Five grams of powdered PuE were homogenized in 100 mL methanol (HPLC grade 99.8%) and centrifuged (SIGMA 3-30K, GmbH, Berlin, Germany) at 10,000 rpm for 10 min. The supernatant was filtered through a 0.2 µm Millipore membrane filter, and approximately 1–3 mL of the filtrate were collected and kept for HPLC application. The phenolic content was analyzed by HPLC (LC-10AS, Shimazu, Japan) equipped with an autosampler, solvent degasser, and quaternary HP pump column (C18, Gemini, 4.60 mm, 5 μm, 35 °C). The mobile phase flow rate of 1 mL/min was determined with a triple-quadruple spectrometer (LCMS 8040, Shimazu, Japan) connected to an electrospray ionization (ESI) source. The gradient solvent system was water (A) and acetonitrile (B) 5–60% in formic acid. Isocratic elution was 95% A and 5% B for the first 5 min, followed by a linear gradient to 50% A and 50% B between 5 and 55 min. It was also 50% A and 50% B between 55 and 65 min, and the liner gradient returned to 95% A and 5% B between 65 and 67 min. The samples were automatically injected using the autosampler SIL-40Cxs (Shimadzu, Kyoto, Japan). The data were managed using LC solution software (Shimadzu), and MS functioned in negative mode. In total, 35% of the collision energy was utilized in MS/MS fragmentation. The ions were discovered using a full-scan method with a mass range of 100 to 1500 m/z [67 (link)].
Lc 10a
Manufactured by Shimadzu
Sourced in Japan, China, United States
The LC-10A is a high-performance liquid chromatography (HPLC) system designed for analytical and preparative applications. It features a high-precision pump, a programmable autosampler, and a UV-Vis detector. The core function of the LC-10A is to provide accurate and reliable separation and analysis of a wide range of chemical compounds.
Automatically generated - may contain errors
Lab products found in correlation
Spd 10a, by Shimadzu (5 mentions)
Ri 10a, by Shimadzu (3 mentions)
Hplc system, by Shimadzu (2 mentions)
Avance 2 400 spectrometer, by Bruker (2 mentions)
Lc 10at, by Shimadzu (2 mentions)
P 2000 polarimeter, by Jasco (2 mentions)
Ods hypersil c18, by Thermo Fisher Scientific (2 mentions)
Spd m20a, by Shimadzu (2 mentions)
Spd 10av, by Shimadzu (2 mentions)
Refractive index detector, by Shimadzu (2 mentions)
10 av lc, by Shimadzu (2 mentions)
Spd 10a vp, by Shimadzu (2 mentions)
Spd 10a, by Phenomenex (2 mentions)
Class vp series software, by Shimadzu (2 mentions)
Ics 5000, by Thermo Fisher Scientific (2 mentions)
Diamonsil c18 column, by Dikma Technologies (2 mentions)
Sil 40c xs, by Shimadzu (1 mentions)
Lc solution software, by Shimadzu (1 mentions)
Spd 10avp ultraviolet detector, by Shimadzu (1 mentions)
Lc 10avp, by Shimadzu (1 mentions)
123 protocols using lc 10a
1
Quantitative HPLC-MS Analysis of Phytochemicals
2
Anthocyanin Analysis by HPLC
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The HPLC (LC-10 AS, Shimazu, Japan) is fully equipped with an auto-sampler, a quaternary pump, a C18 separation column (Gemini, 4.60 mm, 5 μm, 35 °C), a mobile phase flow rate of 1 mL/min, and a multiwavelength detector set at 520 nm to detect anthocyanin compounds following Saad et al. [44 (link)].
3
Controlled Drug Release Evaluation
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The study of drug release was carried out in phosphate buffer solution (PBS, pH 7.4) incubated with collagenase IV (0.3 mg/mL) at 37 °C with mild stirring. The XG-peptide-MMC/DOX (DOX/MMC-peptide), XG-CONH-MMC/DOX (DOX/MMC-CONH), DOX-CONH/MMC-peptide and DOX-peptide/MMC-CONH conjugates were individually immobilized into 10 mL dialyzing bag with molecular weight cutoff (MWCO) 3000 Da and subjected to dialysis against PBS (pH 7.4) at 37 °C The samples (dialysate) were collected in time dependent manner and immediately analyzed by a Shimadzu HPLC system composed of two pumps (LC-10Avp and LC-10AS) and an SPD-10Avp ultraviolet detector (Shimadzu Corporation, Japan) in reverse phase mode at different points of time. Using an Extend-C18 column (4.6 × 250 mm I.D., 5 μm), and the mobile phase used for the analysis was methanol–acetonitrile-phosphate buffer (pH 5.0, 0.2 M) (50:20: 30, v/v/v) and the flow rate was 0.5 mL/min. According to the predetermined standards for each drug, the amount of DOX in the solution was quantified at 245 nm wavelength, and the amount of MMC was determined at 360 nm wavelength.
