HPLC | MPLC | LPLC
Liquid Chromatography is a technique used for separation of a mixture into its individual components. HPLC or High performance liquid chromatography, have various applications as purification, separation, isolation etc. apart from analysis of compounds in chemical and biology research. In the manufacturing process of peptide, insulin or other fermented, synthesized API use chromatography technique, to achieve very pure finish product. High performance liquid chromatography technique is used for analysis of raw material, finish product, in process product etc. impurity isolation on preparative labs and producing kilo scale pure compound through process scale or plant scale chromatography.H PLC is among the very few technologies that play very important role in all size of application such as lab scale, pilot scale and production scale. Consider the purification of peptides weather it is synthesized manually or automatically
using a synthesizer or other small to medium sized organic molecules. Very good separation and pure compound of the components of the mixture is achieved on HPLC. The Liquid Chromatography technique is further classified into High Pressure, Medium Pressure and Low Pressure for various application into normal phase and reverse phase, in the separation and purification of Peptide, insulin, mAB’s, vaccines etc. Though HPLC System play lesser role in the classification of chromatography NP, RP. It is observed the type of mobile phase is different is NP and RP or Low Pressure, High Pressure, Ion Exchange Chromatography. Accordingly we at LC Process ensures the MoC and compatibility of material of seals, metal and other parts that come in contact with mobile phase, sample and solvent used on different type of chromatography. There are many types of stationary phase developed for specific applications such as normal-phase and reversed phase chromatography. The right selection of chromatography media chemistry is particularly important to obtain good separation process. Selection of stationary phase depends on the characteristics of components in the mixture that are to be separated. NP-HPLC or Normal-phase Chromatography is a kind of adsorption chromatography. NP-HPLC method separates analytes based on their affinity for a polar stationary surface such as silica, hence it is based on analyte ability to engage in polar interactions. NP-HPLC uses non-aqueous, non-polar, mobile phase and depends effectively on separating analytes that are readily soluble in non-polar solvents. The separation depends on Hydrogen-bonding or dipole-dipole type of interactions with the sorbent surface playing its role. The analytes are retained on polar stationary surface based on the functional group and other various chemistry factors. The stable gradient flow from LC Process Chromatography system help is good separation and reproducible performance. There are strainers, inline filters to filter mobile phase, load, sample so as to ensure that any undissolved particle or foreign bodies in the mobile phase stuck on the filter and do not flow on the DAC Column, ensure to maintain the DAC Column frits cleaner for longer duration, throughout the process. It is always recommended that Peptide or Recombinant Proteins or anything that goes into the chromatography system should be preferably filtered, through a 0.45 mm filter before using on chromatography system. This also help to improve separation and increase life of stationary phase. In general, A chromatography process will start by washing the column followed by column to re-equilibration to the conditions that will start the chromatography run. The specific separation method is set on the LC Process 21 CFR Part 11 software, the parameters such as pump flow gradient, run time, and the acquisition. LC Process software us user friendly, allows complete automation, partial or semi automation, toggling between automatic and manual mode to run the purification method. Once the equilibrium is achieved, the actual sample loading and fraction collection is initiated. LC Process ensures that the purification is completed with comfort and simple operations. LC Process system is loaded with world class components and modules, with ATEX and Non ATEX Pump, Flow Meter, PLC, ATEX Panel, VFD’s Industrial PC, ATEX Motors, UV Detectors, NIR Sensors, Conductivity Sensor, pH Sensor, Dynamic inline degasser etc. The LC Process HPLC / MPLC / LPLC System is controlled by PLC based 21 CFR Part 11 compliance software.