Configured liquid chromatography (HPLC) system for analysis of carbohydrates, amino acids, peptides and glycoproteins
Equipment description
The HPLC platform will soon be allocated to the Glycoproteomics Laboratory, Dept. of Parasitology, room 09, under the supervision of Prof. Dr. Giuseppe Palmisano.
The platform is composed of two integrated HPLC systems: the first HPLC is an ion chromatography system coupled with an electrochemical detector and the second HPLC is an analytical chromatography system coupled with a UV-Vis and fluorescence detector:
1 – The First HPLC system has a quaternary bomb, capable of reaching 5000 psi, and can operate in isocratic or gradient mode. Different types of gradients can be used: linear, concave, and convex. The system can operate with flow ranging from 0,001 to 10 mL/min. All system piping is made of PEEK, resistant to pH 0 – 14, which is ideal when working with high pH. In addition, the system has a solvent channel that generates KOH as an eluent.
2 – The second HPLC system has a quaternary pump that can reach up to 15000 psi and can operate as HPLC or UPLC. It has an autosampler with temperature control. This system is coupled to a “diode-array” detector and a fluorescence detector.
The equipment will be configured to primarily analyze peptides, glycans, and proteins isolated from different biological matrices. For biomolecules that are not fluorescent, samples must be derivatized. This procedure must be done in the user’s laboratory. Analyzes that require the development of new methods and choice of columns must be discussed with the person responsible for the equipment. Chromeleon 7 software is integrated into the platform and allows data analysis including analyte identification and quantification.
Use of Equipment
All those interested in developing research projects on this equipment should contact the responsible team through the email reservas.hplc.gp@gmail.com and schedule a meeting. The meeting aims to discuss the details of the analysis, such as the column to be used, solvent, temperature, and gradient.
The applicant must be the professor responsible for the research, who is responsible for any and all damages that may occur during the use of the equipment. The applicant must indicate the source of funding for his research project and indicate the qualified users of his group. After the meeting and discussion of the experiment, a date will be scheduled for analysis according to the availability of the calendar. Specific experiments that require more time to use the equipment, as in cases that require method development, should be discussed with the person in charge.
The equipment will be available to users for three days every three weeks: from Wednesday to Friday and may also include Saturday and Sunday if the samples are put to run in the autosampler on Friday. If you give up using the equipment, the user must cancel the appointment at least 48 hours in advance. The Management Committee may deny the use of the equipment if it understands that the user does not comply with correct scheduling/cancellation practices or that it may cause damage to the equipment due to misuse.
Each user must provide their column and analytical standards, and pay an equipment usage fee, to cover expenses for consumables and routine maintenance. The equipment usage fee will be determined based on fees charged by different facilities around the world and will be discussed later by the Management Committee.
Users are responsible for the preparation, execution, and analysis of the data generated by their use. The analysis of the generated data can be done at the Glycoproteomics Laboratory and the time allocated for this must be included in the scheduling time.
Qualified personnel to operate the equipment
The operation of the HPLC system must only be conducted by qualified and trained personnel for its operation. The training will be offered by the Analytical company team or by members of the Glycoproteomics Laboratory group, depending on their availability. The equipment may be used by professors, employees, undergraduate, and graduate students, and researchers from ICB/USP or other teaching and research institutions.
Each user laboratory must be accredited to use the equipment, indicating who would be the people qualified to operate the equipment. Only authorized persons may use the device. Eligibility to use the device will be certified after training.
Equipment Resources and Facilities
The equipment will be allocated in the Glycoproteomics Laboratory in the Dept. of Parasitology, ICB-USP which contains available space and resources for its installation. The consumables necessary for the experiments (injection and collection vials, solvents) are included in the equipment usage fee, being necessary for the user to provide columns and analytical standards. Eventually, the professors responsible for the user laboratories may agree with the Management Committee on the purchase of consumables for replacement and the payment of any necessary maintenance for the proper functioning of the equipment.
Management Committee
The Management Committee is responsible for the equipment, ensuring its proper functioning and access to internal and external users. The following management committee was created for this equipment:
Prof. Dr. Giuseppe Palmisano – Dept. Parasitology – ICB-USP – RESPONSIBLE
Prof. Dra. Graziella Eliza Ronsein IQ-USP
Prof. Dr. Claudio R. F. Marinho – Dept. Parasitology – ICB-USP
Users Committee
The Users Committee is responsible for listening to users and guaranteeing their demands regarding the use of the equipment. It consists of routine users. The committee should take criticisms and suggestions about the equipment’s operating system to the Management Committee.
Prof. Dr. Carlos Winter, Dept. of Parasitology – ICB-USP
Prof. Dr. Carsten Wrenger – Dept. Parasitology – ICB-USP
Prof. Dr. Fernando Rodrigues de Moraes Abdulkader- ICB-USP