4
Characterization of Aloe vera Powder and Its Interaction with Food Components
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De Man, Rogosa, and Sharpe (MRS) media was purchased from BD Biosciences (Franklin Lakes, NJ, USA). A. vera powder was obtained from Morinaga Milk Industry Co. (Tokyo, Japan). ⍺-amylase, pepsin, and bile salt were purchased from Fujifilm Wako Pure Chemical Corporation (Osaka, Japan), and pancreatin was obtained from Sigma (St. Louis, MO, USA). A commercial soft drink and milk (pasteurised) were purchased from a local market. All chemical reagents used in this study were of analytical grade.
Data were collected using a muffle furnace (MFP-300A; IKEDA Scientific Co., Tokyo, Japan), an organic elemental analyser (UNICUBE; Elementar Analysensysteme GmbH Langenselbold, Germany), a scanning electron microscope (JSM-6330F; Tokyo, Japan ), a FT-IR spectrometer (FT-IR 300; JASCO, Tokyo, Japan), a zeta potential instrument (Melles Griot, Carlsbad, CA, USA), an Isoton II diluent instrument with particle size data analyser (Muiltisizer 4, Beckman Coulter, Brea, CA, USA), a high-performance liquid chromatograph (LC-10AS, Shimadzu, Kyoto, Japan), a gas chromatograph (GC-4000, TC-1; GL Sciences, Tokyo, Japan), and a colour measuring instrument (Konica Minolta, Tokyo, Japan).
Samples were prepared using a lyophiliser (Eyela, Tokyo, Japan) and spray drier (pilot-scale SD 1000; Eyela) for further analysis. Double distilled water was used in all experiments.
Data were collected using a muffle furnace (MFP-300A; IKEDA Scientific Co., Tokyo, Japan), an organic elemental analyser (UNICUBE; Elementar Analysensysteme GmbH Langenselbold, Germany), a scanning electron microscope (JSM-6330F; Tokyo, Japan ), a FT-IR spectrometer (FT-IR 300; JASCO, Tokyo, Japan), a zeta potential instrument (Melles Griot, Carlsbad, CA, USA), an Isoton II diluent instrument with particle size data analyser (Muiltisizer 4, Beckman Coulter, Brea, CA, USA), a high-performance liquid chromatograph (LC-10AS, Shimadzu, Kyoto, Japan), a gas chromatograph (GC-4000, TC-1; GL Sciences, Tokyo, Japan), and a colour measuring instrument (Konica Minolta, Tokyo, Japan).
Samples were prepared using a lyophiliser (Eyela, Tokyo, Japan) and spray drier (pilot-scale SD 1000; Eyela) for further analysis. Double distilled water was used in all experiments.
5
HPLC Quantification of Compound 3 in L. floribunda
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For the quantification of compound 3 in L. floribunda, HPLC-DAD was performed on a Shimadzu LC-10 AS (Shimadzu Corp., Tokyo, Japan), equipment with a Phenomenex Prodigy 5 μm ODS 250 × 4.6 mm reversed-phase column, followed by elution with 40% methanol in water acidified with perchloric acid (pH 2.4) as a mobile phase and UV detection at 210 nm and 320 nm. Compound 3 at 1 mg/mL and the extract at 20 mg/mL were prepared prior to use dissolved in ethanol.
6
Tacrolimus Quantification by LC-MS/MS
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Analytical-grade tacrolimus was provided by Astellas Pharma Inc. (Tokyo, Japan). Sirolimus was purchased from LC Laboratories (Woburn, MA, USA) and used as an internal standard. The liquid chromatography-tandem mass spectrometry/mass spectrometry (LC-MS/MS) system was comprised of a high-performance liquid chromatography (HPLC) machine (LC-10AS; Shimadzu Corp., Kyoto, Japan), a column (Inertsil-ODS3, 150 mm × 2.1 mm i.d.; GL Sciences, Inc., Tokyo, Japan), and an MS/MS detector (API4000; Applied Biosystems, Foster City, CA, USA).
