Sample treatment: Extraction

Nutrient extraction in soils, sediments and plants (nitrogenated with CaCl2, phosphorus assimilable by the Olsen method) for further analysis using a segmented autoanalyzer equipment or a UV/VIS spectrophotometer.
Extraction (with ClK or SO4K) of C, N and P microbial biomass in soils and sediments by fumigation with chloroform for its later determination (analyses costs are not included here; see "Contenido en Biomasa" for complete C, N and P determinatios).
Extraction by diffusion of the 15NH4 and 15NO3 dissolved in water for its later determination by IRMS.

Nucleic acids extraction (DNA and RNA) per 10 samples

Additive extraction using sohxlet
Determination of chemical crosslinking in polymers.

Hormone extraction from samples of blood plasma, egg yolk, feces and/or feathers. Different tissues allow us to take the sample in a more or less invasive method and therefore samples can be obtained by directly handling the animal or indirectly from remains. However, results could vary because when using blood we obtain hormone levels at the time of sampling while in feces, eggs or feathers measured hormone levels correspond to a prior period which is variable. Also, on feces and feathers hormones are often modified as metabolites and, therefore, data interpretation may be more complex.
Blood plasma samples and feces will be received in the laboratory as frozen samples and feathers in individual envelopes at room temperature. Egg yolk could be analyzed from whole frozen eggs (before being incubated) or from isolated yolks stored frozen in individual tubes. In the case of blood it is necessary to separate plasma before freezing. Blood will be either collected in tubes containing anticoagulant (heparin or EDTA), or extracted into heparinized capillaries before being decanted into microtubes. Then, tubes must be inverted five or six times and centrifuged (10 min at 1000g). The plasma must be kept frozen at -20 ° C. Similarly, feces will be frozen in vials or bags at -20 ° C. The method for extracting hormones depends on the sample, but usually includes a homogenization and a separation by polar affinity in methanol, ethyl alcohol or ether which will result in a dried sample that will be reconstituted in the appropriate buffer.
The minimum required sample is highly dependent on hormone and species, contact us for more details, before the study when possible. As a general rule a minimum of 100 ul of plasma, 100 mg of feces or egg yolk or 20 mg of feathers per sample is needed.

Treatment of solid samples, to obtain a solution in which we will subsequently determine the corresponding analytes, using the appropriate solvent and the necessary physical methods (agitation, filtration, centrifugation).

EQUIPMENT:
- Shimadzu Nexera - SPDM20A - ABSciex 3200QTRAP
UHPLC Chromatograph (ultra high pressure liquid chromatography) Shimadzu Nexera with ultraviolet absorption photodiode array detector (DAD), coupled to an atmospheric pressure ionization source (ESI and APCI) ABSciex 3200QTRAP® triple quadrupole/ion trap mass spectrometer.
TECHNIQUE:
UHPLC-DAD-MS (ultra high efficiency liquid chromatography with ultraviolet photodiode detector and triple quadrupole mass detector)
PROVISIONS:
A QUALITATIVE ANALYSIS:
Detectors: DAD, MS (singleQ), MS2 (tripleQ) and MS3 (tripleQ-trap)
- Direct infusion of compounds. Detector: mass spectrometer (MS, MS2 and MS3).
- Chromatographic separation of complex mixtures by HPLC and UHPLC. Detector: DAD and mass spectrometer (MS, MS2 and MS3):
- Unknown analytes without structural elucidation
- Unknown analytes with structural elucidation
- Known analytes without standards.
- Known analytes with standards.
B QUANTITATIVE ANALYSIS:
Detectors: DAD, MS (singleQ), MS2 (tripleQ) and MS3 (tripleQ-trap)
- Direct infusion of compounds. Detector: mass spectrometer (MS and MS2). Known analytes with standards.
- Chromatographic separation of complex mixtures by HPLC and UHPLC. Detectors: DAD and mass spectrometer (MS and MS2). Quantification of known analytes with standards.
C OPTIONAL PROVISIONS:
- Sample preparation.
- Development of chromatographic methods: adaptation of an existing one, based on a publication, de novo, ... .
- Data processing: spectra interpretation, signal integration.
- Complete technical report of results, method validation parameters and other actions taken, in addition to the simple direct presentation of the requested results.

Fluorescent in situ hybridization (FISH)