Dried Epimedii Folium were grind and the fine powder was collected by using a 60 mesh sieve. Take 2 g of the fine powder, and then reflux it with 100 mL of ethanol for 60 min. The extract solution was filtered into a round bottom flask for vacuum evaporation and concentration. The residual was dissolved with mobile phase and transferred into a 5 mL volumetric flask and diluted to the exact volume as sample solution. The HPLC equipment used is Shimadzu HPLC system including quaternary pump, a diode array detector (DAD) and a column oven (Tokyo, Japan). Analysis was achieved on a YMC-Pack C18 column (250 × 4.6 mm i.d., 5 μm particle size).The chromatographic conditions was recorded as following: injection volume was 10 μL; column temperature was maintained at 35°C; the detection wavelength was set at 254 nm; the mobile phase was composed of acetonitrile (A) and 0.1% formic acid (B) with gradient elution system (0–25 min, 5–15% A; 25–60 min, 15–35% A; 60-95 min, 35–55% A; 95–115 min, 55–70% A; 115–120 min, 70–5% A) at a flow rate of 1.0 mL/min.
Column oven
Manufactured by Shimadzu
Sourced in Japan, United States
The Column Oven is a laboratory instrument designed to precisely control the temperature of chromatographic columns used in various analytical techniques. It maintains the column at a consistent temperature, enabling accurate and reproducible separation of analytes in the sample.
Automatically generated - may contain errors
Lab products found in correlation
Hplc system, by Shimadzu (1 mentions)
C18 column, by YMC (1 mentions)
Online degasser, by Shimadzu (1 mentions)
Diamonsil c18 reverse phase column, by Dikma Technologies (1 mentions)
Lc 2010a ht liquid chromatograph system, by Shimadzu (1 mentions)
Intelligent autosampler, by Jasco (1 mentions)
High performance liquid chromatography (hplc), by Aminex (1 mentions)
High performance liquid chromatography (hplc), by Jasco (1 mentions)
Zorbax eclipse xdb c18 column, by Agilent Technologies (1 mentions)
Automatic injector, by Shimadzu (1 mentions)
Vacuum degasser, by Shimadzu (1 mentions)
Prominence lc 20a, by Shimadzu (1 mentions)
Sil 20a ht autosampler, by Shimadzu (1 mentions)
Amicon ultra 4, by Merck Group (1 mentions)
Prominence diode array detector, by Shimadzu (1 mentions)
Avance 3 hd, by Bruker (1 mentions)
Absciex 6500 qtrap mass spectrometer, by AB Sciex (1 mentions)
Exionlc ad, by Shimadzu (1 mentions)
Acquity uplc hss t3 column, by Waters Corporation (1 mentions)
Analyst 1, by AB Sciex (1 mentions)
9 protocols using column oven
1
HPLC Analysis of Epimedii Folium
2
Quantification of Psoralen in Samples
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The LC-2010A HT Liquid Chromatograph system (Shimadzu Corporation, Kyoto, Japan) was used to determine the concentration of psoralen in samples. The HPLC system consisted of a double-plunger pump (Shimadzu Corporation), an autosampler (Shimadzu Corporation), an online degasser (Shimadzu Corporation), a Diamonsil C18 reverse phase column (5 μm, 4.6 mm inner diameter × 25 cm; Dikma Technologies, Inc, Lake Forest, CA, USA), a column oven (Shimadzu Corporation), an ultraviolet detector (Hamamatsu Photonics, Hamamatsu, Japan), and a recording integrator (Shimadzu Corporation). The mobile phase was methanol:water (55:45, volume/volume [v/v]) with a flow rate of 1 mL/min. The column temperature was constant at 35°C, and the detection wavelength was 246 nm. Percentage recoveries ranged from 97.4% to 102.8%. The psoralen intra- and interday relative standard deviation values were 0.87% and 2.15%, respectively. Samples from in vitro experiments were filtered through a disposable nylon syringe filter (pore size, 0.45 μm) with a 13-mm diameter (Shanghai Anpel Scientific Instrument Co, Ltd, Shanghai, People’s Republic of China) before automatic injection into the HPLC system. Samples from in vivo microdialysis were directly assayed in a timely manner without any handling.
