的方法操作简单,检测耗时短,具有很好的精密度与准确度,适合鸡肉中磺胺类、喹诺酮类、大环内酯类药物残留的同时检测。     

胶体金试纸条法和液相色谱-串联质谱法检测牛奶中四环素类药物残留比较

采用胶体金试纸条与液相色谱-串联质谱（LC-MS/MS）两种方法检测牛奶中残留的四环素类药物。结果表明：胶体金试纸条的检测限为四环素40μg/L、强力霉素40μg/L、金霉素40μg/L、土霉素80μg/L;LC-MS/MS法在牛奶中四环素、强力霉素、金霉素、土霉素的检测限为50μg/L。采用两种方法对实际样品检测,检测结果一致。与液相色谱-串联质谱法相比,胶体金试纸条具有操作简便、快速、直观的特点,可作为筛选法对样品进行定性筛查,LC-MS/MS法进行阳性样品的确证和精确定量。     

Charting the proteome of Cryptosporidium parvum sporozoites using sequence similarity-based BLAST searching

Cryptosporidium (C.) spp. are important zoonotic parasites causing widespread diarrhoeal disease in man and animals. The recent release of the complete genome sequences for C. parvum and C. hominis has facilitated the comprehensive global proteome analysis of these opportunistic pathogens. The well-known approach for mass spectrometry (MS) based data analysis using the BLAST tool (MS BLAST) is a database search protocol for identifying unknown proteins by sequence similarity to homologous proteins using peptide sequences produced by mass spectrometry. We have used several complementary approaches to explore the global sporozoite proteome of C. parvum with available proteomic tools. To optimize the output of the MS data, a sequence similarity-based MS BLAST strategy was employed for bioinformatic analysis. Most significantly, almost all the constituents of glycolysis and several mitochondrion-related proteins were identified. In addition, many hypothetical Cryptosporidium proteins were validated by the identification of their constituent peptides. The MS BLAST approach was found to be useful during the study and could provide valuable information towards a complete understanding of the unique biology of Cryptosporidium.     

Anti-Inflammatory Potential of Monogalactosyl Diacylglycerols and a Monoacylglycerol from the Edible Brown Seaweed Fucus spiralis Linnaeus

A monoacylglycerol (1) and a 1:1 mixture of two monogalactosyl diacylglycerols (MGDGs) (2 and 3) were isolated from the brown seaweed Fucus spiralis Linnaeus. The structures were elucidated by spectroscopic means (NMR and MS) and by comparison with the literature. Compound 1 was composed of a glycerol moiety linked to oleic acid (C18:1 Ω9). Compounds 2 and 3 contained a glycerol moiety linked to a galactose unit and eicosapentaenoic acid (C20:5 Ω3) combined with octadecatetraenoic acid (C18:4 Ω3) or linolenic acid (C18:3 Ω3), respectively. The isolated compounds were tested for their cytotoxic and anti-inflammatory activity in RAW 264.7 macrophage cells. All of them inhibited NO production at non-cytotoxic concentrations. The fraction consisting of compounds 2 and 3, in a ratio of 1:1, was slightly more effective than compound 1 (IC₅₀ of 60.06 and 65.70 µg/mL, respectively). To our knowledge, this is the first report of these compounds from F. spiralis and on their anti-inflammatory capacity.