HPLC/ESI-MS was used to evaluate putrescine and glutamate as GlnA3 substrates and generated gamma-glutamylated putrescine as a product. Standard reactions typically contained 20 mM HEPES (pH 7.2), 150 mM glutamate sodium monohydrate, 150 mM putrescine dihydrochloride, 20 mM MgCl2 × 6H2O, and 10 mM ATP were mixed with 10 μg of the purified His-GlnA3 (or without GlnA3 as a control) and incubated at 30°C for 5 min. The reaction was stopped by incubation of the reaction mixture at 100°C for 5 min. HPLC/ESI-MS analysis of the glutamylated product generated by GlnA3 was done on an Agilent 1200 HPLC series using a Reprosil 120 C18 AQ column, 5 μm, 200 mm × 2 mm fitted with a precolumn 10 mm × 2 mm (Dr. Maisch GmbH, Ammerbuch, Germany) coupled to an Agilent LC/MSD Ultra Trap System XCT 6330 (Agilent, Waldbronn, Germany). Analysis was carried out using 0.1% formic acid as solvent A and acetonitrile with 0.06% formic acid as solvent B at a flow rate of 0.4 ml min−1. The gradient was as follows: t0 = t5 = 0% B, t20 = 40% B (time in minutes). Injection volume was 2.5 μl, column temperature was 40°C. ESI ionization was done in positive mode with a capillary voltage of 3.5 kV and a drying gas temperature of 350°C.
Agilent 1200 series hplc
Manufactured by Agilent Technologies
Sourced in United States, Germany, United Kingdom, Japan, Switzerland
The Agilent 1200 series HPLC is a high-performance liquid chromatography (HPLC) system designed for analytical and preparative applications. It is a modular system that can be configured with various components, including pumps, autosampler, column compartment, and detectors, to meet the specific needs of the user's application.
Automatically generated - may contain errors
Lab products found in correlation
Biophotometer, by Eppendorf (5 mentions)
Aqua c18 reverse phase resin, by Phenomenex (5 mentions)
Ammonium hydroxide nh3, by Merck Group (3 mentions)
Tskgel amide 80, by Tosoh (3 mentions)
Lichrospher 100 rp 18e column, by Agilent Technologies (3 mentions)
Protein a immunodetection sensor cartridge, by Thermo Fisher Scientific (3 mentions)
Partisphere strong cation exchange resin scx, by Cytiva (3 mentions)
Gemini c18 column, by Phenomenex (2 mentions)
Zorbax c18 column, by Agilent Technologies (2 mentions)
Eclipse xdb c18 column, by Agilent Technologies (2 mentions)
Tsk amide 80, by Tosoh (2 mentions)
Zorbax extend c18 column, by Agilent Technologies (2 mentions)
Chemstation software, by Agilent Technologies (2 mentions)
Chromeleon chromatography management system version 6, by Thermo Fisher Scientific (2 mentions)
Hydro rp, by Phenomenex (2 mentions)
Ltq orbitrap discovery mass, by Thermo Fisher Scientific (2 mentions)
Agilent 6460 qqq mass spectrometer, by Agilent Technologies (2 mentions)
Luna rp c5 column, by Phenomenex (2 mentions)
Ctc pal autosampler, by CTC Analytics (2 mentions)
Quant it picogreen dsdna kit, by Thermo Fisher Scientific (2 mentions)
182 protocols using agilent 1200 series hplc
1
Quantification of Glutamylated Putrescine by HPLC-MS
2
HPLC-MS/MS Analysis of Pesticides
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The HPLC–MS/MS analysis was achieved using an Agilent 1200 HPLC series (Agilent Technologies, Santa Clara, CA, USA) and an Agilent 6410B triple-quadrupole mass spectrometer equipped with an electrospray ionization interface (ESI±). A HPLC reverse-phase C18 column (50 mm × 2.1 mm × 3.5 μm, Agilent, Santa Clara, CA, USA) was employed for the separation of dichlorprop-P, bentazone, 6-hydroxy-bentazone and 8-hydroxy-bentazone at 30 °C. The mobile phase was acetonitrile and 0.1% formic acid water (v/v = 90/10) at a flow rate of 0.25 mL/min, and the injection volume was 5 μL. The total run time was 2.5 min. The HPLC–MS/MS was performed in negative multiple-reaction monitoring (MRM) mode. The desolvation gas (N2) temperature was set at 350 °C with the gas flow at 8.0 L/min, and the nebulizer pressure at 35 psi. The parameters were optimized individually for each target compound (Table 2 ).
