Responsable: Fatima Domínguez Timón - Technician
bioanalisis.cbgp@upm.es 910679185 ( S51)

 

Responsable: Inmaculada Gude Rodríguez - Technician
bioanalisis.cbgp@upm.es 910679185 ( S51)

 

The bioanalysis unit is equipped with equipment for the extraction, prepurification and quantification and/or identification of small molecules.

 

The bioanalysis unit offers service to CBGP members at cost price. The services offered cover the absolute quantification of compounds of interest, such as metals, plant hormones, amino acids, carbohydrates, and their derivatives.

Responsable: Fatima Domínguez Timón - Technician
bioanalisis.cbgp@upm.es 910679185 ( S51)

 

Supervisor científico: Manuel González-Guerrero - Professor
manuel.gonzalez@upm.es 910679135 - 910679134 ( 279 )

RESERVATION FORM

 

contrAA 800 D – Combined device for flame and graphite tube techniques

The contrAA 800 is a high-resolution continuum source atomic absorption spectrometer for flame and graphite tube techniques. It is suited for the sequential detection of metallic and non-metallic traces in liquid and dissolved samples. Combined with an autosampler the contrAA 800 can be used as a multielement automatic unit to be used during routine analysis.

 

Graphite tube system


The autosampler AS-GF is used in the graphite tube technique for feeding liquid samples. The autosampler AS-GF accepts defined volumes of different solutions and places them into the graphite tube. It enables: the addition of up to five modifiers to the sample solution; transport of the sample solution to the thermal pretreatment in the tuve; enrichment of samples; placement of components in the preheated tuve; separate transport of components with intermediate washing; automatic preparation of standards by dilution or by different volumes; fixed, preselected or intelligent sample dilution and fully automatic multi-element mode.

Protocols available for the determination of Fe, Mo, Cu Zn and P. For other determinations, consult in advance.

 

Flame system


Flame atomic absorption spectroscopy is used for the determination of trace elements in the concentration range from µg/L to mg/L and for the determination of main components. It requires a flame with constant properties. The flame composition must also be compatible with the respective element.


The contrAA 800 D can be operated with the following types of flames and their corresponding burners: 

  1. Acetylene-air flame with 50 mm one-slit burner (universal burner)
  2. Acetylene nitrous oxide flame with a 50 mm one-slit (not available)

The acetylene-air flame can be used for most elements.


The autosampler facilitates the automated operation during multiple element analysis. The device parameters for the sample supply are configured with the control software ASpect CS. The autosampler AS-FD: is an automatic autosampler, also has a dilution function. The autosampler has sample tray with 54 positions for 50 mL cups.

 

Responsable: Inmaculada Gude Rodríguez - Technician
bioanalisis.cbgp@upm.es 910679185 ( S51)

 

Supervisor científico: María Garrido Arandia - Assistant Professor
maria.garrido@upm.es 910679129 ( 151/S78)

RESERVATION FORM

 

MicroCal PEAQ-ITC

The MicroCal PEAQ-ITC is a highly sensitive, low volume isothermal titration calorimeter for the label-free in solution study of biomolecular interactions. It delivers direct measurement of all binding parameters in a single experiment and can analyze weak to high affinity binders. 


 

 

Features and benefits


MicroCal PEAQ-ITC is designed for ease-of-use and exceptional sensitivity. The system directly measures heat released or absorbed during biochemical binding events, from which it calculates binding affinity (KD), stoichiometry (n), enthalpy (ΔH), and entropy (ΔS). With minimal sample preparation and system optimization data is generated quickly and easily. The wide affinity range enables analysis of weak to high affinity binders, with excellent reproducibility.

A wide range of applications can be investigated with MicroCal PEAQ-ITC, including characterization of molecular interactions of small molecules, proteins, antibodies, nucleic acids, lipids and other biomolecules. It can also be used to measure enzyme kinetics.

