Empower 2 software

Manufactured by Waters Corporation

Sourced in United States

Empower II software is a chromatography data system that enables the acquisition, processing, reporting, and management of analytical data from various chromatographic instruments. It provides a comprehensive solution for laboratory workflow optimization and data management.

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7 protocols using empower 2 software

Quantitative Amino Acid Analysis by UPLC

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Twenty-one amino acids were separated and quantified using the Waters ACQUITY UPLC analytical system controlled by the Empower II software (Waters). The amino acids were derivatized using AccQ Tag Ultra reagent (6-aminoquinolyl-N-hydroxysuccinimidyl carbamate). The derivatives were separated on a 2.1 × 100 mm AccQ Tag Ultra column (Waters) and detected with Waters ACQUITY Tunable UV detector at 260 nm under the chromatographic conditions described in Cohen (2000). Peak identity and amino acid quantity were determined by comparison to a standard mix containing the 21 amino acids. Results from amino acid determination were expressed as concentrations on DW basis (µmol g⁻¹ DW).

Livingston DP I.I.I., Bertrand A., Wisniewski M., Tisdale R., Tuong T., Gusta L.V, & Artlip T. (2021). Factors contributing to ice nucleation and sequential freezing of leaves in wheat. Planta, 253(6), 124.

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Quantification of Soluble Sugars and LDP Fructans

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Soluble sugars including sucrose, glucose, fructose, raffinose and low degree of polymerization (LDP) fructans were diluted 1:1 ratio with acetonitrile and centrifuged for 3 min at 16,000 × g and kept at 4°C in the sample manager throughout the analysis. Waters ACQUITY Ultra Performance Liquid Chromatography (UPLC) analyzer controlled by the Empower II software (Waters, Milford, MA, USA) was used for the chromatographic analyses. Samples were separated on a BEH Amide Acquity UPLC column (Waters, Milford, MA, USA) and detected on an Electric Light Scattering Detector (ELSD, Acquity, Waters) set to a gas pressure of 45 psi. The chromatic conditions were as follows: 0.25 mL min^-1 with a gradient of eluents A (80% acetonitrile/0.1% NH₄OH) and B (30% acetonitrile/0.1% NH₄OH) described in Piva et al. (2013) (link). The drift tube was adjusted to a temperature of 50°C in the cooling mode. Peak identity and quantity of soluble sugars were determined by comparison to analytical standards following the guidelines of the metabolomics standards initiative (Fiehn et al., 2007 (link)). The degree of polymerization of LDP fructans was established by comparison with retention times (Supplementary Table S1) of purified standards from Jerusalem artichoke (Helianthus tuberosus L.).

Gagné-Bourque F., Bertrand A., Claessens A., Aliferis K.A, & Jabaji S. (2016). Alleviation of Drought Stress and Metabolic Changes in Timothy (Phleum pratense L.) Colonized with Bacillus subtilis B26. Frontiers in Plant Science, 7, 584.

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RP-IP-HPLC Analysis and Purification of Crude RNA

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The crude RNA templates were analyzed by reverse-phase
ion pairing high-performance liquid chromatography (RP-IP-HPLC) to
determine crude purities. Briefly, HPLC analyses (0.1 OD) and purifications
(1 OD) were performed on a Waters 2695 Alliance Separations Module.
Crude RNA templates were dissolved in autoclaved water (1 mL) and
injected into a Waters SymmetryShield RP-8 column (4.6 × 250
mm², 5 μm particle size, 100 Å) heated at 60
°C. HPLC analyses and purifications were conducted using a gradient
of 4–90% eluent B (50% acetonitrile in 0.1 M triethylammnonium
acetate, TEAA) in eluent A (0.1 M TEAA). The HPLC flow rate was set
at 1 mL/min, with run times of 25 min and with dual absorbance detection
at 260 and 488 nm using a Waters 2489 UV/visible detector. Retention
times (min) and peak areas (% area) were integrated with Empower II
software (Waters) and used to confirm RNA purities ≥95% following
sample purifications.

Kozuch S.D., Cultrara C.N., Beck A.E., Heller C.J., Shah S., Patel M.R., Zilberberg J, & Sabatino D. (2018). Enhanced Cancer Theranostics with Self-Assembled, Multilabeled siRNAs. ACS Omega, 3(10), 12975-12984.

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Quantification of Intracellular ATP and Glycolysis

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Concentration of ATP in treated cells was measured using the ATPLite assay kit by PerkinElmer (Waltham, MA, USA). Cultivation media were acquired in preparation for detection of glycolysis function using a cell-based assay kit for quantifying L-lactate, the terminal product generated by cellular glycolysis. Prior to HPLC, cells were washed and resuspended in phosphate-buffered saline (PBS). Nucleotides (ATP and AMP) were isolated via fast lysing the cells with 0.05 M KOH solution, which was adjusted to pH 6 and was used to conduct reverse phase chromatography. The mobile phase (pH 6) included 0.1 M KH₂PO₄, 0.008 M tetrabutylammonium hydrogen sulfate, and acetonitrile (containing 30% solvent B and 2% solvent A). Empower II software (Waters, Milford, MA, USA) was used for analyses and instrument control.

