Quantifications of 4 main flavonoids (calycosin-7-O-β-d -glucoside, ononin, calycosin and formononetin) in WIE and WAE were performed by a Waters ACQUITY-UPLC CLASS system (Waters Corp., Milford, USA) coupled with an ACQUITY UPLC HSS T3 column (150 mm × 2.1 mm, 1.8 µm) maintained at 30 °C. Elution was performed with a mobile phase of A (0.2% phosphoric acid in water) and B (0.2% phosphoric acid in ACN) under a gradient program: 0–1 min, 22% B; 1–10 min, 22–60% B. The flow rate was 0.35 mL/min, and the injection volume was 2 μL. The analytes were monitored at the UV wavelength of 254 nm. Between two injections, the column was washed with 100% B for 2 min and equilibrated with the initial mobile phase for 5 min.
Acquity uplc class system
Manufactured by Waters Corporation
Sourced in United States
The ACQUITY-UPLC CLASS system is a high-performance liquid chromatography (HPLC) instrument designed for analytical separation and detection of compounds. It utilizes ultra-high-pressure liquid chromatography (UPLC) technology to achieve rapid and efficient separation of complex samples. The system is capable of delivering precise and reproducible results through its advanced fluidic and detection capabilities.
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Lab products found in correlation
Acquity uplc hss t3 column, by Waters Corporation (2 mentions)
Acquity uplc beh phenyl column, by Waters Corporation (1 mentions)
Beh phenyl column, by Waters Corporation (1 mentions)
Milli q system, by Merck Group (1 mentions)
Acetonitrile, by RCI Labscan (1 mentions)
Ginsenoside rb1, by Chengdu Pufei De Biotech (1 mentions)
Ginsenoside rg1, by Chengdu Pufei De Biotech (1 mentions)
Masslynx 4, by Waters Corporation (1 mentions)
Xevo tqd, by Waters Corporation (1 mentions)
Acquity uplc beh c18 column, by Waters Corporation (1 mentions)
Unifi 1, by Waters Corporation (1 mentions)
Waters masslynx 4, by Waters Corporation (1 mentions)
6 protocols using acquity uplc class system
2
Quantitative Analysis of Panax notoginseng Extracts
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Extraction of P. notoginseng was prepared and quantitative analysis of PNE and PNS was performed as stated in our previous article [23 (link)]. In brief, the dried P. notoginseng powder was extracted with 95% ethanol and the ethanol was removed by rotary evaporation. Finally, the extract was freeze-dried to obtain PNE. PNS was obtained from Yunnan Yunke Pharmaceutical Co. Ltd. (China). The component analysis of PNS and PNE was determined by Waters ACQUITY-UPLC CLASS system (Waters Corp., USA) with an ACQUITY UPLC BEH phenyl column (150 mm × 2.1 mm, 1.7 μm) maintained at 45 °C to quantify the contents of notoginsenoside R1, ginsenoside Rb1, ginsenoside Re, ginsenoside Rg1, and ginsenoside Rd.
3
Quantitative Analysis of Ginseng Saponins
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The chemical constitutions of PNS and PNE were quantitatively determined using a Waters ACQUITY-UPLC CLASS system (Waters Corp., USA) coupled with an ACQUITY UPLC BEH phenyl column (150 mm × 2.1 mm, 1.7 μm) maintained at 45°C. Elution was performed with a mobile phase of water (A) and Acetonitrile (B) under a gradient program: 0-10 min, 19% B; 10-15 min, 19-35% B; 15-20 min, 35-38% B. The flow rate was 0.4 mL/min, and the injection volume was 2 μL. The analytes were monitored at the UV wavelength of 205 nm. Prior to the next injection, the column was washed with 100% B for 2 min and then equilibrated with the initial mobile phase for 3 min. Notogisenoside R1, ginsenoside Rb1, ginsenoside Re, ginsenoside Rg1, and ginsenoside Rd (the purities of all standards were higher than 98% by HPLC analysis) were purchased from Chengdu Pufei De Biotech Co., Ltd. (China). Acetonitrile was purchased from RCI Labscan Limited (Thailand) of HPLC grade. Milli-Q water was prepared using a Milli-Q system (Millipore, USA).
