Development and validation of a simple analytical method for the determination of 2,4,6-trichloroanisole in wine by GC–MS

A novel method for the residue analysis of wine spoilage compound 2,4,6-trichloroanisole is reported. Wine (60 ml) was extracted with 2 ml toluene in presence of 24 g MgSO₄ and 6 g NaCl. Cleanup of the toluene phase by dispersive solid phase extraction with mixture of 100 mg CaCl₂, 25 mg primary secondary amine and 50 mg MgSO₄ was effective in minimising co-extractives and matrix effects. Time-of-flight and tandem mass spectrometric parameters were optimised to achieve linearity over 0.25–500 ng ml⁻¹ and method detection limit 0.0083 ng ml⁻¹ which is well below the odour threshold of 0.04 ng ml⁻¹. Recoveries at 0.04, 0.2 and 0.8 ng ml⁻¹ were within 80–110% (±8%). The method was reproducible when tested for Argentinean wines with intra-laboratory Horwitz ratios being <0.20 in white and red wines at both the laboratories of India and Argentina. The method could be successfully applied for incurred wine samples.     

Properties of TCA-insoluble peptides in kimoto (traditional seed mash for sake brewing) and conditions for liberation of the peptides from rice protein

It was found that a large amount of TCA (trichloroacetic acid)-insoluble peptides were liberated into the supernatant of kimoto on the 7th-10th day of mashing. These TCA-insoluble peptides had five polypeptide groups (12, 21, 31, 38, and 55 kDa) on SDS-PAGE (SDS-polyacrylamide gel electrophoresis), and a large amount of high molecular weight peptides, higher than 10,000, were observed upon gel filtration chromatography using TSKgel G2000swxl (Tosoh Co.). Four of these peptides (12, 21, 31, and 38 kDa on SDS-PAGE) appeared specifically in kimoto, and were not detected at all either in sokujo-moto or in the main mash for sake brewing. These TCA-insoluble peptides were fractionated from the supernatant of kimoto on the 9th day, and it was revealed that free amino acids were produced abundantly from them in the presence of the enzyme of rice-koji. Therefore, it was assumed that the peptides are related to the abundant production of free amino acids in kimoto. For the liberation of these TCA-insoluble peptides from rice protein, the enzyme of rice-koji was indispensable. The enzyme liberating the TCA-insoluble peptides from rice protein was purified from rice-koji, and was presumed to be identical with acid protease (AP) of rice-koji. The presence of a high concentration of glucose (higher than 20%) was also indispensable for the liberation of the TCA-insoluble peptides. Furthermore, it was revealed that the peptides were liberated from rice protein under a limited pH of around 4.5.     

生熟麦曲在黄酒传统工艺中的应用研究

通过对生麦曲、熟麦曲、生麦曲和熟麦曲混合配比按传统黄酒发酵工艺进行试验,得出生麦曲因液化力、糖化力和蛋白质分解力等3个力都较低,使它起发时间迟,发酵缓慢,不彻底;熟麦曲3个力都较高,使其起发时间早,且发酵较彻底;混合曲由于按生麦曲和熟麦曲一定关系配比,表现出其的3个力处于两者之间,发酵状况也处于两者之间,同时得出可以酿造出与生麦曲所酿的酒风味相媲美的酒,为进一步改良传统工艺打下一定基础.混合曲中生麦曲与熟麦曲按9:1的比例较佳.     

