| 摘要: |
| 为了利用反相高效液相色谱法(RP-HPLC)更准确地测定发酵液中的生物素含量,以安捷伦Zor-bax SB-C18(4.6 cm×250 cm,5μm)为色谱柱,对液相色谱紫外线检测波长和流动相的溶液配比、K2HPO4离子浓度和pH值进行筛选。结果表明,以体积分数91.5%K2HPO4(0.04 mmol/L)与体积分数8.5% CH3CN混合液为流动相,在pH值为2.5和紫外检测波长为215 nm的条件下,获得的生物素吸收峰面积与质量浓度的线性相关系数达0.999 9,生物素加标回收率达99.07%～101.26%,相对标准偏差为0.48%～0.82%。由此可见,在上述条件下高效液相色谱法不仅可用于发酵液中生物素含量的测定,而且操作简便,测定结果可靠。 |
| 关键词: 高效液相色谱 生物素 发酵液 流动相 |
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| Determination of biotin in broth system by reversed-phase high performance liquid chromatography |
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| Abstract: |
| In order to effectually determine the biotin in fermented broth by reversed phase high performance liquid chromatography (RP－HPLC),the Anjielun Zorbax SB－C18(4.6 cm 250 cm,5 μm) was used and the ultraviolet wavelength for RP－HPLC,the compositions and pH of mobile phase and concentration of K2HPO4 in mobile phase were sifted by repeated tests.The results showed that when the complex liquid (pH=2.5) composed of 91.5% K2HPO4(0.04 mol/L) and 8.5% (V/V) was used for mobile phase，the interrelated coefficient between absorption peak and quality concentration of biotin reached 0.9999 with relative standard deviation of 0.48%-0.82% when the ultraviolet wavelength is 215 nm and pH is 2.5.Therefore,the conclusion can be drawn that RP HPLC can not only be used to determine the biotin content in fermented liquid,but also be convenient for operation. |
| Key words: high performance liquid chromatography biotin fermented broth mobile phase |
