摘要
采用高效液相色谱-质谱联用法(HPLC/MS)分析测定植物甾醇侧链降解过程中产物雄甾-1,4-二烯-3,17-二酮(ADD)及雄甾4-烯-3,17-二酮(AD)。其中液相色谱的条件为色谱柱AlltimaC18ODS-2(5μm,250mm×4.6mm);流动相甲醇:水(V/V=7:3);流速1mL/min;柱温室温;紫外检测器的检测波长244nm。质谱为ZMD Micromass电喷雾质谱仪。结果测得ADD与AD标准样品的保留时间分别为9.70min与11.13min,发酵样品的HPLC与MS图谱与ADD与AD标准样品的图谱一致。采用高效液相色谱法定量ADD与AD的线性范围在0.01mg/mL^0.09mg/mL,产物回收率分别为102.6%与105.90%,日内精密度分别为3.02%与3.08%,日间RSD分别为3.50%与3.24%。该方法灵敏度高、选择性好、操作简便、定量准确,适用植物甾醇微生物侧链降解过程中产物的分析及产品质量控制。
The method for analysis and determination the cleavage of soybean sterol, in which the soybean sterol was degraded and the products androst-1,4-diene-, 17-dione （ADD） and androst-4-ene-3,17-dion （AD） were developed by Liquid Chromatography-mass Spectrometry. The HPLC conditions adopted were： a Alltima ODS-2 column （250 mm×4.6 mm, 5 μm）, a mobile phase consisted of menthanol - water （70：30）, a flow rate of 1.0 mL/min, a room column temperature, and the detective wavelength was 244 nm.The ZMD Micromass electrospray ionization （ESI）-mass spectrometer was employed. In such conditions the corresponding HPLC chromatogram and MS spectrum were obtained. The method has a linear ranger of 0.01 mg/mL - 0.09 mg/mL, R^2 =0.9999, the recoveries of ADD and AD were 102.6% and 105.90%, the RSD of ADD and AD were 3.02%, 3.5% and 3.08%, 3.24%. This method showed high sensitivity, accuracy and easy to perform. It is suitable to analysis the process cleavage of soybean sterol as well as quality control of product.
出处
《微生物学通报》
CAS
CSCD
北大核心
2008年第10期1674-1679,共6页
Microbiology China
基金
国家自然科学基金(No.20676053)
国家“863计划”(No.2006AA020103)
江苏省青年科技创新人才(学术带头人)基金(No.BK2006504)
长江学者和创新团队发展计划资助(No.IRT0532)
