Dragendorff s reagent

Standard chromogenic reagents lead acetate, potassium dichromate, ferric chloride, hydrochloric acid, sulfuric acid, Mayer's reagent, Dragendorff's reagent, Wagner's reagent, Hager's reagent, Molisch reagent, Benedict's reagent, and Fehling's solutions used for preliminary phytochemical chemical group test were of reagent grade and purchased from Sigma-Aldrich Co. LLC, Missouri, United States. Vincristine sulfate, used as a standard drug in the cytotoxic assay, was collected from the Techno Drugs Limited, Bangladesh. Methanol supplied by Laboratory Patterson Scientific, UK, was used as solvent for maceration of the plant material. Dimethyl sulfoxide (DMSO, ≥99.9%, BioReagent, for molecular biology; Sigma-Aldrich, India) was used to dissolve the extracts.

Vincristine sulphate and Dragendorff’s reagent were procured from Sigma Aldrich (USA). Dimethyl sulfoxide (DMSO), Dulbecco’s modified eagle medium (DMEM), Sodium dodecyl sulfate (SDS), Tris–HCl, Ethylenediaminetetraacetic acid (EDTA), DAPI, Ethidium bromide, Fetal bovine serum, Propidium iodide, RNase A, Proteinase K, 2,4-DNP were obtained from Sigma Chemicals Company (San Diego, USA). DCFH-DA and Annexin-V stains were obtained from Life Technologies (USA). JC-1 (5, 5′, 6, 6′-tetrachloro-1, 1′, 3, 3′-tetraethylbenzimidazolyl carbocyanine iodide) dye was purchased from Molecular probes (Eugene, OR, USA). All other reagents and chemicals were of analytical grade.

Methanol (MeOH) and ethanol (EtOH) were supplied by QReC (QRec Asia, Rawang, Malaysia). Folin-Ciocalteu reagent, sodium hydroxide (NaOH), sulphuric acid (H₂SO₄), hydrochloric acid (HCl), sodium nitrite (NaNO₂), glacial acetic acid, chloroform, sodium carbonate (Na₂CO₃), aluminium chloride (AlCl₃), gallic acid, quercetin, butylated hydroxyanisole (BHA), 2,2-diphenyl-1-picrylhydrazyl (DPPH), Dragendorff’s reagent, Mayer’s reagent, and ferric chloride hexahydrate (FeCl₃·6H₂O) were supplied by Sigma-Aldrich (Sigma-Aldrich Co, Shanghai, China). All reagents were of analytical grade.

The cell wall peptidoglycan was determined as described by Komagata & Suzuki (1987) . Cellular fatty acids were extracted, methylated, and analyzed using the standard microbial identification system protocol (Sherlock Microbial Identification System, version 6.1), whereas fatty acids were identified using the TSBA6 database of the microbial identification system (Sasser, 1990 ). Polar lipids were analyzed from freeze-dried cells by two-dimensional thin-layer chromatography (TLC), as described by Minnikin et al. (1984) (link). Appropriate detection reagents were used for visualizing TLC bands: phosphomolybdic acid reagent 5% (w/v) solution in ethanol (Sigma-Aldrich, Saint Louis, MO, USA) was used to detect total polar lipids; ninhydrin reagent (0.2% solution; Sigma-Aldrich Saint Louis, MO, USA) was used to detect amino lipids; the Dittmer and Lester reagent (molybdenum blue, 1.3%; Sigma-Aldrich Saint Louis, MO, USA) was used to detect phospholipids; and, Dragendorff’s reagent (Sigma-Aldrich Saint Louis, MO, USA) was used to detect phosphatidylcholine.

Chemicals and reagents used in the study included but not limited to chloroform, ethanol, acetic anhydride, methanol, dichloromethane (Merck BDH, Poole, UK), ferric chloride, sulphuric acid, sodium hydroxide, trifluoroacetic acid, formic acid, ammonia, ammoniacal alcohol solution, and Dragendorff's reagent, (Sigma-Aldrich, London, UK). All other chemicals and reagents used in this study met analytical grade.