Lc 20ad liquid chromatography

Manufactured by Shimadzu

Sourced in Japan

The Shimadzu LC-20AD is a high-performance liquid chromatography (HPLC) system designed for analytical and preparative separations. It features a dual-plunger parallel-flow pump that provides precise and stable solvent delivery. The LC-20AD is capable of operating at a maximum pressure of 40 MPa and a flow rate range of 0.001 to 10.000 mL/min, making it suitable for a variety of liquid chromatography applications.

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6 protocols using lc 20ad liquid chromatography

Quantification of Squalene in Oils

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SQ content was determined after alkaline saponification of the oil by RP-HPLC at 208 nm as described by Grigoriadou et al. [24 (link)]. Briefly, 0.1 g of oil was added in a 25 mL glass stopped tube followed by the addition of 3 mL KOH (600 g/L). Then 2 mL ethanol and 5 mL of an ethanolic pyrogallol solution were added (60 g/L). After alkaline saponification at 75 °C for 30 min, 15 mL of the NaCl solution (10 g/L) was added and the mixture was extracted twice with 15 mL of n-hexane/ethyl acetate (9:1, v/v). After evaporation of the organic phase, the dry matter was diluted in acetone.
Separation was achieved on a C18 column (2504 × 4.6 mm i.d.; 5 mm) (Macherey-Nägel, Düren Germany) maintained at 26 °C on an LC 20AD liquid chromatography (Shimadzu Corporation, Kyoto, Japan) equipped with an SPD-10AV UVVIS detector (Shimadzu Corporation, Kyoto, Japan) using acetonitrile (1.2 mL/min). The injection volume was 10 mL. SQ external calibration curves (10–250 mg/L) were used for quantification. Samples were analyzed in duplicates (CV% = 2.0, n = 5).

Mastralexi A, & Tsimidou M.Z. (2021). On the Squalene Content of CV Chondrolia Chalkidikis and Chalkidiki (Greece) Virgin Olive Oil. Molecules, 26(19), 6007.

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Quantitative Analysis of Chlorfenapyr and Tralopyril

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The content of chlorfenapyr was detect via gas chromatography (GC) using a 7890 N gas chromatograph equipped with an electron capture detector (Agilent Technologies, CA, USA). The content of tralopyril was analyzed via an LC-20AD liquid chromatography (LC) system (Shimadzu Corporation, Kyoto, Japan) with a Sciex 4000 Q TRAP triple quadrupole MS/MS system (Applied Biosystems, Foster City, USA). Detailed analytical parameters are listed in the Supporting Information.

Zhang H., Chen S., Wu S., You Y, & Zhang K. (2024). The fate and potential hazards of chlorfenapyr and one metabolite tralopyril in cabbages: A comprehensive investigation. Food Chemistry: X, 22, 101287.

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Rosemary Extraction and HPLC Analysis

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A total of 50 mg of ground rosemary samples (leaves) was accurately weighed, put into a 50 mL falcon tube, and extracted, as described in the extraction procedure. An aliquot of the extract after centrifugation was filtered through a 0.2 μm syringe filter before the injection of 10 µL in LC-20AD liquid chromatography (Shimadzu, Japan) for the HPLC analysis. An Inertsil ODS 2 column (250 × 4.6 mm, 5 μm particles, GL Sciences Inc, Tokyo, Japan) was used. Mobile phases A and B were water with 0.1% formic acid and acetonitrile, respectively. The column temperature was kept at 30 °C, and the flow rate of the mobile phase was set at 0.5 mL·min⁻¹. The following multistep gradient with different proportions of mobile phase B was applied: 0 min, 20% B; 10 min, 40% B; 15 min, 90% B maintained for 5 min. The initial conditions were maintained for 5 min. The analysis was monitored using an SPD-M20A detector at 210 nm. The quantification was done by comparing the peak areas of the targeted carnosic acid with the abundance of the compound in the corresponding standard used in the calibration curve. All chemical analyses were done in triplicate.

Appiah K.S., Omari R.A., Onwona-Agyeman S., Amoatey C.A., Ofosu-Anim J., Smaoui A., Arfa A.B., Suzuki Y., Oikawa Y., Okazaki S., Katsura K., Isoda H., Kawada K, & Fujii Y. (2022). Seasonal Changes in the Plant Growth-Inhibitory Effects of Rosemary Leaves on Lettuce Seedlings. Plants, 11(5), 673.

