Monitoring survey of patulin in a variety of fruit-based products using a sensitive UHPLC–MS/MS analytical procedure

The mycotoxin patulin is known to be the predominant natural contaminant of apples, apple-based products and a variety of other fruits. Because of its high incidence and harmful health effects, patulin is included with mycotoxins, which are strictly monitored. In this study, a sensitive and reliable ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC–MS/MS) method was developed and validated for determination of patulin in a variety of fruit matrices. A combination of the QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) procedure along with a solid phase extraction (SPE) clean-up strategy enabled an effective removal of sample matrix and pre-concentration of patulin. This resulted in low limits of quantification ranging from 1 to 2.5 μg/kg, depending on fruit type. In our study, quantification of patulin was based on a stable isotope dilution assay (SIDA) using ¹³C₇-patulin as the internal standard. Data showed that the procedure described, in combination with neat solvent internal calibration, can be used for accurate quantification of patulin regardless the type of fruit. Although the SIDA method allowed omission of matrix-matched calibration, matrix-effects were estimated in order to assess suppression of the patulin signal caused by a variety of fruit samples. The method was fully validated for apples, apple baby food, apple juice, peaches, strawberries and blueberries. The recovery values were in the range from 92 to 109%. Repeatability of the method was below 10% for all tested matrices. The method was applied to the monitoring of patulin in 135 samples of fresh fruits and fruit products and can also be used as an efficient tool for routine monitoring of this contaminant in a variety of fruit-based foods.     

A method of assessing essential amino acid availability from microbial and ruminally undegraded protein in lactating dairy cows

The objective of this study was to extend a stable isotope-based assessment of AA absorption from rumen-degradable protein (RDP) sources to include determination of essential AA (EAA) availability from microbial protein (MCP). To demonstrate the technique, a study using a 2 × 2 factorial arrangement of treatments applied in a repeated 4 × 4 Latin square design was undertaken. Factors were high and low rumen-degradable protein and high and low starch. Twelve lactating cows were blocked into 3 groups according to days in milk and randomly assigned to the 4 treatment sequences. Each period was 14 d in length with 10 d of adaption followed by 4 d of ruminal infusions of ¹⁵N-labeled ammonium sulfate. On the last day of each period, a ¹³C-labeled AA mixture was infused into the jugular vein over a 6-h period to assess total AA entry. Rumen, blood, urine, and milk samples were collected during the infusions. Ruminal bacteria and blood samples were assessed for AA enrichment. Total plasma AA absorption rates were derived for 6 EAA from plasma ¹³C AA enrichment. Absorption of 6 EAA from MCP was calculated from total AA absorption based on ¹⁵N enrichment in blood and rumen bacteria. Essential AA absorption rates from total protein, MCP, and rumen-undegradable protein were derived with standard errors of the mean of 6, 14, and 14%, respectively. An average of 45% of absorbed EAA were from MCP, which varied among 6 EAA and was interactively affected by starch and RDP in diets. Microbial AA availability measured by isotope dilution method increased with the high RDP diets and was unaffected by starch level, except for Met, which decreased with high starch. Microbial protein outflow, estimated from urinary purine derivatives, increased with RDP and was not significantly affected by starch. This was consistent with measurements from the isotope dilution method. Total AA absorption rates measured from isotope dilution were similar to estimates from CNCPS (v. 6.55), but a lower proportion of absorbed AA was derived from MCP for the former method. Compared with the isotope and CNCPS estimates, the Fleming model underestimated microbial EAA and total EAA availability. An average of 58% of the absorbed EAA was converted into milk, which varied among individual AA and was interactively affected by starch and RDP in diets. The isotope dilution approach is advantageous because it provides estimates of EAA availability for individual EAA from rumen-undegradable protein and MCP directly with fewer errors of measurement than can be achieved with intestinal disappearance methods.     

