RP-HPLC (Reverse phase-high performance liquid chromatography)

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RP-HPLC (Reverse phase-high performance liquid
chromatography)

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Introduction

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• ???? ???? HPLC ??? ??.
• ?? ???? ?? ??, ??? ???? ??
• ???? ???, ??? ???? ??.
• ??? ??? ?? ??? ??? ??? ??.
• ???, ??????? ?? ??.

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Mobile phase

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• ???? ??? ?? ??? ????? ???(HPLC?? ??? ??? ?).
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• ????? ???????, ???, ?? ???.

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Fig. 1. Mobile phase used in reverse phase HPLC.
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Table 1. Properties of solvent in reverse phase
HPLC
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Fig. 2. Solubility of gas at different polarity
index. (at 25?, 1atm)
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Equipment

• 1. Column (??)
• ?? Octadecyl ??? ??? ??? ???? ??? ??? ?? ??? ??
  ??? ??? ??? ???. ?? ???1030cm, ??? 5 mm ???.
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• ?? ?? ?? ??? ??? ??? ??? ???? ??? ???.

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• 2. Pump (??)
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• ???? ??? ????? ??? ?? ?????? ?? Pulse? ??? ?.
• ?? ?? 15000psi
• ????? ??? ?? 0.52ml ??? ???.

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• 3. Gradient programmer

1. On/Off
2. Numbers
3. Section
4. Time(minute)
5. Gradient()

Fig. 3. Front panel of grdient former.
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• 4. Injector (?? ???)

Fig. 4. Sample injector.
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• 5. Detector (???)

Fig. 5. Front of the detector.
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• 6. Recorder (???)

Fig. 6. Shape of recorder.
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Experimental

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• ??? ???? ??? ???? ? ? 15? ?? ???? ?? ???? ????
  ???.
• ??? ? ???? ??? ?? ???? ?? ?? auto zero? 0??
  ???.
• ??? ??? ? peak? ???? ???? ???? ??? ???? ????.

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Fig.7. Membrane filtration device.
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Fig. 8. filter assembly.
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Fig. 9. Sample filtering.
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Fig. 10. Sample injection and recorded peaks.
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HPLC Data Analysis

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• Peak? ??? ???? ?? ????.
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