The instrumentation of a HPLC apparatus is as shown in the diagram above. A HPLC apparatus is including a solvent reservoir, degasser, pump, injector, HPLC column, detector and data collection device.
Solvent reservoir The solvent reservoir is usually a glassware or reagent bottle which contained the analytic sample. It is located at the upper part of the HPLC. It is also known as mobile phase reservoir since the sample will be moved. The flowing of the sample is depends on gravitational pull. The solvent which needed to be analysed is a liquid which consisted of polar and non-polar components and the concentration of respective components is depends on the contents of the sample used. There are some requirements need to be fulfilled by …show more content…
The pump in the HPLC apparatus is used to create a high pressure in the apparatus. The pump also uses the high pressure to force the mobile phase to deliver it via the HPLC column and detector with a specific rate of flowing. The normal flow rate which induced by the pump in HPLC is in the range of 1ml/min to 2ml/min. In HPLC, the 6000psi to 9000psi of the range of pressure can be generated depending on the rate of flowing of mobile phase, the size of particles of stationary phase and the volume of the column. There are three main types of HPLC pumps which are constant flow reciprocating pump or pistol type pump, pneumatic pump and syringe type. The constant flow reciprocating pump, also known as pistol pump is commonly used in the HPLC instrument due to the constant generation of pressure and can use for both isocratic and gradient pump. There are two types of the ways to operate the pump which are isocratic pump and gradient pump. The isocratic pump is used to move the constant mobile phase configuration and the gradient pump is used to move the variable mobile phase contents and increase the composition of mobile phase. The gradient pump needs dual pump but the isocratic is only needed for single pump. The number of pumps in a HPLC instrument is depends on the flowing conditions. The pump also must be inert to the …show more content…
This is because the analysis of the solvent will only be completed when the solvent passed through the column. The types of HPLC are also determined by the column which the instrument used. The HPLC column is to operate the separation of the solvent. The stationary phase in the column is used to separate the mobile phase by using different types of physical and chemical parameters. There are many types of the HPLC column by differentiating by various dimensions. The length of column is in the range of 50mm to 300mm and the diameter is in the range of 2mm to 5mm. The correct combination of the column length can make up different types of HPLC apparatus and separate the sample in an effective way. The narrow vessel in the column will induce a high pressure in the instrument. The column is always filled with the stationary phase. The stationary phase is commonly filled with silica gel since the size of the silica particle, surface properties and pore structure will lead to good separation results of solvent. The silica gel is also inert to most solvent so it can separate various type of chemical compound with high reproducibility. The materials for constructing a HPLC column are inert to the analytes such as polished stainless steel, glass and PEEK polymers. There are mainly four types of columns used in HPLC which are high performance analytical column, preparation column, capillary column and
Solvent reservoir The solvent reservoir is usually a glassware or reagent bottle which contained the analytic sample. It is located at the upper part of the HPLC. It is also known as mobile phase reservoir since the sample will be moved. The flowing of the sample is depends on gravitational pull. The solvent which needed to be analysed is a liquid which consisted of polar and non-polar components and the concentration of respective components is depends on the contents of the sample used. There are some requirements need to be fulfilled by …show more content…
The pump in the HPLC apparatus is used to create a high pressure in the apparatus. The pump also uses the high pressure to force the mobile phase to deliver it via the HPLC column and detector with a specific rate of flowing. The normal flow rate which induced by the pump in HPLC is in the range of 1ml/min to 2ml/min. In HPLC, the 6000psi to 9000psi of the range of pressure can be generated depending on the rate of flowing of mobile phase, the size of particles of stationary phase and the volume of the column. There are three main types of HPLC pumps which are constant flow reciprocating pump or pistol type pump, pneumatic pump and syringe type. The constant flow reciprocating pump, also known as pistol pump is commonly used in the HPLC instrument due to the constant generation of pressure and can use for both isocratic and gradient pump. There are two types of the ways to operate the pump which are isocratic pump and gradient pump. The isocratic pump is used to move the constant mobile phase configuration and the gradient pump is used to move the variable mobile phase contents and increase the composition of mobile phase. The gradient pump needs dual pump but the isocratic is only needed for single pump. The number of pumps in a HPLC instrument is depends on the flowing conditions. The pump also must be inert to the …show more content…
This is because the analysis of the solvent will only be completed when the solvent passed through the column. The types of HPLC are also determined by the column which the instrument used. The HPLC column is to operate the separation of the solvent. The stationary phase in the column is used to separate the mobile phase by using different types of physical and chemical parameters. There are many types of the HPLC column by differentiating by various dimensions. The length of column is in the range of 50mm to 300mm and the diameter is in the range of 2mm to 5mm. The correct combination of the column length can make up different types of HPLC apparatus and separate the sample in an effective way. The narrow vessel in the column will induce a high pressure in the instrument. The column is always filled with the stationary phase. The stationary phase is commonly filled with silica gel since the size of the silica particle, surface properties and pore structure will lead to good separation results of solvent. The silica gel is also inert to most solvent so it can separate various type of chemical compound with high reproducibility. The materials for constructing a HPLC column are inert to the analytes such as polished stainless steel, glass and PEEK polymers. There are mainly four types of columns used in HPLC which are high performance analytical column, preparation column, capillary column and