Caffeine standard (HPLC standard with a purity of 98% from Sigma-Aldrich GmbH, Steinheim, Germany), HPLC methanol (Chromasolv Gradient, purity ≥99.8%, Sigma-Aldrich GmbH, Steinheim, Germany), formic acid (ACS, purity ≥96%, Sigma-Aldrich GmbH, Steinheim, Germany), and deionized water ddH2O (18.2 MΩ cm−1, 25 °C) were used as standards and for mobile phase preparation.
Hplc standard
Manufactured by Merck Group
Sourced in United States, Germany
HPLC standards are reference materials used to calibrate and validate high-performance liquid chromatography (HPLC) instruments. They provide known concentrations of specific compounds to ensure accurate and reliable HPLC analysis.
Automatically generated - may contain errors
Lab products found in correlation
Folin ciocalteu reagent, by Merck Group (4 mentions)
Gallic acid, by Merck Group (3 mentions)
Hydrochloric acid (hcl), by Merck Group (3 mentions)
Formic acid, by Thermo Fisher Scientific (3 mentions)
Trichloroacetic acid, by Merck Group (2 mentions)
2 2 azinobis 3 ethylbenzothiazoline 6 sulfonic acid (abts), by Merck Group (2 mentions)
Arium 611 uv unit, by Sartorius (2 mentions)
Arium 126 61316 ro, by Sartorius (2 mentions)
Methanol, by Avantor (2 mentions)
Formic acid, by Avantor (2 mentions)
Luciferin luciferase for atp assay, by Merck Group (2 mentions)
Potassium persulfate, by Merck Group (2 mentions)
Acetonitrile, by Thermo Fisher Scientific (2 mentions)
Methanol, by Thermo Fisher Scientific (2 mentions)
Anhydrous sodium sulfate, by Merck Group (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Sonorex digital 10 p dk 156 bp ultrasonic bath, by Bandelin (1 mentions)
Fluostar omega, by BMG Labtech (1 mentions)
Apomorphine, by Merck Group (1 mentions)
6 hydroxydopamine, by Merck Group (1 mentions)
23 protocols using hplc standard
1
Caffeine HPLC Quantification Protocol
2
Encapsulation of Freeze-Dried Elderberry Pulps
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Maltodextrin dextrose equivalent 10 (MD) was from Productos de Maı´z SA, Buenos Aires, Argentina; Promitor TM was from Tate & Lyle, London; Capsul TM from Ingredion Argentina SRL, Buenos Aires, Argentina; and k-carrageenan from Saporiti SACIFIA, Buenos Aires, Argentina, were used for encapsulation of freezedried elderberry pulps. Ethanol and chlorhydric acid used as solvents for juice extraction were from Biopack, Buenos Aires, Argentina. Folin-Ciocalteu reagent was purchased from Merck KgaA Darmstadt, Germany. HPLC standard was obtained from Sigma-Aldrich, USA. Gallic acid used for phenolic standard curve was obtained from Anedra, Buenos Aires, Argentina. All chromatographic solvents were of HPLC grade and the purity of the reagents used was p.a. or similar.
3
Authenticated J. montana Plant Identification
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J. montana was acquired in Puszcza Knyszyńska (Podlasie Province, Poland; 53°15′20.9” N; 23°25’41.5” E). The species has been authenticated based on the scientific literature and verified by Michał Tomczyk using referent literature [19 ]. A voucher specimen (No. JM-15029) has been deposited in the plant collection of the Department of Pharmacognosy, Medical University of Białystok (Poland). The enzymes and the HPLC standards (≥98.5%) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Other reagents and chemicals were of analytical grade. Extraction was performed using Bandelin SONOREX® Digital 10 P DK 156 BP ultrasonic bath (Berlin, Germany). A microplate reader (FLUOstar Omega, BMG Labtech, Ortenberg, Germany) was used for UV-VIS spectroscopic measurements. Agilent 1220 series HPLC instrument equipped with an autosampler, DAD detector, and a Zorbax Eclipse XDB-C18 (5 μm, 25 mm × 4.6 mm) column (Agilent Technologies, Santa Clara, CA, USA) was used for determination of the selected compounds.
4
Comprehensive Chemical Reagents Protocol
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All chemicals were obtained from HiMedia Laboratories Pvt. Ltd. (Mumbai, India), unless otherwise indicated. Spectrophotometric kits to measure serum chemistries were procured from Crest Biosystem; Goa, India. HPLC standards were purchased from Sigma-Aldrich (St. Louis, MO, USA) and ChromaDex (Irvine, CA, USA). MilliQ water (Ω 18.2) from departmental central facility was used for all the experiments.
5
Immunohistochemical Assays Protocol
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6-hydroxydopamine, apomorphine, and HPLC standards were from Sigma-Aldrich (St. Louis, MO, USA); ketamine and xylazine were from König (Santana de Parnaíba, São Paulo, Brazil). Antibodies for immunohistochemistry assays were from Santa Cruz Biotechnology (Dallas, TX, USA) or Merck-Millipore (Darmstadt, Germany). All other reagents were of analytical grade.
