Conjugation and Other Methods in Polymeric Vaccines
Mesut Karahan in Synthetic Peptide Vaccine Models, 2021
Chromatography is known as an important biophysical technique that can separate, identify, and purify the blend segments for subjective and quantitative investigation. Proteins can be purified based on properties such as size and shape, total charge, hydrophobic groups present on the surface, and the ability to bind to the stationary phase. It is based on molecular properties and interaction type use mechanism, four separation technologies, ion exchange, dispersion, surface adsorption, and size exclusion. Column chromatography is one of the most used and common techniques for protein purification methods. This technique is basically used to purify biological molecules. The application of the method can be summarized as follows. The sample is separated on the column (stationary phase) and then the wash buffer is added to the column (mobile phase). It flows through the column material placed on the fiberglass support. With the help of the wash buffer, the samples are accumulated at the bottom of the column chromatography instrument, based on time and volume (Coskun 2016). Column chromatography is a powerful purification and separation process that is closely controlled to the hydrodynamic diameters of the macromolecules depending on the diameter of the pores in the filling material (see in HPLC Method) (Acar 2006; Fornaguera and Solans 2018).
Dictionary
Mario P. Iturralde in Dictionary and Handbook of Nuclear Medicine and Clinical Imaging, 1990
Chromatography. This method of chemical analysis uses a group of methods for separating a mixture of substances into their component parts. Chromatography is a misnomer for the technique since the color of the components is not really used for identifying substances in modern techniques. There are four basic types of chromatography using one phase or substrate which may be liquid or solid through which the test substance (gas or liquid) moves. A common feature of these methods is that the difference in rate of movement of the components of the test sample (plus reagents) is used to identify the separate components. In the clinical laboratory these methods are used to detect complex substances such as drugs and hormones. Examples of chromatographs are the gas-liquid Chromatograph (GLC) and the thin-layer Chromatograph (TLC). Both of these are being superseded by high-performance liquid chromatography (HPLC), although some compounds can only be measured by GLC.
Drug Design, Synthesis, and Development
Nathan Keighley in Miraculous Medicines and the Chemistry of Drug Design, 2020
Once the synthesis of a drug is complete, the compound is not yet ready for consumption. Thorough purification steps are required to clean the organic compound. Chromatography is a commonly adopted technique for separating a target compound from the rest of the matrix. In chromatography, the column is packed with a solid stationary phase consisting of very fine particles and a solvent is selected, referred to as the mobile phase, in which the target compound dissolves. The desired target product will have a different affinity for the stationary phase than any by-products or unreacted material, and therefore will have different retention times on the column. The product elutes from the column at a different time to the contaminants, so can be isolated. The greater the length of the column, the greater the degree of separation, as in the case of high-performance liquid chromatography (HPLC) and gas chromatography (GC), used for volatile compounds, and even separation of stereoisomers is possible.
Using online content uniformity measurements for rapid automated process development exemplified via an X-ray system
Published in Pharmaceutical Development and Technology, 2019
Bernhard Wagner, Thomas Brinz, Johannes Khinast
Machines for the online gravimetric weighing are commercially available from several manufactures and can detect the capsule weight within the RAPD concept. Compared to this, a CU check requires much more effort and equipment. On the one hand, there are the analytical techniques like for example HPLC (high performance liquid chromatography), TLC (thin layer chromatography), or GC (gas chromatography). These techniques require an elaborated sample preparation before the content can be determined. The measurements take a certain time and the content cannot be measured in-line within the capsule production. Thus, an integration into the RAPD would be challenging, as a requirement is a fast measurement of the samples in an automated way. Possibly, the measurements can be automated (e.g. Saitoh and Yoshimori 2008) or executed offline. However, significant time and effort is required.
Comparison of pharmacokinetics of phytoecdysones and triterpenoid saponins of monomer, crude and processed Radix Achyranthis Bidentatae by UHPLC-MS/MS
Published in Xenobiotica, 2020
Liu Yang, Hai Jiang, Meiling Yan, Xudong Xing, Xinyue Guo, Wenjing Man, Ajiao Hou, Bingyou Yang, Qiuhong Wang, Haixue Kuang
Stationary phase and composition of mobile phase were investigated to obtain good chromatographic conditions. Thermo Hypersil GOLD C18 column (100 mm× 2.1 mm, 1.9 μm) was chosen in this study for its good peak symmetry. Different mobile phases (acetonitrile–water and methanol–water or with different concentration of formic acid and acetic acid) were examined to obtain efficient chromatography and relatively short run time for the analytes and IS. It was found that the acetonitrile–water system was better than the methanol–water system for its great resolution of the isomers and the addition of acetic acid in mobile phases could increase the ionization of the analytes and IS. The retention times for β-ecdysterone, 25-S inokosterone, 25-R inokosterone, chikusetsusaponin IVa, ginsenoside R0 and IS were 2.64, 3.00, 3.24, 8.76, 8.64 and 8.51 min, respectively.
A review on human body fluids for the diagnosis of viral infections: scope for rapid detection of COVID-19
Published in Expert Review of Molecular Diagnostics, 2021
Sphurti S Adigal, Nidheesh V Rayaroth, Reena V John, Keerthilatha M Pai, Sulatha Bhandari, Aswini Kumar Mohapatra, Jijo Lukose, Ajeetkumar Patil, Aseefhali Bankapur, Santhosh Chidangil
Several studies have reported the use of different spectroscopic techniques for early diagnosis of various diseases using different body fluids as clinical specimen [91]. High-performance liquid chromatography (HPLC) is considered as a versatile tool for separation and analysis of the biological and pharmaceutical compounds. Chromatography column is the heart of chromatography technique and hence a column selection is important to separate the mixture of components in a sample of interest [92]. Generally, chromatography-based techniques are used as standard techniques by industrial sectors, federal agencies, academies, and food and drug administration (FDA). Using chromatography-based-techniques the limit of detection (LOD) for melamine was achieved to the ppb level [93]. The combination of ultrasensitive optical technique Laser-Induced Fluorescence (LIF), with highly efficient separation technique such as HPLC, detection of ultra-trace quantities of individual biomolecules in complex, multicomponent physiological systems is feasible [94]. Our earlier studies demonstrated the capability of HPLC-LIF for protein profile analysis of micro-quantities of clinical samples such as saliva [95], serum [96], cellular samples [16] and tissue homogenates [97] for the diagnosis of cancers of different types.
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