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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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DOI: |
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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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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