High‐fat Diet‐induced Intestinal Hyperpermeability is Associated with Increased Bile Acids in the Large Intestine of Mice

Metabolic syndrome is characterized by low‐grade chronic systemic inflammation, which is associated with intestinal hyperpermeability. This study examined the effects of 3 high‐fat diets (HFDs) composed of different fat sources (soybean oil and lard) on the intestinal permeability, tight junction (TJ) protein expression, and cecal bile acid (BA) concentrations in mice, and then analyzed their interrelations. C57/BL6 mice were fed the control diet, HFD (soybean oil), HFD (lard), and HFD (mix; containing equal concentrations of soybean oil and lard) for 8 wk. Glucose tolerance, intestinal permeability, TJ protein expression, and cecal BA concentration were evaluated. Feeding with the 3 HDFs similarly increased body weight, liver weight, and fat pad weight, and induced glucose intolerance and intestinal hyperpermeability. The expression of TJ proteins, zonula occludens‐2 and junctional adhesion molecule‐A, were lower in the colons of the 3 HFD groups than in the control group (P < 0.05), and these changes appeared to be related to intestinal hyperpermeability. Feeding with HFDs increased total secondary BA (SBA) and total BA concentrations along with increases in some individual BAs in the cecum. Significant positive correlations between intestinal permeability and the concentrations of most SBAs, such as deoxycholic acid and ω‐muricholic acids, were detected (P < 0.05). These results suggest that the HFD‐induced intestinal hyperpermeability is associated with increased BA secretion. The abundance of SBAs in the large intestine may be responsible for the hyperpermeability.     

高效液相色谱测定番茄制品中的三种功能性脂肪酸

通过高效液相色谱-质谱技术和高效液相色谱技术对番茄制品中9-羟基-10(E),12(Z)-十八碳二烯酸、9-羰基-10(E),12(Z)-十八碳二烯酸和9-羰基-10(E),12(E)-十八碳二烯酸三种功能性脂肪酸进行定性定量分析。结果表明,在色谱柱为Eclipse XDB-C18(4.6 mm×250 mm,5 μm),流动相为0.005%的三氟乙酸水溶液和0.005%的三氟乙酸乙腈溶液,检测波长为234、280 nm的条件下,番茄制品中的三种脂肪酸在20 min内得到较好分离,重复性好(RSD ≤ 2.13%),精密度高(RSD ≤ 0.70%),稳定性好(RSD ≤ 0.98%),加标回收结果准确可靠(平均回收率分别为99.78%、99.70%、99.39%,RSD ≤ 3.38%)。该色谱方法快速、准确、可靠;测试结果表明,新鲜番茄中不含有这三种功能性脂肪酸,其仅存在于番茄制品中,且加工工艺对其含量影响较大;此外9-羟基-10(E),12(Z)-十八碳二烯酸为首次在番茄制品中发现。     

Fermentation dynamics of Lactobacillus helveticus H9 revealed by ultra‐performance liquid chromatography quadrupole time‐of‐flight mass spectrometry

Lactobacillus (L.) helveticus H9 is a probiotic strain that can produce antihypertensive peptides during milk fermentation. This study analysed the dynamics of skim milk fermentation by L. helveticus H9 by ultra‐performance liquid chromatography coupled to quadrupole time‐of‐flight mass spectrometry (UPLC/Q‐TOF MS). A total of 1992 metabolites were detected from all of the fermented samples in the LC‐MS analysis by multivariate statistical analysis. Metabolites with variable importance in projection (VIP) values ≥2 were considered differentially abundant among samples and were responsible for the unique taste and nutritional and functional qualities of fermented milk. Valine, threonine, l ‐methionine, tyrosine, asparagine and leucine were the predominant amino acids produced during fermentation, and their quantities changed remarkably during the fermentation process. Citric acid and uric acid were the major, and only detectable, organic acids. Some intermediate metabolites, such as N‐acryloylglycine and nicotinamide‐N‐oxide, were also detected. Moreover, certain oligopeptides such as Val‐Leu, Lys‐Gly, Ala‐Glu, Asp‐Ser, Leu‐Pro and Val‐Phe‐Ala were not detected until the middle and late fermentation periods. This study demonstrated dynamic metabolic changes, providing a strong foundation for deciphering the physiological and biochemical mechanisms of the fermentation process.     

