The Basic Principles Of high performance liquid chromatography
The Basic Principles Of high performance liquid chromatography
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. The working pump and the equilibrating pump Each and every Have got a piston whose back and forth motion maintains a continuing circulation charge of nearly several mL/min and supplies the high output pressure required to thrust the cellular section through the chromatographic column.
각각 다른 산업 분야에 대한 자세한 정보 및 다양한 카테고리는 다음 써모 피셔 사이언티픽 학습 센터에서 산업 및 응용 과학 페이지를 확인하세요.
Adsorption chromatography includes the conversation of chemicals with the surface in the stationary phase. A compound’s affinity for the stationary phase establishes its diploma of retention. In reverse-section HPLC, by way of example, nonpolar molecules are held by a polar stationary section.
Recall, consulting your instrument handbook and also the producer's technical guidance may also be beneficial methods when troubleshooting particular issues together with your HPLC system.
The a few crimson circles are binary cell phases made by combining equivalent volumes in the pure cell phases. The ternary mobile stage demonstrated via the purple circle incorporates all 3 of the pure cellular phases.
Peak spots: The area underneath Just about every peak from the chromatogram is proportional to the amount of analyte present, permitting for quantification.
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Building an optimized HPLC technique consists of strategically modifying numerous parameters to realize the absolute best separation to your distinct analytes. Vital parameters for optimization contain:
-hydroxybenzoic acid—over a nonpolar C18 column utilizing an aqueous buffer of acetic acid and sodium acetate as the cellular more info period. The retention periods for these weak acids are shorter when using a a lot less acidic cell section due to the fact Each individual solute is present in an anionic, weak base variety that is certainly less soluble while in the read more nonpolar stationary section.
). If the detector is often a diode array spectrometer, then we can also Display screen The end result as A 3-dimensional chromatogram that shows absorbance for a function of wavelength and elution time.
- 분석물의 분리여부는 고정상(컬럼)과 이동상의 조합에 의해 결정합니다.(실제 시료 측정에서는 시료 중에 분석물 이외의 오염물질에 존재하는 경우가 많아 분석자는 그 시료의 측정에 최적인 분석 조건의 검토가 필요합니다.
, for instance, exhibits retention times for 4 weak acids in two mobile phases with just about identical values for (P^ primary ). Even though the purchase of elution is identical for the two cell phases, Every single solute’s retention time is afflicted in different ways by the choice of organic and natural solvent.
The sample injector introduces the sample into your HPLC system. Precise and correct sample injection is crucial for acquiring trusted outcomes.
이 검량 곡선을 바탕으로 실제 시료 분석으로 얻은 피크 면적에서 시료 중의 존재량을 산출하여 정량화를 실시합니다.