The polyphenolic compounds present in the most active fractions were analyzed by high-performance liquid chromatography (UPLC Dionex Ultimate 3000 RS, Waltham, MA, USA) coupled to a Hybrid Quadrupole-Orbitrap Mass Spectrometer (QExactive) with HESI (Heated Electrospray Ionization source in negative mode. Prior to direct injection, the samples were filtered through a 0.45 μm nylon filter (Sigma Aldrich, St. Louis, MO, USA). Separation of the compounds was achieved with a Xbridge BEH C18 column (100 mm × 2.1 mm, 2.5 μm) through a binary solvent system consisting of a phase A (H2O at 0.1% in trifluoroacetic acid) and phase B (methanol at 0.1% in trifluoroacetic acid) [24 (link)]. The following gradient was applied: 5% B (1 min), 5–100% B (10min), 100% B (2 min), 5% B (3min); the flow rate was 0.5 mL/min. The column temperature was 40 °C. The injection volume for all selected fractions was 20 µL. The instrument was operated in negative ion mode using a scan range from m/z 50 to 1000. Nitrogen was used as the dry gas at a flow rate of 4.5 mL/min, the temperature was set at 250 °C and the collision energy (HCD cell) was operated at 30 kv. Detection was based on calculated exact mass and on retention time of target compounds.
0.45 μm nylon filter
Manufactured by Merck Group
Sourced in United States
The 0.45 μm nylon filter is a laboratory filtration device designed to remove particulates and contaminants from liquids. It has a pore size of 0.45 micrometers, which allows the passage of small molecules and ions while retaining larger particles and suspended solids. The filter is made of nylon material, providing durability and chemical compatibility for a variety of applications.
Automatically generated - may contain errors
Lab products found in correlation
Xr ods c18 column, by Shimadzu (1 mentions)
Nexera x2 uhplc, by Shimadzu (1 mentions)
Eluent a, by Merck Group (1 mentions)
Eluent b, by Merck Group (1 mentions)
Lcms 9030 qtof, by Shimadzu (1 mentions)
Polaris c18 a column, by Agilent Technologies (1 mentions)
Pda 2996 photodiode array detector, by Waters Corporation (1 mentions)
Diclofenac, by Merck Group (1 mentions)
Gpc malls, by Wyatt Technology (1 mentions)
2690d separations module, by Waters Corporation (1 mentions)
2414 refractive index detector ri, by Wyatt Technology (1 mentions)
Dawn eos malls detector, by Wyatt Technology (1 mentions)
Gf c filter, by Merck Group (1 mentions)
9 protocols using 0.45 μm nylon filter
1
UPLC-QExactive Analysis of Polyphenols
2
UHPLC-QTOF Analysis of Betalains
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A Shimadzu UHPLC Nexera X2 attached to a Shimadzu (Tokyo, Japan) LCMS-9030 QTOF with ESI source was used. Once samples were filtered (0.45 μm nylon filter, Sigma-Aldrich, St. Louis, MO, USA), betalains were separated using a Shim pack XR–ODS C18 column (150 × 2 mm, 2.2 μm particle size), at 7500 psi maximum working pressure and maximum flow rate of 0.4 mL/min, and using 1% formic acid in water (v/v, eluent A) and a mixture of acetonitrile/water/formic acid (80:19:1) (eluent B) (Sigma-Aldrich, St. Louis, MO, USA). The volume injection was 5 μL. The chromatographic method started with 100% of A, followed by a linear gradient from 0 to 20% of B in 35 min and then a linear gradient from 20 to 100% of B in 5 min [6 (link)]. To re-establish the initial conditions, a linear gradient from 100% of B to 100% of A was used for 10 min. The identification of the individual betalains was performed in positive mode using a sweeping range of m/z 100 to 1000 u. An amount of 4.5 mL/min of nitrogen was used as the drying gas, the temperature was set at 300 °C, the nebulizer gas flow was 3 L/min, the interface temperature was 526 °C, the sampling rate was 5 µ/s, the heater gas flow was 10 L/min, and the detector voltage was 0.20 kV.
3
Diclofenac Quantification by HPLC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
During the testing (always before and after the bath change), samples for the determination of Diclofenac concentration were regularly withdrawn from test tanks. Diclofenac determination in water samples was performed by high performance liquid chromatography (HPLC) with photometric detection. Water samples were filtered through a 0.45 μm nylon filter (Millipore, Billerica, MA) and used for analysis. The sample volume injected into the HPLC system was 20 μL. Diclofenac was separated by an isocratic elution method with acetonitrile/water 50/50 (v/v) on a Polaris C18-A column (3 μm, 150 × 4.6 mm, Varian, Inc., Palo Alto, CA). The mobile phase flow rate was 1 mL min−1, the column temperature was 25°C, and UV detection was performed at 310 nm. Chromatographic analysis was accomplished by means of an Alliance 2695 chromatographic system (Waters, Milford, MA) with a PDA 2996 photodiode array detector (Waters, Milford, MA). Diclofenac was purchased from Sigma-Aldrich (St. Louis, MO). All solvents were of HPLC-grade purity (Chromservis, s.r.o., CZ). The detection limit for Diclofenac was 11 ng mL−1. The limit of quantification for Diclofenac was 37 ng mL−1. The coefficient of variation was 4.5%.