3
Analytical Quantification of Metabolites
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Ethane-1,2-diol, glycerol, 1,2-propanediol, 1,2-butanediol, 1,2-hexanediol, acetaldehyde, propionaldehyde, butyraldehyde, pentanaldehyde, hexanaldehyde, ethanol, 1,3-propanediol, 1-propanol, 1-butanol, 1-pentanol, 1-hexanol, acetic acid, 3-hydroxypropionic acid, propionic acid, butyric acid, pentanoic acid and hexanoic acid were measured via HPLC (Jasco, Tokyo, Japan). For HPLC, an Aminex HPX-87H chromatographic column was used with an upstream pre-column (Biorad, Richmond, CA, USA) and a refractive index detector (Perkin Elmer, Series 200a, Waltham, Massachusetts, USA) and an intelligent autosampler (Jasco, Tokyo, Japan). Temperature of the column was maintained at 65°C by a column oven (Shimadzu, Tokyo, Japan). Samples from the bioreactor were diluted with MilliQ water and mixed with 20% v/v sulfuric acid and injected into 0.5 mM sulfuric acid mobile phase.
For quantification of 3-HPA, the modified photometric method according to Circle et al. [34 ], with acrolein as standard was used. Briefly, 200 μL of the sample (diluted to fit in the range of the assay) was mixed with 150 μL of DL-tryprophan, followed by addition of 600 μL of hydrochloric acid (37%), and incubating the mixture for 20 min at 37°C. The resulting purple colour was measured spectrophotometrically at 560 nm and the absorbance was compared with the standard curve.
For quantification of 3-HPA, the modified photometric method according to Circle et al. [34 ], with acrolein as standard was used. Briefly, 200 μL of the sample (diluted to fit in the range of the assay) was mixed with 150 μL of DL-tryprophan, followed by addition of 600 μL of hydrochloric acid (37%), and incubating the mixture for 20 min at 37°C. The resulting purple colour was measured spectrophotometrically at 560 nm and the absorbance was compared with the standard curve.
4
Determination of Amylose Content via HPSEC
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The analysis method for determination of amylose contents of starch samples by high performance size exclusion chromatography (HPSEC, separations module, Waters Corporation, Milford, MA, USA) was modified according to [24 (link)]. A total of 3 Ultrahydrogel HPSEC columns were connected in series. The columns including two Ultrahydrogel 120 and an Ultrahydrogel linear were maintained at 40 °C by a column oven (Shimadzu, Kyoto, Japan) and a mobile phase of deionized water was controlled at 0.8 mL min−1. Starch samples were gelatinized in boiling water at 0.4% (w v 1) for 30 min and gelatinized completely by an ultrasonic processor (Model VC 501, Sonic & Material Inc.,Newtown, CT, USA). After that, the solutions were filtered through a Millipore filter (8.0 µm) before injecting into the HPSEC system equipped with an auto-injector and refractive index detector.
5
HPLC Analysis of POH and PA Compounds
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The chromatography setup comprised of Shimadzu Prominence HPLC system modules, a degasser, an auto-sampler, a UV detector (wavelengths set to 210 nm and 217 nm for POH and PA detection respectively), and a column oven (Shimadzu Scientific Instruments Inc., Columbia, MD, USA). The separation was performed using a Zorbax Eclipse XDB-C18 column (4.6 × 150 mm, 5 µm; Agilent Technologies, Santa Clara, CA, USA) and a Zorbax C18 guard (8 × 4 mm, 5 µm; Agilent Technologies, Santa Clara, CA, USA). Mobile phase composition for both analytes was isocratic, comprising of water–acetonitrile mixture (60:40, v/v, pre-mixed, 1.0 mL/min) for POH analysis and 0.05 M ammonium acetate (pH 5)–acetonitrile mixture (64:36, v/v, pre-mixed, 2.0 mL/min) for PA analysis. Both phases were passed through 0.45 µm filters (Millipore-Sigma, St. Louis, MO, USA) before use.