HPLC/Q‐TOF‐MS‐Based Identification of Absorbed Constituents and Their Metabolites in Rat Serum and Urine after Oral Administration of Cistanche deserticola Extract

As a famous health food in China, Cistanche deserticola (C. deserticola) suggested an estrogenic activity according to our previous study. However, no one clarifies its active material basis to date. To find more potentially active constituents and elucidate metabolic pathways of metabolites, a method to simultaneously analyze multiple absorbed constituents and metabolites from C. deserticola in rat serum and urine was established using high‐performance liquid chromatography/quadrupole time‐of‐flight mass spectrometry (HPLC/Q‐TOF‐MS). Based on HPLC/Q‐TOF‐MS method, a total of 24 components, involving 9 prototype constituents and 15 metabolites in rat serum and urine samples, were tentatively identified based on retention time, ultraviolet spectrum, MS data, compound fragmentation laws, published literatures, and reference substances. Most of the compounds existed in the form of metabolites. The proposed metabolic pathways of main metabolites were discussed, including methylation, demethylation, hydrolysis, hydroxylation, acetoxylation, glucuronidation, dehydrogenation, sulfation, esterification, and so on. Phenylethanoid glycosides were extensively metabolized and mutually transformed in vivo. This investigation provided valuable information for further study of the active ingredients and action mechanism of C. deserticola.     

染发剂中对苯二胺等32种染料检测结果分析

目的了解染发剂中染料使用情况和染发剂类化妆品安全风险点,为监管提供技术支撑。方法根据《化妆品安全技术规范》(2015年版),采用高效液相色谱法和超高效液相色谱-串联质谱法对湖南地区14个市、州地区抽样的50批染发剂类化妆品进行检测,分析其中染料种类、含量及使用情况。结果 50批染发剂共检出对苯二胺、对氨基苯酚等14种染料成分,其中对苯二胺、间氨基苯酚、间苯二酚3种染料的使用频率最高,均超过了60%。有2批染料的苯基甲基吡唑啉酮超出规定限量,其余均未超标。19批染发剂产品批件标识成分与实际检出结果不一致,不合格率为38%。结论染发剂使用情况及批件标识有待规范,应加强对染发剂类产品的监管。     

UPLC-MS/MS分析紫薯酒发酵前后花色苷种类和含量变化

采用超高效液相色谱—串联四极杆质谱（UPLC-MS/MS）法对紫薯酒关键生产环节中花色苷的种类及含量进行分析。结果表明：共鉴定出8种结构差异较大的花色苷,各种花色苷在发酵前后均有不同的变化;高温蒸煮后,紫薯中花色苷种类未发生变化,但花色苷总含量减少;发酵为紫薯酒后,8种花色苷得率约为30%。     

超高效液相色谱-串联质谱法同时测定蜂蜜中8种生物胺

建立了同时测定蜂蜜中色胺(TR)、2-苯乙胺(PHE)、腐胺(PUT)、尸胺(CAD)、组胺(HIS)、酪胺(TYR)、亚精胺(SPD)和精胺(SP)8种生物胺的超高效液相色谱串联质谱柱前衍生的分析方法。样品经5%三氯乙酸溶液提取后用丹磺酰氯柱前衍生,采用Agilent Poroshell 120 SB-C₁₈(50 mm×2.1 mm,2.7 μm)柱,以0.1%甲酸水溶液-0.1%甲酸乙腈溶液为流动相进行定性和定量分析。采用本方法对22种市售蜂蜜进行检测,结果显示8种生物胺在不同的蜂蜜中检出,所有蜂蜜样品中均检测到2-苯乙胺和腐胺,其中2-苯乙胺也是蜂蜜样品中平均含量最多的生物胺。本方法中8种生物胺的线性相关系数均大于0.99,检出限为0.01~0.51 μg/L。在添加水平为100、500、1000 μg/kg时,样品的平均回收率在76%~112%之间,日内精密度小于等于9.8%,日间精密度小于等于13.1%。该方法快速简单,灵敏准确,且回收率稳定,适用于蜂蜜中生物胺的检测。     

