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    高效液相色谱法测定手性生物转化中肾上腺素与肾上腺酮

    HPLC Determination of Adrenalin and Corticosterone in Chiral Biotransformation

    • 摘要: 应用离子对反相离子对高效液相色谱法测定了手性生物转化中肾上腺素和肾上腺酮的含量,将活化的菌株接种至液体培养基中,培养24 h后加入肾上腺酮(910 mg·L-1),在pH 5.5振荡培养并定时取样,取发酵液离心10 min(转速10 000 r·min-1),取上层清液经0.2 μm滤膜过滤后供HPLC测定.用色谱柱(ODS C18)作固定相,流动相由甲醇及25 mmol·L-1乙酸-乙酸钠缓冲液按15比85体积比混合后组成.此缓冲液中含4 mmol·L-1庚烷磺酸钠,流速为1 mL·min-1,测定时采用紫外检测,波长为233 nm.肾上腺素及肾上腺酮分别在质量浓度为28~1 800 mg·L-1及36~1 350 mg·L-1范围内与相应的峰面积之间呈线性关系,信噪比S/N为3时,两者的检出限分别为1.4,2.8 mg·L-1,回收率试验所得结果在99.3%~100.3%之间.

       

      Abstract: The ion-pair reversed phase HPLC was applied to the determination of adrenalin and corticosterone in chiral biotransformation.Activated bacterin was inoculated to the liquid culture medium and cultured for 24 h.Then corticosterone (910 mg·L-1) was added and cultured at pH 5.5 with constant agitation.Samples from the fermented liquor were taken at definite intervals and centrifuged for 10 min at 10 000 r·min-1.The supernatant liquor was taken,filtered through 0.2 μm menbrane and used for HPLC determination.ODS C18 column was used as stationary phase,and a mixture of methanol and 25 mmol·L-1 NaOAc-HOAc buffer containing 4 mmol·L-1 sodium octanesulfonate (15+85) was used at mobile with a flow-rate of 1.0 mL·min-1.UV-detection at 233 nm was used in the determination.Linear relationships between the peak area and the mass concentration of adrenalin and corticosterone were obtained in the ranges of 28-1 800 mg·L-1 and 36-1 350 mg·L-1with their detection limits (S/N=3) of 1.4 mg·L-1 and 2.8 mg·L-1 respectively.Results of recovery test were in the range of 99.3%-100.3%.

       

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