Following injection into the trapping column (µPAC trapping column, Pharma Fluidics, Gent, Belgium), the peptides were separated by nano-reverse-phase (µPAC, 200 cm separation column, Pharma Fluidics, Gent, Belgium) using an UltiMate3000 nano rapid separation liquid chromatography HPLC (Thermo Fisher, Germering, Germany) separation system. Both, the trap- and separation columns were operated at 50 °C, and the UV peptide detection at 214 nm served as quality control for the HPLC separation. The samples were loaded onto the trap column using a loading solvent of 2% acetonitrile (ACN, VWR, Vienna, Austria) in an aqueous mix of 0.1% trifluoroacetic acid (TFA)/0.01% heptafluorobutyric acid (HFBA), both purchased from Sigma-Aldrich, Vienna, Austria, at 30 μL/min and precooled to 3 °C [36 (link)]. Nano separation was performed in gradient mode at 600 nL/min. A user defined injection program was used for sample injection and additional injector and trap column wash. Every sample injection was followed by two blank runs with injections of 2,2,2-trifluoroethanol (Alfa-Aeser, Vienna, Austria) for the removal of possible sample remains in the injector or on the trap column and for the prevention of carryover in the separation system. In order to perform the label-free quantitation (LFQ), equimolar amounts of peptides were injected.
Heptafluorobutyric acid hfba
Manufactured by Merck Group
Sourced in United States, Australia, Italy, Germany
Heptafluorobutyric acid (HFBA) is a colorless, volatile liquid chemical compound. It is used as a mobile phase additive in high-performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS) applications to improve the separation and detection of analytes.
Automatically generated - may contain errors
Lab products found in correlation
Spermine, by Merck Group (5 mentions)
Putrescine, by Merck Group (4 mentions)
L arginine, by Merck Group (3 mentions)
L ornithine hydrochloride, by Merck Group (3 mentions)
Spermidine hydrochloride, by Merck Group (3 mentions)
Cadaverine hydrochloride, by Merck Group (3 mentions)
N acetylputrescine hydrochloride, by Merck Group (3 mentions)
N acetylspermine trihydrochloride, by Merck Group (3 mentions)
N acetylspermidine dihydrochloride, by Merck Group (3 mentions)
Agmatine sulfate salt, by Merck Group (2 mentions)
Deuterated histamine, by Merck Group (2 mentions)
Trichloroacetic acid tca, by Merck Group (2 mentions)
Milli q system, by Merck Group (2 mentions)
Ammonium acetate, by Merck Group (2 mentions)
Methanol, by Merck Group (2 mentions)
Kinetex c18 column, by Phenomenex (2 mentions)
Mass hunter quantitative software, by Agilent Technologies (2 mentions)
Formic acid, by Merck Group (2 mentions)
Spermidine, by Merck Group (2 mentions)
Trifluoroacetic acid, by Merck Group (1 mentions)
23 protocols using heptafluorobutyric acid hfba
1
Peptide Separation by Nano-RPLC
2
Protein Digestion Protocol
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Trypsin, for protein digestion, was purchased from Promega Inc. (Vienna, Austria). Methanol (MeOH), ACN, 2,2,2‐trifluoroethanol, formic acid (FA), heptafluorobutyric acid (HFBA), iodoacetamide (IAA), triethylammonium bicarbonate (TEAB), and dithiothreitol were purchased from Sigma‐Aldrich (Vienna, Austria). Digestion of hCG formulations was performed using the routine approach described in earlier publications 10 .
3
Quantitative Analysis of Indospicine Compounds
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Indospicine (1 ) and 2-aminopimelamic acid (2 ) (>99% pure), both external standards, as well as D3-l -indospicine (4 ) (>99% pure) as internal standard, were synthesized and provided by Prof. James De Voss and Dr. Robert Lang, The University of Queensland [16 (link),25 (link)]. Another external standard, 2-aminopimelic acid (3 ) (>99% pure), and heptafluorobutyric acid (HFBA), ion chromatography grade, were purchased from Sigma Aldrich (Castle Hill, NSW, Australia). External (0.005–2 mg/L) and internal (1 mg/L) standard solutions were prepared in 0.1% HFBA in Milli-Q water.
4
Quantitative Analysis of Biogenic Amines
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The reference standards of 1,3-diaminopropane, putrescine, cadaverine hydrochloride, spermidine hydrochloride, spermine, agmatine sulfate salt, N-acetylputrescine hydrochloride, N-acetylspermine trihydrochloride, N-acetylspermidine dihydrochloride, l -ornithine hydrochloride, lysine, l -arginine, S-adenosyl-l -methionine, γ-aminobutyric acid and 1, 6-diaminohexane (used as an internal standard) were all obtained from Sigma-Aldrich (St. Louis, MO, USA). HPLC grade methanol was purchased from Fisher Chemicals (Fair Lawn, NJ, USA). Heptafluorobutyric acid (HFBA) was obtained from Sigma-Aldrich. All the other reagents were of analytic grade. Redistilled and deionized water was used throughout the study.
