The methods used for the characterization of the samples were described in our previous study. 15 (link) The different phenolic compounds detected in the samples of wine and must were identified by their UV-Vis spectra and chromatographic comparison with authentic commercial markers. 17, 18 For the determination of OHTyr and its metabolites, a UHPLC coupled to a 6460 QqQ-MS/MS (Agilent Technologies, Waldbronn, Germany) was used, following the method designed by Domínguez-Perles et al. (2015). 19 (link) For the analysis of MEL and serotonin, the extraction procedure for the wines and musts was as previously described. 20 The separation and determination of MEL and serotonin were performed using a UHPLC coupled to a 6460 QqQ-MS/MS (Agilent Technologies, Waldbronn, Germany) and a Waters Acquity UHPLC BEH C 18 1.7 µm 2.1 × 50 mm column.
6460 qqq ms ms
Manufactured by Agilent Technologies
Sourced in Germany, United States
The 6460 QqQ-MS/MS is a triple quadrupole mass spectrometer designed for high-performance liquid chromatography (HPLC) and ultra-high-performance liquid chromatography (UHPLC) applications. It is a sensitive and selective analytical instrument capable of performing quantitative and qualitative analysis of a wide range of analytes in complex matrices.
Automatically generated - may contain errors
Lab products found in correlation
1290 series uhplc, by Agilent Technologies (3 mentions)
Zorbax eclipse plus c18 column, by Agilent Technologies (2 mentions)
Masshunter software version b 04, by Agilent Technologies (2 mentions)
Spe cartridge, by Waters Corporation (1 mentions)
Agilent sb c18 column, by Agilent Technologies (1 mentions)
Whatman filter paper, by Cytiva (1 mentions)
Zorbax sb c18 column, by Agilent Technologies (1 mentions)
Slgvr04nl, by Merck Group (1 mentions)
C18 solid phase extraction cartridge, by Waters Corporation (1 mentions)
Acquity beh c18 column, by Waters Corporation (1 mentions)
Zorbax eclipse plus c18, by Agilent Technologies (1 mentions)
13 protocols using 6460 qqq ms ms
1
Quantification of Phenolic, Melatonin, and Serotonin Compounds in Wine and Must
2
Phenolic Compounds Analysis in Wines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The methods used for the characterization of the samples were described in our previous reports. 28, 29 The different phenolic compounds detected in the samples of wine and must were identified by their UV-Vis spectra and chromatographic comparison with authentic commercial markers. 18, (link)30 Anthocyanins were quantified at 520 nm by comparison with malvidin-3glucoside (LOD = 0.074 ppm, LQD = 0.240 ppm) and flavonols were quantified at 360 nm by comparison with quercetin-3-rutinoside (LOD = 0.123 ppm, LQD = 0.409 ppm). Hydroxycinnamic acid derivatives were quantified at 320 nm by comparison with caffeic acid (LOD = 0.064 ppm, LQD = 0.212 ppm) and finally, E-stilbenes were quantified by comparison with trans-resveratrol (LOD = 0.024 ppm, LQD = 0.074 ppm).
For the determination of OHTyr and its metabolites, a UHPLC coupled to a 6460 QqQ-MS/MS (Agilent Technologies, Waldbronn, Germany) was used, following the method designed by Domínguez-Perles et al. (2015). 31 (link) For the analysis of MEL and serotonin, the extraction procedure for the wines and musts was as previously described. 32 The separation and determination of MEL and serotonin was performed using a UHPLC coupled to a 6460 QqQ-MS/MS (Agilent Technologies, Waldbronn, Germany) and a Waters Acquity UHPLC BEH C 18 1.7 µm 2.1 × 50 mm column.
For the determination of OHTyr and its metabolites, a UHPLC coupled to a 6460 QqQ-MS/MS (Agilent Technologies, Waldbronn, Germany) was used, following the method designed by Domínguez-Perles et al. (2015). 31 (link) For the analysis of MEL and serotonin, the extraction procedure for the wines and musts was as previously described. 32 The separation and determination of MEL and serotonin was performed using a UHPLC coupled to a 6460 QqQ-MS/MS (Agilent Technologies, Waldbronn, Germany) and a Waters Acquity UHPLC BEH C 18 1.7 µm 2.1 × 50 mm column.
