[18F]FSPG formulated in PBS (a 10-μL aliquot) was mixed with OPA reagent solution (10 μL) and stirred for 1 min at room temperature. The resultant sample was analysed by HPLC using a Waters Spherisorb C18, 3-µm column (250 × 4.6 mm) eluted at 1.0 mL min−1 with a mixture of 10-mM NaH2PO4 (pH adjusted to 7.0 using 5.0-M NaOH) and methanol (60:40). The eluate was monitored continuously for radioactivity and absorbance at 338 nm. A sample of authentic FSPG (100 µM, 50 µL) when incubated at room temperature for 1 min with 50 µL of OPA reagent gave a single peak with a retention time of 4.6 min. The derivatised18F-fluorinated product gave one radioactive peak and one stable peak both with the same retention time (4.6 min) as the OPA derivative of authentic FSPG. Typically, the radiochemical purity was 95–97%, and the average molar activity was 66 GBq µmol−1 (range, 44–73 GBq µmol−1) at the end of synthesis (EOS).
Spherisorb c18
Manufactured by Waters Corporation
Sourced in United States
Spherisorb C18 is a reversed-phase HPLC packing material composed of high-purity silica particles chemically bonded with octadecylsilane (C18). It is designed for the separation and analysis of a wide range of organic compounds in liquid chromatography applications.
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Lab products found in correlation
2996 photodiode array detector, by Waters Corporation (1 mentions)
Waters 2487 dual λ absorbance detector, by Waters Corporation (1 mentions)
Waters 717 plus autosampler, by Waters Corporation (1 mentions)
Milli q purification system, by Merck Group (1 mentions)
Waters 1525 binary pump, by Waters Corporation (1 mentions)
Model 2998, by Waters Corporation (1 mentions)
Pda 2998 series, by Waters Corporation (1 mentions)
Elix 03, by Merck Group (1 mentions)
6 protocols using spherisorb c18
1
Radiolabeling and Purity Analysis of [18F]FSPG
2
Homogeneity Analysis of Purified Proteins
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Homogeneity of purified proteins (A1, B1, B2, C1) was determined by performing reverse phase HPLC using Waters Spherisorb® C18 (5 μm, 4.6 X 250 mm) column with solvent A (0.1% TFA in water) and solvent B (100% acetonitrile containing 0.1% TFA) in a linear gradient of acetonitrile (20–70%) over a period of 50 minutes at a flow rate of 1 mL/min. The sample injection volume was 20 μL and column was washed with solvent A and brought to 20% acetonitrile in 5 minutes. The protein peak was detected at 280 nm using Waters 2996 photodiode array detector [25 (link)].
3
Quantitative Analysis of Essential Oils
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All 22 standards (GR grade, 1-22) of respective essential oil components were purchased from following sources: BA (100%), BNZLD (99%) from Merck, Germany, TCNMA (99%), CNMOH (98%), EGL (99%), CVL (97%), ATPNOL (90%), GRNL (98%), MYR (95%) from Aldrich, Germany, SFRL (97%), TCRPHLN (98.5%) from Sigma, Germany, CMN (90%), TCNM (99%), CVN(98%), DHCVN (98%), LNL (97%), BBZT (99%), LMN (90%) from Across Organics, ATPA (90%), PNN (98%) from Fluka. Methanol and acetonitrile gradient grade for HPLC analysis were obtained from Merck (Germany). High purity water was prepared using Milli Q purification system from Millipore (Peenya, Bangalore, India); all commercial oils were purchased from International Flavors and Fragrances India Limited, India.
The HPLC system consisted of Waters 1525 binary pump, Waters 717 plus auto sampler and Waters 2487 dual λ absorbance detector (Waters, Milford, MA, USA). Reversed phase columns, Hypersil ODS C18 (100×4.6 mm, i.d. 3μ) from Thermo Fischer, USA, Spherisorb C18 (125×4.6 mm, i.d. 3μ) from Waters, USA and Wakosil-II C18 (150×4.6 mm, i.d. 3μ) from SGE, Australia were used. Free statistics software, version 1.1.23-r7m Wessa, P. (2013) was used for statistical calculation.
The HPLC system consisted of Waters 1525 binary pump, Waters 717 plus auto sampler and Waters 2487 dual λ absorbance detector (Waters, Milford, MA, USA). Reversed phase columns, Hypersil ODS C18 (100×4.6 mm, i.d. 3μ) from Thermo Fischer, USA, Spherisorb C18 (125×4.6 mm, i.d. 3μ) from Waters, USA and Wakosil-II C18 (150×4.6 mm, i.d. 3μ) from SGE, Australia were used. Free statistics software, version 1.1.23-r7m Wessa, P. (2013) was used for statistical calculation.
4
Rat Plasma Corticosterone Quantification
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Rat plasma corticosterone levels were measured by high-performance liquid chromatography (HPLC)/ultraviolet (Waters, USA) according to Woodward and Emery[15 (link)] with slight changes. Dexamethasone was used as an internal standard. Five hundred microliters of plasma comprising a 50 μL of dexamethasone was extracted with 5 mL of dichloromethane. The dichloromethane extract was allowed to dry and dissolved in 100 μL of the mobile phase. Twenty microliters of the sample was inserted into the HPLC for quantification purpose. Mobile phase contained methanol and water in the ratio of 70:30. The flow rate was 1.0 mL/min and analytical column used was Waters Spherisorb® C 18 (250 mm × 4.6 mm, 5 μm). Plasma corticosterone was identified at 250 nm wavelength by photodiode array detector (Model 2998, Waters, USA). The chromatogram was documented and evaluated by “Empower” software.
5
Quantitative Analysis of Withanolides
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For the extraction of withaferin A, withanolide A, and 12 deoxywithastramonolide from the roots and leaves, reverse HPLC analysis was carried out at the analytical chemistry division of the institute. LC-10A gradient HPLC instrument with two LC-10AD pumps controlled by a CBM-10 interface module, a model 7725 I manual injector valve, a 20 μL sample loop, and a multidimensional UV-visible detector, SPD-10 was used for analysis. For peak purity tests of the compound, a SPDM10AVP photodiode array detector was used. The solvents were filtered using a filtering system (Millipore, Bedford, MA, USA). As per HPLC method [22] (link), the different accessions of Withania were analyzed with slight modifications [23] (link) using a Waters Spherisorb C18 analytical column (4.6 mm × 250 mm, 10 μm ODS); mobile-phase acetonitrile; 0.1% tri-flouro acetic acid in water (40:60; flow rate 1.0 mL/min; column temperature 26°C, detector wavelength 220 nm.
6
Colchicine Quantification in Medicinal Plants
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The analysis of the colchicine was done by using HPLC system called WATERS (Milford, USA) having Waters PDA 2998 series Eur J Biol 2020; 79(2): 67-74 Maqsood et al. Colchicine Quantification in Salt Treated Colchicum luteum photodiode array detector set at wavelength range 190-800 nm. The column from the Waters Spherisorb® C18 bonded with 5 µm (4.6 x 250 mm) accompanied with EMPOWER-2 software was utilized for collection and processing of chromatographic data. Ultrasonic cleaner (Steryl medi-equip systems) and water purification system ELIX 03 (MILLIPORE, USA) was also used.
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