The total carbohydrate content was determined by the anthrone test using glucose as a standard and the total protein by the Lowry method using bovine serum albumin (BSA) as a standard [8 (link),12 (link)]. The monosaccharide composition was analyzed by the 1-phenyl-3-methyl-5-pyrazolone (PMP)-HPLC method as reported previously [14 (link)]. In brief, the polysaccharide samples (1–2 mg) were completely hydrolyzed with 2 M trifluoroacetic acid (TFA) at 110 °C for 4 h. The hydrolysate was dried by evaporation under vacuum and then derivatized in 450 μL PMP solution (0.5 M in methanol) and 450 μL of 0.3 M NaOH at 70 °C for 30 min. The reaction was stopped by neutralization with 450 μL of 0.3 M HCl, followed by extraction with chloroform (1 mL, 3 times). The extract solution was applied to HPLC analysis with an Agilent ZORBAX Eclipse XDB-C18 column (5 μm, 4.6 × 150 mm) on an Agilent 1100 instrument with a G1312A Bin pump and a UV detector.
G1312a binpump
Manufactured by Agilent Technologies
Sourced in Ireland, United States
The G1312A BinPump is a binary high-performance liquid chromatography (HPLC) pump designed for analytical applications. It is capable of delivering a stable and accurate flow of mobile phase solvents to the HPLC system. The pump features two independent solvent channels, allowing for the creation of binary gradients.
Automatically generated - may contain errors
Lab products found in correlation
G1313a als, by Agilent Technologies (5 mentions)
G13795 degasser, by Agilent Technologies (4 mentions)
G1316a column oven, by Agilent Technologies (4 mentions)
1100 hplc system, by Agilent Technologies (3 mentions)
Kinetex phenyl hexyl column, by Phenomenex (2 mentions)
Allure pfp propyl column, by Restek (2 mentions)
Zorbax eclipse xdb c18 column, by Agilent Technologies (1 mentions)
1100 instrument, by Agilent Technologies (1 mentions)
Single quadrupole msd, by Agilent Technologies (1 mentions)
G1379a degasser, by Agilent Technologies (1 mentions)
1100 series hplc system, by Agilent Technologies (1 mentions)
7 protocols using g1312a binpump
1
Carbohydrate and Protein Analysis of Polysaccharides
2
Quantification of Analytes by LC-MS
3
Quantitative LC-MS Analysis of Analytes
4
Extraction and Quantification of Soybean Glycosides
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Soybeans (GM, 3000 g) were powdered and made circumfluent with 30% ethanol (3 times the weight of the beans) for 3 h at 90 °C. The ethanol supernatants were percolated and lyophilized to yield the final extract (362 g, 12.1%). GE was supplied by Sigma–Aldrich. (Sigma–Aldrich, Saint Louis, MO, USA).
High-performance liquid chromatography (HPLC) was performed using Agilent 1100 series G1379A Degasser, G1312A Bin Pump, G1313A ALS, G1316 column, and G1313A VWD (Agilent Technologies, Santa Clara, CA, USA). As the HPLC column, a Luna (18, 150 × 4.6 mm, 5 μm) column was used, and the wavelength of the ultraviolet absorption detector was 254 nm. As the moving phase, an ammonium sulfate solution (1 → 100) and acetonitrile mixture solution (91.5:8.5, v/v) were used. Thereafter, 10 μL of the sample was injected.
Using this method, the concentration of GE in GM was calculated to be 7.74 µg/g, as described inFigure 6 .
High-performance liquid chromatography (HPLC) was performed using Agilent 1100 series G1379A Degasser, G1312A Bin Pump, G1313A ALS, G1316 column, and G1313A VWD (Agilent Technologies, Santa Clara, CA, USA). As the HPLC column, a Luna (18, 150 × 4.6 mm, 5 μm) column was used, and the wavelength of the ultraviolet absorption detector was 254 nm. As the moving phase, an ammonium sulfate solution (1 → 100) and acetonitrile mixture solution (91.5:8.5, v/v) were used. Thereafter, 10 μL of the sample was injected.
Using this method, the concentration of GE in GM was calculated to be 7.74 µg/g, as described in
5
LC-MS Analysis of Organic Compounds
6
HPLC-ESI-MS Analysis of Dihydroquinidine
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The HPLC-ESI-MS system consisted of an Agilent 1100 series HPLC system equipped with a G1312A Bin pump, a G1379A Degasser (Agilent, San Jose, CA, USA) and a G1316A automatic column temperature control box. Chromatographic separation was performed on a HiQ sil-C18 reversed-phase column (4.6 mm×250 mm, 5 µm, KYA TECH). An API3000 Triple tandem quadrupole mass spectrometer with a Turbolon-Spray interface from Applied Biosystems (Foster City, CA, USA) was operated in the positive electrospray ionization (ESI+) source mode. All mass spectra were acquired in multiple reaction monitoring transitions.