7
HPLC Analysis of Phenolic Compounds in Fruit
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In order to identify phenolic compounds in fruit extracts, a high-performance liquid chromatography (HPLC) method was used (Shimadzu LC–10AS chromatograph equipped with a C18 RP column and SPD-10AV UV–Vis detector). Signal detection was set at the wavelengths of 325 and 265 nm. Chromatographic separation was carried out at 33 ± 1 °C. using the following solvents: (A) water (Sigma–Aldrich) with acetic acid (0.1%), (B) methanol (Sigma–Aldrich, ultra pure) with acetic acid (0.1%) and applying the gradient: 90% A, 10% B for 20 min; 75% A, 25% B for 30 min; 65% A, 35% B for 40 min; 55% A, 54% B for 50 min; 50% A, 50% B for 60 min; 30% A, 70% B for 62 min; 100% B to 80 min; 80% A, 10% B up to 85 min. The flow rate was 1 mL/min. The identification of phenolics was based on the retention times of chlorogenic, caffeic, p-hydroxy-benzoic acid, p-coumaric, ferulic, protocatechuic, syringic acids (Sigma–Aldrich), salicylic acid (LGC Standards) as well as (+)-catechin (LGC Standards) andkaempferol (Sigma–Aldrich).
8
MTT Cytotoxicity Assay and Spectroscopic Analysis
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3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was purchased from Sigma-Aldrich CO (St Louis, MO, USA). Doxorubicin hydrochloride 99.8% (DOX, Synbias Pharma Ltd.) was purchased from Nanox Release Technology (Buenos Aires, Argentina).
Gentamicin sulfate (potency: 550–590 μg/mg) and erythromycin (potency: 863 μg/mg) were provided by Laboratorio Fabra S.A, Buenos Aires, Argentina, and Unifarma, Buenos Aires, Argentina, respectively.
1H- and 13C-NMR and two-dimensional spectra were recorded with a Bruker AVANCE II 400 spectrometer (Bruker Corporation, Ettlingen, Germany) with tetramethylsilane (TMS) as the internal reference. HPLC was performed on a Shimadzu LC-10 AS (Shimadzu Corp., Tokyo, Japan), equipped with a Phenomenex Prodigy 5 μ ODS (4.6 mm i.d. × 250 mm) reversed-phase column. The mobile phase was water/methanol/trifluoroacetic acid 65 : 35 : 1 with detection at 365 nm.
Flow cytometry analysis was performed in a Becton-Dickinson (BD) FACS Canto II flow cytometer (BD Biosciences, USA).
Gentamicin sulfate (potency: 550–590 μg/mg) and erythromycin (potency: 863 μg/mg) were provided by Laboratorio Fabra S.A, Buenos Aires, Argentina, and Unifarma, Buenos Aires, Argentina, respectively.
1H- and 13C-NMR and two-dimensional spectra were recorded with a Bruker AVANCE II 400 spectrometer (Bruker Corporation, Ettlingen, Germany) with tetramethylsilane (TMS) as the internal reference. HPLC was performed on a Shimadzu LC-10 AS (Shimadzu Corp., Tokyo, Japan), equipped with a Phenomenex Prodigy 5 μ ODS (4.6 mm i.d. × 250 mm) reversed-phase column. The mobile phase was water/methanol/trifluoroacetic acid 65 : 35 : 1 with detection at 365 nm.
Flow cytometry analysis was performed in a Becton-Dickinson (BD) FACS Canto II flow cytometer (BD Biosciences, USA).
9
Polyphenol Analysis of Lemongrass Extract
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HPLC analyses for polyphenol compounds of lemongrass extract was performed on LC-10AS (Shimadzu, Kyoto, Japan) system equipped with Shimadzu LC-10AT pumps, and CBM-20A communication bus module. Chromatography was carried out in a gradient system using 250 × 4.60 mm, 5μm C18 column (Gemini, Phenomenex, California, USA). A flow rate of 1 mL/min and the injection volume of 10μL was employed. The mobile phases consisted of A (99.9% acetonitrile and 0.1% acetic acid) and B (0.1% acetic acid in MQ water). The gradient elution was 0–15min (8% A and 92% B), 30min (22% A and 78% B), 45min (78% A and 22% B), 55min (8% A and 92%B), and 60min (8% A and 92% B). A UV-Visible DAD detector was used, and the wavelengths detected at 280 and 320nm. Pure standards were injected at different concentrations and the calibration curves obtained. Polyphenols were identified by comparing their retention times with those of pure standards and the polyphenol concentration of each compound calculated with respect to the calibration curves of the standards (Sonmezdag et al., 2017 (link)).
10
Amino Acid Profiling via HPLC
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The amino acid profile was determined using high-performance liquid chromatography. The processed leaf and grain samples were applied to an LC-10AS Shimadzu liquid chromatography system, which operated under the following conditions: oven temperature, 60 °C; excitation wavelength, 350 nm; emission wavelength, 450 nm; sample cooler temperature, 4 °C; eluent solution flow, 0.60 m2 min−1; injection volume, 5 μL; run time, 45 min [47 ].
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