A pump provides the mobile phase by way of a column filled with a stationary section. An autosampler injects the sample onto the column. The stationary stage separates the sample compounds or analytes. A detector steps the analytes just after separation and elution through the column.
高效液相色谱仪工作原理 高效液相層析儀根據各種各樣的相互作用力來分離混合物。這種相互作用力通常是分析物及分析管柱之間的一種非共價性質。使用高效液相色谱时，液体待检测物在不同的時間被注入色谱柱，通过压力在固定相中移动，由于被测物中不同物质与固定相的相互作用不同，不同的物质顺序离开色谱柱，通过检测器得到不同的峰信号，每個峰頂都代表一個另外化合物的種類，最后通过分析比对这些信号来判断待測物所含有的物质。
Chromatographers can use 3 exceptional sorts of second-LC techniques to help improve sample resolution by employing many column selectivity.
The separated factors are then detected on the exit of the column by a detector that actions their amount of money. Output from this detector known as a “liquid chromatogram.”
Molecules migrate into pores inside of a porous media and so are segregated primarily based on their dimensions compared to the pore sizing. Huge molecules elute to start with, accompanied by more compact kinds.
The column is packed with a stationary period substance. The selection of column and stationary section is determined by the nature on the compounds becoming analyzed and also the separation objectives.
Therefore, owing to interactions Together with the stationary phase, the constituent components of a mixture migrate with the column at distinct speeds.
Beside the pump, get more info There may be an injector. The easiest way is to make use of a syringe to insert the sample in the eluent stream. Sampling loops are by far the most extensively utilised injection system.
An internal common is necessary when employing HPLC–MS because the interface amongst the HPLC plus the mass spectrometer doesn't make it possible for to get a reproducible transfer of your column’s eluent get more info in to the MS’s ionization chamber.
A little volume of sample for being analyzed is introduced to the cell phase stream which is retarded by distinct chemical or physical interactions With all the stationary phase.
Next, some of the compounds in the serum may soak up too strongly on the stationary section, degrading the column’s performance. Last but not least, Even though an HPLC is able to separating and examining complex mixtures, an Examination should still be hard if the amount of constituents exceeds the column’s peak capacity.
Gradient separations typically give exceptional performance about isocratic modes but are more intricate and require State-of-the-art pump hardware.
Analyte detection. Detection of goal analytes according to an electrical signal produced by particular Attributes.
These analyses are often paired with mass spectrometry a result of the inverse partnership amongst stream amount and electrospray ionization effectiveness, considerably improving technique sensitivity.