液相色谱-串质谱法测定大米中异噁唑草酮及代谢物残留量

目的建立大米中异噁唑草酮及代谢物残留量的液相色谱-串联质谱法测定方法。方法试样中残留的异噁唑草酮除草剂及代谢物用乙酸乙腈溶液(1:99,V:V)高速匀浆提取,提取液经N-丙基乙二胺(PSA)、十八烷基硅烷(ODS)净化,经色谱柱为ACQUITY UPLC BEH Phenyl分离和流动相为1.5%乙酸的乙酸铵(1mmol/L)-乙腈进行梯度洗脱,用配有大气压化学电离源(APCI)的液相色谱-串联质谱仪检测和确证。结果异噁唑草酮及代谢物在0.0025~1.000μg/m L浓度范围内呈现良好线性关系(r=0.9998~0.9999)。当样品加标浓度在5~200μg/kg范围内时,其样品加标平均回收率为70.2%~110.6%,相对标准偏差为7.38%~12.84%。异噁唑草酮及代谢物的最低检出限分别为10μg/kg、5μg/kg。结论本方法适用于大米中异噁唑草酮及代谢物残留的同时测定。     

Diffusion and accumulation in cultivated vegetable plants of di-(2-ethylhexyl) phthalate (DEHP) from a plastic production factory

Di-(2-ethylhexyl) phthalate (DEHP) concentrations in the atmosphere and in four vegetable crops including Brassica chinensis L. (bok choy), Brassica campestris L. (field mustard), Vigna unguiculata Walp. (cowpea), and Solanum melongena L. (eggplant) cultivated on land surrounding a plastic production factory were determined. The air DEHP concentrations (means) at the sites 0.2, 0.4, 0.8, and 1.6 km away from the plastic production building were about 9.4–12.8, 5.8–9.6, 1.6–5.0, and 0.04–0.27 µg m^?3 dry weight (DW), respectively. Wind direction is a key factor influencing the measurable DEHP concentration of the air, which was highest in the downwind direction and lowest in the upwind direction, and thus the vegetables accumulated the highest DEHP contents in the downwind direction and the lowest quantities in the upwind direction. The highest DEHP accumulations content of bok choy, field mustard, eggplant, and cowpea were 52.0 ± 3.1, 43.1 ± 2.2, 36.2 ± 2.8, and 19.4 ± 0.47 mg kg^?1 DW, respectively. Safety estimation on the basis of the daily intake limit referenced by the US Environmental Protection Agency (USEPA) led to the conclusion that eating vegetables cultivated 0.2 km away from the plastic production building is not a food safety problem under normal conditions. A strong positive linear correlation between atmospheric DEHP concentration and DEHP content of the vegetable crops was found. The limits for air DEHP concentration for safe vegetable cultivation are 24.0, 34.8, 40.8, and 82.8 µg m^?3 for bok choy, field mustard, cowpea, and eggplant, respectively, by calculating from the equation of linear regression between air DEHP concentration and vegetable DEHP content.     

超高效液相色谱-串联质谱法测定食品包装材料中全氟辛酸及其盐类物质

建立了超高效液相色谱-串联质谱(UPLC-MS/MS)结合加速溶剂萃取测定食品包装材料中全氟辛酸及其盐类物质(PFOA)的方法.采用甲醇作为溶剂,加速溶剂提取法提取食品包装材料中的PFOA,提取液经0.2 μm有机滤膜过滤后,以甲醇、水为流动相,梯度洗脱,经UPLC分离后用多级反应监测(MRM)方式测定.用两个子离子的...     

Identification of steroid hormones in pomegranate (Punica granatum) using HPLC and GC–mass spectrometry

Although it has been known that pomegranate (Punica granatum L.) contains several steroid hormones, concrete experimental proofs about that have not been published until now. In order to identify and evaluate the contents of steroid hormones including estrone in pomegranate, we analyzed pomegranate seed, fruit juice and commercial preparations. We developed a reproducible and sensitive method for separation and identification of steroid hormones in pomegranate samples using both high performance liquid chromatography (HPLC)–PDA and gas chromatography (GC)–MS. In case of HPLC, an isocratic elution method using 35% aqueous acetonitrile solution at 1.0 ml/min with photodiode-array (PDA) detection at 225 nm and 254 nm was found to optimally separate and identify the steroid hormones from the pomegranate samples with a run time of less than 30 min. The pomegranate samples were comparatively analyzed to the HPLC results by GC/FID or GC/MS detection on a HP-1 (30 m length, 0.32 mm I.D.) with helium as carrier gas under the oven temperature control as follows: start 220 °C for 5 min, raising 5 °C per min, final 280 °C for 10 min. The HPLC and GC methods were successfully applied to the identification of steroid hormones in pomegranate samples. Our results suggested that there were no steroid estrogens including estrone, estradiol and testosterone in pomegranate seed, fruit juice and preparations. Consequently, we assumed that the previously reported analysts of pomegranate were misunderstood their analytical results according to either the estrogen-like effects or similarity of peak retention time and Rf values in experiments.     

