Samplejet autosampler

Manufactured by Bruker

Sourced in Germany, United States

The SampleJet autosampler is a versatile laboratory instrument designed to automate sample handling and introduction for various analytical techniques. Its core function is to efficiently and precisely deliver samples to the appropriate measurement device, enabling consistent and reliable data acquisition.

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SampleJet (36 citations)

25 protocols using samplejet autosampler

Plasma Metabolite Profiling by NMR Spectroscopy

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After all subjects had fasted for 10 hours, venous blood samples were drawn into vacuum tubes and stored at room temperature for 30 min before centrifugation (1,200 ×g for 10 min). Plasma samples were collected in vacuum tubes, stored at −80 ℃, and transported on dry ice. Samples were collected in the blood collection room of the hospital. Plasma samples were prepared and tested using Bruker IVDr Standard Operating Procedures (19 (link)) at ProteinT Biotechnology Co., Ltd. (Tianjin, China). Samples were thawed at room temperature, and 400-µL plasma samples were mixed thoroughly with 400 µL buffer (phosphate buffer pH 7.4 containing TSP-D4; Bruker, Rheinstetten, Germany), of which 600 µL was transferred to a 5-mm NMR tube for analysis.
The test was performed on a 600 MHz NMR AVANCE III HD spectrometer equipped with a BBI probe head and a SampleJet autosampler, adjusted to 6 ℃ during the test (Bruker Biospin). Before acquisition of sample data, each sample was automatically tuned and shimmed. The free induction decay signals (FIDs) were presented in the form of a Fourier-transformed spectrum, and automatic phase and baseline correction were performed in Toppin software as Bruker IVDr. The concentrations of metabolites were expressed as mmol/L (Figure S1).

Gu W., Pan Y., Zhao W., Liu J, & Meng Y. (2023). Metabolic signatures of lymphangioleiomyomatosis in biofluids: nuclear magnetic resonance (NMR)-based metabonomics of blood plasma: a case-control study. Annals of Translational Medicine, 11(2), 76.

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Metabolic Profiling of CSF Samples

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CSF samples was mixed (5:1) with phosphate buffer solution containing 1.5 M KH₂PO₄, 580 µM TSP-d4, NaN₃, D₂O, pH 7.4 and transferred to 3 mm NMR tubes. The samples were analyzed at the Swedish Nuclear Magnetic Resonance (NMR) Centre in Gothenburg using an Oxford 800 Mhz magnet equipped with a Bruker Avance III HD console, 3 mm TCI cryoprobe and a cooled Sample Jet auto sampler (Bruker BioSpin, Fällanden, Switzerland), as described previously [16 (link)]. Generated data from NMR spectrometer were processed in TopSpin3.5pl7 (Bruker BioSpin, Fällanden, Switzerland). Chenomx 9.0 (Chenomx Inc., Edmonton, AB, Canada) and the Human Metabolite Database [24 (link)] were used for identification of the metabolite signals.

Bäckryd E., Thordeman K., Gerdle B, & Ghafouri B. (2023). Cerebrospinal Fluid Metabolomics Identified Ongoing Analgesic Medication in Neuropathic Pain Patients. Biomedicines, 11(9), 2525.

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NMR Analysis of Chicken Serum Metabolites

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¹H-NMR spectroscopic analysis of chicken serum samples was performed by the NMR Laboratory (nmr@chemistry.mcmaster.ca) in the Department of Chemistry and Chemical Biology, McMaster University (Hamilton, Ontario, Canada). One hundred microlitres of 0.9% saline in D₂O with 1 mg/ml sodium [2,2,3,3-d4]3-trimethylsilyIpropanoate (TSP-d4) was mixed with 500 µl serum in high quality 5-mm NMR tubes. ¹H NMR spectra of serum were recorded on a Bruker Avance III 700 MHz NMR spectrometer (Bruker Biospin, Rheinstetten, Germany) equipped with a 5 mm QNP cryo-probe and SampleJet autosampler, and operating at a proton frequency of 700.17 MHz. A carr-Purcel-Meiboom-Gill (CPMG) spin-echo pulse sequence [recycle delay−90°− (τ–180°–τ)_n–acquisition] was used to emphasize resonances from low molecular-weight metabolites^{14 (link),28 (link),61 (link)}. ¹H NMR data for each sample were acquired using 128 scans (64k data points) with a 2.5 second relaxation delay. Chemical shifts were referenced to the internal reference (TSP-d₄: 0.00 ppm). ¹H NMR data with water suppression using excitation sculpting with gradients were subsequently acquired using the same number of scans and time of relaxation delay as that for ¹H NMR data.

