The HPLC-UV-DAD analysis was performed on F2-UAE, F3-UAE, F2-MHG, and F3-MHG using a 1260 Infinity II LC System (Agilent, Santa Clara, CA, USA) equipped with an Agilent G7111A quaternary pump and a WR G7115A diode array detector. Poroshell 120 EC-C18 was used for the separation (150 × 4.6 mm i.d., 4.0 μm particle size, Agilent, Santa Clara, CA, USA) with the column at 30 °C, using water (mobile phase A) and acetonitrile (mobile phase B), both with 0.02% trifluoroacetic acid. The elution condition involved the following linear gradient: 0–2 min, 0→0% B; 2–14 min, 0→18% B; 14–24 min, 18→25% B; 24–60 min, 25→58% B; 60–65 min, 58→100% B. Phase B then returned to the initial conditions and was re-equilibrated for 1 min. The total analysis time was 66 min, the flow rate was 0.350 mL/min, and the injection volume was 10 μL. Three wavelengths (280, 320, and 520 nm) were chosen for UV detection. Retention times and spectral data were compared to standards to perform identification.
Wr g7115a
Manufactured by Agilent Technologies
Sourced in United States
The WR G7115A is a laboratory equipment product from Agilent Technologies. It is a specialized device designed for scientific and technical applications, but a detailed and unbiased description of its core function cannot be provided while maintaining conciseness.
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4 protocols using wr g7115a
1
HPLC-UV-DAD Analysis of Plant Extracts
2
HPLC Analysis of Phytochemical Compounds
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The analysis was performed on a 1260 Infinity II LC System (Agilent, Santa Clara, CA, USA) equipped with an Agilent G7111A quaternary pump and a WR G7115A diode array detector. The separation was done with Poroshell 120 EC-C18 (150 × 4.6 mm i.d., 4.0 μm particle size, Agilent, Santa Clara, CA, USA) column at 30 °C, using water (mobile phase A) and acetonitrile (mobile phase B), both with 0.02% trifluoroacetic acid. The elution condition involved a linear gradient as follows: 0–2.5 min, 5→20% B; 2.5–5 min, 20→30% B; 5–12 min, 30→45% B; 12–17 min, 45→60% B; 17–21 min, 60→80% B; held at 80% B for other 6 min. Phase B reached 95% and held at 95% for 3 min; then returned to the starting conditions and re-equilibrated for 2 min. The total analysis time was 32 min, the flow rate was 0.5 mL/min, and the injection volume was 20 μL. UV detection was set at four different wavelengths (220, 280, 320, and 360 nm). Identification was carried out by comparing the retention times and spectral data with those of standards.
3
UV-DAD Analysis of Fatty Acid Isomers
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To achieve UV-DAD information of fatty acids isomers, separation was also performed by using a 1260 Infinity II LC System (Agilent, Santa Clara, CA, USA) equipped with an Agilent G711A quaternary pump and a WR G7115A diode array detector. The instrument was equipped with Kinetex® PS C-18 (50 × 2.1 mm i.d., 2.6 μm particle size) with a linear gradient in which the percentage of B increases as the following: 0–5 min, 5%→55% B; 5–10 min, 55%→75% B; 10–11% min, 75%→95% B; 11–12 min, and 95% B. The wavelengths were 205, 268, and 282 nm. The flow was 0.4 mL/min.
4
Biogenic Amines Quantification in Cheese
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The eight biogenic amines (TRP, PHE, PUT, CAD, HIS, TYR, SPD, and SPM) were detected and quantified using high-performance liquid chromatography equipped with a diode array detector (WR G7115 A, Agilent Technologies, Santa Clara, CA, USA) at 254 nm, as described in Eerola et al. [38 (link)]. The extraction of BA’s was carried by homogenizing each cheese sample (5 g) with 20 mL of 0.4 M perchloric acid followed by centrifugation (3000 rpm, 10 min, 4 °C). A second extraction was performed again with 20 mL 0.4 M perchloric acid and both extracts were pooled together and the total volume was made up to 50 mL with 0.4 M perchloric acid. The extract was further filtered through Whatman paper No.1 (0.2 μm).
Before derivatization, the extract/standard solutions (1 mL) were initially alkalinized by adding 200 μL 2M NaOH and 300 μL saturated sodium bicarbonate. To this, 2 mL of dansyl chloride (1% w/v in acetone) was added and allowed to react in a dark room at 40 °C, 45 min with intermittent stirring. For removal of the residual dansyl chloride, 100 μL of 25% ammonium hydroxide was added to the mixture and kept in room temperature for 30 min. The sample volume was made up to 5 mL with acetonitrile and centrifuged at 3000 rpm for 10 min at 4 °C. The final supernatant was filtered using 0.2 μm membrane filter (Sartorius, Goettingen, Germany) and stored at −25 °C until HPLC analysis.
Before derivatization, the extract/standard solutions (1 mL) were initially alkalinized by adding 200 μL 2M NaOH and 300 μL saturated sodium bicarbonate. To this, 2 mL of dansyl chloride (1% w/v in acetone) was added and allowed to react in a dark room at 40 °C, 45 min with intermittent stirring. For removal of the residual dansyl chloride, 100 μL of 25% ammonium hydroxide was added to the mixture and kept in room temperature for 30 min. The sample volume was made up to 5 mL with acetonitrile and centrifuged at 3000 rpm for 10 min at 4 °C. The final supernatant was filtered using 0.2 μm membrane filter (Sartorius, Goettingen, Germany) and stored at −25 °C until HPLC analysis.
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