The nine compounds (1 –9 ) were determined using HPLC-MS to be present in CareVidTM as described previously [1 (link)]. Extracts were prepared initially from 20 g each of CareVidTM Tea powder. The extracts were methylene chloride, ethanol, methanol, water, acidified water (0.1 M HCl), and steeping in hot water (>90 °C). Metabolic profiling of the extracts was performed by UHPLC-UV-HRMS/MS on a Vanquish UHPLC system (Thermo Scientific, Waltham, MA, USA) coupled to a 100 Hz photodiode array detector (PDA) and an Orbitrap Fusion Tribrid (Thermo Scientific) high-resolution tandem mass spectrometer (Thermo Scientific, Waltham, MA, USA). A detailed protocol that was followed is described in Rotich et al., 2021 (Figure 1 ). For the assay, pure standards of pellitorine (1 ), oleuropein (2 ) and ellagic acid (9 ), urolithin A (10 ), and urolithin B (11 ) were bought from Sigma Aldrich, whereas pure standards of magnoflorine (3 ) earlier isolated from the seed cake of Croton megalocarpus, crotocorylifuran (6 ) earlier isolated from Croton megalocarpoides, crotepoxide (4 ) earlier isolated from Croton alienus, ent-kaurane-16β,17-diol (5 ) earlier isolated from Croton haumanianus, and lupeol (7 ) and betulin (8 ) isolated in substantial quantities from CareVidTM were obtained in the Jodrell Laboratory, Kew, UK.
Magnoflorine
Manufactured by Merck Group
Sourced in United States
Magnoflorine is a laboratory equipment product manufactured by Merck Group. It is designed to perform magnetic separation and concentration of biomolecules and particles. The core function of Magnoflorine is to enable efficient isolation, purification, and enrichment of target analytes from complex samples through the application of magnetic fields.
Automatically generated - may contain errors
Lab products found in correlation
Berberine, by Merck Group (4 mentions)
Gallic acid, by Merck Group (4 mentions)
Palmatine, by Merck Group (3 mentions)
Ellagic acid, by Merck Group (2 mentions)
Jatrorrhizine, by Merck Group (2 mentions)
Rutin, by Merck Group (2 mentions)
Berberine chloride, by Merck Group (2 mentions)
Formic acid, by Merck Group (2 mentions)
Acetonitrile, by Merck Group (2 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Folin ciocalteu reagent, by Merck Group (2 mentions)
Lupeol, by Merck Group (1 mentions)
Oleuropein, by Merck Group (1 mentions)
Pellitorine, by Merck Group (1 mentions)
Vanquish uhplc system, by Thermo Fisher Scientific (1 mentions)
Orbitrap fusion tribrid, by Thermo Fisher Scientific (1 mentions)
Urolithin a, by Merck Group (1 mentions)
Urolithin b, by Merck Group (1 mentions)
Acetonitrile, by Thermo Fisher Scientific (1 mentions)
Lc ms grade methanol, by Thermo Fisher Scientific (1 mentions)
13 protocols using magnoflorine
1
Profiling Compounds in CareVid™ Tea Powder
2
Quantitative Analysis of Herbal Compounds in XLGB Samples
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Thirty-four batches of XLGB samples (batch number: 091104, 100142, 100317, 100324, 100506, 100509, 100523, 100832, 100840, 100846, 100901, 100929, 101038, 101123, 101151, 101157, 101165, 101171, 101214, 101220, 110111, 110115, 110125, 110220, 110319, 110353, 110354, 110358, 110361, 110362, 110955, 111031, 1112009, and 1203061) that were manufactured by Guizhou Tongjitang Pharmaceutical Co., Ltd. (Guiyang, China) were collected from drugstores in Guangzhou of China. Epimedin A, epimedin B, asperosaponin VI, isobavachin, neobavaisoflavone, icariside II, bavachinin, isobavachalcone, sweroside, and corylifol A were purchased from Shanghai Winherb Medical Technology Co., Ltd. (Shanghai, China). Psoralen, timosaponin BII, isopsoralen, epimedin C, icariin, psoralidin, and tanshinone II A were obtained from Guangzhou PI&PI Inc. (Guangzhou, China). Magnoflorine was obtained from Sigma-Aldrich (St. Louis, USA). Their purities were all determined to be over 98% by HPLC.
LC-MS grade methanol, acetonitrile and water were all purchased from Fisher Scientific (Pittsburgh, PA, USA). LC-MS grade formic acid was purchased from Sigma-Aldrich (St. Louis, MO, USA). Other reagents were of analytical grade.
