【摘要】：建立了人生長激素(h GH)純化分析和高效液相色譜(HPLC)與同位素稀釋質(zhì)譜(IDMS)聯(lián)用高準確度絕對定量方法。采用快速蛋白液相色譜來分離純化h GH,以傅里葉變換離子回旋共振質(zhì)譜儀(FTICR-MS)準確地測定蛋白質(zhì)的分子質(zhì)量。純化的h GH經(jīng)酸水解后,采用KINETEX C18色譜柱為分析柱,以水(含0.1%三氟乙酸)和乙腈為流動相,等梯度分離,流速0.2 m L/min,溫度40℃,采用電噴霧正離子模式進行電離,選擇多反應(yīng)監(jiān)測模式進行檢測,內(nèi)標法定量分析。結(jié)果表明,FTICR-MS所測得的分子量實測值與理論值僅相差0.31 Da,脯氨酸、纈氨酸和苯丙氨酸在液相條件下5 min內(nèi)達到基線分離,在最優(yōu)條件下,h GH含量測定結(jié)果為186.80"g/g,相對標準偏差為0.52%。利用本方法參加國際比對,比對結(jié)果與參考值等效一致。本方法具有簡易、實用的特點,并且準確可靠,可作為h GH純品標準物質(zhì)的定值方法,為h GH的日常檢測提供參考。
[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，

本文編號：2364571