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MB Sample ID: SA086293
| Local Sample ID: | selenium-5 |
| Subject ID: | SU001289 |
| Subject Type: | Invertebrate |
| Subject Species: | Apis mellifera |
| Taxonomy ID: | 7460 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001289 |
| Subject Type: | Invertebrate |
| Subject Species: | Apis mellifera |
| Taxonomy ID: | 7460 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| selenium-5 | SA086293 | FL012865 | Selenate | Factor |
Collection:
| Collection ID: | CO001283 |
| Collection Summary: | We sampled three bees from 13 cages after four days of continuous exposure to the treatments and immediately placed the samples on dry ice, followed by long-term storage at -80 °C. |
| Sample Type: | Insect tissue |
| Storage Conditions: | Described in summary |
Treatment:
| Treatment ID: | TR001304 |
| Treatment Summary: | Once the bees had an established microbiome, we prepared treatment feeding solutions of 50% sucrose (as a no metal/metalloid control), 50% sucrose spiked with 0.6 mg/L sodium selenate or 50% sucrose with 0.24 mg/L cadmium chloride (Alfa Aesar, Ward Hill, MA) and pollen patties spiked with either 6.0 mg/L selenium or 0.46 mg/L cadmium and allowed the bees to feed ad libitium. |
Sample Preparation:
| Sampleprep ID: | SP001297 |
| Sampleprep Summary: | Samples were freeze-dried, homogenized, and extracted with 30:30:20:20 acetonitrile:methanol:water:isopropanol. Samples were then sonicated, vortexed, and centrifuged before analysis. |
Combined analysis:
| Analysis ID | AN002035 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Waters Acquity I-Class |
| Column | SeQuant ZIC-pHILIC (150 x 2.1mm,5um) |
| MS Type | ESI |
| MS instrument type | Triple quadrupole |
| MS instrument name | Waters Xevo-TQ-S |
| Ion Mode | POSITIVE |
| Units | peak area |
Chromatography:
| Chromatography ID: | CH001475 |
| Chromatography Summary: | We analyzed targeted, polar primary metabolites on a Xevo TQ-XS triple quadrupole mass spectrometer (Waters) coupled to an I-class UPLC system (Waters) in the UC Riverside Metabolomics Core Facility. We used a ZIC-pHILIC column (2.1 x 150 mm, 5 µM) (EMD Millipore) for separations with the following mobile phases:(A) water with 15 mM ammonium bicarbonate adjusted to pH 9.6 with ammonium hydroxide and (B) acetonitrile. We set the flow rate to 200 µL/min and held the column 50° C with an injection volume of 1 µL. We used the following gradient: 0 min, 90% B; 1.5 min, 90% B; 16 min, 20% B; 18 min, 20% B; 20 min, 90% B; 28 min, 90% B. |
| Instrument Name: | Waters Acquity I-Class |
| Column Name: | SeQuant ZIC-pHILIC (150 x 2.1mm,5um) |
| Column Temperature: | 50 |
| Flow Gradient: | 0 min, 90% B; 1.5 min, 90% B; 16 min, 20% B; 18 min, 20% B; 20 min, 90% B; 28 min, 90% B |
| Flow Rate: | 200 ul/min |
| Solvent A: | 100% water; 15 mM ammonium bicarbonate, pH 9.6 |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS001887 |
| Analysis ID: | AN002035 |
| Instrument Name: | Waters Xevo-TQ-S |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | We operated the MS in selected reaction monitoring mode with source and desolvation temperatures at 150° C and 500° C, respectively, with the desolvation gas set to 1000 L/hr, cone gas set to 150 L/hr and collision gas set to 0.15 mL/min. We used nitrogen gas for each step, except for the collision gas, which was argon, and we set capillary voltage to 1 kV in positive ion mode and 2 kV in negative ion mode. Similar to the untargeted methods, we generated a quality control sample by pooling equal aliquots of each sample and analyzed the QC sample every 3-4 injections to monitor system stability and performance. We processed the metabolite data (peak picking, alignment, deconvolution, integration, normalization, and spectral matching) with Progenesis Qi software (Nonlinear Dynamics, Durham, NC). We used Skyline software (MacLean et al. 2010) for data processing our targeted metabolites. |
| Ion Mode: | POSITIVE |
