动物营养学报 >

2020 , Vol. 32 >Issue 6: 2824 - 2830

DOI: https://doi.org/10.3969/j.issn.1006-267x.2020.06.043

实验方法与实验动物 EXPERIMENTAL METHOD AND ANIMAL

气相色谱-质谱联用法测定山羊血浆中甘露醇和乳果糖含量

  • 许丽卫 ,
  • 陈闻 ,
  • 耿梅梅 ,
  • 张丽萍 ,
  • 贺珍 ,
  • 袁红朝 ,
  • 彭灿 ,
  • 李春勇 ,
  • 贺志雄 ,
  • 王久荣
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  • 中国科学院亚热带农业生态研究所, 亚热带农业生态过程重点实验室, 长沙 410125
许丽卫(1983-),女,河北定州人,高级工程师,博士,从事色谱分离与分析研究。E-mail:xlw818@isa.ac.cn

收稿日期: 2019-12-02

  网络出版日期: 2020-06-16

基金资助

国家自然科学基金项目（31772631）；中国科学院率先行动"百人计划"

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Determination of Mannitol and Lactulose Contents in Goats’ Plasma by Gas Chromatography-Mass Spectrometry Method

  • XU Liwei ,
  • CHEN Wen ,
  • GENG Meimei ,
  • ZHANG Liping ,
  • HE Zhen ,
  • YUAN Hongzhao ,
  • PENG Can ,
  • LI Chunyong ,
  • HE Zhixiong ,
  • WANG Jiurong
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  • Key Laboratory for Agro-Ecological Processes in Subtropical Region, Institute of Subtropical Agriculture, Chinese Academy of Sciences, Changsha 410125, China

Received date: 2019-12-02

  Online published: 2020-06-16

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摘要

本试验旨在建立一种基于糖腈乙酰酯衍生反应的测定山羊血浆中甘露醇和乳果糖含量的气相色谱-质谱联用法。首先对山羊血浆用2倍体积的乙腈进行处理，离心取其上清液；再用1倍体积的水对上述沉淀进行清洗，离心取其上清液，合并2次上清液，并对其进行离心浓缩干燥，再进行糖腈乙酰酯衍生反应；衍生后的样品采用气相色谱-质谱联用法测定甘露醇和乳果糖含量。以蜜二糖一水作为内标物，HP-5毛细管柱进行分离，内标物、甘露醇和乳果糖在22.75 min内达到基线分离。结果表明：甘露醇和乳果糖的保留时间和浓度的相对标准偏差分别小于0.5%和5.0%。甘露醇和乳果糖含量在0.18~10.00 μg/mL和0.23~10.00 μg/mL范围内线性关系良好，相关系数均在0.998以上，检出限分别为0.052和0.071 μg/mL，加标回收率分别为71.3%~72.5%和89.8%~110.0%。综上可知，本试验建立的测定血浆中甘露醇及乳果糖含量的气相色谱-质谱联用法专一性强、灵敏度高，可应用于动物肠道通透性评价中。

关键词： 乳果糖; 甘露醇; 内标法; 糖腈乙酰酯衍生; 气相色谱-质谱联用

本文引用格式

许丽卫 , 陈闻 , 耿梅梅 , 张丽萍 , 贺珍 , 袁红朝 , 彭灿 , 李春勇 , 贺志雄 , 王久荣 . 气相色谱-质谱联用法测定山羊血浆中甘露醇和乳果糖含量[J]. 动物营养学报, 2020 , 32(6) : 2824 -2830 . DOI: 10.3969/j.issn.1006-267x.2020.06.043

Abstract

A gas chromatography-mass spectrometry method based on aldononitrile acetate derivatization reaction was developed to determine the goats’ plasma mannitol and lactulose contents. Firstly, the goats’ plasma was treated with two times volume of acetonitrile, and the supernatant was centrifuged. The precipitate was cleaned with one times volume of water, and the supernatant was centrifuged. The two supernatants were combined and dried for aldononitrile acetates derivation. The contents of mannitol and lactulose in derivatives were analyzed by the gas chromatography-mass spectrometry method. The internal standard melibiose, mannitol and lactulose were separated by HP-5 capillary column within 22.75 min. The results showed that the relative standard derivations of retention time and concentration for mannitol and lactulose were less than 0.5% and 5.0%, respectively. Mannitol and lactulose contents showed a good linear relationship at a range of 0.18 to 10.00 μg/mL and 0.23 to 10.00 μg/mL, respectively, and the correlation coefficients of them both more than 0.998. The detection limits of mannitol and lactulose were 0.052 and 0.071 μg/mL, and the recovery of standard addition were 71.3% to 72.5% and 89.8% to 110.0%, respectively. In conclusion, the gas chromatography-mass spectrometry method developed in this experiment to determine the goats’ plasma mannitol and lactulose contents has strong selectivity and high sensitivity, which can be used for the intestinal permeability evaluation in animals.