4
Quantification of HCA Levels via LC-MS
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The samples were subjected to LC-MS on a Waters Acquity UPLC CLASS system coupled with a triple quadrupole mass spectrometer (Xevo TQD) with positive electrospray ionisation (ESI+) (Waters, Milford, MA, USA). Waters ACQUITY UPLC BEH C18 column (1.7 μm, 50 mm × 2.1 mm i.d) with mobile phases consisting of mobile phase A (10 mM ammonium formate, pH 6.80) and mobile phase B (acetonitrile) was employed to achieve separation. The gradient elution was delivered using a flow rate of 0.3 mL min−1. The mobile phase A was as follows: 0–0.2 min, 90%; 0.2–1.0 min, 90–70%; 1.0–3.0 min, 70–40%; 3.0–3.5 min, 40–10%. The MS system was operated in multi-reaction monitoring (MRM) scan mode; mass spectrometric parameters were: capillary voltage, 0.5 kV; ion source temperature, 120 °C; desolvation temperature, 450 °C; cone gas (nitrogen, purity: 99.99%) flow, 50 L h−1 and desolvation gas (nitrogen, purity: 99.99%) flow, 1000 L h−1, collision (argon, purity: 99.99%). Three standard HCA levels were determined according to standard curves, and the data was acquired using Waters MassLynx 4.1 software (Waters).
5
UPLC-based Quantification of Phytochemicals
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Quantification of four main compounds (rutin, isochlorogenic acid A, isochlorogenic acid C and darutoside) in the active D101 fraction was performed using a Waters ACQUITY-UPLC CLASS system (Waters Corp. United States) coupled with an ACQUITY UPLC HSS T3 column (150 mm × 2.1 mm, 1.8 µm) maintained at 40°C. Samples were eluted with a mobile phase of A (0.2% H3PO4 in water) and B (0.2% H3PO4 in ACN) under a gradient program: 0–2 min, 14% B; 2–8 min, 14%–18% B; 8–10 min, 18–20%; 10–14 min, 20–25%; 14–20 min, 25–35%. The flow rate was 0.4 ml/min and the injection volume was 2 µL. Analytes were monitored at the UV wavelength of 220 nm. Between two injections, the column was washed with 100% B for 2 min and equilibrated with the initial mobile phase for 5 min.
6
UPLC-MS/MS Analysis of Organic Compounds
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The analysis was performed
on a Waters ACQUITY UPLC-Class system (Waters, USA). Separation was
carried out on an ACQUITY CORTECS UPLC C18 column (1.6 μm, 2.1
× 100 mm, Milford, MA, USA) at 40 °C. The flow rate was
0.3 mL/min. The mobile phase consisted of 0.1% (v/v) formic acid in
water (A) and acetonitrile (B) with a simple gradient program: 0–15
min, 5–52% B; 15–18 min, 52–95% B; 18–20
min, 95% B; 20–23 min, 95–5% B; 23–25 min, 5%
B. The injection volume was 2 μL. MS data were recorded using
the Waters SYNAPT G2-SI MS systems (Waters, USA) equipped with an
electrospray ion source and Q-TOF/MS with the MSE model.
The source parameters were set as follows: capillary voltage, 2.5
kV; cone voltage, 45 V; source temperature, 120 °C; desolvation
temperature, 300 °C; desolvation gas flow, 800 L/h. The low collision
energy was 6 eV, and the high collision energy was 30–70 eV
for the negative ion mode. Mass spectra were recorded across the range m/z 50–1200 under negative ion modes, and 3D data
were collected in the continuum mode. MS data was acquired by Waters
MassLynx 4.1 and processed using UNIFI 1.7 software (Waters).
on a Waters ACQUITY UPLC-Class system (Waters, USA). Separation was
carried out on an ACQUITY CORTECS UPLC C18 column (1.6 μm, 2.1
× 100 mm, Milford, MA, USA) at 40 °C. The flow rate was
0.3 mL/min. The mobile phase consisted of 0.1% (v/v) formic acid in
water (A) and acetonitrile (B) with a simple gradient program: 0–15
min, 5–52% B; 15–18 min, 52–95% B; 18–20
min, 95% B; 20–23 min, 95–5% B; 23–25 min, 5%
B. The injection volume was 2 μL. MS data were recorded using
the Waters SYNAPT G2-SI MS systems (Waters, USA) equipped with an
electrospray ion source and Q-TOF/MS with the MSE model.
The source parameters were set as follows: capillary voltage, 2.5
kV; cone voltage, 45 V; source temperature, 120 °C; desolvation
temperature, 300 °C; desolvation gas flow, 800 L/h. The low collision
energy was 6 eV, and the high collision energy was 30–70 eV
for the negative ion mode. Mass spectra were recorded across the range m/z 50–1200 under negative ion modes, and 3D data
were collected in the continuum mode. MS data was acquired by Waters
MassLynx 4.1 and processed using UNIFI 1.7 software (Waters).
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