同位素 ›› 2010, Vol. 23 ›› Issue (4): 221-224.DOI: 10.7538/tws.2010.23.04.0221

• 特约专栏:稳定同位素制备及应用技术 • 上一篇    下一篇

15N标记S-苄基-L-半胱氨酸含量及其同位素丰度检测

宋明鸣;杜晓宁   

  1. 上海化工研究院,上海200062
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2010-11-20 发布日期:2010-11-20

Detection of Content and Isotope Abundance of 15N-S-benzyl-L-cysteine

SONG Ming-ming;DU Xiao-ning   

  1. Shanghai Research Institute of Chemical Industry, Shanghai 200062, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2010-11-20 Published:2010-11-20

摘要: 建立了一种测定发酵液中15N标记S-苄基-L-半胱氨酸(SBC)含量的高效液相色谱法(HPLC),该方法采用Cosmosil C18(5 μm,150 mm×4.6 mm)色谱柱为分离柱,2,4-二硝基氟苯为衍生剂,0.05 mol/L 乙酸钠缓冲溶液(pH=6.5,含10 mL/L N,N二甲基甲酰胺)与V(乙腈)∶V(水)=3∶1为流动相,梯度洗脱,流速1.0 mL/min,检测波长为350 nm,柱温为30 ℃;并采用微量高温燃烧-质谱法对15N标记SBC样品的同位素丰度进行了检测。结果显示,SBC浓度为0.2~1.0 g/L时,与其峰面积呈良好的线性关系,y=3×106x-4.16×104,R2=0.998 6,平均加标回收率97.30%,检测限为3.95×10-4 g/L,测定结果的相对标准偏差为0.55%;15N标记SBC丰度检测时,最小称样量为2 mg,最佳反应温度为530 ℃,反应时间为3 h;15N丰度测定值的相对标准偏差为0.17%。

关键词: 15N, S-苄基-L-半胱氨酸(SBC), 同位素丰度, 检测

Abstract: A HPLC method for determination of S-benzyl-L-cysteine in fermentation broth was established. Cosmosil C18 column(5 μm,150 mm×4.6 mm) was used with 2,4-dinitrofluorobenzene as derivatization reagent and 0.05 mol/L sodium acetate buffer (pH=6.5,including 10 mL/L N,N-dimethylformamide) and V(acetonitrile)∶V(water)=3∶1 as mobile phase. The content was detected at a flow rate of 1.0 mL/min(gradient), the detection wavelength was 350 nm, column temperature was 30 ℃. The calibration curves were linear in the range of 0.2~1.0 g/L(R2=0.998 6), the average recovery rates were 97.30%, the minimum limit was 3.95×10-4 g/L, the relative standard deviation of results was 0.55%. Microscale high temperature combustion-MS method was also described to detect the isotope abundance of 15N-SBC. The minimum consumption of 15N-SBC product was 2 mg, the optimal combustion temperature was 530 ℃ and reaction time was 3 h. The RSD of results was 0.17%.

Key words: 15N, S-Benzyl-L-cysteine(SBC), isotope abundance, detection