Ectoine was detected by high-performance liquid chromatography (HPLC) analysis using an Agilent 1260 Infinity LC system (Agilent, Waldbronn, Germany) and a GROM-SIL Amino 1PR column (GROM, Rottenburg-Hailfingen, Germany) essentially as described (Kuhlmann and Bremer, 2002 (link)) with the exception that a 1260 Infinity Diode Array Detector (DAD) (Agilent) was employed, instead of the previously used UV/Vis system. The ectoine content of samples was quantified using the OpenLAB software suite (Agilent). The individual values are given as intracellular ectoine concentration (μM) in cultures of S. salinus M19-40 that correspond to an OD600 of 1.
1260 infinity diode array detector
Manufactured by Agilent Technologies
Sourced in United States
The 1260 Infinity Diode Array Detector is a high-performance liquid chromatography (HPLC) detector from Agilent Technologies. It is designed to provide accurate and precise measurements of absorbance spectra for a wide range of analytes. The detector uses a diode array technology to simultaneously measure the absorbance of the sample across a broad wavelength range, enabling the identification and quantification of unknown compounds.
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Lab products found in correlation
1260 infinity binary pump, by Agilent Technologies (2 mentions)
1260 infinity quaternary pump, by Agilent Technologies (2 mentions)
Openlab software suite, by Agilent Technologies (1 mentions)
Agilent 1260 infinity lc system, by Agilent Technologies (1 mentions)
1260 infinity standard autosampler, by Agilent Technologies (1 mentions)
1260 infinity hplc system, by Agilent Technologies (1 mentions)
Eclipse plus c18 column, by Agilent Technologies (1 mentions)
Hplc system, by Agilent Technologies (1 mentions)
C18 reversed phase column, by Agilent Technologies (1 mentions)
Tissue tearor, by Biospec (1 mentions)
Agilent chemstation software, by Agilent Technologies (1 mentions)
Ferulic acid, by Agilent Technologies (1 mentions)
Accuri c6, by BD (1 mentions)
Zorbax sb c18 column, by Agilent Technologies (1 mentions)
Agilent hplc system, by Agilent Technologies (1 mentions)
Agilent 1260 infinity autosampler, by Agilent Technologies (1 mentions)
1260 infinity fraction collector, by Agilent Technologies (1 mentions)
Kinetex column, by Phenomenex (1 mentions)
Nucleodur c18 isis column, by Macherey-Nagel (1 mentions)
1260 infinity manual injector, by Agilent Technologies (1 mentions)
8 protocols using 1260 infinity diode array detector
1
Quantification of Ectoine in Salinibacter Salinus
2
Targeted LC-MS Metabolic Profiling
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Low-resolution ESi-MS data were acquired on an Agilent 1260 Infinity HPLC system (Agilent® 1260 Infinity Binary Pump, Agilent® 1260 Infinity Diode Array Detector (DAD), Agilent® 1290 Infinity Column Compartment, and Agilent® 1260 Infinity Standard Autosampler) coupled to an Agilent 6120 Quadrupole MS system and Peak Scientific® Genius 1050 nitrogen generator. A Phenomenex Kinetex® 2.6 μm EVO C18 100 Å (30 × 2.1 mm) reverse-phase analytical column was used. The chromatographic method included a column temperature of 40°C, an injection volume of 2 μL, a flow rate of 0.7 mL/min, and maximum column backpressure set at 600 bars. The mobile phase consisted of 10 mM NH4OAc in water (A) and 10 mM NH4OAc in methanol (B). The LC run is a 4.50 min duration beginning with 15% a of B from 0 to 0.30 min, followed by a speedy increase in the gradient to 100% B over 0.90 min. The mobile phase composition at 100% B was maintained at 4.50 min. Mass spectra were collected from m/z 100–800 in both the negative and positive modes.
3
HPLC Analysis of Phenolic Compounds
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The identification analyses of phenolic compounds were conducted using a HPLC unit (Agilent Technologies, California, US) that consisted of a Agilent 1260 Infinity Quaternary Pump, a 1260 Infinity Diode-Array Detector, a Agilent 1260 Infinity Standard Autosampler Injector with a loop of 20 μL and a reversed phase Eclipse Plus C18 column (250 mm × 4.6 mm × 5 μm). An isocratic elution consisting of ultrapure water with 0.1% orthophosphoric acid and HPLC grade methanol in the ratio of 80:20 over 30 min was used. The flow rate, wavelength and column temperature were set at 1.0 mL min−1, 210 nm and 30°C, respectively. This method was modified from Wang et al. (2000) (link).
4
HPLC Quantification of Analyte X
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First, tissue homogenates were prepared by homogenization of 250 mg tissue in 1 mL normal saline (0.9%) using a Tissue Tearor (BioSpec Products, Bartlesville, OK, USA). Upon 200 µL plasma or tissue homogenates, 200 µL of acetonitrile was added and vortexed for 1 minute. After standing for 5 minutes, the mixture was centrifuged at 3,500 rpm for 5 minutes. A 50 µL of supernatant filtered earlier through 0.45 µm filters was automatically injected concurrently with the standard solutions into an Agilent HPLC system. The HPLC instrument (1260 Infinity LC systems; Agilent Technologies, Santa Clara, CA, USA) was equipped with a reversed-phase C18 column (25 cm ×4.6 mm; PS =5 µm) and 1260 Infinity Diode Array Detector. The system was also equipped with Agilent ChemStation® Software. A previously validated HPLC method was utilized for the determination of AE, with slight modification.7 The samples were eluted isocratically using acetonitrile–water–phosphoric acid (64:36:0.1, v/v/v) at a flow rate of 1 mL/min at column temperature of 30°C and wavelength of 254 nm. The calibration curve of peak area against AE concentration was y =188.45x +39.105, R2=0.9961, under concentration range of 0.15–50 µg/mL. Retention time was 5.8±0.1 minutes.