LC-MS grade methanol, acetonitrile and water were all purchased from Fisher Scientific (Pittsburgh, PA, USA). LC-MS grade formic acid was purchased from Sigma-Aldrich (St. Louis, MO, USA). Other reagents were of analytical grade.
3
Simultaneous Quantification of Phytochemicals in Dengue Medicine
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The HPLC and LCMS grade solvents acetonitrile and methanol were obtained from J. T. Baker (USA) and Honeywell (Germany). Analytical grade solvents, glacial acetic acid, and formic acid, toluene and ethyl acetate were purchased from Rankem, India. Deionized water, purified by a Milli-Q system (Millipore, USA), was used throughout the study. Reference standards with defined purity in parenthesis, for gallic acid (97.3%), magnoflorine (99.0%), berberine (88.4%), ellagic acid (99.6%), rosmarinic acid (98.0%) were procured from Sigma Aldrich (USA). 5-HMF (97.3%), methyl gallate (99.5%) from Tokyo Chemical Industries (Japan) and protocatechuic acid (99.5%), rutin (98.7%) from Natural Remedies (India) and β-ecdysone (99.9%) were purchased from PHY-proof (Germany), respectively. Divya-Denguenil-Vati (DNV) batches were sourced from Divya Pharmacy, Haridwar (India), and were stored in airtight bottles. DNV (batch # 015) was used for UPLC/MS-QToF identification, as well as UHPLC and HPTLC validation studies. Twenty-six commercial batches of DNV, # 014, # 015, # 029 to # 034, and # 036 to # 053 were analyzed to evaluate quality consistency across different DNV batches.
4
Analytical Standards Preparation and HPLC
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Analytical standards (magnoflorine, AA-I, and AA-II) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Milli-Q water (Millipore, Bedford, MA, USA) was used for preparing standard solutions, extraction buffers, and eluents. Formic and acetic acids were of high-performance liquid chromatography (HPLC)-grade and obtained from Merck (Darmstadt, Germany). HPLC-grade acetonitrile (ACN) was obtained from PanReac AppliChem (Darmstadt, Germany). All the other chemicals were of analytical-grade.
5
Neuroprotective Effects of Magnoflorine and Berberine in MCAO Model
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The rats were randomly divided into five groups (with 18 rats for each group): the sham group, MCAO group, MCAO + Magnoflorine 10 mg/kg group,MCAO + Magnoflorine 20 mg/kg group, and MCAO + berberine 50 mg/kg group. The rats in the MCAO models were subjected to brain ischemia for 90 min and reperfusion for 24 h. Magnoflorine and berberine (intragastric; Sigma-Aldrich, MO, USA) were dissolved with dimethyl sulfoxide (DMSO) as a stock solution and diluted with saline (DMSO final concentration <0.5%). The sham and MCAO groups received equal volumes of saline with the same amount of DMSO. The described treatment groups were administered agents once a day for 14 days before MCAO and 1 day after surgery. The dose of Magnoflorine was chosen according to a previous study with minor modifications [20 ]. Behavioral tests were performed on day 1 following MCAO (Figure 1 ). All the rats were anesthetized with phenobarbital sodium (40 mg/kg, i.p.), and blood was collected from the aorta abdominalis. The rats were then euthanized by decapitating, and brain tissues were quickly obtained for subsequent experiments.
6
Analytical Standards for Phytochemical Analysis
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Standard compounds, berberine, jatrorrhizine, palmatine, magnoflorine, choline, ecdysteroids, lidocaine, D-camphor and 5,7-isoflavone were purchased from the Sigma-Aldrich, St. Louis, MO, USA. Acetonitrile, formic acid and water of LC-MS grade were purchased from the Sigma-Aldrich.
7
Quantitative Analysis of Giloy Ghanvati Bioactives
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Giloy Ghanvati is available as tablets and for this analysis, the tablets were finely powdered and 0.5 gm powder was diluted with 10 ml methanol: water (70:30) and sonicated for 30 min. The extract was centrifuged at 8000 rpm for 5 min at 4°C and filtered using 0.45 µm nylon filter. This was labeled as a working solution and was used for the analysis of Cordifolioside A, Palmatine and Beta-Ecdysone. The working solution was further diluted to 10 times with same solvent as above and used for the analysis of Magnoflorine.
Standards used for comparative analysis were dissolved in methanol to get the desired concentrations. Palmatine hydrochloride hyrate (Potency-97.0%) and Magnoflorine (Potency-99.0%) were procured from Sigma-Aldrich (St. Louis, MO, United States), Cordifolioside A (Potency-98.6%, ChemFaces, Wuhan, Hubei Province, China), and Beta-Ecdysone (Potency-99.0%, Tokyo Chemical Industries, Tokyo, Japan) were used as standards.