How it Works


Isothermal titration microcalorimeters measure the heat change that occurs when two molecules interact. Heat is released or absorbed as a result of the redistribution and formation of non-covalent bonds when the interacting molecules go from the free to the bound state. ITC monitors these heat changes by measuring the differential power, applied to the cell heaters, required to maintain zero temperature difference between the reference and sample cells as the binding partners are mixed. The reference cell usually contains water, while the sample cell contains one of the binding partners (the sample, often but not necessarily a macromolecule) and a stirring syringe which holds the other binding partner (the ligand). The ligand is injected into the sample cell, typically in 0.5 to 2 μL aliquots, until the ligand concentration is two- to three-fold greater than the sample. Each injection of ligand results in a heat pulse that is integrated with respect to time and normalized for concentration to generate a titration curve of kcal/mol vs molar ratio (ligand/sample). The resulting isotherm is fitted to a binding model to generate the affinity (KD), stoichiometry (n) and enthalpy of interaction (ΔH).







 
ITC-TEORIA Y PRACTICA

 

Responsable: Inmaculada Gude Rodríguez - Technician
bioanalisis.cbgp@upm.es 910679185 ( S51)

 

Supervisor científico: Antonio Molina Fernández - Professor
antonio.molina@upm.es 910679213 - 910679201 ( 235)


RESERVATION FORM


Dionex HPIC System ICS-6000


The Dionex ICS 6000 chromatography system is a compact, modular, and versatile system designed to perform various types of isocratic and gradient ion exchange separations. The entire system is inert and metal-free, ensuring that no metallic components come into contact with the sample or eluent, allowing operation at extreme pH levels. It is a high-pressure system designed for the determination of ions and charged molecules in diverse applications.


 

 

ELUENT DELIVERY SYSTEM


The Dionex ICS-6000 system features a dual-piston, high-pressure quaternary pumping system with internal vacuum degassing for eluent delivery. It has the capability to perform unlimited gradients of varying concentrations using up to four different mobile phases.

 

AUTOSAMPLER MODULE


It includes an autosampler for 50 vials of 5 mL or 0.5 mL, incorporating a sample filtration system vial by vial. It has individual plugs and filters for each vial to prevent cross-contamination.

DETECTOR MODULE


The system incorporates a column oven that can heat guard and analytical columns within a temperature range from 10°C to 70°C (at least 5°C above ambient temperature). The bioanalysis unit provides CarboPac PA1, PA20 and PA100 columns.

Furthermore, the detector compartment can operate within a temperature range of 18 to 40°C to enhance analyte signals and improve sensitivity. 

The electrochemical detector contains a gold electrode, which is essential for the analysis of inorganic ions, carbohydrates, and contaminant determination.

APPLICATIONS


The Dionex ICS-6000 HPIC system is a high-pressure ion chromatography equipment designed for the separation and quantification of ions and charged molecules in various applications, ranging from water and wastewater analysis to biological research and environmental studies.

 

 

Responsable: Inmaculada Gude Rodríguez - Technician
bioanalisis.cbgp@upm.es 910679185 ( S51)

 

Supervisor científico: Araceli Díaz Perales - Professor
araceli.diaz@upm.es 910679129 ( 151/S78)

RESERVATION FORM

 

Agilent HPLC System (1260 Infinity II Quaternary System)

The LC 1260 Infinity II is a versatile HPLC system that offers the highest functionality and operational convenience for analytical and preparative HPLC.

The Agilent 1260 Infinity II quaternary pump is ideal for a wide range of applications and columns. The formation of quaternary gradients with low-pressure mixing makes it the ideal pump for applications requiring maximum flexibility in solvent combination, as well as for UHPLC separations (including the use of STM columns) to reduce analysis times by over 70%. With a maximum pressure of 600 bars, a vacuum degasser for four channels, and flow rates of up to 5 ml/min, it allows for injection overlapping to increase productivity.

 

The column compartment has two independently temperature-controlled zones using Peltier technology. It allows for preheating of the solvent in the precolumn. With a temperature range of 10⁰C below room temperature (minimum of 4⁰C) up to 85⁰C. The bioanalysis unit has a Poroshell 120 EC-C18 column.

The 1260 Infinity II system is equipped with two detectors:

 

  1. Variable and programmable ultraviolet/visible wavelength detector. Dual-beam photometer and deuterium lamp for high sensitivity, low detection limits with very low noise and baseline drift.
  2. Fluorescence detector with a long-lasting pulsating xenon lamp that stabilizes in microseconds, reducing costs by allowing it to be turned off when data is not being acquired.

Additionally, the system includes a fraction collector that allows us to recover the separated and detected analytes by the HPLC, with the minimum possible dead volume to avoid peak dispersion. The equipment allows us to trace the different samples and their fractions, being able to select them by retention times, peaks, or analyte mass of the required fractions.