Key words： lactulose; mannitol; internal standard method; aldononitrile acetates derivation; gas chromatography-mass spectrometry

参考文献

[1] JAYALAKSHMI K,GHOSHAL U C,KUMAR S,et al.Assessment of small intestinal permeability using 1H-NMR spectroscopy[J].Journal of Gastrointestinal and Liver Diseases,2009,18(1):27-32.
[2] FARHADI A,KESHAVARZIAN A,HOLMES E W,et al.Gas chromatographic method for detection of urinary sucralose:application to the assessment of intestinal permeability[J].Journal of Chromatography B,2003,784(1):145-154.  
[3] BRUET V,BOURDEAU P,BIZZARRI M,et al.Rapid blood sampling method for measuring intestinal permeability by gas chromatography in dogs[J].Revue de Médecine Vétérinaire,2008,159(5):276-281.  
[4] 王和兴,黎源倩,李磊.保健食品中甘露醇的毛细管气相色谱分析[J].四川大学学报(医学版),2006,37(3):480-483.
[5] 陈丹,姜春燕,马辰.HPLC-ELSD法测定尿液中甘露醇和乳果糖浓度[J].感染、炎症、修复,2007,8(2):79-82.
[6] 瞿平,胡燕,黎海芪.高效液相色谱法检测尿样中甘露醇和乳果糖浓度[J].儿科药学杂志,2005,11(2):1-3.
[7] LOSTIA A M,LIONETTO L,PRINCIPESSA L,et al.A liquid chromatography/mass spectrometry method for the evaluation of intestinal permeability[J].Clinical Biochemistry,2008,41(10/11):887-892.
[8] DONG Y,YAN K,MA Y H,et al.A modified LC-MS/MS method to simultaneously quantify glycerol and mannitol concentrations in human urine for doping control purposes[J].Journal of Chromatography B,2016,1022:153-158.
[9] 胡强,徐红兵,李水军,等.液相色谱-串联质谱法测定尿中乳果糖、甘露醇和乳糖含量[J].中国现代医学杂志,2008,18(13):1810-1813,1817.
[10] GILANI S,HOWARTH G S,KITESSA S M,et al.Intestinal permeability induced by lipopolysaccharide and measured by lactulose,rhamnose and mannitol sugars in chickens[J].Animal,2017,11(7):1174-1179.  
[11] GILANI S,HOWARTH G S,TRAN C D,et al.Reduced fasting periods increase intestinal permeability in chickens[J].Journal of Animal Physiology and Animal Nutrition,2018,102(1):e486-e492.
[12] GILANI S,HOWARTH G S,TRAN C D,et al.Effects of delayed feeding,sodium butyrate and glutamine on intestinal permeability in newly-hatched broiler chickens[J].Journal of Applied Animal Research,2018,46(1):973-976.  
[13] 曾道艳.高效液相色谱在食品质量检测中的应用[J].现代食品,2019(5):173-175.
[14] 段秀辉,李露,薛淑静,等.杏鲍菇、香菇及其预煮液中可溶性糖的GC-MS分析[J].食品工业科技,2015,36(17):281-285.
[15] 陈晓,张可冬,赵广才,等.气相色谱-质谱联用技术测定虫草类口服液中甘露醇的含量[J].分析仪器,2015(6):38-42.
[16] ZHANG W,HE H B,ZHANG X D.Determination of neutral sugars in soil by capillary gas chromatography after derivatization to aldononitrile acetates[J].Soil Biology and Biochemistry,2007,39(10):2665-2669.  
[17] 李雪莹,林晨,王李平,等.衍生化-气相色谱法在食品安全检测中的应用[J].广州化工,2016,44(8):21-23.
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