Standards used for comparative analysis were dissolved in methanol to get the desired concentrations. Palmatine hydrochloride hyrate (Potency-97.0%) and Magnoflorine (Potency-99.0%) were procured from Sigma-Aldrich (St. Louis, MO, United States), Cordifolioside A (Potency-98.6%, ChemFaces, Wuhan, Hubei Province, China), and Beta-Ecdysone (Potency-99.0%, Tokyo Chemical Industries, Tokyo, Japan) were used as standards.
8
Comprehensive Phytochemical Characterization of Madhugrit
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Madhugrit (batch #1MDT-210081) was sourced from Divya Pharmacy, India. The standards for HPLC analysis namely gallic acid, magnoflorine, piperine, rutin, ellagic acid, coumarin, cinnamic acid, and palmatine were obtained from Sigma-Aldrich, USA; protocatechuic acid, corilagin from Natural remedies, India, and methyl gallate from TCI chemicals, India. Reagents namely RPMI 1640 (no glucose), DMEM (low glucose), DPBS, antibiotic-antimycotic solution, starch, Malondialdehyde (MDA), Trichloroacetic acid (TCA), LPS, and 2′,7′-Dichlorofluorescin diacetate (H2DCFDA) were procured from Sigma-Aldrich, USA. Heat-inactivated FBS, TPVG, Bovine serum albumin (BSA), Nile red, and Alamar blue were obtained from HiMedia, India. Horse serum was bought from Gibco, USA. Chemicals, 2-thiobarbituric acid (TBA), D-glucose, Metformin, and dexamethasone were obtained from TCI chemicals, India. Phorbol 12-myristate 13-acetate (PMA) was purchased from Alfa Aesar, UK. Mitomycin C was obtained from Roche, Germany. Pierce BCA protein assay kit and sodium azide were obtained from Thermo Fisher Scientific, USA. Human TNF-α and IL-6 ELISA kits were obtained from BD Biosciences, USA. QUANTI-Blue reagent was purchased from InvivoGen, USA.
9
Therapeutic Evaluation of Berberine and Magnoflorine
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The compounds tested were:
Berberine (2.5, 5, 10, 20, 50 mg/kg) from methanolic extract of Berberis siberica herb, and Magnoflorine (5, 10, 50 mg/kg)—obtained from methanolic extract of Berberis cretica root.
Scopolamine hydrochloride (1 mg/kg) was produced by Sigma-Aldrich (St. Louis, MO, USA).
Magnoflorine, berberine and scopolamine used for the study were dissolved in 0.9% NaCl with an addition of 0.2% of DMSO (vehicle). The tested compound was administered i.p. at a constant volume of 10 mL/kg. Each day prior to the experimentation, the drug solutions were prepared. In parallel, a control group of mice was given i.p. injections of saline with an addition of 0.2% DMSO (later referred to as vehicle), at the very same volume as tested drugs.
Berberine (2.5, 5, 10, 20, 50 mg/kg) from methanolic extract of Berberis siberica herb, and Magnoflorine (5, 10, 50 mg/kg)—obtained from methanolic extract of Berberis cretica root.
Scopolamine hydrochloride (1 mg/kg) was produced by Sigma-Aldrich (St. Louis, MO, USA).
Magnoflorine, berberine and scopolamine used for the study were dissolved in 0.9% NaCl with an addition of 0.2% of DMSO (vehicle). The tested compound was administered i.p. at a constant volume of 10 mL/kg. Each day prior to the experimentation, the drug solutions were prepared. In parallel, a control group of mice was given i.p. injections of saline with an addition of 0.2% DMSO (later referred to as vehicle), at the very same volume as tested drugs.
10
Antioxidant Determination Protocol
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Reagents used to perform the antioxidant determinations—2,2-diphenyl-1-picrylhydrazyl (DPPH) and Folin-Ciocalteu reagent, the standards of gallic acid, chlorogenic acid, berberine, palmatine, jatrorrhizine, magnoflorine, quercetin, D-saccharic acid (glucaric acid), glucaric acid potassium salt and rutoside at purity exceeding 95% were purchased from Sigma-Aldrich (St. Louis, MO, USA). Spectroscopic grade solvents used for the LC-MS analyses—acetonitrile, water and formic acid, were manufactured by Merck (Darmstadt, Germany). The reagent grade chemicals used for the preparation of extracts and spectrophotometric determinations were purchased from Avantor Performance Materials (Gliwice, Poland).
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