5 Simple Techniques For HPLC working
5 Simple Techniques For HPLC working
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ディテクター（検出器）としては目的とする物質の性質に応じて光学的性質（吸光度、屈折率、蛍光等）、電気化学的性質、質量分析法などを利用する装置がある。
각각 다른 산업 분야에 대한 자세한 정보 및 다양한 카테고리는 다음 써모 피셔 사이언티픽 학습 센터에서 산업 및 응용 과학 페이지를 확인하세요.
機械的に高い圧力をかけることによって移動相溶媒を高流速でカラムに通し、これにより分析物が固定相に留まる時間を短くして分離能・検出感度を高くすることを特徴とする。
The best way to take pleasure in the theoretical and the practical aspects talked about During this section would be to carefully analyze an average analytical technique.
. Illustration of a normal high-performance liquid chromatograph with insets displaying the pumps that go the cell stage with the system along with the plumbing accustomed to inject the sample in to the cell section.
분석물의 피크 면적 값(=검출기의 응답)은 정량화를 위해 사용됩니다. 분석자는 분석을 수행하기 전, 분석물의 표준 용액(기지 농도의 시액)을 몇 가지 측정하고, 시료 농도와 획득한 피크 면적 값에 의해 도표된 검량선을 그립니다.
Incorporate a recognized amount of the antidepressant protriptyline, which serves as an internal regular, to each serum sample and to every exterior normal. To eliminate matrix interferents, go a 0.five-mL aliquot of each serum sample or standard via a C18 strong-section extraction cartridge. Immediately after washing the cartridge to eliminate the interferents, elute the remaining constituents, such as the analyte and The inner common, by washing the cartridge with 0.
The pump is the guts with the HPLC system. It delivers the mobile phase at a relentless and high strain (as much as 400 atm) through the column. Consistent movement level is essential for attaining optimum separation and maintaining reproducibility. Components to look at when deciding on a flow rate include:
Very poor resolution implies analytes elute as well shut together, building them hard to tell apart. This is tips on how to troubleshoot:
System contamination: Soiled HPLC strains, injectors, or detectors can introduce contaminants that display up as ghost peaks. Flush the system with suitable solvents to eliminate any gathered website contaminants.
Altering the cell section’s polarity index variations a solute’s retention element. As we uncovered in Chapter twelve.three, nonetheless, a improve in k is just not a successful way to improve resolution once the First price of k is larger than 10.
The realm less than Each and every peak is proportional to the amount of the corresponding analyte. The information acquisition system permits the Assessment of peak retention instances, peak locations, plus the calculation of analyte concentrations.
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To influence an even better separation among two solutes we must Increase the selectivity issue, (alpha). There are two frequent techniques for rising (alpha): introducing a reagent to the mobile period that reacts With all the solutes within a secondary equilibrium reaction or website switching to a unique cellular period.