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Determination of Polyamines in Human Serum by Liquid Chromatography-Tandem Mass Spectrometry
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實驗結果顯示，以1 mL含0.1 % HFBA的乙腈/水(70/30, v/v)混合溶液，對200 μL的血清樣品進行萃取，震盪並離心後取1 mL的上層澄清溶液，加入15 mg的C18粉末進行淨化後可得良好的萃取效果。Putrescine的偵測線性範圍為5 ~ 500 ng/mL，spermidine和spermine為2 ~ 250 ng/mL，線性相關係數皆在0.9922以上，偵測極限在0.1 ~ 0.9 ng/mL之間。於健康自願者的空白血清中可以測得putrescine 17.0 ng/mL、spermidine 14.1 ng/mL及spermine 8.0 ng/mL，而病人的血清樣品中可測得多胺類含量為一般健康人的約2 ~ 15倍。故本技術可作為人體血清樣品中多胺類確認檢驗技術之參考依據，及病人血清中多胺類含量異常之資訊。|
In the study, a simple and sensitive method using liquid-liquid extraction (LLE)/ dispersive solid-phase extraction (d-SPE) combined with liquid chromatography-mass spectrometry for determination and quantification of polyamines in human serum was evaluated. The target polyamines are putrescine, spermidine, and spermine. Liquid-liquid extraction was used to extract trace analytes from serum matrix and then dispersive solid-phase extraction was used to clean up the solution of extraction. Polyamines were extracted from serum by using 1 mL acetonitrile/water solution (70/30, v/v) which was contained 0.1% HFBA then using 15 mg C18 sorbent to clean up the solution of extraction. Polyamines were then analysed by ion-pair chromatography-mass spectrometry. The linear range was 2 to 500 ng/mL. Limits of detection(LOD) of three polyamines were between 0.1 to 0.9 ng/mL. The coefficients of determination above 0.9922 were obtained. The polyamines concentration profiles in serum of patients were all determined. The concentrations of polyamines were detected to range from 6.5 to 235.2 ng/mL which are higher than those in health people. The results of this study reveal the proposed method offered the ng/mL level sensitivity and high selectivity for determination of polyamines in human serum.
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