5
Quantitative Analysis of Lupine Alkaloids
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Lupanine (≥98%),
angustifoline (≥98%), and 13-hydroxylupanine (≥98%)
were purchased from ChemFaces (China); sparteine (>98%) was purchased
from Santa Cruz Biotechnology, Inc. (USA). Lupinine and heptafluorobutyric
acid (HFBA, ≥98%) were purchased from Sigma-Aldrich (USA).
Methanol (HPLC grade, ≥99.8%) was purchased from J.T. Baker
(USA). Deionized water (≥18.2 MΩ·cm) was prepared
using an ultrapure water system (arium pro, Sartorius, Germany).
For analysis, an Acquity 1 UPLC system (Waters, Millford, MA, USA)
was employed with a Xevo TQ-S triple quadrupole mass spectrometer
(Waters, Millford, MA, USA). These instruments were controlled using
the MassLynx software program (Waters).
angustifoline (≥98%), and 13-hydroxylupanine (≥98%)
were purchased from ChemFaces (China); sparteine (>98%) was purchased
from Santa Cruz Biotechnology, Inc. (USA). Lupinine and heptafluorobutyric
acid (HFBA, ≥98%) were purchased from Sigma-Aldrich (USA).
Methanol (HPLC grade, ≥99.8%) was purchased from J.T. Baker
(USA). Deionized water (≥18.2 MΩ·cm) was prepared
using an ultrapure water system (arium pro, Sartorius, Germany).
For analysis, an Acquity 1 UPLC system (Waters, Millford, MA, USA)
was employed with a Xevo TQ-S triple quadrupole mass spectrometer
(Waters, Millford, MA, USA). These instruments were controlled using
the MassLynx software program (Waters).
6
Comprehensive Polyamine Metabolite Analysis
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The reference standards of putrescine, cadaverine hydrochloride, spermidine hydrochloride, spermine, agmatine sulfate salt, N-acetyl-putrescine hydrochloride, N-acetyl-spermine trihydrochloride, N-acetyl-spermidine dihydrochloride, L-ornithine hydrochloride, lysine, L-arginine, S-adenosyl-1-methionine, aminobutyric acid, deuterated histamine, heptafluorobutyric acid (HFBA), and methanol LC/MS grade were purchased from Sigma-Aldrich (St. Louis, MO, USA). Water for LCMS was purchased from Fisher Scientific (Fair Lawn, NJ, USA).
7
Multi-class Aminoglycoside Residue Analysis
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HPLC-grade acetonitrile, methanol, and isopropanol were purchased from J.T. Baker (Deventer, The Netherlands). Ethylenediaminetetraacetic acid (EDTA), sodium chloride, and sodium hydroxide were from POCH (Gliwice, Poland), and potassium hydrogen phosphate (K2HPO4) was from Chempur (Piekary Śląskie, Poland). Ammonium acetate, heptafluorobutyric acid (HFBA), and trichloroacetic acid (TCA) were obtained from Sigma–Aldrich, (St. Louis, MO, USA). Formic acid was from Fluka (Charlotte, NC, USA). Syringe 0.22 µm hydrophilic polyvinylidene fluoride (PVDF) membrane filters were purchased from Restek (Bellefonte, PA, USA). Ultra-pure water was generated by a Millipore Milli-Q System (Millipore, Molsheim, France).
The analytical reference standards of DHSTR, GEN, KAN, NEO, PAR, STR, and SPC were purchased from Dr Ehrenstorfer (Augsburg, Germany), and AMI, APR, HYG, RIB, SIS, TOB from Sigma–Aldrich (St. Louis, MO, USA). Strata X (100 mg, 6 mL), Strata X-CW (100 mg, 6 mL), and Strata X-AW (100 mg, 6 mL) cartridges were from Phenomenex (Torrance, CA, USA). Oasis HLB (60 mg, 3 mL) cartridges were from Waters (Milford, MA, USA). All matrices were obtained from local supermarkets or were originating from the Polish official residue control program. Samples were analyzed to ensure the absence of aminoglycoside residues and kept frozen at −18 °C until use.
The analytical reference standards of DHSTR, GEN, KAN, NEO, PAR, STR, and SPC were purchased from Dr Ehrenstorfer (Augsburg, Germany), and AMI, APR, HYG, RIB, SIS, TOB from Sigma–Aldrich (St. Louis, MO, USA). Strata X (100 mg, 6 mL), Strata X-CW (100 mg, 6 mL), and Strata X-AW (100 mg, 6 mL) cartridges were from Phenomenex (Torrance, CA, USA). Oasis HLB (60 mg, 3 mL) cartridges were from Waters (Milford, MA, USA). All matrices were obtained from local supermarkets or were originating from the Polish official residue control program. Samples were analyzed to ensure the absence of aminoglycoside residues and kept frozen at −18 °C until use.
