Ultrasonic assisted enzymatic digestion (USAED) coupled with high performance liquid chromatography and electrothermal atomic absorption spectrometry as a powerful tool for total selenium and selenium species control in Se-enriched food supplements

The suitability of hyphenated USAED with HPLC separation and ET-AAS determination as a new rapid methodology for Se control in Se-enriched food supplements is demonstrated. Total Se determination and Se speciation are accomplished in a single sample treatment using low sample amounts (ca. 10 mg), and low extracting volume (1 mL). The total Se content in seven of the 10 Se-enriched supplements studied was in agreement with the values obtained after microwave pressurized acid digestion, MW, (test t, p = 0.05). The Se species studied were Se(IV), Se(VI), SeMet, SeMeSeCys, and SeCys₂, being some of the most common found in the 10 supplements studied. Although SeMet was the Se species expected to be present at the highest concentration in most Se-enriched food supplements, we detected it in only three of the 10 samples studied. In the other seven samples, two of them had Se(IV) as the main Se species. The other five supplements had Se species that did not match with any of the five standards selected by us. We have also systematically demonstrated that ultrasonication does not alter the following Se species: Se(IV), Se(VI), SeMet, SeMeSeCys, and SeCys₂. The new procedure can be easily adapted to more Se species and can be routinely used for Se control in Se-enriched food supplements. Concerning the supplements studied, our results suggest that stricter control on the Se content in enriched food supplements in terms of Se species will need to become mandatory.     

富硒酵母中硒蛋氨酸的GC-MS/MS测定

建立酵母中硒蛋氨酸含量的气相色谱——串联质谱法(GC-MS/MS)分析方法。比较3种富硒酵母中硒蛋氨酸检测样品的提取方法,优化硒蛋氨酸的酶解提取条件;以氯甲酸乙酯为衍生化试剂,2-氯苯丙氨酸为内标物,采用选择离子模式对衍生物进行GC/MS/MS检测。结果表明,硒蛋氨酸的回收率为90.0%～97.0%,检测限为4μg/L,方法精密度(RSD)为7.8%。该方法简单快捷、定量准确、灵敏度高。     

Selenomethionine increases proliferation and reduces apoptosis in bovine mammary epithelial cells under oxidative stress

The decline in mammary epithelial cell number as lactation progresses may be due, in part, to oxidative stress. Selenium is an integral component of several antioxidant enzymes. The present study was conducted to examine the effect of oxidative stress and selenomethionine (SeMet) on morphology, viability, apoptosis, and proliferation of bovine mammary epithelial cells (BMEC) in primary culture. Cells were isolated from mammary glands of lactating dairy cows and grown for 3 d in a low-serum gel system containing lactogenic hormones and 0 or 100 μM H₂O₂ with 0, 10, 20, or 50 nM SeMet. Hydrogen peroxide stress increased intracellular H₂O₂ to 3 times control concentrations and induced a loss of cuboidal morphology, cell-cell contact, and viability of BMEC by 25%. Apoptotic cell number more than doubled during oxidative stress, but proliferating cell number was not affected. Supplementation with SeMet increased glutathione peroxidase activity 2-fold and restored intracellular H₂O₂ to control levels with a concomitant return of morphology and viability to normal. Apoptotic BMEC number was decreased 76% below control levels by SeMet and proliferating cell number was increased 4.2-fold. These findings suggest that SeMet modulated apoptosis and proliferation independently of a selenoprotein-mediated reduction of H₂O₂. In conclusion, SeMet supplementation protects BMEC from H₂O₂-induced apoptosis and increased proliferation and cell viability under conditions of oxidative stress.     

Se in Se‐enriched peanut,and losses during peanut protein preparation

This study determined the Se species in Se‐enriched peanut, and Se losses during peanut protein processing by enzymatic extraction, high‐performance liquid chromatography (HPLC) separation and inductively coupled plasma‐mass spectrometry determination. The study revealed that mixed enzymes (protease and lipase, 2:1 w/w) in Na₂S₂O₃, assisted by 1 h ultrasonic processing, could effectively extract Se speciation from defatted peanut powder. Separation of organic Se by HPLC was optimised using pentafluoropropionic anhydride at a concentration of 0.1% in 2% methanol as mobile phase. Selenomethionine is the dominant Se species in peanut, accounting for 65% of the total Se. During the peanut protein preparation, nearly 37% of Se losses were due to the complexity of the multistage process. The loss can be ascribed to volatilisation, dissolution, degradation or other physical modes of transfer or loss.     