4
Molecular Weight Distribution of UPF
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
GPC-MALLS (Wyatt Technology Corporation, Santa barbara, CA, USA) was employed for evaluating the distributions of molecular weight and mass of UPF. This system employs a Waters 2690D separations module, a Waters 2414 refractive index detector (RI) and a Wyatt DAWN EOS MALLS detector. Ten milligrams of UPF was dissolved in 1 mL of 0.1 M NaNO3 solution that was rolled overnight so that the entire amount was dissolved. Post a four-fold dilution, a 0.45 μm nylon filter (Millipore Corp., Billerica, MA, USA) was utilized to filter the solution of which 100μL was injected. A series of OHpak SB-805 HQ, OHpak SB-806 HQ, and OHpak SB-803 HQ columns were used. The collection and analysis of data was done using Astra 4.90.08 software (Wyatt Technology Corporation, Santa barbara, CA, USA).
5
Quantification of Gymnemic Acid in Herbal Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The quantification of gymnemic acid, a method described previously by Manohar et al. (2009), was followed with minor modifications.[12 ] Five hundred milligrams of dried leaf sample was put into a 100-mL round bottom flask and 50 mL of extraction solvent was added (volume ratio of methanol to water is 1:1) with 10 mL of 11% potassium hydroxide solution. The mixture was refluxed for an hour. The 9 mL concentrated HCl was added and refluxed again for 1 h and the mixture was cooled to room temperature. The extract was filtered through 0.45 μm nylon filter (Millipore), the volume was made up to 100 mL with extraction solvent, and the clean supernatant was used for HPLC analysis. The isocratic mobile phase composition (acetonitrile-A and water-B, 80:20 [v/v]) with elution rate of 1 mL/min, data acumination and quantification was done at 210 nm. Gymnemagenin standard was purchased from Sigma Aldrich, USA. The conversion of gymnemagenin to gymnemic acid was done using the equation C = X(809.0 / 506.7), where C is the content of gymnemic acid in the sample, X is the content of gymnemagenin present in the sample, 506.7 is the molecular weight of gymnemagenin, and 809.0 is the molecular weight of gymnemic acid. The chromatogram of gymnemic acid standard shown in Figure 1 , with a retention time of 1.83 min.
6
Extraction and Analysis of PM2.5 Particles
7
Extraction of Gallotannin-rich Phenolics from Red Sword Bean Coats
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Bean coats were manually separated from the dry red sword beans, and milled to fine powder using a Kenwood Multi-Mill (Kenwood, Havant, UK), thoroughly mixed and stored at 4°C before further used. For the extraction of gallotannin-rich phenolics, the powders of red sword bean coats were extracted with 70% ethanol (1:20, w/v) for 1 h in a water bath shaker (200 rpm) at room temperature (22 ± 1°C), and then centrifuged (2,000 × g, 15 min, 4°C). The supernatant was collected and the extraction was repeated twice more. After the extractions, the supernatants were combined, filtered through 0.45 μm nylon filter (Millipore, Bedford, MA), concentrated by rotary evaporation at 40°C under vacuum, and freeze-dried. The crude extract of red sword bean coats were stored at −20°C for further separation process.
8
Fungal Biomass Air-Pulsed Bioreactors
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Two fungal biomass air-pulsed fluidized bed glass bioreactors (1.5 L) (Blánquez et al. 2006) (link) were set up in parallel for each experiment (batch and continuous), one inoculated with T. versicolor (B-I and C-I for batch and continuous, respectively) and the other non-inoculated as a control (B-NI and C-NI). They were filled with 1.5 L of VHW, and temperature was set up at 25 °C, and pH was controlled to be constant at 4.5±0.5 by HCl 1 M or NaOH 1 M addition. Samples of approximately 250 mL, collected along bioreactors operation, were filtered by vacuum with Whatman GF/C filters and a 0.45-μm nylon filter (Millipore) from which 200 mL were then frozen until pharmaceutical characterization, and approximately 50 mL were used immediately for the other routine analyses (check the BRoutine analyses^section). In the batch experiment, samples for microbial community analyses were also collected and frozen at -20 °C in 2 mL eppendorfs without any processing until DNA extraction.
9
HPLC Analysis of Hydrolyzed Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HPLC analysis of hydrolyzed samples was performed using a Metacarb 87H carbohydrate analysis column (300 × 7.8 mm, Varian, USA) at 60 °C. Sugars and acetic acid were analyzed with a refractive index (RI) detector and furfural and hydroxymethylfurfural (HMF) contents with a UV detector, both in a Jasco chromatograph. The mobile phase was 0.005 M H 2 SO 4 in ultrapure water filtered through 0.45 μm nylon filter (Millipore) and degassed. The flow rate was 0.7 mL/min.
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!