Yang X., Yin F., Yang Y., Lepp D., Yu H., Ruan Z., Yang C., Yin Y., Hou Y., Leeson S, & Gong J. (2018). Dietary butyrate glycerides modulate intestinal microbiota composition and serum metabolites in broilers. Scientific Reports, 8, 4940.

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Brain Metabolite Extraction and NMR Analysis

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All brain samples were prepared using a modified version of the protocol previously described by Graham et al. (2013) ²⁶ and Bahado-Singh et al. (2016) ²⁷. In brief, all brain specimens were weighed and defrosted on ice. Subsequently, samples were milled and extracted in 50 % methanol/water (1 g/ml) in a sterile 2 ml Eppendorf tube. Samples were mixed for 20 min, sonicated for 15 min and the protein removed via centrifugation at 13,000 g at 4 ºC for 30 min. Supernatants were collected and dried under vacuum using a Savant DNA Speedvac (Thermo Scientific, USA) and reconstituted in 285 μL of 50 mM potassium phosphate buffer (pH 7.0), 30 μL of Sodium 2,2-dimethyl-2-silapentane-5-sulfonate (DSS) and 35 μL of D₂O ²⁸. 200 μL of the reconstituted sample was transferred to a 3 mm Bruker NMR tube for NMR analysis. All samples were housed at 4°C in a thermostatically controlled SampleJet autosampler (Bruker-Biospin, USA) and heated to room temperature over 3 min prior to analysis by NMR.

Graham S.F., Rey N.L., Yilmaz A., Kumar P., Madaj Z., Maddens M., Bahado-Singh R.O., Becker K., Schulz E., Meyerdirk L.K., Steiner J.A., Ma J, & Brundin P. (2018). Biochemical profiling of the brain and blood metabolome in a mouse model of prodromal Parkinson’s disease reveal distinct metabolic profiles. Journal of proteome research, 17(7), 2460-2469.

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NMR Analysis of Hydrolyzed Biomass

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Collected biomass (5 mg) was resuspended in 6 M HCl and hydrolyzed for 12 h at 110 °C. The obtained hydrolysate was dried and washed twice with D₂O (dried again between each washing step), resuspended in 200 μL DCl [0.1% (v/v) in D₂O], and transferred into 3 mm NMR tubes for analysis. The biomass hydrolysates were analyzed in duplicate by 1D ¹H NMR, 1D ¹³C NMR, 2D ZQF-TOCSY NMR and 2D HSQC [21 (link)]. For the analysis, a spectrometer Bruker Ascend™ 800 MHz (Bruker, Billerica, Massachusetts, USA) equipped with a 5 mm CQPI (¹H, ¹³C, ³¹P, ¹⁵N) cryoprobe and a Sample Jet auto sampler (Bruker, Billerica, Massachusetts, USA) was used.

Schulze D., Kohlstedt M., Becker J., Cahoreau E., Peyriga L., Makowka A., Hildebrandt S., Gutekunst K., Portais J.C, & Wittmann C. (2022). GC/MS-based 13C metabolic flux analysis resolves the parallel and cyclic photomixotrophic metabolism of Synechocystis sp. PCC 6803 and selected deletion mutants including the Entner-Doudoroff and phosphoketolase pathways. Microbial Cell Factories, 21, 69.