8
Quantification of Neurotransmitters by HPLC
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Dopamine (DA), norepinephrine hydrochloride (NE), 5-hydroxytryptamine (5-HT), acetyl- choline (Ach), l -tryptophan (Trp), γ-aminobutyric acid (GABA), glutamic acid (Glu) and aspartic acid (Asp) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Isoproterenol hydrochloride (internal standard, IS) was supplied by the National Institute for Food and Drug Control (Beijing, China). Heptafluorobutyric acid (HFBA) was obtained from Sigma-Aldrich. The purity of these reference standards were all more than 98%. Methanol and acetonitrile (HPLC grade) were purchased from Fisher Scientific (Fair Lawn, NJ, USA). Distilled water prepared with demineralized water was used throughout the study. Other reagent and solvent of analytical grade, were provided by the department of pharmaceutics, Shenyang Pharmaceutical University (Shenyang, China).
9
LC-HRMS Analysis of Polyamines and Derivatives
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The reference standards of putrescine, cadaverine hydrochloride, spermidine hydrochloride, spermine, agmatine sulphate salt, N-acetyl-putrescine hydrochloride, N-acetylspermine trihydrochloride, N-acetylspermidine dihydrochloride, l-ornithine hydrochloride, lysine, l-arginine, aminobutyric acid, deuterated histamine, heptafluorobutyric acid (HFBA) and methanol were purchased from Sigma-Aldrich (St. Louis, MO, USA) for analytical type analysis. Water for LCMS was purchased from Fisher Scientific (Fair Lawn, NJ, USA).
Liquid chromatography–high-resolution mass spectrometry (LC-HRMS) analysis was performed using an UPLC Ultimate 3000 (Thermo Fisher-Dionex San Jose, CA, USA) system equipped with a HESI-II electrospray source to a Q-Exactive-Orbitrap™-based mass spectrometer (all from Thermo Scientific, San Jose, CA, USA). Chromatographic separation was carried out on the C18 column of the Gemini C18 (Phenomenex, Torrance, CA, USA), 100 mm × 2 mm, particle size 3 µm, the column was held at 37 °C. Chromatographic separation was achieved with gradient elution using a mobile phase composed of 0.05% heptafluorobutyric acid (HFBA) in water (A) and 0.05% HFBA in methanol (B).
Liquid chromatography–high-resolution mass spectrometry (LC-HRMS) analysis was performed using an UPLC Ultimate 3000 (Thermo Fisher-Dionex San Jose, CA, USA) system equipped with a HESI-II electrospray source to a Q-Exactive-Orbitrap™-based mass spectrometer (all from Thermo Scientific, San Jose, CA, USA). Chromatographic separation was carried out on the C18 column of the Gemini C18 (Phenomenex, Torrance, CA, USA), 100 mm × 2 mm, particle size 3 µm, the column was held at 37 °C. Chromatographic separation was achieved with gradient elution using a mobile phase composed of 0.05% heptafluorobutyric acid (HFBA) in water (A) and 0.05% HFBA in methanol (B).
10
RRLC-MRM Analysis of Biological Samples
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Targeted analysis was performed on a RRLC 1260 system coupled to a Triple Quadrupole 6410 (Agilent Technologies) equipped with an electrospray source operating in positive mode. The gas temperature was set to 350°C with a gas flow of 12 l/min. The capillary voltage was set to 3.5 kV.
10 μl of sample were injected on a Column Kinetex C18 (150 mm x 2.1 mm particle size 2.6 µm) from Phenomenex, protected by a guard column C18 (5 mm × 2.1 mm) and heated at 40°C by a Pelletier oven. Heat the column more than the room temperature allowed rigorous control of the column temperature.
The gradient mobile phase consisted of water with 0.1% of Heptafluorobutyric acid (HFBA, Sigma-Aldrich) (A) and acetonitrile with 0.1% of HFBA (B) freshly made. The flow rate was set to 0.2 ml/min, and gradient as follows: initial condition was 95% phase A and 5% phase B. Molecules were then eluted using a gradient from 5% to 40% phase B over 10 min. The column was washed using 90% mobile phase B for 2.5 minutes and equilibrated using 5% mobile phase B for 4 min. The autosampler was kept at 4°C.
The collision gas was nitrogen. The scan mode used was the MRM for biological samples. Peak detection and integration of analytes were performed using the Agilent Mass Hunter quantitative software (B.07.01).
10 μl of sample were injected on a Column Kinetex C18 (150 mm x 2.1 mm particle size 2.6 µm) from Phenomenex, protected by a guard column C18 (5 mm × 2.1 mm) and heated at 40°C by a Pelletier oven. Heat the column more than the room temperature allowed rigorous control of the column temperature.
The gradient mobile phase consisted of water with 0.1% of Heptafluorobutyric acid (HFBA, Sigma-Aldrich) (A) and acetonitrile with 0.1% of HFBA (B) freshly made. The flow rate was set to 0.2 ml/min, and gradient as follows: initial condition was 95% phase A and 5% phase B. Molecules were then eluted using a gradient from 5% to 40% phase B over 10 min. The column was washed using 90% mobile phase B for 2.5 minutes and equilibrated using 5% mobile phase B for 4 min. The autosampler was kept at 4°C.
The collision gas was nitrogen. The scan mode used was the MRM for biological samples. Peak detection and integration of analytes were performed using the Agilent Mass Hunter quantitative software (B.07.01).
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