For qualitative and quantitative analysis of the prepared 5-KF and D -fructose solution, a Knauer Smartline HPLC-system (Knauer GmbH, Berlin, Germany) was used. The system was composed of a degasser (Knauer Smartline manager 5000), a pump (Knauer Smartline pump 1000), an autosampler (Knauer Smartline autosampler 3800), a column oven (Knauer Column-Thermostat Jetstream 2 Plus), an RI detector (AZURA RID 2.1L), and an ultraviolet (UV) detector (Knauer Smartline UV detector 2600), which measured the absorbance at a wavelength of 210 nm. Sample separation was achieved using the Eurokat H column (300 × 8 mm; Knauer GmbH) and a precolumn (30 × 8 mm; Knauer GmbH) heated to 65°C. The mobile phase, 5 mM H2SO4, was applied at a flow rate of 0.6 mL min–1. The injection volume was set to 20 μL. Data evaluation and control of the HPLC system were accomplished using ClarityChrom® 8.2.3 (Knauer GmbH). Peak assignment and quantification were performed by applying external standards of D -fructose and 5-KF in concentrations of 0.5, 1, and 2 mM.
Smartline hplc system
Manufactured by Knauer
Sourced in Germany
The Smartline HPLC system is a high-performance liquid chromatography (HPLC) instrument designed for analytical and preparative applications. It features modular components that can be configured to meet specific research or analytical requirements. The system is capable of performing various chromatographic techniques, including reverse-phase, normal-phase, and ion-exchange separations.
Automatically generated - may contain errors
Lab products found in correlation
Model 5050, by Eckert & Ziegler (3 mentions)
Model 1000, by Eckert & Ziegler (3 mentions)
B fc 1000, by Bioscan (3 mentions)
Bfc 4100, by Bioscan (3 mentions)
Gemini c18 column, by Phenomenex (2 mentions)
Sec diol columns, by YMC (2 mentions)
Ri detector 2300, by Knauer (2 mentions)
Precolumn, by Knauer (1 mentions)
Eurokat h column, by Knauer (1 mentions)
Azura rid 2.1l, by Knauer (1 mentions)
Chromquest 5, by Thermo Fisher Scientific (1 mentions)
Smartline 2600, by Knauer (1 mentions)
Kinetex, by Phenomenex (1 mentions)
Centrifugal vacuum concentrator, by Labconco (1 mentions)
Milli q water, by Merck Group (1 mentions)
Chromgate software, by Knauer (1 mentions)
Merlintm high resolution field emission scanning electron microscope, by Zeiss (1 mentions)
Agilent zorbax sb c18, by Agilent Technologies (1 mentions)
13 protocols using smartline hplc system
1
HPLC Analysis of 5-KF and D-Fructose
2
Quantitative Analysis of Microbial Metabolites
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the analysis of substrate consumption and product formation analysis, 1-ml samples were taken at different time points and centrifuged at 13,000×g for 1 min. The supernatants were diluted 1:10 with H2O. HPLC analysis (Knauer Smartline HPLC system, Knauer GmbH, Berlin, Germany) was performed using an Aminex HPX-87H 300 mm × 7.8 mm column (Biorad, Munich, Germany) with 5 mM H2SO4 at 65 °C and a flow rate of 0.6 ml min–1. Substrates and products were quantified by a refraction index detector (RI detector; Azura RID2.1 L, Knauer GmbH, Berlin, Germany) and a UV detector (Smartline 2600, Knauer, Berlin, Germany) at 210 nm by comparison to calibration curves. For product analysis of 5-KF reduction enzyme assays samples were analyzed by a SpectraSYSTEM HPLC system (Thermo Fisher Scientific Inc., Waltham, USA) using an amino phase column Eurospher II 100 NH2 (250 × 3.0 mm; 5 μm particle size) (Knauer GmbH, Berlin, Germany) with integrated precolumn, 90% acetonitrile as solvent, at 40 °C and a flow rate of 0.6 ml min–1. Products were quantified by a refraction index (RI) detector (Shodex RI-101) (Showa Denko Europe GmbH, Munich, Germany) and evaluated by the external standard method with ChromQuest 5.0 (Thermo Fisher Scientific Inc.).