For HPLC-ESI-MS analysis, acetonitrile-water-acetic acid (18∶82∶0.1, v/v/v) was used at a flow rate of 1.0 ml/min with a run time of 65 min. The injection volume was 10 µl and the column temperature was maintained at 25°C. The ion source was operated at a temperature of 250°C. The nebulizing gas, curtain gas and collision gas were set at 12, 10 and 6 a.u., respectively. The ion spray voltage was 5500 V. The entrance potential and focusing potential were set at 10 and 400 V, respectively. Analyst software (version 1.4, Ab Sciex, Framingham, MA, USA) installed on a Dell computer was used for data acquisition and processing. The LC-ESI-MS chromatogram of DHQ is shown inFigure 1(d) .
For HPLC-ESI-MS analysis, acetonitrile-water-acetic acid (18∶82∶0.1, v/v/v) was used at a flow rate of 1.0 ml/min with a run time of 65 min. The injection volume was 10 µl and the column temperature was maintained at 25°C. The ion source was operated at a temperature of 250°C. The nebulizing gas, curtain gas and collision gas were set at 12, 10 and 6 a.u., respectively. The ion spray voltage was 5500 V. The entrance potential and focusing potential were set at 10 and 400 V, respectively. Analyst software (version 1.4, Ab Sciex, Framingham, MA, USA) installed on a Dell computer was used for data acquisition and processing. The LC-ESI-MS chromatogram of DHQ is shown in
7
Quantification and Characterization of Clonazolam and Nitrazolam
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
This was used for the quantification of clonazolam in the tablets and to obtain product ions spectra for clonazolam and nitrazolam utilizing in-source CID.
Analyses were performed on an Agilent 1100 HPLC system equipped with a G13795 degasser, G1312A BinPump, a G1313A ALS and G1316A column oven (COLCOM) (Agilent, Little Island, Cork). Separation was obtained on an Allure PFP Propyl column (5 μm, 50 x 2.1 mm) Restek (Bellefonte, PA, USA). Mobile phase A consisted of 0.1% formic acid in water, whereas mobile phase B consisted of 0.1% formic acid in acetonitrile. The flow rate was set at 0.80 mL/min, with an injection volume of 10 µL, and the column temperature was 30°C. The following gradient elution program was used: 0-2 min 2% B, followed by an increase to 60% B within 15 min, followed by another increase to 80% B within 18 min before returning to 2% B within 25 min (total run time was 25 min.). The Agilent LC-MSD mass spectrometer settings were as follows: positive electrospray mode, capillary voltage 3500 V, drying gas (N 2 ) 12 L/min at 350 °C, nebulizer gas (N 2 ) pressure 50 psi, and scan mode m/z 70-500.
To obtain product ions spectra, samples were dissolved in acetonitrile/water (1:1, containing 0.1% formic acid) and the fragmentor voltage was set at 150 V. For the quantification of clonazolam in tablets, the fragmentor voltage was set at 50 V.
Analyses were performed on an Agilent 1100 HPLC system equipped with a G13795 degasser, G1312A BinPump, a G1313A ALS and G1316A column oven (COLCOM) (Agilent, Little Island, Cork). Separation was obtained on an Allure PFP Propyl column (5 μm, 50 x 2.1 mm) Restek (Bellefonte, PA, USA). Mobile phase A consisted of 0.1% formic acid in water, whereas mobile phase B consisted of 0.1% formic acid in acetonitrile. The flow rate was set at 0.80 mL/min, with an injection volume of 10 µL, and the column temperature was 30°C. The following gradient elution program was used: 0-2 min 2% B, followed by an increase to 60% B within 15 min, followed by another increase to 80% B within 18 min before returning to 2% B within 25 min (total run time was 25 min.). The Agilent LC-MSD mass spectrometer settings were as follows: positive electrospray mode, capillary voltage 3500 V, drying gas (N 2 ) 12 L/min at 350 °C, nebulizer gas (N 2 ) pressure 50 psi, and scan mode m/z 70-500.
To obtain product ions spectra, samples were dissolved in acetonitrile/water (1:1, containing 0.1% formic acid) and the fragmentor voltage was set at 150 V. For the quantification of clonazolam in tablets, the fragmentor voltage was set at 50 V.
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!