G4226a autosampler

Manufactured by Agilent Technologies

Sourced in United States

The G4226A autosampler is a laboratory instrument designed to automate the sample introduction process for various analytical instruments. It features a robotic arm that can precisely transfer liquid samples from sample vials to the appropriate analysis system. The G4226A autosampler is capable of handling a wide range of sample volumes and types, enabling efficient and consistent sample handling in analytical workflows.

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5 protocols using g4226a autosampler

Quantifying L-DOPA in Coffee Samples

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To verify the occurrence of L-DOPA in Coffea, sample extracts in triplicate from C. arabica and C. canephora leaves, flowers, and fruits were analyzed by LC–MS/MS. Those analyses were performed in an Agilent Technologies system consisting of a binary pump, a degassing unit, a G4226A autosampler, a column oven, and a triple-quadrupole mass spectrometer (QqQ G6420A). The system was controlled by MassHunter Workstation Software (Version B.08.00). The separation was carried out on Zorbax Eclipse XDB-C18, 4.6 × 250 mm × 5 µm, thermostated at 30 °C, using a mobile phase composed of acetic acid 1% in water (Solvent A) and methanol (Solvent B) at a ratio of 95:5 (v/v) and a flow rate of 1.0 mL·min⁻¹. Full scan spectra were acquired from m/z 10 to 500. Identification of L-DOPA was performed in multiple reaction monitoring (MRM) mode, detecting the following transitions: m/z 198 → m/z 152, m/z 198 → m/z 107, and m/z 198 → m/z 135 [51 (link),52 (link),53 (link)].

Cherubino Ribeiro T.H., de Oliveira R.R., das Neves T.T., Santiago W.D., Mansur B.L., Saczk A.A., Vilela de Resende M.L, & Chalfun-Junior A. (2023). Metabolic Pathway Reconstruction Indicates the Presence of Important Medicinal Compounds in Coffea Such as L-DOPA. International Journal of Molecular Sciences, 24(15), 12466.

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Quantitative LCMS Analysis of Fosetyl

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A 1290 Infinity HPLC pump system (Agilent Technologies, Santa Clara, CA, USA) with an Agilent G4226A autosampler was coupled to an Agilent G6460 triple quadrupole mass spectrometer equipped with the electrospray jet stream interface (ESI). In order to perform MS and MS/MS analyses in full scan (mass range m/z 50–500) and in product ion mode, the acquisition parameters were optimized by infusion at a flow rate of 10 μL·min⁻¹ of a solution of fosetyl in water without additives (10 ng·μL⁻¹). Nitrogen was used as a nebulizing and collisional gas. The fragmentor potential was optimized in order to maximize the parent ion intensities, and, by operating in product scan mode, the collision energy (CE) was optimized (Table 1). Finally, all the analyses were carried out by LC-MS/MS in MRM mode, acquiring diagnostic product ions from the chosen precursor to obtain high specificity and sensitivity. Three main fragments 81, 63, and 79 m/z were formed from the precursor m/z 109; the ion m/z 81 was chosen as a quantifier for the definitive MRM analyses. MassHunter Software was used for the acquisition and the elaboration of the data set. Figure 2 shows MS/MS spectrum obtained with the electrical parameters in Table 1.

Buiarelli F., Di Filippo P., Riccardi C., Pomata D., Marsiglia R., Console C, & Puri D. (2018). Hydrophilic Interaction Liquid Chromatography-Tandem Mass Spectrometry Analysis of Fosetyl-Aluminum in Airborne Particulate Matter. Journal of Analytical Methods in Chemistry, 2018, 8792085.

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Leaf ABA Content Determination by LC-MS

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The leaf ABA content was determined by LC-MS analysis according to the protocol reported in Rong Zhou et al. [26 (link)]. The analyses were performed on an Agilent 1290 infinity series (Agilent Technologies, Milano, Italy) equipped with a Model G4220A binary pump, G6410B mass detector, G4226A autosampler and a G1316C column compartment (Agilent Technologies, Milano, Italy). All mass spectra were obtained by Mass Hunter workstation data acquisition software (v B.08.02, Agilent Technologies, Milano, Italy) and were analyzed by Mass Hunter workstation software (v B.08.02, Agilent Technologies, Milano, Italy) for qualitative and quantitative analysis.

Petrozza A., Summerer S., Melfi D., Mango T., Vurro F., Bettelli M., Janni M., Cellini F, & Carriero F. (2023). A Lycopene ε-Cyclase TILLING Allele Enhances Lycopene and Carotenoid Content in Fruit and Improves Drought Stress Tolerance in Tomato Plants. Genes, 14(6), 1284.

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HPLC-MS/MS Quantification of Analytes

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Experiments were carried out on an Agilent 1290 series high-performance liquid chromatography, including a G4220A quaternary pump, G4212A DAD detector and a G1316C column heater and a G4226A autosampler, a tandem Agilent 6460A triple-quadrupole mass spectrometer equipped with an electrospray ionization source (Agilent Inc., MA, United States) and operated with MassHunter Version B.06.00 workstation software. The separation was performed on an Agilent Zorbax SB-C18 column (3.5 μm, 2.1*100 mm) with the column temperature maintained at 55°C and the sample injection volume was 10 μl. The mobile phase consisted of a mixture of phase A (10 mM ammonium acetate buffer containing 0.1% formic acid) and phase B (methanol with 10 mM ammonium acetate buffer and 0.1% formic acid) with the flow rate at 0.25 mL min⁻¹. The gradient program was set as follows: 0 min, 80% B; 0–2 min, 80–100% B; 2–10 min, 100% B.
Quantification was performed using electrospray in the positive mode with the spray voltage set at 4,000 V. Nitrogen was used as nebulizer gas and nebulizer pressure was set at 20 psi. Desolvation gas (nitrogen) was heated to 325°C and delivered at a flow rate of 12 L/min.

Qu J., Bian R., Liu B., Chen J., Zhai J., Teng F., Guo W, & Wei H. (2022). The pharmacokinetic study of tacrolimus and Wuzhi capsule in Chinese liver transplant patients. Frontiers in Pharmacology, 13, 956166.

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Quantification of Phenolic Compounds and Proanthocyanidins

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Previously optimized chromatographic conditions [43 ] were applied with some modifications. The LC hardware was an Agilent 1290 UPLC equipped with a G4226A autosampler, a G4220 A binary pump, a G1316 TCC, and a G4212 DAD detector. The column was a Core-shell Kinetex pentafluorophenyl (PFP) (100 mm × 2.1 mm id), with 2.6 μm particle size (Phenomenex, Torrance, CA). The mobile phases were water:formic acid (FA) (99:1, v/v) as solvent A and water:acetonitrile (ACN):FA (9:90:1, v/v/v) as solvent B. The same solvent gradient was employed as previously reported (Table 3) [43 ]. Although some retention times were slightly earlier than in our previous study, this was readily attributed to the smaller mixing volume of the Agilent 1290 UPLC compared with the Agilent 1100 HPLC. Relative quantification was determined for phenolic compounds (Table 1) using selected reaction monitoring (SRM) and for proanthocyanidins (Table 2) using single ion monitoring (SIM). Peak areas of each analyte were integrated with Thermo Xcalibur 2.1 software and normalized to the peak area of a related internal standard (IS). Values are reported per mg of dry sample.

Mirali M., Purves R.W., Stonehouse R., Song R., Bett K, & Vandenberg A. (2016). Genetics and Biochemistry of Zero-Tannin Lentils. PLoS ONE, 11(10), e0164624.

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