Guo L., Chen D., Yin X, & Shu Q. (2019). GSK-3β Promotes Cell Migration and Inhibits Autophagy by Mediating the AMPK Pathway in Breast Cancer. Oncology Research, 27(4), 487-494.

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Quantification of Cellular Metabolism by ATP and Lactate Analysis

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After treatment, cells were harvested and the ATP levels were determined using the ATPLite^™ assay kit obtained from PerkinElmer (Boston, MA) following the protocol provided by the manufacturer. Cell culture media were collected for the determination of glycolysis activity with a cell-based assay kit, which was designed to detect extracellular levels of L-lactate, the end-product of cellular glycolysis (Catalog #600450, Cayman Chemical, Ann Arbor, MI).
For HPLC-based measurement, cells were harvested, washed, and re-suspended in PBS. Nucleotides (ATP and AMP) were extracted by fast lysing the cells in 0.05 M KOH solution, and then immediately neutralized to pH 6 with 0.1 M KH₂PO₄. After centrifuge, the supernatant was analyzed by a gradient HPLC method on a Waters e2695 HPLC with UV detection at 254 nm and 340 nm (Waters e2,489 diode array UV detector, Waters, MA). The reversed-phase chromatography was performed with an XBridge^™ C18 column 3.5 μm (Waters, Milford, MA). Mobile phase (pH 6) contained acetonitrile (2% for Solvent A and 30% for Solvent B), 0.1 M KH₂PO₄, and 0.008 M Tetrabutylammonium hydrogen sulfate. The Empower II software (Waters, MA) was used for instrument control and data analysis. All values were normalized to the protein content of whole homogenaties using the bicinchoninic acid method (Pierce Biotechnology, IL).

Sun A., Li C., Chen R., Huang Y., Chen Q., Cui X., Liu H., Thrasher J.B, & Li B. (2015). Gsk-3β Controls Autophagy by Modulating LKB1-AMPK Pathway in Prostate Cancer Cells. The Prostate, 76(2), 172-183.

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Quantifying Leaf Carbohydrates in Stressed Plants

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One hundred (100) mg of freeze-dried ground leaf tissues of inoculated or not plants subjected to drought or not were pooled and reduced to fine powder in liquid nitrogen. Soluble sugars were extracted with methanol/chloroform/water solutions and analyzed as described in Bertrand and coworkers, [68 ] using a Waters High Performance Liquid Chromatography (HPLC) analytical system controlled by the Empower II software (Waters, Milford, MA, USA). Peak identity and quantity of raffinose, sucrose, glucose and fructose were determined by comparison to standards. Total starch was extracted from the non-soluble residue left after the methanol/chloroform/water extraction and quantified as a glucose equivalent following enzymatic digestion with amyloglucosidase (Sigma-Aldrich) and colorimetric detection with ρ-hydrobenzoic acid hydrazide method of Blakeney and Mutton [69 ].

Gagné-Bourque F., Mayer B.F., Charron J.B., Vali H., Bertrand A, & Jabaji S. (2015). Accelerated Growth Rate and Increased Drought Stress Resilience of the Model Grass Brachypodium distachyon Colonized by Bacillus subtilis B26. PLoS ONE, 10(6), e0130456.

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Quantifying Foliar Amino Acid Profiles

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The concentrations of free amino acids in leaves, roots, and nodules were analyzed by HPLC from the same extract used for water-soluble carbohydrate determination. Twenty-one amino acids were analyzed: alanine, arginine, asparagine, aspartate, glutamate, glutamine, glycine, γ-aminobutyric acid (GABA), α-aminobutyric acid (AABA), histidine, proline, methionine, lysine, serine, leucine, isoleucine, ornithine, phenylalanine, threonine, tyrosine, and valine. These amino acids were separated and quantified using Waters ACQUITY UPLC analytical system controlled by the Empower II software (WATERS, Milford, MA, USA) as described by [25] (link). The results from amino acid determinations were expressed as concentrations on a dry matter (DM) basis (µmol g -1 DM). The total free amino acids was the sum of each individual concentration of the 21 free amino acids.

Bertrand A., Gatzke C., Bipfubusa M., Lévesque V., Chalifour F., Claessens A., Rocher S., Tremblay G.F., & Beauchamp C.J. (2020). Physiological and Biochemical Responses to Salt Stress of Alfalfa Populations Selected for Salinity Tolerance and Grown in Symbiosis with Salt-Tolerant Rhizobium. Agronomy, 10(4), 569-569.

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