富硒酵母菌株的选育

以酿酒酵母Saccharomyces cerevisiae L-11为出发菌株,通过DES多次诱变,Na2SeO3和乙硫氨酸抗性平板选择,得到一株高硒和高蛋氨酸含量的菌株L-11-7-2,其蛋氨酸含量达到1.37%,是L-11的169%.对L-11-7-2进行富硒培养,细胞内总硒和硒蛋氨酸分别为2452μg/g和1650μg/g,分别为出发株的280.2%和369.1%.     

富硒大豆中蛋白提取工艺优化及HPLC-MS联用测定硒代蛋氨酸

根据Box-Behnken试验设计原理和响应面分析法确定了富硒大豆中硒蛋白提取的最佳工艺;在此条件下,获取可溶性硒蛋白中的硒代氨基酸,并采用高效液相色谱-质谱联用技术对富硒大豆可溶性蛋白中的硒代蛋氨酸进行定性定量分析。结果表明,液料比11∶1（mL/g）、提取温度 44 ℃、提取时间96 min时,可溶性硒蛋白的提取率可达到76.03%。对4 种自行种植的富硒大豆中可溶性蛋白分析可知：硒-L-蛋氨酸中硒含量与可溶性蛋白中硒含量、富硒大豆中总硒量呈正相关;实验范围内硒代蛋氨酸相对于可溶性蛋白中硒含量最高可达28.46%;说明富硒大豆中不仅含有硒代蛋氨酸,还存在其他形态的硒代氨基酸,有待于进一步研究探索。对大豆进行富硒种植,不仅能提高大豆中含硒总量,而且能通过大豆的生物代谢将其转化成安全无毒营养的有机硒。     

硒代蛋氨酸的合成方法研究进展

综述了硒代蛋氨酸的主要化学合成方法：2-氨基-4-溴丁酸氢溴酸盐取代反应法,2-氨基-4-苄基硒代蛋氨酸取代反应法,三锅煮法,Na/L iqNH3法,丙烯醛法,丁酸内酯法及蛋氨酸法。其中重点介绍了各个方法的优缺点,并确定蛋氨酸法是实现工业化的首先方法。     

硒代蛋氨酸的合成及其改善猪肉和鸡肉品质的研究进展

硒代蛋氨酸是一种含硒的α-氨基酸,与人类多种病等密切相关,在动物生产中有重要的作用,文章仅综述了硒代蛋氨酸的合成及其在改善猪肉和鸡肉质方面的研究进展。     

气相色谱串联质谱法测定加工工艺对毛豆硒蛋氨酸含量的影响

采用气相色谱串联质谱(GC-MS/MS)方法,研究硒氨基酸衍生物的一级质谱以及二级碰撞离子裂解规律,建立硒蛋氨酸的GC-MS/MS 测定方法,并以此为基础研究在加工和贮藏过程中毛豆中硒蛋氨酸的含量变化。结果显示：在4℃条件下冷藏4 周,以硒蛋氨酸计的硒的损失约为13%;在常温保存过程中,脱水毛豆的硒蛋氨酸降解率最小,而毛豆汁的硒蛋氨酸降解率却显著升高,说明产品的形态对于硒蛋氨酸的影响是非常重要的;毛豆烫漂过程中硒蛋氨酸的损失较小,烫漂210s 硒蛋氨酸的保存率仍然在85% 以上,毛豆脱水工艺中冷冻干燥处理对硒蛋氨酸的保存率最高,其次是真空干燥和热风干燥,硒蛋氨酸的保存率均可达80% 以上,而喷雾干燥因接触空气面积较大,造成有机硒的损失较大,会造成硒蛋氨酸的损失率达到30%～40%。