基于氡同位素的闸塘渗漏检测研究

为拓展水利工程渗漏研究方法,研究利用氡同位素识别地下水入渗点,建立了氡箱模型估算地下水入渗量,并对模型参数进行了敏感性分析。针对盐城市东台养殖区闸塘渗漏进行了实例研究。结果表明:利用氡同位素进行示踪,可有效地识别地下水入渗点,同时可定量计算地下水入渗量;计算结果的不确定性主要来自于入渗地下水的氡活度。氡同位素在地下水入渗的勘查及定量计算中具有较好的应用前景。     

Quantification of 1,8‐cineole and of its metabolites in humans using stable isotope dilution assays

The metabolism of 1,8‐cineole after ingestion of sage tea was studied. After application of the tea, the metabolites 2‐hydroxy‐1,8‐cineole, 3‐hydroxy‐1,8‐cineole, 9‐hydroxy‐1,8‐cineole and, for the first time in humans, 7‐hydroxy‐1,8‐cineole were identified in plasma and urine of one volunteer. For quantitation of these metabolites and the parent compound, stable isotope dilution assays were developed after synthesis of [²H₃]‐1,8‐cineole, [9/10‐²H₃]‐2‐hydroxy‐1,8‐cineole and [¹³C,²H₂]‐9‐hydroxy‐1,8‐cineole as internal standards. Using these standards, we quantified 1,8‐cineole by solid phase microextraction GC‐MS and the hydroxyl‐1,8‐cineoles by LC‐MS/MS after deconjugation in blood and urine of the volunteer. After consumption of 1.02 mg 1,8‐cineole (19 μg/kg bw), the hydroxycineoles along with their parent compound were detectable in the blood plasma of the volunteer under study after liberation from their glucuronides with 2‐hydroxycineole being the predominant metabolite at a maximum plasma concentration of 86 nmol/L followed by the 9‐hydroxy isomer at a maximum plasma concentration of 33 nmol/L. The parent compound 1,8‐cineole showed a low maximum plasma concentration of 19 nmol/L. In urine, 2‐hydroxycineole also showed highest contents followed by its 9‐isomer. Summing up the urinary excretion over 10 h, 2‐hydroxycineole, the 9‐isomer, the 3‐isomer and the 7‐isomer accounted for 20.9, 17.2, 10.6 and 3.8% of the cineole dose, respectively.     

Quantitation of estragole by stable isotope dilution assays

Various calibration strategies for the quantitation of the phenylpropane estragole by gas chromatography-mass spectrometry were developed and compared. For application in stable isotope dilution assays, two deuterium labelled estragole isotopologues were synthesized. Of these, [3′,3′-²H₂]estragole was prepared by Wittig reaction of 4-methoxy-phenylacetaldehyde with [²H₃]methyl-triphenyl-phosphonium bromide, whereas [1″,1″,1″-²H₃]estragole was obtained by demethylation of estragole and deuteromethylation of the resulting 4-allylphenole.Besides estragole isotopologues, 1,2,4-trimethoxybenzene and 4-propylanisole were also tested as internal standards (I.S.) for the determination of estragole in fennel tea.[1″,1″,1″-²H₃]Estragole, 1,2,4-trimethoxybenzene, and 4-propylanisole revealed linear calibration functions and, therefore, were suitable for estragole quantitation. In contrast to this, [3′,3′-²H₂]estragole could only be applied as I.S. if it was added to the extracts in stoichiometric deficiency compared to unlabelled estragole. Moreover, due to its different chemical and physical properties, 1,2,4-trimethoxybenzene showed a recovery as low as 77%, whereas the other I.S. revealed recovery rates close to 100%. Considering the “real” values of estragole in fennel tea, the choice of the I.S. obviously is less important than the way of preparing the tea. In contrast to the common method for tea preparation, squeezing of the teabags increased the estragole content significantly by 50%.     

11C‐ and 18F‐Labeled Radioligands for P‐Glycoprotein Imaging by Positron Emission Tomography

