PDP-23 was mixed with human pooled male serum (Sigma-Aldrich) to a final concentration of 50 μg mL−1 and incubated at 37 °C. Aliquots (100 μL) were taken at different time points (0, 2, 4, 6 and 24 h) and quenched with 100 mM ammonium acetate, pH 3 (900 μL), followed by incubation on ice for 30 min. To separate the peptide from the serum components, Oasis HLB 3 cc 60 mg cartridges (Waters) were activated with 6 mL methanol washes, preconditioned with 3 mL 70% acetonitrile, 1% formic acid and equilibrated with 3 mL 1% formic acid. The serum samples were then loaded onto the column and further washed with 3 mL 5% acetonitrile, 1% formic acid before elution with 35% acetonitrile, 1% formic acid.41 (link) Samples were lyophilized and re-dissolved in 100 μL 1% formic acid before LC-MS/MS analysis on an API2000 (SCIEX) and quantification by MultiQuant (SCIEX). The serum stability tests were performed in triplicate and peptide stability was calculated using a non-linear fit of one phase decay in GraphPad Prism 6.
Oasis hlb 3 cc 60 mg cartridges
Manufactured by Waters Corporation
Sourced in United States
The Oasis HLB 3 cc 60 mg cartridges are a type of lab equipment used in various analytical and purification processes. They are designed to facilitate solid-phase extraction (SPE) procedures. The cartridges contain a sorbent material that allows for the selective separation and concentration of analytes from liquid samples.
Automatically generated - may contain errors
Lab products found in correlation
Multiquant, by AB Sciex (1 mentions)
Api 2000, by AB Sciex (1 mentions)
Prism 6, by GraphPad (1 mentions)
Acetonitrile, by Thermo Fisher Scientific (1 mentions)
Formic acid, by Thermo Fisher Scientific (1 mentions)
Methanol, by Thermo Fisher Scientific (1 mentions)
13c34 fb1, by Romer Labs (1 mentions)
Milli q system, by Merck Group (1 mentions)
Mixed cellulose ester membrane filter, by Cytiva (1 mentions)
Acquity hss t3 column, by Waters Corporation (1 mentions)
Acquity beh c18 column, by Waters Corporation (1 mentions)
Ultra performance liquid chromatography system, by Waters Corporation (1 mentions)
4 protocols using oasis hlb 3 cc 60 mg cartridges
1
Serum Stability Assay for Peptide PDP-23
2
Determination of Fumonisins in Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Acetonitrile, methanol, and formic acid were of LC/MS grade (Fisher Scientific, Loughborough, UK). All other chemicals were of analytical grade or better. Deionized water (18.2 MΩ cm) was collected from a Milli-Q system (Millipore Corp., Bedford, MA, USA). Oasis MAX 3cc 60 mg cartridges and Oasis HLB 3cc 60 mg cartridges were purchased from Waters (Milford, MA, USA). Standard solutions of mixed FB1, FB2, and FB3 with certified concentrations of each analyte were purchased from Biopure (Tulln, Austria). [13C34]-FB1, [13C34]-FB2, and [13C34]-FB3 standard solutions with certified concentrations were also purchased from Biopure (Tulln, Austria).
3
Wastewater Solid Phase Extraction
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Wastewater samples were vacuum-filtered through a glass microfiber filter 1.6 µm GF/A (Whatman, Kent, U.K.) and a 0.45 µm mixed cellulose ester membrane filter (Whatman, Kent, U.K.) before extraction, according to the procedures of each laboratory. SW was not filtered. The method is described in detail elsewhere (Bade et al., 2015c) . Briefly, solid phase extraction (SPE), using OASIS HLB 3 cc/60 mg cartridges (Waters Corp., Milford, MA, USA), was applied to all water samples in order to extract the analytes. Cartridges were conditioned with 6 mL of MeOH and equilibrated with 6 mL of Milli-Q water. Then 100 mL of the samples (IWW was diluted four times with Milli-Q water, i.e. 25 mL sample in 100 mL) were passed through the cartridges by gravity and vacuum-dried for approximately 15 min. The analytes were eluted with 6 mL of MeOH and the extracts were evaporated to dryness at 35°C under a gentle nitrogen stream and finally reconstituted in 1 mL of 10 % MeOH aqueous solution.
4
Analytical Procedure for VHW2 Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The analytical procedure performed for the VHW2 samples was based on Gros et al. (2012) .
Briefly, 50 mL of sample was pre-concentrated by solid-phase extraction (SPE) using Oasis HLB (3 cc, 60 mg) cartridges (Waters Corp. Mildford, MA, USA), which were previously conditioned with 5 mL methanol and 5 mL HPLC grade water. Elution was performed with 6 mL of pure methanol. The extracts were evaporated under nitrogen stream and reconstituted with 1 mL of methanol-water (10:90 v/v). Lastly, 10 µL of internal standard mix at 1 ng µL -1 was added to the extracts for internal standard calibration. Chromatographic separation was performed with an ultra-performance liquid chromatography system (Waters Corp. Mildford, MA, USA), using an Acquity HSS T3 column (50 mm x 2.1 mm i.d. 1.7 μm particle size) for the compounds analysed under positive electrospray ionisation (PI) and an Acquity BEH C18 column (50 mm × 2.1 mm i.d., 1.7 μm particle size) for compounds analysed under negative electrospray ionisation (NI), both from Waters Corporation. The UPLC instrument was coupled to a 5500 QqLit, triple quadrupole-linear ion trap mass spectrometer (5500 QTRAP, Applied Biosystems, Foster City, CA, USA) with a Turbo V ion spray source. Two MRM 11 31 32 transitions per compound were recorded using the Scheduled MRM TM algorithm, and the data were acquired and processed using Analyst 2.1 software.
Briefly, 50 mL of sample was pre-concentrated by solid-phase extraction (SPE) using Oasis HLB (3 cc, 60 mg) cartridges (Waters Corp. Mildford, MA, USA), which were previously conditioned with 5 mL methanol and 5 mL HPLC grade water. Elution was performed with 6 mL of pure methanol. The extracts were evaporated under nitrogen stream and reconstituted with 1 mL of methanol-water (10:90 v/v). Lastly, 10 µL of internal standard mix at 1 ng µL -1 was added to the extracts for internal standard calibration. Chromatographic separation was performed with an ultra-performance liquid chromatography system (Waters Corp. Mildford, MA, USA), using an Acquity HSS T3 column (50 mm x 2.1 mm i.d. 1.7 μm particle size) for the compounds analysed under positive electrospray ionisation (PI) and an Acquity BEH C18 column (50 mm × 2.1 mm i.d., 1.7 μm particle size) for compounds analysed under negative electrospray ionisation (NI), both from Waters Corporation. The UPLC instrument was coupled to a 5500 QqLit, triple quadrupole-linear ion trap mass spectrometer (5500 QTRAP, Applied Biosystems, Foster City, CA, USA) with a Turbo V ion spray source. Two MRM 11 31 32 transitions per compound were recorded using the Scheduled MRM TM algorithm, and the data were acquired and processed using Analyst 2.1 software.
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!