3
Quantification of Betalains by HPLC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Agilent 1200 HPLC series (Agilent Technologies, Palo Alto, CA, USA) connected to a photodiode array detector was used for the quantification of betalains. The separation was carried out on a Gemini C18 Column (250 mm × 4.6 mm i.d.) (Phenomenex, Torrance, CA, USA) at 30 °C. The mobile phase consisted of 0.3 Mphosphoric acid (solvent A) and acetonitrile (solvent B). At a flow rate of 0.8 mL/min, different betalains were monitored at 540, 480, and 420 nm and identified by elution order and UV spectra from published papers [8 (link)]. Different concentrations (31.25–500 μg/mL) of purified betanin were injected into the HPLC and a calibration curve was prepared. Samples of 10 μL were injected and the betalain content was expressed as μg betanin equivalent/g of fresh weight [18 (link)] using the formula BC (mg/g) = [A(DF)(MW)Vd/ƐLWd], where A is the absorbance at the λmax of 535 and 477 nm for betacyanins and betaxanthins, respectively, DF is the dilution factor, MW is the molecular weight, Vd is the solution volume (mL), Ɛ is the extinction coefficient, Wd is the beet weight (g), and L is the path length (1 cm) of the cuvette. The MW of betanin is 550 g/mol and Ɛ = 60000L/(mol cm); for betacyanin, Ɛ = 48,000 L/(mol cm).
4
Quantitative HPLC-MS/MS Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The chromatographic system used is of Agilent 1200 HPLC series, consisting of a binary pump (Model G1312B), a vacuum degasser (Model G1379B), an autosampler (Model G1367C), and a column oven (Model G1316B). The mass spectrometer used was an Applied Biosystems Sciex 4000 Q-Trap® mass spectrometer (Foster, CA, USA) combined with a Source 5000 LC/MS gas generator (Parker Balston Analytical Gas Systems). Data acquisition was carried out by Analyst® 1.4.2 software on a Dell computer.
5
Adsorption of Antibiotics and Pesticides by GDY
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To verify the adsorption performance of GDY, four tetracycline antibiotics (oxytetracycline, doxycycline, tetracycline and chlorotetracycline) and five neonicotinoid pesticides (acetamiprid, clothianidin, thiacloprid, imidacloprid and nitenpyram) were tested. The concentration of the adsorption material GDY and the above mixed solution in centrifuge tubes was 1 g L−1 and 2 mg L−1, respectively, which was oscillated at a speed of 170 rpm for 2 h until equilibrium was reached. All samples were analyzed by HPLC-MS/MS (Agilent Technologies, USA) equipped with an Agilent 1200 HPLC series and an Agilent 6410B triple-quadrupole mass spectrometer.
6
Isocratic HPLC Quantification of Doxycycline
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An isocratic method for detecting doxycycline hyclate dissolved in aqueous solutions was conducted on a modular HPLC system (Agilent HPLC 1200 series, Agilent Technologies, Edinburgh, UK) using a 15 cm × 4.6 nm, 5 μm, RP-amide column (Supelco Analytical, Poole, UK). The mobile phase consisted of 75% (v/v) di-iodinated water (with 0.1% [v/v] trifluoroacetic acid) and 25% (v/v) HPLC-grade acetonitrile (Sigma-Aldrich). System parameters were set with a pump flow rate of 1.0 ml/min, sample injection volume of 10 ml, a method stop time of 10 min and detection wavelength of 273 nm. This method produced a chromatogram with a major analyte peak corresponding to doxycycline at 6 min. A calibration curve was created from serial dilutions of doxycycline solution. This was used to calculate doxycycline concentration from solutions of unknown concentration using the area under the chromatogram peak.
