Simultaneous determination of 18 different amino acids in in vitro fertilization medium by ultra-high performance liquid chromatography–tandem mass-spectrometry

doi:10.3969/j.issn.2095-4344.0860

Abstract

Abstract:

BACKGROUND: At present, in the quality control file or technical standards of in vitro fertilization medium, indicators for component contents and detection methods have not yet been defined. To ensure the safety and effectiveness of these products, we should try to establish and improve the quality standards.

OBJECTIVE: To establish a method for simultaneously testing 18 kinds of amino acids by ultra-high performance liquid chromatography–tandem mass-spectrometry (UHPLC-MSMS), and to analyze the difference in the content of different amino acids in the medium for different uses.

METHODS: Using the UHPLC-MSMS, we detected the indicator components of 18 different amino acids in the in vitro fertilization medium. These amino acids included glycine, leucine, methionine, tyrosine, histidine, threonine, alanine, isoleucine, tryptophan, cystine, lysine, aspartic acid, valine, phenylalanine, valine, serine, glutamic acid, arginine. The UHPLC separation was performed on a SUPELCO Discovery HS F5-3 column (15 cm×2.1 mm, 3 μm) in a gradient elute mode with acetonitrile and water (both containing 0.1% formic acid) as the mobile phase at a flow rate of 0.35 mL/min. The column temperature was 40 ℃. MS detection was performed with multiple-reaction monitoring mode using negative electro spray ionization.

RESULTS AND CONCLUSION: The linearity was achieved in the range of 0.25-12.5 mmol/L for the 18 different amino acids. The average recovery rate of these amino acids ranged 86.3% to 125.3%. The relative standard deviation of the precision experiment and the repeatability experiment was less than 4.7%. These findings indicate that this is a sensitive, rapid, accurate, and specific method that can be used for the quality control of in vitro fertilization medium.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

Key words: Amino Acids, Fertilization in Vitro;, Chromatography, Liquid, Quality Control, Tissue Engineering

CLC Number:

R318

Huang Yuan-li, Ke Lin-nan, Li Chang-kun, Wang Chun-ren . Simultaneous determination of 18 different amino acids in in vitro fertilization medium by ultra-high performance liquid chromatography–tandem mass-spectrometry[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(18): 2915-2920.

Figures/Tables 4

References

[1] 庄广伦,卢光琇,石一复,等.辅助生育技术进展[J].中国实用妇科与产科杂志,2001,17(1):6-31.

[2] 韩堃,郑源强,石艳春.人类辅助生殖技术研究进展[J].中国妇幼健康研究, 2008,19(6):606-609.

[3] 彭靖,卢大儒.试管婴儿技术的发展与探讨[J].自然杂志, 2010,32(6): 338-343.

[4] 丁萍,张爱青,孟凡会,等.影响助生殖技术的因素分析[J].青岛大学医学院学报,2001,37(4):277-278.

[5] 梅志强.人类辅助生殖技术的发展历史与现状[J].中国计划生育学杂志, 2008,16(10):584-586.

[6] 庄广伦.辅助生殖技术的发展历程[J].中国实用妇科与产科杂志, 2010, 26(10):729-731.

[7] 陈贵安.北京大学第三医院开展辅助生育技术近10年回顾[J].北京大学学报:医学版,2002,34(5):623-626.

[8] 张婷,王晓民.体外受精与试管婴儿—外受精与年诺贝尔生理学或医学奖简介[J].首都医科大学学报,2010,31(5):678-682.

[9] 乔杰,马彩虹.中国辅助生殖技术发展现状[J].国际生殖健康/计划生育杂志,2012,31(5):332-333.

[10] 甄璟然,郁琦.体外培养中胚胎发育潜能的评价[J].发育医学电子杂志, 2014,2(2):117-121.

[11] Alpha Scientists in Reproductive Medicine and ESHRE Special Interest Group of Embryology.The Istanbul consensus workshop on embryo assessment: proceedings of an expert meeting.Hum Reprod. 2011;26(6):1270-1283.

[12] Guerif F,Gouge A,Giraudeau B,et al.Limited value of morphological assessment at days 1 and 2 to predict blastocyst development potential: A prospective study based on 4042 embryos.Hum Reprod. 2007;22(7):1973-1981．

[13] Nicholson JK,Lindon JC,Holmes E.'Metabonomics': understanding the metabolic responses of living systems to pathophysiological stimuli via multivariate statistical analysis of biological NMR spectroscopic data. Xenobiotica.1999;29(11):1181-1189.

[14] Pumel ET,Warner CM,Kort HI,et al.Influence of the preimplantation embryo development (Ped) gene on embryonic platelet-activating factor (PAF) levels.J Assist Reprod Genet.2006;23:269-273．

[15] Sher G,Keskintepe L,Fisch JD,et al.Soluble human leukocyte antigen G expression in phase I culture media at 46 hours after fertilization predicts pregnancy and implantation from day 3 embryo transfer.Fetil Steril.2005;83:1410-1413.

[16] Zheng P,Rita Vassena R,Latham KE.Effects of in vitro oocyte maturati on and embryo culture on the expression of glucose transporters，glucose metabolism and insulin signaling genes in rhesus monkey oocytes and preimplantation embryos.Mol Hum Reprod. 2007;13: 361-371．

[17] Bedin Pasqualotto E.Oxidative stress in follicular fluid [C]．ESHRE Campus 2005 Copenhagen，Denmark 29 and 30 April,2005:115-123.

[18] Houghton FD,Hawkhead JA,Humpherson PG,et al. Non-invasive amino acid turnover predicts human embryo developmental capacity. Hum Reprod.2002;17(4):999-1005．

[19] Brison DR,Houghton FD,Falconer D,et al.Identification of viable embryos in IVF by non-invasive measurement of amino acid turnover. Hum Reprod.2004;19(10):2319-2334．

[20] Sturmey RG,Hawkhead JA,Barker EA,et al.DNA damage and metabolic activity in the preimplantation embryo.Hum Reprod. 2009;24(1):81-91．

[21] Seli E,Sakkas D,Scott R,et al.Noninvasive metabolomic profiling of embryo culture media using Raman and near－infrared spectroscopy correlates with repraduefive potentlal of embryos in women undergoing in vitro fertilization.Fenil Stefil. 2007; 88:1350-1357．

[22] Seli E,Botros L,Sakkas D,et al.Non－invasive metabolomic profiling of embryo culture media using proton NMR correlates with reproductive potential of embryos in women undergoing in vitro fertilization．Fenil Steril. 2008;90:2183-2189．

[23] 王增艳,何方方,孙正怡,等.冷冻环玻璃化法冷冻小鼠卵母细胞的效果评价[J].生殖医学杂志,2008,17(1):38-42.

[24] 李宗武,朱传金,刘庆勇.人血浆及精浆中钙、镁、锌、铜对精液成分的影响[J].国外医学:医学地理分册, 2004,25(3):124-126.