3
HPLC Analysis of Metabolites
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The analysis of substrate and product concentrations was performed by HPLC (Knauer Smartline HPLC system, Knauer GmbH, Berlin, Germany) with an Aminex HPX-87H column (BioRad, Munich, Germany, 300 mm × 7.8 mm) using 5 mM H2SO4 as mobile phase. One milliliter of culture was harvested by centrifugation (10,000 × g, 2 min and 10 °C) at different OD600 in the exponential growth phase. Metabolic end products and the substrate glucose were separated at a column temperature of 65 °C with a flow rate of 0.6 ml min−1 and detected by a refractive index detector. Concentrations were calculated by comparison to corresponding calibration curves.
4
Radiolabeled Tracer Identification and Purification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Analytical radio-HPLC was used to identify the product of each synthesis (via co-injection with reference standard) and to isolate pure products to confirm the Rf value of the product bands in radio-TLC. The radio-HPLC system setup comprised a Smartline HPLC system (Knauer, Berlin, Germany) equipped with a degasser (Model 5050), pump (Model 1000), UV detector (254 nm; Eckert & Ziegler, Berlin, Germany) and gamma-radiation detector, and counter (B-FC- 4100 and BFC-1000; Bioscan, Inc., Poway, CA, USA). All HPLC separations used a C18 Gemini column (Kinetex, 250 × 4.6 mm, 5 µm, Phenomenex, Torrance, CA, USA). Using a mobile phase of 3:1 H2O:MeCN with 0.1% TFA (v/v) and a flow rate of 1.0 mL/min, the retention time of [18F]Flumazenil was 11 min. For [18F]PBR06, the retention time was 8 min using a mobile phase of 60:40 (v/v) MeCN:20 mM sodium phosphate buffer (pH = 5.8) with a flow rate of 1.5 mL/min. [18F]Fallypride samples were separated with a mobile phase of 60% MeCN in 25 mM HN4HCO2 with 1% TEA (v/v) and a flow rate of 1.5 mL/min resulting in a retention time of 4.5 min. [18F]FEPPA samples were separated with a mobile phase of 70:30 v/v H2O:EtOH with 0.1% H3PO4 at 0.8 mL/min, giving a retention time of 15.5 min.
5
HPLC Analysis of Carbohydrates
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The supernatant was centrifuged (20,000 g, 2 min), diluted in di-deionized water and transferred into HPLC vials. The analysis was performed in the smartline HPLC system of Knauer (Berlin, Germany). As stationary phase the column VA300/7.8 Nucleogel sugar 810-H (300 mm) and precolumn CC 30/4 Nucleogel Sugar 810-H (3 × 4 mm) of Macherey-Nagel (Düren, Germany) was used, heated to 72°C. As mobile phase an isocratic flow of 0.8 mL⋅min–1 2.5 mM H2SO4 was applied. Twenty microliter of each sample was injected and separated in a 25 min run. Detection of substrates was carried out with a refraction index detector of ERC GmbH (Riemerling, Germany) and quantified from the peak areas of the calibration curve of standards (1–20 g⋅L–1). The chromatograms were evaluated with the Chromastar software (SCPA GmbH, Weyhe, Germany).
6
HPLC-ELSD Analysis of Plant Sugars
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Sugar analysis was conducted with HPLC-ELSD according to a protocol described in O’Donoghue et al. [16 (link)], with minor modifications. Briefly, 20 mg of pulverized material was extracted in three independent replicates with 2 mL of 75% methanol (v/v) for 15 min. After centrifugation at 14,500× g for 10 min, 750 µL of the supernatant was completely evaporated using a centrifugal vacuum concentrator (Labconco, Kansas City, MO, USA) at 40 °C. Upon reconstitution in 750 µL of water and subsequent filtration through a 0.22 µm nylon syringe filter (Chromservis), 10 µL of the sample was injected on a WATREX polymer IEX Ca-form 8 × 300 mm, 8 µm particle size column (WATREX, Prague, Czech Republic). The Smartline HPLC system (Knauer, Berlin, Germany) was coupled with an Alltech 3300 ELSD detector (Thermo Fisher). Sucrose, glucose, and fructose were identified and quantified using authentic standards (Lach-Ner, Neratovice, Czech Republic). MilliQ water (Milipore Sigma, Billerica, MA, USA), at a flow rate of 0.5 mL min−1, was used as the mobile phase. The ELSD detector operated at a temperature of 80 °C and a nitrogen flow rate of 2 L h−1. The analysis had a total run time of 20 min.
