Optimization of mobile phase conditions of bioactive compounds in RP-HPLC

Optimization of mobile phase conditions of bioactive compounds in RP-HPLC
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The use of mobile phase additives enhances the separation and resolution of the target compounds on a commercial C18 column. Acetic acid, triethylamine, inorganic salts and several ionic liquids were added into mobile phase methanol/water (40:60, v/v). Chlorogenic acid, caffeic acid, rutin and scoparone from Herba Artemisiae Scopariae were used as the target compounds to investigate the effect and the result revealed that ionic liquid [BMIM][BF4] with the concentration of 0.01 mol/L was quite increase the resolution of the target compounds. An efficient optimization method was used to separate flavonoid compounds from celery by HCI program software. Two interfering species, tentatively considered the target compounds, were separated successfully by the HCI program with the optimum composition of mobile phase for RP-HPLC separation obtained. The elution profiles were calculated by the plate theory based on the linear and quadratic equations of the retention factor: lnk = lnkw + SF, k = A + B/F, lnk = L + MF + NF2, where F was the volume percentage of the organic modifier. In the binary solvents model, the calculated results of the mobile phase conditions suggested that methanol/water (60/40, v/v) or acetonitrile/water (34/66, v/v) was the optimized component, as was acetonitrile/ methanol/water (10/30/60, v/v/v) when changed to a ternary solvents model. Under the experimental conditions, the agreement between the experimental elution profiles and the calculated values of the eluted concentration were relatively good.
ABSTRACT iii Table of Contents v List of Tables vii List of Figures viii 1. Introduction 1 1.1 Mobile Phase Additives 1 1.2 Optimization of Mobile Phase Condition by Using HCI Program 4 2. Theoretical Background 7 3. Experimental 9 3.1 Chemicals 9 3.1.1 HAS 9 3.1.2 Celery 10 3.2 HPLC analysis 10 3.3 Experimental Methods 11 3.3.1 HAS 11 3.3.2 Celery 11 4. Results and Discussions 12 4.1 Mobile Phase Additives 12 4.1.1 Retention Factor Estimation 12 4.1.2 Effect of the pH on the Resolution 13 4.1.3 Effect of the Concentration of Inorganic Salt 15 4.1.4 Effect of Ionic Liquid on the Retention and Resolution of Target Compounds 17 4.2 Optimization of Mobile Phase Condition by Using HCI Program 24 4.2.1 Optimum Mobile Phase Determination 24 4.2.2 Separation of Target Compounds from Extract 31 5. Conclusion 34 Summary in Korean 35 Summary in Chinese 36 References 37 Appendix 1. Molecularly Imprinted Monolithic Material for Extraction of Three Organic Acids from Salicornia herbacea L. 41 Appendix 2. Optimum Separation Condition of Five Bioactive Compounds from Marine Plant by HCI Program in HPLC 43 Appendix 3. Retention Mechanism of Some Solutes Using Ionic Liquids as Mobile Phase Modifier in RP-HPLC 44 List of Publications 45 List of Tables Table 1. Effect of concentration of different mobile phase additives on retention factors. Table 2. Effect of concentrations of different ionic liquids on retention factors Table 3. Parameters of the Snyder, Langmuir and Binary-poly equations on content of methanol mobile phase (standard sample) Table 4. The parameters of Binary-poly equation (methanol) Table 5. The parameters of Binary-poly equation (acetonitrile) Table 6. The parameters of Binary-poly equation (methanol and acetonitrile) List of Figures Figure 1. The chemical structures of the four target compounds Figure 2. Chemical Structures of (a) Luteolin and (b) Apigenin Figure 3. Effect of concentration of
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College of Engineering(공과대학) > Chemical Engineering (화학공학) > Theses(화학공학 석박사 학위논문)
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