 

 

Responsable: Fatima Domínguez Timón - Technician
bioanalisis.cbgp@upm.es 910679185 ( S51)

 

RESERVATION FORM

 

ASE 350 Accelerated Solvent Extractor

The ASE® 350 Accelerated Solvent Extractor (see Figure 1-1) is an automated system for extracting organic compounds from a variety of solid and semisolid samples. The DioniumTM internal pathway allows extraction of mildly acidic or basic samples.


The ASE 350 uses a combination of elevated temperature and pressure with common solvents to increase the efficiency of the extraction process. The result is faster run times and a significant reduction in solvent use. To further reduce solvent use, the ASE 350 supports a unique extraction method called the solvent saver mode. There are two solvent saver options:



  1. The pressure mode is a static mode of extraction that minimizes solvent usage by  maintaining a fixed cell pressure and eliminating the introduction of fresh solvent during the heating and static cycles. This mode uses less solvent than any other ASE 350 extraction mode.
  2. The flow mode is a continuous dynamic extraction mode in which small aliquots of solvent are delivered to the cell at the specified Flow rate throughout the static step. The cell temperature remains constant throughout the run.

Solvent saver methods are recommended for:

  1. Modifying and enhancing existing extraction methods
  2. Exploring new extraction methods
  3. Simulating extraction on a small scale before scale-up to a plant scale extraction module.

During a run, the cell is filled with solvent and then heated and pressurized to maintain the solvent in a liquid state. After the run, the extract is rinsed from the cell into a collection vessel (a bottle or vial). If adsorbents are used in the sample cell, post-extraction cleanup may not be required. If adsorbents are not used, sample cleanup may be required.

Theory of ASE


Responsable: Inmaculada Gude Rodríguez - Technician
bioanalisis.cbgp@upm.es 910679185 ( S51)

 

Supervisor científico: Stephan Pollmann - Professor
stephan.pollmann@upm.es 910679143 ( 134 )

 

 

GC-QqQ-MS/MS System (SCION TQ)

The GC-QqQ-MS/MS system is a triple quadrupole mass spectrometer coupled to a gas chromatograph. With its high precision and mass resolution, this equipment allows the determination of analytes from 50 to 600D that can be volatilized. The bioanalysis unit has protocols for the determination of plant hormones: abscisic acid, 3-idolacetic acid, jasmonic acid, and salicylic acid. For other determinations, consult in advance.

 

The equipment consists of:

Ion Source


In the ion source, sample components are exposed to conditions that generate ions. These conditions can be gentle or harsh enough to fragment molecules.

 

  1. Electron Ionization: It is the traditional ionization technique in GC/MS and is suitable for all compounds in the gas phase.
  2. Chemical Ionization: It is a good strategy when no molecular ion is observed in electron ionization and to confirm the mass-to-charge ratio of the molecular ion.

Mass Analyzer

Ions exit the ion source and enter the mass analyzer, which consists of four parts:
  1. The first section provides collimation of the ion beam and synchronization of the ions with the quadrupole field. This provides a more focused ion stream that improves sensitivity, eliminates neutrals, and reduces noise.
  2. The second section separates ions according to their mass-to-charge ratio.
  3. The third section can operate in two modes:
    1. In MS/MS mode, it is used as a collision or gas cell to provide additional fragmentation of the precursor ion(s) that have been filtered. The generated fragments are directed to the next filtering stage.
    2. In MS mode, it is used as a transmission quadrupole to send ions to the detector. Neutrals are lost to reduce noise.
  4. The last section separates ions according to their mass-to-charge ratio. Ions are moved from here to the detector.
Positive and negative ions can be detected with similar efficiency.



Responsable: Fatima Domínguez Timón - Technician
bioanalisis.cbgp@upm.es 910679185 ( S51)

 

Supervisor científico: Antonio Molina Fernández - Professor
antonio.molina@upm.es 910679213 - 910679201 ( 235)

 

 

Low-Pressure Chromatography System BioLogic LP (Bio-Rad)

This is an appropriate system for protein separation using conventional chromatographic techniques such as ion exchange, gel filtration, etc. The system includes a peristaltic pump capable of delivering flows from 0.1 to 40 mL/min at a maximum pressure of 30 psi (2 bars). Additionally, it features a gradient mixer capable of using two buffers simultaneously and a collector for up to 80 fractions.