一种水包油包胶型乳液的制备及其在乳化肠中的应用

以结冷胶和无水氯化钙为内水相凝固剂,酪蛋白酸钠为外水相乳化剂,制备一种水包油包胶（S/O/W）型 乳液。以多重乳液粒径和分布为指标,研究酪蛋白酸钠添加量对S/O/W型多重乳液加工适应性的影响。结果表明： 正交试验得到S/O型单重乳液最佳制备条件为：内水相中结冷胶添加量0.2%、无水氯化钙添加量0.5%;内水相乳化 剂聚甘油蓖麻醇酯添加量2.5%;油相为精炼猪油,油水体积比3∶2;剪切速率17 500 r/min,剪切时间1.5 min。将制 得的S/O型单重乳液与不同添加量酪蛋白酸钠混合制得S/O/W型多重乳液。当酪蛋白酸钠添加量0.1%时,S/O/W型 多重乳液粒径符合加工要求,且贮藏、热处理、剪切稳定性较好。以多重乳液替代猪脂肪制备的低脂乳化肠与高脂 （精炼猪油含量20%）乳化肠外观不存在明显差异;微观结构观察结果表明,多重乳液在乳化肠中包裹良好、分布 均匀。     

UHPLC-DAD-ESI-MS/MS法分析紫山药块根和茎叶中酚类物质

本文采用微波辅助法,以80%甲醇作为溶剂提取经冻干后紫山药块根和地上茎叶中的多酚类物质,提取物经超高效液相色谱分离、电喷雾串联三重四级杆质谱正和负离子方式检测分析。根据液相保留时间、紫外吸收、相对分子质量和二级碎片离子等信息,结果表明,紫山药块根和茎叶中酚类物质主要为酚酸、花色苷及黄酮类化合物。分析得到的29种酚类化合物中,5-O-咖啡酰奎宁酸、阿魏酰奎宁酸(tR:12.289)、迷迭香酸、芦丁和槲皮素为块根和茎叶共有组分,块根中另含有芥子酸、芥子酸葡萄糖苷、丁香酸衍生物、香豆酸衍生物、阿魏酸衍生物、芥子酸二葡萄糖苷和花色苷等11种酚类物质。茎叶中除5种共有组分,还含有咖啡酸、咖啡酰奎宁酸甲酯、咖啡酰莽草酸、对香豆酰奎宁酸、香豆酰奎宁酸甲酯、山奈酚-3-O-芦丁糖苷与迷迭香酸甲酯等13种酚类化合物。     

Identification of 5-hydroxymethyl-2-furfural (5-HMF) in Cava sparkling wines by LC-DAD-MS/MS and NMR spectrometry

The aim of this study was to elucidate the structure of a potential ageing marker for Cava sparkling wine. In order to clarify the structure of this compound, NMR spectroscopy and hyphenated UHPLC-DAD-MS/MS techniques were used. We identified the hitherto unknown compound as 5-hydroxymethyl-2-furfuraldehyde (5-HMF). This is the first time that this compound has been reported in sparkling wines. A survey, based on the analysis of 80 commercial sparkling wines, showed that 5-HMF is present between 0.25 and 12.81 mg/L, and is compared with those reported for other types of wine. Hypothetical origin of 5-HMF in Cava sparkling wine is discussed.     