5
Quantification of Fermentation Compounds
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Ferulic acid, 4-vinyl guaiacol (4-VG) and cinnamic acid were measured using an Agilent 1260 LC equipped with an 1260 Infinity Diode Array Detector measuring at 214 nm, and an Agilent Zorbax SB-C18 Column (4.6 × 5.0, 3.5 micron) operated at 30°C (Koopman et al., 2012 (link); Vos et al., 2015 (link)). A gradient of acetonitrile and 20 mM KH2PO4 (pH 2) with 1% (v/v) acetonitrile was used as eluent at a flow rate of 0.8 mL⋅min-1, increasing from 0–10% acetonitrile in 6 min. followed by an increase to 40% acetonitrile until 23 min. From 23 to 27 min, 100 % 20 mM KH2PO4 (pH 2) with 1 % acetonitrile was used as eluent. Ferulic acid, 4-VG and cinnamic acid standards for calibration were obtained from Sigma Aldrich. Stock solutions were prepared in 100 % ethanol. Dilutions were made in milliQ water.
Staining of cells with SYTOX Green Nucleic Acid Stain was performed as described by (Haase and Reed, 2002 (link)). Stained cells were analyzed on a flow cytometer equipped with a 488 nm laser (BD Accuri C6, BD Biosciences). The hybrids were compared with strains of known ploidy (n/2n, S. cerevisiae CEN.PK113-7D; 2n/4n, S. cerevisiae CEN.PK122; 3n/6n, S. cerevisiae FRY153) (van den Broek et al., 2015 (link)).
Staining of cells with SYTOX Green Nucleic Acid Stain was performed as described by (Haase and Reed, 2002 (link)). Stained cells were analyzed on a flow cytometer equipped with a 488 nm laser (BD Accuri C6, BD Biosciences). The hybrids were compared with strains of known ploidy (n/2n, S. cerevisiae CEN.PK113-7D; 2n/4n, S. cerevisiae CEN.PK122; 3n/6n, S. cerevisiae FRY153) (van den Broek et al., 2015 (link)).
6
Quantitative Analysis of Synovial and Plasma Metabolites
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The synovial washing fluid and plasma samples were analyzed using an Agilent HPLC system (Agilent Technologies, Palo Alto, CA, USA) equipped with an automated solvent delivery system, which has an integrated degasser (1260 Infinity Quaternary Pump, Agilent Technologies), an Agilent 1260 Infinity autosampler (Agilent Technologies) and a 1024-element diode array detector (1260 Infinity Diode Array Detector, Agilent Technologies). The system control and data acquisition were performed with Agilent ChemStation B.04.03 software (Agilent Technologies). The chromatographic parameters and the sample extraction procedure have been provided in the Supplementary materials.
7
Preparative HPLC Isolation of Pure Compounds
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For the isolation of pure compounds, a preparative high-performance liquid chromatography (HPLC) device equipped with a 1260 Infinity binary pump, a 1260 Infinity manual injector, a 1260 Infinity fraction collector, a 1260 Infinity diode array detector (all Agilent Technologies, Santa Clara, USA) and a Kinetex® column (Biphenyl, 100 Å, 5 μm, 21.2 × 250 mm, Phenomenex, Aschaffenburg, Germany) at a flow rate of 21 mL/min or—for fractions M1.2C6 and M1.2C7—a NucleodurTM C18 Isis column (RP18, 5 µm, 10 × 250 mm, Macherey-Nagel, Düren, Germany) at a flow rate of 5 mL/min was used. Separation was achieved by gradients consisting of acetonitrile (A)/water (B) and peaks were detected at 200 nm. After the elimination of acetonitrile via evaporation, the water fractions were partitioned four times with diethyl ether or ethyl acetate (M1.2C6 and M1.2C7) and organic phases were dried under a nitrogen stream. The gradients used for the separation can be found in Table 5 .
8
Indoor Air Carbonyl Sampling and Analysis
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Air was collected at a constant sampling rate of 200 mL/min using a low flow sampling pump (Universal PCXR8, SKC) equipped with a single adjustable low flow tube holder (224-26-01, SKC). The pump was calibrated before and after each air sampling as described above. Packed glass cartridges with two sections of silica gel impregnated with 2,4-dinitrophenyl hydrazine (2,4-DNPH) with built-in ozone scrubber (226-120, SKC) were used to collect formaldehyde and other carbonyl compounds. The indoor air was sampled (48 L) every 4 h and in duplicate, at a 1.60-m height from the floor. All three sections were separated by glass wool. After sampling, the cartridges and field blanks were closed with polypropylene plugs and stored at ~4 °C protected from sunlight. The cartridges were analyzed by eluting each of the sections of silica gel-DNPH with 3 mL of acetonitrile, discarding the section of the ozone scrubber. The 2,4-DNPH derivatized carbonyls were analyzed by reverse-phase liquid chromatography (1260 Infinity Quaternary LC System, Agilent Technologies) with an ultraviolet detector operated at 360 nm (1260 Infinity Diode Array Detector, Agilent Technologies).
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