利用高效液相色谱检测发酵液中S-腺苷蛋氨酸含量  

Determination of S-adenosylmethionine Contentsin in Culture Broth by HPLC

作　　者：杨秋明[1,2,3] 许海琴 倪辉[1,2,3] 蔡慧农[1,2,3] 肖安风[1,2,3] 

机构地区：[1]集美大学生物工程学院,福建厦门361021 [2]福建省高校食品微生物与酶工程研究中心,福建厦门361021 [3]厦门市食品生物工程技术研究中心,福建厦门361021

出　　处：《集美大学学报（自然科学版）》2012年第6期421-427,共7页Journal of Jimei University：Natural Science

基　　金：福建省自然科学基金资助项目(2011J01224);集美大学中青年创新团队专项基金资助项目(2010A006);集美大学科研基金资助项目(ZQ2006043)

摘　　要：建立了利用Waters2695高效液相色谱测定毕赤酵母发酵液中S-腺苷蛋氨酸(S-adenosylmethi-onine,SAM)含量的方法,对HPLC-UV法检测SAM含量的色谱条件进行优化,最佳色谱条件如下:All-tima C18柱,150 mm×4.6 mm,5μm,流动相为V(0.01 mmol/L甲酸铵)∶V(甲醇)=97∶3,流速0.6 mL/min,柱温30℃,检测波长260 nm.优化后的色谱分离条件可使样品中的SAM与杂质达到较佳分离,稳定性、精密度及重复性都较好,其RSD均小于1.5%.利用SAM标品制作标准曲线Y=32408606.0721 X+130181.2824,线性范围为0.012～1.2 g/L,R2达到0.9998,平均回收率为100.027%～102.533%.此方法可有效地检测发醇液中S-腺苷蛋氨酸含量.An HPLC determination method was developed for measuring S-adenosylmethionine （SAM） contents in culture broth of Pichia pastoris. The optimized HPLC conditions for SAM analysis were 0. 01 mmol/L ammonium formate solution （ pH = 3.5 ） - methylalcohol （ V： V = 97 ： 3 ） as mobile phase, flow rate 0. 6 mL/min, column temperature 30℃, and detection wavelength 260 nm. The separation conditons described above achieved good separation effect, together with high stability, sensitivity, and reproducibility. The standard curve of SAM was Y = 32408606. 0721 X ＋ 130181.2824 , and the linear range was 0. 012 × 1.2 g/L, R2 was 0. 9998, the average recycle rate was 100. 027% - 102. 533%. This method would be useful for SAM analysis during Pichia pastoris fermentation.

关 键 词：高效液相色谱 S-腺苷蛋氨酸 检测 

分 类 号：R917[医药卫生—药物分析学]