高效液相色谱-示差折光检测法测定辅助生殖培养液中果糖及葡萄糖的含量  

Determination of fructose and glucose in assisted reproductive culture medium by high performance liquid chromatography-differential refractive index detector

作　　者：孙雪[1] 黄元礼[1] 柯林楠[1] 

机构地区：[1]中国食品药品检定研究院,生物材料和组织工程室,北京市100050

出　　处：《中国组织工程研究》2014年第30期4873-4877,共5页Chinese Journal of Tissue Engineering Research

基　　金：国家科技支撑计划课题(2012BAI22B01)~~

摘　　要：背景:目前国内外辅助生殖培养液的质量技术标准中对成分含量的指标以及检测方法未进行明确要求,亟待建立相关的质量标准弥补这一空缺,从而对辅助生殖培养液的质量进行有效的控制。目的:建立辅助生殖培养液中果糖及葡萄糖含量的评价方法,为其质量技术标准提供制定依据。方法:采用高效液相色谱法,Rezex RCM-Monosaccharide Ca2+(8%)色谱柱(300.0 mm×7.8 mm),流动相为超纯水,流速:0.6 mL/min,柱温80℃,示差折光检测器。结果与结论:葡萄糖峰与果糖峰的分离度为3.2。葡萄糖浓度和果糖质量浓度分别在30.1-502 mg/L(r=0.999 8)和102-408 mg/L(r=0.999 8)内,线性关系良好,重复性实验的相对标准偏差分别为0.17%和0.40%,稳定性实验的相对标准偏差分别为0.22%和0.73%。葡萄糖组和果糖组的平均加样回收率分别为1.22%和1.38%。方法学验证符合要求。供试品1的葡萄糖和果糖含量分别为96.7 mg/L和208.5 mg/L,供试品2的葡萄糖和果糖含量分别为99.6 mg/L和197.4 mg/L。文章填补了国内外辅助生殖培养液质量技术标准中关于组分含量的空缺,对辅助生殖培养液的质量控制具有重要的实际意义。BACKGROUND： Nowadays, the component content of assisted reproductive culture medium and their test methods are unclear in the quality standards. We need to establish the test methods to fill this vacancy, so as to control the quality of assisted reproductive culture medium effectively. OBJECTIVE： To establish an evaluation method for determination of fructose and glucose in assisted reproductive culture medium, thereby based on which to establish the quality standards. METHODS： High performance liquid chromatography-differential refractive index detector was adopted and ride Ca2＋ （8%） （300.0 mmx7.8 mm） column was used. The mobile phase was ultrapure water at the flow rate of 0.6 mL/min and the temperature of 80 ℃. RESULTS AND CONCLUSION： Resolution of the peaks of glucose and fructose was 3.2. The linear ranges of glucose and fructose were 30.1-502 mg/L （r=0.999 8） and 102-408 mg/L （r=0.999 8）, respectively. The relative standard deviation of reproducibility test was 0.17% and 0.40%, respectively. The relative standard deviation of stability test was 0.22% and 0.73%, respectively. The glucose group and fructose group average recovery rateswere 1.22% and 1.38%, respectively. The methodology of High performance liquid chromatography-differential refractive index detector accorded with the requirements. The glucose contents of samples 1 and 2 were 96.7 mg/L and 99.6 mg/L, respectively. The fructose contents of samples 1 and 2 were 208.5 mg/L and 197.4 mg/L, respectively. A reliable, simple, and accurate method was provided for the quality control of assisted reproductive culture medium, which fills the domestic vacancy in the quality standards for assisted reproductive culture medium.

关 键 词：生物材料 材料相容性 高效液相色谱法 生物材料基础实验 辅助生殖 培养液 果糖 葡萄糖 示差折光 含量 质量控制 

分 类 号：R318[医药卫生—生物医学工程]