3
Melatonin Extraction and Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Melatonin and melatonin-derivatives extraction was done through a modification of the methods described by Riga et al [28 (link)] and Li et al. [33 (link)]. Briefly, 1 g of frozen leaves were ground into powder with liquid nitrogen and homogenized in a mix of acetone:methanol:water (89:10:1) containing 2.5 mM trichloroacetic acid. The homogenates were shaken for 30 min at RT and centrifuged at 10,000× g at 4 °C for 15 min. The supernatants were centrifuged again and subsequently filtered with Whatman filter paper (0.4 µm). The filtered supernatants were purified using an SPE cartridge (Waters, Milford, MA, USA). The cartridge was then washed with 10 mL 5% methanol, and melatonin was finally eluted at a natural flow rate with 2 mL 80% methanol. The extracts were subsequently filtered through a Whatman filter paper (0.20 μm) before UHPLC-ESI-MS/MS analysis. Melatonin, 3OH-Mel, AFMK and AMK determination and quantification was analyzed using a UHPLC-ESI-MS/MS (UHPLC-1290 Series and a 6460 QqQ-MS/MS; Agilent Technologies, Waldbronn, Germany) with an Agilent SB-C18 column (4.6 × 50 mm; 1.8 μm; Agilent Technologies, Santa Clara, CA, USA). The data reported are the mean ± SE of 3 biological replicates per treatment.
4
Quantification of Anthocyanins in Dried Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The anthocyanins of samples at five timepoints including the start, the end, the inflection point, the midpoint from start to the inflection point, and the midpoint from the inflection point to the end of the drying curves were identified and quantified by ultra-high-pressure liquid chromatography (UPLC, Agilent, Santa Clara, CA, USA), and triple quadruple mass spectrometry (6460 QqQ-MS/MS, Agilent, Santa Clara,, CA, USA) equipped with an electrospray ionization source (ESI). The ZORBAX SB-C18 column (100 mm × 2.1 mm i.d., 1.8 µm, Agilent, Santa Clara, CA, USA) was used to isolate the compounds. The mobile phase consisted of a gradient of 0.1% aqueous formic acid (A) and acetonitrile with 0.1% formic acid (B) at a flow rate of 0.2 mL/min. The gradient elution was set as follows: 0–6.8 min from 90% to 10% A, 6.8–7.0 min with 10% A, 7.0–7.5 min from 10% to 90% A, and 7.5–9.5 with 90% A. The sample injection volume was 5 μL, and the column temperature was 35 °C. The dynamic MRM (Multiple Reaction Monitoring) data were acquired, and the compounds were identified by comparing the dynamic MRM information with reference standards.
5
UHPLC-MS/MS Analysis of Analytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Chromatographic analyses were carried out with a UHPLC coupled to a 6460 QqQ-MS/MS (Agilent Technologies, Waldbronn, Germany) equipped with an electrospray ionization (ESI) source, following the analytical methodology previously reported [33 (link),34 (link),35 (link)].
6
Quantitative Analysis of Metabolites by UHPLC-MS/MS
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Chromatographic separation coupled with mass spectrometry was performed with an ultra‐HPLC system (Agilent) and triple quadruple mass spectrometry (6460QqQ‐MS/MS; Agilent) equipped with an electrospray ionization source. For chromatographic elucidation, samples were injected onto a ZORBAX Eclipse Plus C18 column (50 mm × 2.1 mm i.d., 1.8 µm, Agilent) reversed phase packing column. The mobile phase consisted of 10 mmol/L ammonium formate (A) and methanol (B) at a flow rate of 0.4 ml/min, and the injection volume was 5 μl at 30°C column temperature. A gradient elution was this as follows: 90%–10% B in 6 min, 10% B in 7 min, 90% B in 7.5 min, and 90%–90% B in 10 min.
ESI in negative ionization mode was performed with the following parameters: the drying gas flow rate was 10.0 ml/min, gas temperature was set at 350°C, and capillary voltage was 3,500 V. A multiple reaction monitoring (MRM) mode was used, and all the transitions (Table1 ) were monitored over the run time with dwell times of 20 ms.