The porcine tribbles homolog 3 (TRIB3) gene: Identification of a missense mutation and association analysis with meat quality and production traits in Italian heavy pigs

TRIB3 plays an important role in energy metabolism. This work aimed to study the porcine tribbles homolog 3 (TRIB3) gene and to evaluate its association with meat quality and carcass traits in pigs. By sequencing a portion of the porcine TRIB3 gene two single nucleotide polymorphisms (SNPs) in the first coding exon (one synonymous SNP: c.132 T > C; and one missense mutation: c.146C > T, p.P49L) were identified. The two polymorphisms were in complete linkage disequilibrium. In silico analysis of the p.P49L mutation suggested that it could have functional effects. Association studies in four groups of pigs (651 animals in total) indicated that this gene marker was associated with back fat thickness in Italian Large White and Italian Duroc pigs in two different experimental designs (P < 0.1 and P < 0.05). This polymorphism tended to be associated with lactate content of the semimembranosus muscle (P < 0.1). Among several other tissues, TRIB3 is expressed in fat and skeletal muscle.     

Identification of xyloglucan endotransglucosylase/hydrolase genes (XTHs) and their expression in persimmon fruit as influenced by 1-methylcyclopropene and gibberellic acid during storage at ambient temperature

Xyloglucan endotransglucosylase/hydrolase (XTH) is thought to contribute to fruit softening by degrading xyloglucan that is a predominant hemicellulose in the cell wall. In this study, two full-length XTH genes (DKXTH1 and DKXTH2) were identified from ‘Fupingjianshi’ persimmon fruit, and the expression level of both XTH genes was investigated during softening for 18–24 d using RT-qPCR. Sequence analysis showed that DKXTH1 and DKXTH2 contained a putative open reading frame of 861 and 876 bp encoding polypeptides of 287 and 292 amino acid residues, respectively, which contained the conserved DEIDFEFLG motif of XTH, a potential N-linked glycosylation signal site. RT-qPCR analysis showed that DKXTH1 and DKXTH2 in untreated fruit had different expression patterns during fruit softening, in which maximum expression occurred on days 3 and 12 of ripening, respectively. 1-Methylcyclopropene (1-MCP) and gibberellic acid (GA₃) treatments delayed the softening and ethylene peak of persimmon fruit, as well as suppressed the expression of both XTH genes, especially DKXTH1. These results indicated that the expression of both XTH genes might be ethylene dependent action, and closely related to softening of persimmon in the early (DKXTH1) and later (DKXTH2) ripening stages.     

Identification,characterisation, and quantification of phenolic compounds in the antioxidant activity-containing fraction from the seeds of Korean perilla (Perilla frutescens) cultivars

The present research was the first to investigate phenolic compound profiles and antioxidant properties in the seeds of various perilla (Perilla frutescens) cultivars. The 80% methanol extract (50 μg/ml) of this species showed potent antioxidant activities against 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radicals. Phenolic compounds were characterised by nuclear magnetic resonance (NMR) spectroscopy, and ultra performance liquid chromatography with photodiode array detector and electrospray ionisation/mass (UPLC-PDA-ESI/MS) analysis. Nine compounds were elucidated as caffeic acid-3-O-glucoside (1), caffeic acid (2), luteolin-7-O-glucoside (3), apigenin-7-O-glucoside (4), rosmarinic acid-3-O-glucoside (5), rosmarinic acid (6), luteolin (7), apigenin (8), and chrysoeriol (9). The individual and total phenolic contents were remarkably different, especially rosmarinic acid-3-O-glucoside (5) and rosmarinic acid (6) which were the predominant compounds (>95%) in all perilla cultivars. Additionally, Yeupsil cultivar exhibited the highest phenolic content (5029.0 μg/g) and antioxidant activity, whereas the lowest was shown by Dasil (2138.7 μg/g). Therefore, these results suggest that antioxidant effects of perilla seeds are correlated with phenolic contents.     

