基于微孔板比色法的微生物胞内油脂快速检测方法的建立及应用
Establishment and application of a rapid microbial intracellular lipid determination method based on colorimetric microplate format
  
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中文关键词:  单细胞油脂  磷酸香草醛反应  称重法  裂殖壶菌  隐甲藻
英文关键词:single-cell lipid  sulfo-phospho-vanillin reaction  macro-gravimetric method  Schizochytrium sp.  Crypthecodinium cohnii
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Author NameAffiliation
HU Yuanwei  
REN Lujing  
SUN Guannan,etc  
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中文摘要:
      对磷酸香草醛显色法的反应条件进行优化研究,建立了磷酸香草醛显色法快速检测微生物胞内油脂的方法,并探讨该方法在微生物油脂测定上的应用。经过实验确定最佳比色条件为:将油脂样品蒸干溶剂后加入2 mL 98%硫酸,于90℃水浴20 min,冷却至室温后添加3 mL 0.25 g/L香草醛磷酸溶液(34%的磷酸溶液配制),摇匀室温显色20 min,扣除非油脂成分干扰,在530 nm波长下测定吸光度,经验证该反应体系稳定性良好。在裂殖壶菌和隐甲藻两种产DHA油脂微生物中,通过称重法与磷酸香草醛显色法比较,进一步证实建立的方法分析结果准确可靠,分析过程简便、快速,稳定性好,适合作为常规分析检测方法。
英文摘要:
      To achieve a rapid and accurate determination method for single-cell lipid, sulfo-phospho-vanillin(SPV) reaction conditions were optimized and the application of SPV reaction in determination of microbial intracellular lipid was discussed. The optimal SPV reaction conditions were that oil samples were added with 2 mL of 98%H2 SO4 keeping in 90℃ water bath for 20 min, after they were cooled to room temperature, 3 mL of vanillin-phosphoric acid solution(prepared with 34%phosphoric acid solution) was added for color development and absorbance measurement was made at 530 nm after 20 min of color reaction to eliminate the non-oil components interference. Under the optimal conditions, lipid contents of Schizochytrium sp. and Crypthecodinium cohnii were detected, and there was no significant difference in lipid determination comparing with macro-gravimetric method. Thus, SPV reaction was proved to be a simple, accurate, rapid method for detecting microbial intracellular lipid which could be used as a routine analytical method for the determination of single-cell lipid.
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