高效液相色谱-同位素稀释质谱法定量分析人生长激素
发布时间：2018-11-29 09:02
【摘要】：建立了人生长激素(h GH)纯化分析和高效液相色谱(HPLC)与同位素稀释质谱(IDMS)联用高准确度绝对定量方法。采用快速蛋白液相色谱来分离纯化h GH,以傅里叶变换离子回旋共振质谱仪(FTICR-MS)准确地测定蛋白质的分子质量。纯化的h GH经酸水解后,采用KINETEX C18色谱柱为分析柱,以水(含0.1%三氟乙酸)和乙腈为流动相,等梯度分离,流速0.2 m L/min,温度40℃,采用电喷雾正离子模式进行电离,选择多反应监测模式进行检测,内标法定量分析。结果表明,FTICR-MS所测得的分子量实测值与理论值仅相差0.31 Da,脯氨酸、缬氨酸和苯丙氨酸在液相条件下5 min内达到基线分离,在最优条件下,h GH含量测定结果为186.80"g/g,相对标准偏差为0.52%。利用本方法参加国际比对,比对结果与参考值等效一致。本方法具有简易、实用的特点,并且准确可靠,可作为h GH纯品标准物质的定值方法,为h GH的日常检测提供参考。
[Abstract]:An accurate and accurate method for the purification and analysis of human growth hormone (h GH) and the combination of high performance liquid chromatography (HPLC) and isotope dilution mass spectrometry (IDMS) was established. Rapid protein liquid chromatography was used to separate and purify h GH,. The molecular weight of protein was determined accurately by Fourier transform ion cyclotron resonance mass spectrometer (FTICR-MS). The purified h GH was hydrolyzed by acid. The column of KINETEX C18 was used as analytical column, the mobile phase was water (containing 0.1% trifluoroacetic acid) and acetonitrile as mobile phase, the flow rate was 0.2 mL / min, and the temperature was 40 鈩，
本文编号：2364571
[Abstract]:An accurate and accurate method for the purification and analysis of human growth hormone (h GH) and the combination of high performance liquid chromatography (HPLC) and isotope dilution mass spectrometry (IDMS) was established. Rapid protein liquid chromatography was used to separate and purify h GH,. The molecular weight of protein was determined accurately by Fourier transform ion cyclotron resonance mass spectrometer (FTICR-MS). The purified h GH was hydrolyzed by acid. The column of KINETEX C18 was used as analytical column, the mobile phase was water (containing 0.1% trifluoroacetic acid) and acetonitrile as mobile phase, the flow rate was 0.2 mL / min, and the temperature was 40 鈩，
本文编号：2364571
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