在线固相萃取-高效液相色谱-高分辨质谱法测定植物中11种水溶性有机酸

采用在线固相萃取-高效液相色谱-四极杆-静电场轨道阱高分辨质谱建立植物中11种水溶性有机酸(乳酸、富马酸、琥珀酸、苹果酸、酒石酸、香豆酸、没食子酸、柠檬酸、香草酸、阿魏酸、新绿原酸)的分析方法。样品经匀浆、超声、过滤、稀释后,经在线Oasis MAX柱(20 mm×2.1 mm,30 μm)富集净化,采用BEH C₁₈ AX色谱柱(100 mm×2.1 mm,1.7 μm)分离,以含10 mmol/L甲酸铵的0.9%甲酸和0.9%甲酸-乙腈为流动相,梯度洗脱。在负离子扫描模式下,以Full MS/dd-MS²模式进行分析,内标法定量。结果表明,11种有机酸可在9 min内完成分离分析,各组分的精确质量数偏差小于5.00×10^-6,在1～100 μg/L质量浓度范围内线性关系良好,相关系数(R²)大于0.999,方法检出限为0.1～3.2μg/kg,定量限为0.3～9.6μg/kg。以芡实和小白菜为基质样品,在2个不同加标水平下,11种有机酸的平均添加回收率分别为92.2%～111%和94.2%～11...     

基于超高效液相色谱-四极杆-静电场轨道阱高分辨质谱快速测定水果中18种游离氨基酸

本文采用超高效液相色谱-四极杆-静电场轨道阱高分辨质谱(UPLC-Q-Exactive Orbitrap)建立了水果中18种游离氨基酸的快速测定方法.水果样品经粉碎、超声提取、离心、过滤后,使用SHISEIDO CAPCELL PAK BB-H C18色谱柱(2.1 mm×150 mm,3 μm)分离,以0.2％甲酸水...     

UHPLC‐ESI‐MS/MS for the Quantification of Eight Major Gingerols and Shogaols in Ginger Products: Effects of Ionization Polarity and Mobile Phase Modifier on the Sensitivity

We developed and validated an improved ultra high performance liquid chromatography (UHPLC)‐electrospray ionization (ESI)‐tandem mass spectrometry (MS/MS) method for the fast separation and quantification of 8 major gingerols and shogaols (4‐, 6‐, 8‐, 10‐, and 12‐gingerols, and 6‐, 8‐, and 10‐shogaols) in gingerol products. The ionization polarity and mobile phase modifier greatly affected the mass ion profiles and sensitivity of the analytes. A mobile phase modifier of 0.05 mM ammonium formate dramatically decreased the sodium adduct ions and greatly increased the protonated ions of the gingerols and shogaols. Positive ion UHPLC‐MS/MS with the ammonium formate in mobile phase showed greatly higher (4.5‐ to 15.7‐fold) sensitivities than negative ion UHPLC. The positive ion UHPLC‐MS/MS method showed excellent linearities (r² > 0.999), low limits of detection (LOD = 2.5 to 8.2 pg), high accuracy and precision, and no considerable matrix effect. Baseline separation of the 8 target compounds was achieved in 1 min by the UHPLC with a short C18 core‐shell column. The method was successfully applied to analyze the compounds in fresh and dried powdered gingers, and dietary supplements. The total contents of the major compounds in the fresh gingers, dried powdered gingers, and dietary supplements were in the range of 1114 to 1478, 4380 to 11324, and 2915 to 29998 mg/kg, respectively. There was about 10‐fold variation of the content in the dietary supplements with the commercial brands. This represents the 1st report on the ionization polarity and mobile phase modifier on the sensitivity of gingerols and shogaols in LC‐mass spectrometry. Furthermore, the established method provided great improvement in chromatographic separation of the target gingerols.     

