Modification of Retention Factor of Mononucleotides by Compositions of Buffers and Methanol in RP-HPLC

RP-HPLC에서 Buffer와 메탄올의 조성에 의한 Mononucleotides 체류인자의 조절

  • Published : 2000.10.01

Abstract

Due to the advantage of RP-HPLC with a variety of compositions of mobile phases, experiments on water-soluble charged species were examined. The samples were mononucleotides (5-CMP, 5-UMP, 5-GMP, 5-IMP, 5-AMP), and the buffers used were sodium phosphate monobasic and acetic acid. The concentrations of buffers ranged from 0.01 to 10 mM, while that of the methanol, an additive to the mobile phase was 5 to 20 vol.%. To predict the retention factor of a sample in terms of its methanol composition (M, vol.%) and buffer(C(sub)B, mM), the following nonlinear equation is suggested, k= $\frac{a+b C_B}{(1+c C_B) M^d}($ where a, b, c, and d were experimentally determined constants. The regression coefficients were above 0.96, and the agreement between experimental and calculated retention factors were relatively good.

Buffer의 특성에 맞는 체류인자에 관한 모텔식을 얻기 위 해서는 buffer 9} 양이온의 농도에 관한 관련식을 실험으로 결 정해야 한다 또한 model 식에서 비이온과 음이온의 체류인 자를 나타내는 ko와 k의 값이 일정한 경향을 나타내고 있기 때문에 buffer의 농도와 modifier의 농도를 동시에 고려한 model 식이 가능하게 된다. 본 연구에서 제안한 모델식을 이용하여 임의의 buffer의 농도에 따른 시료의 체류시간을 예측할 수 있다. 이러한 모텔식을 이용하여 buffer를 이 용한 RP-HPLC의 분석 및 분리조건을 예측할 수 있고 시행 오차적인 방법보다 빠른 시간 내에 최적의 분석 및 분리조건 을 얻을 수 있다

Keywords

References

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