6
Shimadzu Prominence HPLC Procedure
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The DAD chromatography procedure utilized a Shimadzu LC 20 A Prominence® (Kyoto, Japan) liquid chromatograph equipped with a photodiode array (PDA) Shimadzu® (Kyoto, Japan) detector, Shimadzu® column oven, Shimadzu® automatic injector, Shimadzu Lab Solutions integration system® (Kyoto, Japan), and a Shimadzu vacuum degasser® (Kyoto, Japan).
7
Efficient Encapsulation Evaluation of LF-CA Nanoparticles
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The EE (%) of the LF-CA NPs was determined according to a reported method [32 (link)] by the HPLC system comprising a binary LC-20C D pump, a SIL-20AHT autosampler, and a column oven (Shimadzu, Tokyo, Japan). To efficiently separated free CA, we used the Amicon Ultra-4 centrifugal filter devices (Millipore Co., Billerica, MA, USA) made up of a centrifuge tube and a filter unit with a low-binding Ultracel membrane (MWCO 3000) [33 (link)]. The 1 mL samples were dissolved in 2 mL methanol containing 2 mL 0.5% phosphoric acid and then sonicated for 15 min. Finally, needle syringes and 0.45 μM microporous membranes were used to filter the samples. The mobile phase (water containing 0.2% phosphoric acid: acetonitrile at a ratio of 75:25 v/v) was delivered with a flow rate of 1.0 mL/min at 35 °C, and separation was performed using an Agilent Eclipse column XDB-C18 (5 mm, 4.6 × 150 mm) with a sample injection volume of 20 μL. The wavelength for detection of CA was 327 nm. Results (EE (%)) were calculated using the following equation:
8
HPLC and NMR Analysis of Compounds
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HPLC purification of described compounds were carried out on a Shimadzu Prominence Nexera XR HPLC coupled to LC-6AD pumps, SPD-M20A Prominence Diode Array Detector, CTO-20A Prominence Column Oven, CBM-20A Communication Bus Module, and Class VP software. 1D and 2D NMR spectra were acquired on Bruker (R) – DRX500-Ultra Shield (R) (1H: 500.13 MHz, 13C: 125.77 MHz) and Bruker Avance III HD 600-MHz spectrometers.
9
LC-MS/MS Analysis of Chemical Compounds
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LC and MS conditions LC-MS/MS analysis was conducted on ExionLC AD consisting of binary pumps, an on-line degassing unit, an autosampler, and a column oven (Shimadzu Corporation, Kyoto, Japan), which is coupled with an AB Sciex 6500 + QTRAP mass spectrometer consisting of an electrospray ionization (ESI) source (AB SCIEX, Framingham, MA, USA). Chromatographic separation was achieved on Waters Acquity UPLC HSS T3 Column, 100Å, 1.8 μm, 2.1 mm X 100 mm maintained at 40 °C, at a flow rate of 0.3 mL/min. The mobile phases consisted of Solution A (0.1% Formic acid in water) and Solution B (100% acetonitrile). The following gradient was used: 0–1 min, 98% A; 1–5 min, 98%-45% A; 5–8 min, 45%-0% A; 8–13 min, 100% A; 13-13.1 min, 100%-2% A; 13.1–18 min, 98% A, with a total run time of 18 min. The ion source was operated in mix mode: curtain gas, 35 psi; nebulizer gas 50 psi; auxiliary gas 50 psi; ion spray voltage, 5500 V/-4500 V (positive/negative); and temperature 500 °C. Multiple reaction monitoring (MRM) transitions were identified for all analyses and isotope-labeled standard. Data acquisition and analysis were all performed with Analyst 1.6.3 software (AB SCIEX) and OS software (AB SCIEX).
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