6
Neurotoxin-induced Parkinson's Model
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6-hydroxydopamine (6-OHDA), apomorphine, and HPLC standards were from Sigma-Aldrich (St. Louis, MO, USA); ketamine and xylazine were from Konig do Brasil (Santana de Parnaíba, São Paulo, Brazil). The BDNF kit for ELISA was from Abcam (Cambridge, UK), and antibodies for immunohistochemistry assays were from Santa Cruz Biotechnology (Dallas, TX, USA) or Merck Millipore (Darmstadt, Germany). All other reagents were of analytical grade.
7
Characterization of Polyphenols and Antioxidants
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Analytical, HPLC and LCMS-grade chemicals were used in this study. Gallic acid, Folin–Ciocalteu reagent, vanillin, L-ascorbic acid, iron(III) chloride hexahydrate (Fe[III]Cl3•6H2O), sodium phosphate dibasic hepta-hydrate (Na2HPO4•7H2O), hexahydrate aluminium chloride sodium phosphate, sodium phosphate monobasic monohydrate (Na2HPO4•H2O), trichloroacetic acid (TCA), hydrochloric acid (HCl), ferrozine, hydrated sodium acetate, ethylene diamine tetra acetic acid (EDTA), potassium ferrocyanide (III), iron (II) and iron (III) chloride, 3-hydrobenzoic acid (C7H6O3), ammonium molybdate, iron (II) sulphate heptahydrate, catechin, quercetin, DPPH, 2,4,6-tripyridyl-s-triazine (TPTZ) and ABTS were purchased from the Sigma Aldrich (Castle Hill, Australia) for the estimation of polyphenols and antioxidant potential. Hydrogen peroxide (30%) and Sodium carbonate anhydrous were purchased from Chem-Supply Pty Ltd. (Adelaide, Australia) and 98% sulfuric acid was purchased from RCI Labscan (Rongmuang, Thailand). HPLC and LC-MS grade reagents include ethanol, methanol, acetonitrile, formic acid, glacial acetic acid and iron (III) chloride anhydrous were purchased from Thermo Fisher Scientific Inc (Scoresby, Australia). HPLC standards were also purchased from Sigma Aldrich.
8
Polysaccharide Extraction and HPLC Analysis
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About 10 mg of dry cell wall mass was used for polysaccharide extraction, in accordance with François [32 (link)]. After extraction, 1 mL from the final preparation was concentrated 10× by lyophilization. The sugars were analyzed using HPLC (Young Lin YL9100 series system) by a refractive index (RI) detector at 40 °C. The samples were loaded in the REZEX ROA (Phenomenex, Torrance, CA, USA) column (300 × 7.8 mm) at 85 °C and eluted with 0.05 M sulfuric acid at a flow rate of 1.5 mL/min [33 (link)]. The saccharides were quantified based on a standard curve (5–100 mM) using HPLC standards from Sigma-Aldrich (St. Louis, MO, USA): Tetraacetylchitotetraose (Chit4), triacetylchitotriose (Chit3), diacetyl-β-D-chitobiose (Chit2), N-Acetil-β-D-glucosamina (nag), and glucose (gluc).
9
Sensitizing MCF-7 Cells to Doxorubicin
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The MCF-7 breast cancer line was used in this study (ATCC, USA). The cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Corning, Corning, USA) supplemented with 10% fetal bovine serum (Corning, Corning, USA), incubated at 37 °C with 5% CO2 in air atmosphere. The tested cells were treated for 48 h with 1 μM DOX (EBEWE Pharma, Unterach, Austria) and 50 μM of following HPLC standards (Sigma-Aldrich, USA): apigenin, hesperidin or combined (1 μM DOX + 50 μM single HPLC standard). The tested concentration of DOX was based on observed cytotoxicity for MCF-7 cell (IC50) and was consistent with plasma concentrations in patients treated with this drug. Doxorubicin is a standard drug used in breast cancer therapy [71 (link)]. Apigenin and hesperidin were used in optimal concentrations that sensitized the MCF-7 cells on DOX treatment in preliminary study.
10
Comprehensive Antioxidant Evaluation Protocol
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Ultra-pure water (<5 µg/L TOC) was obtained from the water purification systems Arium 126 61316-RO, plus an Arium 611 UV unit (Sartorius, Goettingen, Germany). Formic acid (MS grade) and methanol (HPLC grade) were purchased from J.T. Baker (Phillipsburg, NJ, USA). Folin–Ciocalteu (FC) reagent, 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric chloride hexahydrate, 2,4,6-tris(2-pyridyl)-s-triazine, quercetin, gallic acid, Amberlite® resin (XAD4), phosphate buffer, trichloroacetic acid, ferric chloride, hydrochloric acid, ascorbic acid, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,4,6-tris(2-pyridyl)-s-triazine (TPTZ), gallic acid, potassium hexacyanoferrate(III), 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), and dimethylsulfoxide (DMSO) were obtained from Sigma-Aldrich; sodium carbonate, ferrous sulfate, sodium persulfate, sodium acetate, sodium sulfate anhydrous, and ethanol were obtained from Merck (Lima, Peru). HPLC standards (with purity higher than 95% by HPLC) were purchased from Sigma Aldrich Chem. Co. (St. Louis, MO, USA) or Extrasynthèse (Genay, France).
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