7
Quantification of γ-Oryzanol in Rice Powder
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
γ-Oryzanol in the rice powder samples was extracted in the rice samples and determined as described by Islam and Becerra (10 ). Briefly, 1.00 g of pulverized rice samples was extracted with 3.0 mL of HPLC-grade methanol. The mixture was shaken using a rotator mixer (Chang Shin Scientific, Co., Pusan, Korea) for 2 h. After the extraction, the samples were centrifuged at 825 rpm, 4°C for 10 min. The extraction process was repeated three times and the supernatants were then collected and filtered through 0.45-µM microfilter (Chromdisc®). Fifty microliters of the extracts were injected into Agilent HPLC (1200 Series, Japan) using C18 column (4.6×250 mm, 5 µm). The HPLC was equipped with a UV–Vis photodiode array detector set at 330 nm wavelength. Methanol/acetonitrile/dichloromethane/acetic Acid (50:44:3:3) were used as the mobile phase at room temperature. The flow rate was set at 1 mL/min. γ-Oryzanol standard was used for to calibrate and calculate the concentration in the rice samples.
8
HPLC-UV Quantification of Albendazole
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The concentration of ABZ was determined using HPLC-UV. The analysis was carried out using an Agilent HPLC 1200 series, consisting of a binary pump, degasser, column oven, and variable wavelength UV detector, from Agilent Technologies, UK. Samples were analyzed using a Zorbax C 18 column (100 mm Ã-4.6 mm with 3.5 μm packing) from Agilent Technologies, UK, at 228 nm. The mobile phase consisted of 4.35 mM monobasic ammonium phosphate, pH 4.8 (40%) and methanol (60%). The flow rate was set constant at 1 mL/min, with the column compartment maintained at 30 °C throughout the entire time frame of all analyses. The obtained chromatograms were analyzed using Agilent OpenLab software. Using the described assay, a linear relationship (y = 33,904x) was established between the integral intensity of the identified main analyte peak and ABZ concentration, and it was validated for good specificity, linearity (R 2 > 0.99), inter-and intraday variations, and precision.
9
Quantification of NAD+ Metabolome in Liver
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
NAD+ and its related metabolites and end-products, Nam, NMN, Na, Nr, NaMN, NaAD, nicotinamide n-oxide, nicotinuric acid, n-methylnicotinamide, and Trp, were measured in the liver samples using an LC-QqQ-MS/MS system consisting of an Agilent HPLC 1200 Series (Agilent Technologies, Palo Alto, USA). Briefly, 60 mg of the lyophilized liver samples were vigorously vortexed in 0.5 mL of physiological saline for 30 seconds, followed by 30 seconds of ultrasonication in a Vibra Cell (Sonics, Newton, USA). Then, 0.5 mL of acetone was added and the samples were centrifuged at 10,000 × g for 15 min at 20 °C. This procedure was repeated twice; the upper phases were mixed and evaporated under nitrogen flow to dryness. Finally, the dried samples were dissolved in 100 μL of the mobile phase at the initial conditions and injected into the liquid chromatograph (LC). A Scherzo SM-C18 (3 μm; 150 mm × 2 mm i.d.; Imtakt, Japan) was used to perform the analysis. The LC was coupled to a triple quadrupole (QqQ) mass spectrometer (MS) 6410 (Agilent Technologies, Palo Alto, USA).
10
Fungal Extract Analysis by HPLC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fungal extracts were analyzed on an Agilent HPLC series 1200 (Agilent Technologies) by using an Eclipse XDB-C18 column (Agilent Technologies, 5 μm, 4.6 × 150 mm) and ACN/H2O as elution solvents. A linear gradient from 10 to 90% ACN in H2O containing 0.1% (v/v) HCOOH in 20 min was used. After washing with 100% ACN for 5 min, the column was equilibrated with 10% ACN for another 5 min. A photodiode array detector was used for detection and the absorptions at 254 nm are illustrated in this study.
For product isolation, a Multospher 120 RP-18 column (5 μm, 10 × 250 mm) was used on the same HPLC system, with the same elution solvents at a flow rate of 2 mL min−1. Separation was performed by isocratic elution with 45–70% ACN in H2O containing 0.1% (v/v) HCOOH for 10–20 min, and.
For product isolation, a Multospher 120 RP-18 column (5 μm, 10 × 250 mm) was used on the same HPLC system, with the same elution solvents at a flow rate of 2 mL min−1. Separation was performed by isocratic elution with 45–70% ACN in H2O containing 0.1% (v/v) HCOOH for 10–20 min, and.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!