超高效液相色谱-串联质谱法检测汤料食品中罂粟壳成分

目的建立超高效液相色谱-串联质谱法(ultra performance liquid chromatography tandem mass spectrometry, UPLC-MS/MS)定性定量检测市售麻辣烫、火锅底料及牛肉汤等汤料食品中罂粟碱、吗啡、可待因、那可丁和蒂巴因的分析方法。方法样品经0.1 mol/L HCl提取、混合阳离子(mixed-cation, MCX)固相萃取柱净化。选用Waters BEH HILIC(2.1 mm×100 mm, 1.7μm)色谱柱进行分离,以0.1%(V/V)甲酸-乙腈和0.1%甲酸-10mmol/L甲酸铵溶液为流动相,进行梯度洗脱。采用正离子源模式(positive electrospray ionization, ESI+)和多反应检测(multi reaction monitor, MRM)模式进行检测,内标法定量。结果在0.1～20μg/L(吗啡和可待因)以及0.1～4.0μg/L(蒂巴因、那可丁和罂粟碱)的浓度范围内, 5种罂粟壳物质线性良好,相关系数均大于0.995,检出限为0.1～0.5μg/L,方法回收率的范围在77.9%～95.5%之间,相对标准偏差1.75%～9.49%(n=6)。结论方法简单易行可操作,且精密度及准确度均较高,适用于汤料食品中罂粟壳的检测。     

超高效液相色谱-串联飞行时间质谱法测定绿茶中9种拟除虫菊酯类农药

目的建立绿茶中9种拟除虫菊酯农药残留量检测的方法。方法采用超高效液相色谱电喷雾串联飞行质谱对9种拟除虫菊酯进行定量和裂解途径研究。结果样品经QuEChERS方法处理后上机,正离子模式下氟氯氰菊酯、氯氰菊酯、溴氰菊酯、氰戊菊酯、氟氰戊菊酯、四溴菊酯易形成[M+NH_4]~+加合离子,苯醚氰菊酯、甲氰菊酯、氟胺氰菊酯易形成[M+H]~+加合离子。采用一级母离子定量,9种拟除虫菊酯1～20μg/L范围内线性关系良好(r~20.99);9种拟除虫菊酯的最低检出限均为1μg/kg;2～10μg/kg范围内回收率为85.2%～106.3%;相对标准偏差(RSD)≤6.1%。结论方法简便、高效适用于绿茶中拟除虫菊酯类农药残留的快速筛查。     

超高效液相色谱-串联质谱法测定乳饼中的氯霉素

目的建立一种超高效液相色谱-串联质谱法(ultra high performance liquid chromatography tandem mass spectrometry, UPLC-MS/MS)检测乳饼中氯霉素的分析方法。方法样品经乙酸乙酯提取,分析了分别经C18(100 mm×2.1 mm, 1.7μm)及MCS固相萃取柱净化、富集的结果;以乙腈-0.05%氨水为流动相,经C18色谱柱分离,采用多反应检测负离子模式进行定性及定量分析。结果 MSC固相萃取柱具有较好的回收率,氯霉素的方法检出限为0.004μg/kg,方法定量限为0.01μg/kg,在0.10、0.50、1.00、10.00 ng 4个加标水平下回收率分别为76.4%、101%、62.4%、61.1%。结论本方法灵敏度高、重复性好,可满足乳饼样品中痕量氯霉素残留的测定。     