Identification and quantification of flavonol aglycons in cactus pear (Opuntia ficus indica) fruit using a commercial pectinase and cellulase preparation

The applicability of pectinases and cellulases as soft hydrolysing agents on flavonol glycosides was investigated for identification and quantification of flavonol aglycons in cactus pear fruit. Freeze-dried samples of cactus pear fruit’s peel (cactus pear peels) and onions were treated with commercial pectinase and cellulase preparations at 50 °C for different time periods (up to 16 h). Additionally isorhamnetin-3-O-rutinoside and quercetin-3,4′-O-diglucoside were used as model compounds. In parallel, samples of cactus pear peels and onions were treated by the traditional acidic hydrolysis using HCl. Following hydrolysis, flavonols were characterised using HPLC–DAD. Enzymatically, all model compounds and plant material were highly hydrolysable. Flavonol glycosides were completely hydrolysed after 16 h (cactus pear) and 4 h (onion), respectively. While the acidic hydrolysis caused degradation of the flavonols and produced protocatechuic acid as a degradation product, the enzymatic hydrolysis gave gentle effects and did not produce any protocatechuic acid at all.     

Residual determination of pyrethrins in Lycium barbarum (goji) by GC-MS/MS and a dietary risk assessment of Chinese goji consumption

ABSTRACT

An accurate and simple analytical approach for the determination of residues cinerin I, cinerin II, jasmolin I, jasmolin II, pyrethrin I and pyrethrin II (six active ingredients of pyrethrins) in fresh and dried goji berries was developed and validated for analysis by gas chromatography with tandem mass spectrometry. Good linearity (determination coefficient >0.99), accuracy (average recoveries of 88.3%–111.5%) and precision (intra- and inter-day relative standard deviations of 0.4%–8.3%) were obtained with the optimised determination method. The LODs and LOQs of the six analytes in two goji matrices were 0.24–2.1 µg/kg and 0.8–7 µg/kg, respectively. In a field trial, the terminal residual levels of pyrethrins (the sum of the concentrations of the six target analytes) in fresh and dried goji berry samples were <20–304 µg/kg at harvest, which could provide some information for the establishment of a maximum residue limit of pyrethrins on goji berries in China. Moreover, the risk assessment results indicated that because the risk quotient values were ?100%, the potential dietary risk of pyrethrins on goji berries could be negligible for Chinese consumers. These detection and field results could provide some supporting data for the determination of pyrethrin residues in other crops and the proper application and safety assessment of pyrethrins in goji plants.     

Detection and quantification of 47 antibiotic residues in farmed European sea bass (Dicentrarchus labrax) using a multi-class and multi-residue UHPLC-MS/MS method

In the present study, a multi-class multi-residue method for the simultaneous detection and determination of antibiotics in European sea bass (Dicentrarchus labrax) was developed and validated. The method based on UHPLC-MS/MS proved to be a rapid, highly selective and sensitive tool, requiring minimum sample preparation, for screening and detection of 47 compounds from eight different classes. The validation was performed according to EU regulation 2002/657/EC, proving the method’s suitability for application in routine analysis. The method was applied to the analysis of 30 samples of farmed European sea bass purchased in different supermarkets in Portugal. Antibacterial residues were detected in 6 of the 30 analysed samples, namely enrofloxacin and oxytetracycline, in concentrations ranging from 0.1 to 12 µg kg^?1.     

Identification and sensory evaluation of flavour enhancers in Japanese traditional dried herring (Clupea pallasii) fillet

Flavour-enhancing components of dried herring fillet (migaki-nishin in Japanese) were isolated and evaluated for their effects on sensory perception. Sensory evaluation revealed that addition of dried herring fillet water-soluble extracts to Japanese noodle soup significantly (P < 0.05) enhanced soup flavour characters, such as thickness, mouthfulness and continuity. The extracts were fractionated by dialysis and chromatography. Fractions containing flavour enhancers were isolated by sensory perception. Results from instrumental analyses showed that the kokumi flavour enhancers in dried herring fillet were creatine and creatinine.