Acetonitrile, methanol water and dichloromethane (Merck; Darmstadt, Germany) were used for liquid chromatography diode array detection (LC-DAD) analysis and liquid chromatography tandem mass spectrometry (LC-MS/MS). Ethanol absolute and chloroform were obtained from VWR Chemicals (Radnor, PA). Hexane, butylated hydroxytoluene (BHT), formic acid (99% for mass spectrometry) along with analytical standards (chicoric acid, chlorogenic acid, lutein, β-carotene, violaxanthin, neoxanthin, β-cryptoxanthin, and cyanidin) were purchased from Sigma-Aldrich (St. Louis, MO). Ultrapure water was obtained from a Milli-Q Gradient A10 water purification system.
Methanol water
Manufactured by Merck Group
Sourced in Germany
Methanol water is a laboratory reagent used as a solvent and diluent in various analytical and experimental procedures. It is a clear, volatile liquid composed of a mixture of methanol and water. Methanol water is commonly used in applications that require a polar, protic solvent with specific properties, such as controlled polarity and miscibility with other compounds.
Automatically generated - may contain errors
Lab products found in correlation
Cyanidin, by Merck Group (1 mentions)
Chloroform, by Avantor (1 mentions)
Absolute ethanol, by Avantor (1 mentions)
Neoxanthin, by Merck Group (1 mentions)
Chicoric acid, by Merck Group (1 mentions)
Violaxanthin, by Merck Group (1 mentions)
Butylated hydroxytoluene bht, by Merck Group (1 mentions)
β cryptoxanthin, by Merck Group (1 mentions)
Formic acid, by Merck Group (1 mentions)
Hexane, by Merck Group (1 mentions)
Chlorogenic acid, by Merck Group (1 mentions)
β carotene, by Merck Group (1 mentions)
Lutein, by Merck Group (1 mentions)
Acetonitrile, by Merck Group (1 mentions)
Dichloromethane, by Merck Group (1 mentions)
Methanol, by Merck Group (1 mentions)
4 protocols using methanol water
1
Multimodal Analysis of Bioactive Compounds
2
HPLC Analysis of Active Compounds in Samples
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The collected samples were analyzed via reverse-phase HPLC, using the Agilent 1620 Infinity II LC System (Waldbronn, Germany) equipped with a quaternary pump (G7111B), autoinjector (G7167A), multicolumn thermostat (G7116A), and WR diode-array detector (G7115A). The software was OpenLab (version 2.2.0). A LiChrocart250-4/LiChrosorb RP-18 (5 µm) column (Merck, Darmstadt, Germany) was used to quantify the active substances, at 25 °C with an injection volume of 40 µL. The elution conditions were Methanol/water (Merck, Darmstadt, Germany) at a gradient from 60:40 to 90:10 in 15 min, constant at 90:10 for 15 min and the last 60:40 in 10 min with a flow of 1 mL/min. Detection for caffeine (CAF) was 280 nm, for ibuprofen (IBU) was 221 nm and for dexamethasone (DEX) and ivermectin (IVE) was 240 nm. Methanol (Merck, Darmstadt, Germany) was used as an extraction solvent for all active substances. All analytical procedures were validated following the guidelines developed by the International Conference on Harmonization (ICH) [36 ]. The calibration curve, limit of quantification (LoQ), and limit of detection (LoD) were obtained and are detailed in Table 5 .
3
Mouse Brain Metabolite Extraction
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Five milligrams of fresh mouse brain tissue were mixed with 500 μL methanol-water (Sigma-Aldrich Trading Co., Ltd, dilution of 80:20), and 10 μL of 100 μg/mL CA-d4 (CDN, Inc., CAN) internal standard was added. After that, the sample was swirled for 1 min and centrifuged at 13,000 rpm for 5 min at 4 °C. Then, 400 μL of supernatant was collected into a new centrifuge tube and the procedure described above was repeated. The supernatant was then blow-dried with nitrogen and then poured into the 100 μL ultrapure water, followed by centrifugation at 13,000 rpm for 5 min at 4 °C. The supernatant was then collected for subsequent analysis.
4
Lacrimal Gland Organoid Culture and Stimulation
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Lacrimal gland organoids cultured for 21 days and 45 days (stimulated with forskolin for 5 min) were washed with .9% NaCl solution (Sigma-Aldrich) and fixed with methanol:water (9:1, v/v) (Sigma-Aldrich). Samples were frozen in liquid nitrogen and scraped into 2 ml Eppendorf tubes. Tubes were centrifuged at 15,000 rpm for 15 min and supernatants were transferred into 2 ml Eppendorf tubes and stored at −80°C until analysis. Culture media samples were collected on days 1, 7, 14, 21, 28, 35, and 45 and stored at −80°C.
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