超高效液相色谱-串联质谱法检测蜂蜜中的内源性酚酸和黄酮类物质

本研究建立了超高效液相色谱-串联质谱法测定蜂蜜中18种内源性酚酸、黄酮类物质检测方法。样品经盐酸水溶液提取,HLB固相萃取柱净化,采用Acquity UPLC HSS T3(2.1*100 mm,1.8μm)色谱柱进行分离,以乙腈-10 mmol/L醋酸铵溶液为流动相进行梯度洗脱,电喷雾电离,负离子多反应监测模式检测,外标法定量。实验结果表明,所建立的方法线性范围宽,能适用于各类蜂蜜中酚酸、黄酮类物质的定量分析。各物质线性关系良好,相关系数(r2)≥0.991;各分析物检出限在20~200μg/kg之间,加标回收率范围为69.2%~94.1%,RSD为0.9%~9.5%。通过对150批次天然成熟蜂蜜、市售洋槐蜜及以不同比例(20%、40%、60%、80%)糖浆混合的洋槐蜜进行检测,发现不同蜜源蜂蜜中,酚酸、黄酮的种类与含量差异较大。其中,枣花蜜中阿魏酸平均含量最高,为550μg/kg;洋槐蜜中芹菜素平均含量最高,含量为3910μg/kg;咖啡酸在荆条蜜含量最高,为1721μg/kg;糖浆掺假蜂蜜中目标物含量明显低于真正蜂蜜。该方法前处理操作简便、分析速度快、准确度高,可用于蜂蜜中多种内源性酚酸、黄酮类物质的同时测定。该方法为通过检测不同蜜源中酚酸及黄酮类物质含量建立蜂蜜指纹图谱,提供技术参考。     

超高效液相色谱-线性离子阱/静电场轨道阱质谱对化妆品中抗生素类成分的快速筛查和确证

目的建立化妆品中抗生素快速筛查和确证的超高效液相色谱-线性离子阱/静电场轨道阱高分辨质谱法(ultra high performance liquid chromatography-linear ion trap/orbitrap high resolution mass spectrometry)。方法采用超声波辅助提取化妆品中的抗生素类化合物,提取液经ACQUITY BEH C18色谱柱分离,以水(含5 mmol/L乙酸铵和0.1%甲酸)-乙腈为流动相,梯度洗脱分离。静电场轨道阱一级全扫描得到的准分子离子的精确质量数用于快速筛选分析,保留时间和离子阱的二级质谱数据用于进一步定性确证。结果 36种抗生素的精确质量相对偏差小于5×10-6(5 ppm),线性相关系数r2大于0.994,方法检出限(limit of detection)均≤10μg/kg,方法的加标回收率为54.15%~117.55%,相对标准偏差(relative standard deviation)为1.19%~13.25%。结论应用该方法对不同剂型的50余种化妆品实际样品筛查的分析结果良好,说明该方法是化妆品中抗生素类成分快速筛选和确证的有效方法。     

Determination of Estrogens in Milk Samples by Magnetic‐Solid‐Phase Extraction Technique Coupled With High‐Performance Liquid Chromatography

A magnetic‐solid‐phase extraction method coupled with high‐performance liquid chromatography and diode array detection has been developed for simultaneous determination of 3 estrogens in milk samples. In this work, Fe₃O₄ NPs were synthesized by a simple chemical co‐precipitation reaction, and the surface of Fe₃O₄ was modified with cetyltrimethyl ammonium bromide (CTAB; designed as Fe₃O₄@CTAB). The synthesized Fe₃O₄@CTAB NPs were characterized by Fourier transformed infrared spectroscopy, scanning electron microscopy, and transmission electron microscope. Fe₃O₄@CTAB NPs have high binding affinity toward estrone (E1), 17β‐estradiol (17β‐E2), and diethylstilbestrol (DES). Fe₃O₄@CTAB NPs can be easily separated from sample solutions using an external magnet due to the high super‐paramagnetic property. The separation, preconcentration procedure is fast and will be completed in 2 min. Estrogens linear dynamic ranges were achieved in the range of 10 to 1000 ng/mL with regression coefficients (R²) higher than 0.9992. The limits of detection were between 0.26 and 0.61 ng/mL. Parameters influencing the recoveries were investigated and optimized. The proposed method was used for the determination of E1, 17β‐E2, and DES in milk samples, and recoveries were ranged from 91.3% to 105.0%, with the relative standard deviations in the range of 2.7% to 4.0%.     