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NMR Metabolomics Profiling of Fecal Samples

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NMR data were recorded using Bruker 600MHz AVANCE III Spectrometer equipped with a BBFO + probe and a Sample Jet autosampler, which enabled the storage of 5 racks of 96 NMR tubes at 5 °C.
The sample temperature was controlled at 300 K during experiments. Spectra were recorded using the 1D Nuclear Over Hauser Effect spectroscopy pulse sequence (trd-90°-t1-90°-tm-90°-taq) with a relaxation delay (trd) of 24 s, a mixing time (tm) of 4 ms, and a t1 of 4 μs. The sequence enables optimal suppression of the water signal that dominates the spectrum. We collected 128 free induction decays (FIDs) of 65,532 data points using a spectral width of 12,019.230 kHz and an acquisition time of 2.726 s. The spectra were automatically phased and baseline corrected and referenced to the internal standard (TSP; δ = 0.0 ppm).
The relaxation delay was set at 24 s in order to reach the complete relaxation of all the metabolites between scans; this is a mandatory step in NMR when absolute concentration of the metabolites is calculated.
Two-dimensional (2D) NMR spectra were obtained to aid the assignment of fecal metabolites. The set of 2D experiments included ¹H-¹H correlation spectroscopy (COSY); ¹H-¹H total correlation spectroscopy (TOCSY), and ¹H-¹³C heteronuclear single quantum correlation (HSQC) using the standard parameters implemented in Topspin 3.5pl7 (Bruker Biospin GmbH, Karlsruhe, Germany).

Zidi O., Souai N., Raies H., Ben Ayed F., Mezlini A., Mezrioui S., Tranchida F., Sabatier J.M., Mosbah A., Cherif A., Shintu L, & Kouidhi S. (2021). Fecal Metabolic Profiling of Breast Cancer Patients during Neoadjuvant Chemotherapy Reveals Potential Biomarkers. Molecules, 26(8), 2266.

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Optimized Metabolomics NMR Protocol

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¹H-NMR experiments were performed on Bruker Avance 600 MHz equipped with a SampleJet autosampler using a noesypr1d sequence, mixing time of 100 ms, a spectral window of 12 ppm, acquisition time of 2 s, relaxing time of 3 s, 516 scans, 4 dummy scans, and T = 298 K. This sequence has become the best choice for NMR-based metabolomics studies [42 (link)] for several reasons. Firstly, the quality of water suppression is very high without the need for extensive optimization. Secondly, an increasing number of well-established groups utilize the sequence, reflecting its consistency [43 (link)]. Finally, the library of Chenomx used in this study to quantify metabolite concentrations is optimized for this sequence and compensates for incomplete relaxation.

Petrella G., Montesano C., Lentini S., Ciufolini G., Vanni D., Speziale R., Salonia A., Montorsi F., Summa V., Vago R., Orsatti L., Monteagudo E, & Cicero D.O. (2021). Personalized Metabolic Profile by Synergic Use of NMR and HRMS. Molecules, 26(14), 4167.

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NMR Analysis of Brown Fat Metabolites

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Brown fat tissue samples (~20 mg) were homogenized and extracted with a mixture of ice-cold solvents water/methanol/chloroform (H₂O/MeOH/CHCl₃ 1:1:1, v:v:v). The water/methanol phase was separated and dried in a speedvac. The extract was reconstituted in a 550 μl mixture containing 500 μl of phosphate buffer pH 7.4 and 50 μl of 1mM of TSP-d₄ in D₂O, vortexed for 20 seconds and transferred to 5 mm NMR tubes. The NMR spectra were acquired on a Bruker 600 MHz Avance III HD spectrometer equipped with a BBI room temperature probehead and SampleJet auto sampler (Bruker Biospin, Rheinstetten, Germany). ¹H NMR spectra were recorded using 1D noesy pulse sequence with presaturation (noesygppr1d), collecting 256 scans with calibrated 90 degree pulse (~11 μs), 4.55 second acquisition time, and 4 second relaxation delay. Metabolites were identified and quantified using Chenomx NMR suite 8.2 software, by fitting the spectral lines of library compounds into the recorded NMR spectrum of tissue extract. The quantification was based on peak area of TSP-d₄ signal. The metabolite concentrations are exported as μM in NMR sample and normalized to wet tissue mass (nmol/mg of tissue).