7
Radio-HPLC Analysis of Radiopharmaceuticals
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Radio-HPLC was used to analyze crude radiopharmaceuticals and to perform tests for radiochemical and chemical purity and radiochemical identity of TLC-purified batches of radiopharmaceuticals. The radio-HPLC system setup comprised a Smartline HPLC system (Knauer, Berlin, Germany) equipped with a degasser (Model 5050), pump (Model 1000), UV detector (254 nm; Eckert & Ziegler, Berlin, Germany), gamma-radiation detector (BFC-4100, Bioscan, Inc., Poway, CA, USA), and counter (BFC-1000; Bioscan, Inc., Poway, CA, USA). A C18 Gemini column was used for separations (250 × 4.6 mm, 5 µm, Phenomenex, Torrance, CA, USA). [18F]PBR-06 samples were separated with a mobile phase of 60:40 (v/v) MeCN:20 mM sodium phosphate buffer (pH = 5.8) at a flow rate of 1.5 mL/min resulting in a retention time for [18F]PBR-06 of 6.5 min. [18F]Fallypride samples were separated with a mobile phase of 60% MeCN in 25 mM NH4HCO2 with 1% TEA (v/v) at a flow rate of 1.5 mL/min resulting in a retention time for [18F]Fallypride of 5.8 min.
8
Polysaccharide Molecular Weight Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The MW distribution of the polysaccharides was analyzed via high-performance size exclusion chromatography (HP-SEC) with refractive index (RI) detection (Houben et al., 2011 (link)). AIR (100 mg) was suspended in 5 mL water and heated at 50 °C for 5.5 h under stirring. The solution was dialyzed (MWCO: 12–14 kDa) for 24 h against demineralized water. Subsequently, 2 mL of the retentate was centrifuged (11,000g, 10 min) and the pH and sample concentration were adjusted using 1 M NaOH, 1 M HCl, and 500 mM NaNO3 + 0.025% NaN3. The dialysis and the HP-SEC analysis were repeated five times. Three different polymer mixtures (Pullulan ReadyCial-Kit) with known molecular weight (9.6–708 kDa) were used for calibration. The analyses were conducted on a Smartline HPLC system with a RI detector 2300 (Knauer, Berlin, Germany) equipped with two different, connected SEC-Diol columns (300 and 120 Å, 3 μm; YMC, Kyoto, Japan). Samples (20 μL) were injected and eluted with 50 mM sodium nitrate and 0.025% sodium azide (w/w) at pH 7 for 30 min at a flow rate of 0.3 mL min−1 and isocratic conditions. The HP-SEC chromatograms were divided into three segments: high (>208 kDa), medium (37–208 kDa), and low (19–37 kDa) molecular weight (HMW, MMW, LMW) fractions (supplemental data, Figure A.1).
9
Polysaccharide Molecular Weight Profiling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
High-performance size exclusion chromatography (HPSEC) on a Smartline HPLC system with a RI detector 2300 (Knauer, Berlin, Germany) equipped with two SEC-Diol columns (300 and 120 Å, 3 μm; YMC, Kyoto, Japan) was used to determine the molecular weight (MW) distribution of the soluble polysaccharides as described by Larsen et al. (2019) (link). Samples were dissolved in water (50 °C) and dialyzed against demineralized water (MWCO 12–14 kDa). Polysaccharides were eluted using water with 50 mM sodium nitrate and 0.25% sodium azide. MWs were calculated with eight pullulan standards ranging from 0.504 to 708 kDa (ReadyCal-Kit Pullulan, PSS- Polymer Standards, Mainz, Germany). The chromatograms were divided into three representative fractions: High molecular weight fraction (15–708 kDa), medium molecular weight fraction (5.5–15 kDa), and low molecular weight fraction (<5.5 kDa). The proportions of the fractions relative to the total area were calculated. Raw data of the MW distribution determined with SEC is shown in table A1 in the supplemental data.
10
HPLC Analysis of Dexamethasone Content
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HPLC measurements for the analysis of dexamethasone content was performed on a Knauer Smartline-HPLC system with an internal UV absorption detector (λ = 254 nm), equipped with a Gemini RP C18 column (Phenomenix, 250 nm × 4.6 mm, particle size: 5 μm) and run with an acetonitrile-water (40:60) mixture as the mobile phase at a flow rate of 1.0 mL·min−1 under isocratic regime. The data were analyzed with Chromgate software (Knauer, Berlin, Germany). A calibration curve of dexamethasone was obtained by measuring dexamethasone in an acetonitrile-water (40:60) mixture in the concentration range of 0.004–2 mg·mL−1.
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!