Analysis of josamycin in three kinds of feed using ultra high performance liquid chromatography with tandem mass spectrometry

ABSTRACT

In this study, a new and accurate method based on ultra-high performance liquid chromatography (UHPLC) with electrospray ionisation tandem triple quadrupole mass spectrometry (MS/MS) was developed and validated for the determination of josamycin in three kinds of feeds (compound, concentrated and premix feeds). The optimised sample pretreatment procedure included extracting from the matrix with a mixture of methanol and acetonitrile (1:1, v/v) followed by drying under nitrogen and redissolving in phosphate buffer solution (pH = 8, 0.1 mol/L) followed by purifying using HLB cartridges. Validation of the method was carried out in three different matrices (compound, concentrated and premix feed) by recovery experiments, using samples spiked at concentration levels of 20, 100 and 200 μg/kg. Satisfactory recoveries from 82.4% to 93.0%, with RSDs lower than 17.4% under intermediate precision conditions were obtained in all feed matrices tested. The developed method was applied to commercially samples and josamycin was found frequently in feed containing kitasamycin.     

UPLC-MS/MS法测定酒中甜蜜素含量的研究

目的 本文采用UPLC-MS/MS方法对市售30种酒中甜蜜素进行测定。方法 UPLC－MS/MS采用Acquity UPLC@ BEH C18, (1.7祄,2.1?00mm)色谱柱分离,甲醇-0.1％乙酸（90+10）作为流动相洗脱,运用三重四极杆质量分析器定性定量分析。结果 该方法测定酒中甜蜜素具有良好的线性关系,测定方法检出限为0.010 μg/mL,定量限为0.021 μg/mL,加标回收率为75.0-105%;精密度RSD值为10.5-12.1%。30种酒样品中,有13个样品检测出甜蜜素,含量水平为0.0027-891 礸/ml.。结论 该方法测定酒中甜蜜素,具有较高的灵敏度和选择性,样品测定结果表明存在滥用甜蜜素现象。     

Comparison of ginsenosides in radix and rhizome of wild Panax species using LC‐ELSD and LC‐Q‐TOF‐MS

High‐performance liquid chromatography (HPLC) with evaporative light scattering detector (ELSD) and quadrupole time‐of‐flight mass spectrometry (Q‐TOF‐MS) were used for qualitative and quantitative analysis of wild Panax species, especially wild P. vietnamensis, which is an important medicinal plant. We determined the types and concentrations of ginsenosides in radix and rhizome of wild P. vietnamensis. Identification of ginsenosides was achieved using Q‐TOF‐MS; concentrations were determined by ELSD. The most abundant ginsenosides in wild Vietnamese ginseng were of the ocotillol type, accounting for more than 50% of the total. Compared to the rhizome, the radix had 31% higher ginsenoside content due to variation in protopanaxatriol‐type ginsenoside contents. We also found an unusual difference in the chromatograms of the two parts of wild P. vietnamensis. This difference did not appear in other wild species such as P. ginseng and P. quinquefolius. Our study provides an opportunity for further in‐depth study of the distinctive characteristics of P. vietnamensis.     

直接进样法-超高效液相色谱串联质谱法测定黑龙江流域水中10种农药残留

目的建立直接进样-超高效液相色谱串联质谱法(ultra high performance liquid chromatography tandem mass spectrometry,UPLC-MS/MS)测定黑龙江流域水中10种农药残留的分析方法。方法水样无需富集,经0.22μm微孔滤膜过滤后,超高效液相色谱分离,动态多反应监测模式(dynamic multi reaction monitoring,DMRM)质谱分析,基质匹配标准曲线外标法定量。结果 10种农药的检出限(S/N=3)在0.1～3.0μg/L之间,定量限(S/N=10)在0.2～5.0μg/L之间,在各自线性范围内,具有良好的线性关系(r≥0.997),在5.0、10.0和50.0μg/L 3个添加水平下平均回收率在78.0%～109.9%之间,相对标准偏差(relative standard deviation,RSD)小于20%(n=6)。结论该方法简单、快速、灵敏度高,在10 min内完成10种农药的检测分析,可用于流域水中多种农药残留的快速检测。