Kazak L., Chouchani E.T., Lu G.Z., Jedrychowski M.P., Bare C.J., Mina A., Kumari M., Zhang S., Vuckovic I., Laznik-Bogoslavski D., Dzeja P., Banks A.S., Rosen E.D, & Spiegelman B.M. (2017). Genetic depletion of adipocyte creatine metabolism inhibits diet-induced thermogenesis and drives obesity. Cell metabolism, 26(4), 660-671.e3.

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Fungal Metabolite Extraction for NMR Analysis

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Metabolites were extracted from 5 mg freeze-dried fungal mycelia and dried in a speed vacuum as described previously (28 (link)). Extracts were reconstituted in 50 μl of deuterated sodium phosphate buffer (100 mM, pH 7.0) containing 0.5 mM trimethylsilylpropanoic acid (TMSP), 3 mM sodium azide, and 100% D₂O. Each sample was sonicated for 10 min and vortexed briefly, before a volume of 35 μl was transferred into 1.7-mm nuclear magnetic resonance (NMR) tubes.
Spectra were acquired on a Bruker 600 MHz spectrometer equipped with a TCI 1.7-mm z-PFG cryogenic probe and a Bruker SampleJet autosampler. One-dimensional (1D) ¹H NMR spectra and 2D ¹H-¹³C HSQC (heteronuclear single quantum coherence) spectra were recorded and analyzed for each sample as previously described (63 (link)).

Ries L.N., Alves de Castro P., Pereira Silva L., Valero C., dos Reis T.F., Saborano R., Duarte I.F., Persinoti G.F., Steenwyk J.L., Rokas A., Almeida F., Costa J.H., Fill T., Sze Wah Wong S., Aimanianda V., Rodrigues F.J., Gonçales R.A., Duarte-Oliveira C., Carvalho A, & Goldman G.H. (2021). Aspergillus fumigatus Acetate Utilization Impacts Virulence Traits and Pathogenicity. mBio, 12(4), e01682-21.

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Metabolic Profiling of Brown Fat

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Brown fat tissue samples (~20 mg) were homogenized and extracted with a mixture of ice-cold solvents water/methanol/chloroform (H₂O/MeOH/CHCl₃ 1:1:1, v:v:v). The water/methanol phase was separated and dried in a speedvac. The extract was reconstituted in a 550 μl mixture containing 500 μl of phosphate buffer pH 7.4 and 50 μl of 1mM of TSP-d₄ in D₂O, vortexed for 20 seconds and transferred to 5 mm NMR tubes. The NMR spectra were acquired on a Bruker 600 MHz Avance III HD spectrometer equipped with a BBI room temperature probehead and SampleJet auto sampler (Bruker Biospin, Rheinstetten, Germany). ¹H NMR spectra were recorded using 1D noesy pulse sequence with presaturation (noesygppr1d), collecting 256 scans with calibrated 90-degree pulse (~11 μs), 4.55 second acquisition time, and 4 second relaxation delay. Metabolites were identified and quantified using Chenomx NMR suite 8.2 software, by fitting the spectral lines of library compounds into the recorded NMR spectrum of tissue extract. The quantification was based on peak area of TSP-d₄ signal. The metabolite concentrations are exported as μM in NMR sample and normalized to wet tissue mass (nmol/mg of tissue).

Kazak L., Rahbani J.F., Samborska B., Lu G.Z., Jedrychowski M.P., Lajoie M., Zhang S., Ramsay L.C., Dou F.Y., Tenen D., Chouchani E.T., Dzeja P., Watson I.R., Tsai L., Rosen E.D, & Spiegelman B.M. (2019). Ablation of adipocyte creatine transport impairs thermogenesis and causes diet-induced obesity. Nature metabolism, 1(3), 360-370.

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