ESI in negative ionization mode was performed with the following parameters: the drying gas flow rate was 10.0 ml/min, gas temperature was set at 350°C, and capillary voltage was 3,500 V. A multiple reaction monitoring (MRM) mode was used, and all the transitions (Table
7
Quantification of 6mA DNA Methylation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Zebrafish embryonic gDNA (20−300 ng) or tissue gDNA (1−2 μg) in 26 μl of nuclease-free H2O was denatured at 100 °C for 5 min, chilled on ice for 2 min and digested by 1 μl nuclease P1 (1 U μl−1, Wako USA, 145-08221) in 10 mM NH4OAc pH 5.3 (adding 3 μl 100 mM NH4OAc) at 42 °C overnight. This process was followed with the addition of 3.4 μl NH4HCO3 (1 M) and 1 μl of phosphodiesterase I from crotalus adamanteus venom (0.001 U, Sigma, P3243-1VL) at 37 °C for 2 h and finally by addition of 1 U of alkaline phosphatase from E. coli (Sigma, P5931-500UN) at 37 °C for 2 h. Digested DNA was diluted twofold with nuclease-free H2O and was filtered through 0.22 μm filter (Millipore, SLGVR04NL). A portion of 10 μl sample was injected into LC–MS/MS and the nucleosides were separated by reverse-phase UHPLC on a C18 column (Agilent, 927,700-092), with online MS detection using Agilent 6,460 QQQ–MS/MS set to multiple reaction monitoring in positive electrospray ionization mode. Nucleosides were quantified using the nucleoside precursor ion to base ion mass transitions of 266.1−150.0 for 6mA and 252.1−136.0 for A. Quantification of the ratio 6mA/A was performed using the calibration curves obtained from nucleoside standards running at the same time.
8
Melatonin Extraction and Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Melatonin was extracted using the method described by Pothinuch and Tongchitpakdee [69 (link)]. Embryo samples (0.2 g) were pulverized with liquid nitrogen and homogenized in 5 mL of methanol. The homogenates were centrifuged at 10,000× g for 15 min at 4 °C, after ultrasonication (80 Hz) at 45 °C for 40 min. The extracts were dissolved in 1 mL of 5% methanol and purified using a C18 solid phase extraction cartridge (Waters). The sample solution was eluted through a 0.1 μm syringe filter by 1 mL of 80% methanol, and then assayed by UHPLC-ESI-MS/MS (UHPLC-1290 Series and a 6460 QqQ-MS/MS; Agilent Technologies). The excitation and emission wavelengths were at 285 and 345 nm, respectively. Melatonin content was calculated by comparing the peak area (% fluorescence) of the sample with that of its standard curve.
9
Ultra-high pressure liquid chromatography analysis of tea polyphenol profile
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The tea polyphenol profile was analyzed by an ultra-high pressure liquid chromatograph (UPLC) (Agilent, Santa Clara, California, USA) equipped with an electrospray ionization source (ESI) and a triple quadrupole mass spectrometer (6460QqQ-MS/MS, Agilent). The ZORBAX Eclipse Plus C18 column (100 mm × 2.1 mm i.d., 1.8 µm, Agilent, USA) was used to separate the polyphenol compounds. The mobile phase consisted of a gradient of 0.1 % aqueous formic acid (A) and methanol with 0.1 % formic acid (B) at a flow rate of 0.4 mL/min. The gradient elution was set as follows: 0–11.5 min from 95 % to 5 % A, 11.5–12 min from 5 % to 95 % A, 12–14.5 min with 95 % A. The sample injection volume was 5 μL, and the column temperature was 35 °C.
All the compounds were exactly identified by comparing the dynamic MRM information including retention time, fragmentor voltages, collision energies and transitions with reference standards.
All the compounds were exactly identified by comparing the dynamic MRM information including retention time, fragmentor voltages, collision energies and transitions with reference standards.
10
Urinary F4-NeuroPs and F2-dihomo-IsoPs Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The separation of the F 4 -NeuroPs and F 2 -dihomo-IsoPs in the urine was performed using a UHPLC coupled with a 6460 QqQ-MS/MS (Agilent Technologies, Waldbronn, Germany), using the set-up described previously Medina, et al. [27] . Data acquisition and processing was performed using MassHunter software version B.04.00 (Agilent Technologies, Walbronn, Germany). The qualitative and quantitative analysis of F 4 -NeuroPs and F 2 -dihomo-IsoPs was performed using the authentic markers synthesized by Durand's team. Three deuterated anal ytes were used as internal standards (Fig. 2).
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!