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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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