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Synthesis and Characterization of Polymer Nanoparticles

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All reagents were purchased from Sigma-Aldrich and used as received unless stated otherwise. CH₂Cl₂ was distilled over calcium hydride. Styrene was washed with 1 N NaOH, followed by water to remove inhibitors, dried over MgSO₄ and then purified by vacuum distillation over calcium hydride. AIBN was recrystallized from MeOH twice prior to use. Lactose-oxime 8 was synthesized according to previously reported procedures.^{[25 (link)]} The NMR spectra were recorded on a Varian Mercury 300 MHz or Varian Inova 500 MHz spectrometers using CDCl₃ as a solvent unless stated otherwise. ¹H NMR-based M_n of dodecyl trithiocarbonate-terminated polymers was calculated by comparing the integral areas under CH_ar peaks of repeating units with integrals of CH₂–S signal of ω-chain end. The weight of CTA (566 g mol⁻¹) was then added to the sum of weights of repeating units. GPC analyses were performed on Shimadzu LC-20AD liquid chromatography instrument, equipped with RI detector. Two Waters Styragel columns (HR3 and HR4) were placed in series. THF was used as eluent at 1 mL min⁻¹ flow rate; the column oven was set to 40 °C. Molecular weights were calculated against polystyrene standards. TEM images were obtained on Philips/FEI Technai 20 instrument using copper TEM grids. A 2% (wt/vol) uranyl acetate solution was used to stain the polymeric nanoparticles before recording.

Ledin P.A., Kolishetti N., Hudlikar M.S, & Boons G.J. (2014). Exploring Strain-Promoted 1,3-Dipolar Cycloadditions of End Functionalized Polymers. Chemistry (Weinheim an der Bergstrasse, Germany), 20(28), 8753-8760.

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Size Exclusion Chromatography of DEC-205 ECD

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50 μl of DEC-205 ECD at 1 mg ml⁻¹ was loaded onto a Superdex 200 5/150 column (GE Healthcare) in either TBS, HBS, or BIS-TRIS buffer at a flow rate of 0.3 ml min⁻¹. The system comprises of a DGU-20A5 degasser, LC-20AD liquid chromatography, SIL-20ACHT autosampler, CBM-20A communications bus module, SPD-20A UV-visual detector, and CTO-20AC column oven (Shimadzu) coupled with a DAWN HELIOS-II light scattering detector and Optilab T-rEX refractive index detector (Wyatt). Detector number 12 was substituted for a WyattQELS detector installed at a 90° angle. The system was controlled using LC solutions (Shimadzu), and data collection and analysis were performed in ASTRA6 (Wyatt Technology Corp).

Gully B.S., Venugopal H., Fulcher A.J., Fu Z., Li J., Deuss F.A., Llerena C., Heath W.R., Lahoud M.H., Caminschi I., Rossjohn J, & Berry R. (2020). The cryo-EM structure of the endocytic receptor DEC-205. The Journal of Biological Chemistry, 296, 100127.

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HPLC Quantification of Resveratrol and BD

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Resveratol and BD were quantified using a validated HPLC method. A Shimadzu Prominence UFLC system equipped with a DGU-20 A5R Prominent degasser unit, LC-20 AD Liquid chromatography, SIL-20A HT Autosampler and SPD-20A UV-Vis detector was used (all Shimadzu Corporation, Japan). For both, RES and BD, the volume of sample injection was 100 μl and the detection wavelength for BD and RES were 243 nm and 306 nm, respectively. Quantification of BD was conducted using methanol:water 80:20 (%v/v) as the mobile phase at flow rate of 1 ml/min in isocratic mode with a Luna C18 column (3 µm, 4.6 × 150 mm) (Phenomenex, Sydney, Australia). The linearity was confirmed between 0.2 and 50 μg/ml (R² = 0.999). For quantification of RES, the mobile phase consisted of methanol:water 60:40 (%v/v) with 0.5% acetic acid (%v/v). Samples were analysed using a XbridgeTM column (5 μm, 4.6 × 150 mm), (Waters, Massachusetts, USA) at a flow rate of 0.7 ml/min. The content of resveratrol was quantified from the peak area correlated with the predetermined standard curve between 0.2 and 50 μg/ml (R 2 =0.999).

Trotta V., Lee W.H., Loo C.Y., Young P.M., Traini D, & Scalia S. (2016). Co-spray dried resveratrol and budesonide inhalation formulation for reducing inflammation and oxidative stress in rat alveolar macrophages. European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences, 86.

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