P‐Glycoprotein (P‐gp) is an efflux transporter widely expressed at the human blood–brain barrier. It is involved in xenobiotics efflux and in onset and progression of neurodegenerative disorders. For these reasons, there is great interest in the assessment of P‐gp expression and function by noninvasive techniques such as positron emission tomography (PET). Three radiolabeled aryloxazole derivatives: 2‐[2‐(2‐methyl‐(¹¹C)‐5‐methoxyphenyl)oxazol‐4‐ylmethyl]‐6,7‐dimethoxy‐1,2,3,4‐tetrahydroisoquinoline ([¹¹C]‐ 5 ); 2‐[2‐(2‐fluoromethyl‐(¹⁸F)‐5‐methoxyphenyl)oxazol‐4‐ylmethyl]‐6,7‐dimethoxy‐1,2,3,4‐tetra‐hydroisoquinoline ([¹⁸F]‐ 6 ); and 2‐[2‐(2‐fluoroethyl‐(¹⁸F)‐5‐methoxyphenyl)oxazol‐4‐ylmethyl]‐6,7‐dimethoxy‐1,2,3,4‐tetrahydroisoquinoline ([¹⁸F]‐ 7 ), were tested in several in vitro biological assays to assess the effect of the aryl substituent in terms of potency and mechanism of action toward P‐gp. Methyl derivative [¹¹C]‐ 5 is a potent P‐gp substrate, whereas the corresponding fluoroethyl derivative [¹⁸F]‐ 7 is a P‐gp inhibitor. Fluoromethyl compound [¹⁸F]‐ 6 is classified as a non‐transported P‐gp substrate, because its efflux increases after cyclosporine A modulation. These studies revealed a promising substrate and inhibitor, [¹¹C]‐ 5 and [¹⁸F]‐ 7 , respectively, for in vivo imaging of P‐gp by using PET.     

湖南沃溪金锑钨矿床中成矿物质来源的硫同位素地球化学证据

沃溪AuSbW矿床是湖南湘西前寒武纪地层中最具有代表性的金矿床之一,位于扬子准地台江南地轴南缘的雪峰弧形构造隆起带的中段。矿石以石英-金-硫化物型为主,与金共存的矿石矿物为辉锑矿、黑钨矿、白钨矿、黄铁矿等。文章对沃溪矿床深部矿段中辉锑矿和黄铁矿的硫同位素地球化学进行了研究,结果表明:黄铁矿的δ~(34)S变化范围-4. 05‰～0.42‰,极差4. 47‰,平均值-1. 54‰;辉锑矿的δ~(34)S变化范围-4. 99‰～-1. 52‰,极差3. 47‰,平均值-2. 87‰。δ~(34)S变化范围小,说明其来源的均一性,同时与赋矿的马底驿组富硫围岩的δ~(34)S很相近,表明硫来源于赋矿地层。同时,δ~(34)S具有从浅部向深部增大的趋势。     

天然气中稀有气体浓度与同位素比值联测技术及应用

研制了天然气中稀有气体纯化富集的前处理装置,并与稀有气体同位素质谱仪联用,构建了气体样品中He、Ne、Ar、Kr、Xe浓度和同位素比值联测的分析系统,通过1次进样可得到5种稀有气体组分浓度和同位素比值的共计23个数据。稀有气体纯化效果可达99.9%,质谱分析He、Ne、Ar、Kr浓度的相对标准偏差小于10%,同位素比值分析的标准偏差普遍小于5%。将该方法用于分析塔里木盆地天然气样品,得到了丰富的地质地球化学信息,通过稀有气体分布特征进一步明确了塔河和雅克拉气田区域构造活动的差异,以及二者气源岩特性的不同。     

同位素稀释-气相色谱-串联质谱法同时测定肉制品中16种欧盟优控多环芳烃

目的 基于分子印迹特异性净化及同位素稀释-气相色谱-串联质谱法建立同时检测肉制品中16种欧盟优控多环芳烃的分析方法。方法 样品中加入氘代同位素内标,经正己烷-二氯甲烷(7:3, V:V)混合溶液提取,特异性分子印迹柱净化,采用DB-EUPAH色谱柱分离,在多反应监测(multiple reaction monitoring, MRM)模式下采集,内标法定量。结果 为降低基质效应,采用基质匹配标准曲线的定量方法。在0.2～200.0 ng/mL范围内,16种欧盟优控多环芳烃均有良好的线性关系,线性相关系数(r²)均大于0.9990,在1.0、5.0、10.0μg/kg3个浓度水平下进行加标回收实验, 16种欧盟优控多环芳烃的回收率在85.9%～118.3%之间,相对标准偏差(n=6)在0.4%～6.6%之间,方法的检出限为0.03～0.10μg/kg,定量限为0.10～0.30μg/kg。结论 该方法前处理简便高效、灵敏度高、准确度高、抗干扰能力强,可同时实现肉制品中16种欧盟优控多环芳烃的测定。