Magnesium chloride

All the chemicals used in this work were of analytical reagent grade. Millipore water having a resistivity of 18.2 MΩ cm obtained from Merck Millipore water unit was used for the preparation of all the aqueous solutions employed for electrochemical studies. Sodium chloride (Merck), lactic acid (Merck) and urea (Fischer Scientific) were used for the preparation of sweat bio-mimic compositions. Similarly, methyl-p-hydroxybenzoate (Avra), sodiumcarboxymethyl cellulose (Sigma Aldrich), potassium chloride (Alfa Aesar), magnesium chloride (Merck), calcium chloride (Fischer Scientific), dipotassium hydrogen phosphate (Merck) and potassium dihydrogen phosphate (Hi-Media) were utilized for the preparation of saliva bio-mimic solutions. Electrochemical studies were carried out using potassium ferricyanide, K₃[Fe(CN)₆] (Fischer Scientific) and potassium ferrocyanide, K₄[Fe(CN)₆] (Hi-Media) as the redox probe. Finally Triton X-100 (Sisco Research Laboratories) was used to prepare the hexagonal lyotropic liquid crystalline medium. All these chemicals were used as received without any further purification.

The activity of the intracellularly generated ALP was analyzed after 1, 3, 7, 14, 21, and 28 days of stimulation, each 20 h after the last stimulation interval started. The cells were washed twice using TRIS buffer (50 mM, pH = 8.0) lysed with 1% Triton X and 1% phenylmethylsulfonyl fluoride (both: Merck, Darmstadt, Germany) for 10 min. A solution containing 10 mM 4-Nitrophenylphosphat (AppliChem, Darmstadt, Germany), 100 mM 2-amino-2-methyl-1,3-propanediol (Sigma-Aldrich, Munich, Germany), and 5 mM magnesium chloride (Merck, Darmstadt, Germany) was added to the lysate and incubated over 1 h at 37°C and 5% CO₂. The reaction was stopped with a 2 M sodium hydroxide solution, and the absorption was measured at 405 nm with multimode plate reader Infinite 200 pro (Tecan Group Ltd., Maennedorf, Switzerland). A blank served as an internal control and was subtracted from each value.

Acetylcholine chloride (ACh), verapamil, isoprenaline hydrochloride, carbachol (CCh) and phenylephrine hydrochloride (PE) were purchased from Sigma Chemicals Co. MO, USA. Various kinds of chemical were used for the preparation of physiological solutions like: magnesium sulfate,sodium dihydrogen phosphate, calcium chloride, glucose, sodium bicarbonate ,magnesium chloride and potassium dihydrogen phosphate were purchased from Merck, Germany, while potassium chloride (KCl), ethylenediamine tetraacetic acid from Sigma Chemical Co, USA. Similarly Sodium chloride was purchased from BDH Laboratory, England. Analytical grade chemical were used for performing the experiments.

Acetophenone, aliphatic ketones, 2-propanol, triethanolamine (TEA), citric acid, dipotassium phosphate, and methyl tert-butyl ether (MTBE) were obtained from Sigma-Aldrich, Seelze, Germany. Magnesium chloride, potassium dihydrogen phosphate, mandelonitrile, glucose, and concentrated hydrochloric acid were purchased from Merck, Darmstadt, Germany. Sodium cyanide was received from Fluka Chemika AG, Buchs, Switzerland. In addition, mandelonitrile was stored at -18°C to minimize decomposition to benzaldehyde and hydrogen cyanide. Sodium hydroxide was purchased from VWR International, Darmstadt, Germany and calcium carbonate was obtained from Fisher Scientific, Loughborough, United Kingdom. NOA81 was obtained from Thorlabs, Dachau, Germany. All chemicals were obtained in highest available purity and used as received. The hydroxynitrile lyase from Manihot esculenta was a gift from Jülich Fine Chemicals (now Codexis). Alcohol dehydrogenase from Lactobacillus kefir (LkADH) and glucose dehydrogenase (GDH) were purchased from evocatal, Monheim, Germany. Deionized water was produced with an Ultra Clear Reinstwassersystem by SG Water (now Evoqua, Guenzburg, Germany) and used throughout this study.

All chemicals of research grade were used for experimental work. Magnesium chloride, potassium dihydrogen phosphate, sodium bicarbonate, potassium chloride, magnesium sulphate, glucose, sodium dihydrogen phosphate (Merck, Darmstadt, Germany), and sodium chloride (BDH Laboratory Supplies, Poole, England) were used for physiological solution preparation. Carbachol (CCh), acetylcholine chloride (Ach) and loperamide (Sigma Chemical Co., St. Louis, MO) were utilized as standards and castor oil (Marhaba Laboratories, Lahore, Pakistan) for the induction of diarrhoea. Whereas chloroform, hydrochloric acid, benzene, sulphuric acid, iodine, picric acid (BDH Laboratory Supplies, Poole, England), ammonium hydroxide, ferric chloride (Sigma Chemical Co., St. Louis, MO), ethanol, ammonia solution (Saba Saleem Industries and Fine Chemicals, Karachi, Pakistan), sodium hydroxide (Sigma Aldrich Laborchemikalien GmbH, Steinheim am Albuch, Germany), benzene, lead acetate, and potassium iodide (Merck, Darmstadt, Germany) were used in phytochemical analysis of crude extract.
Distilled water was used for the preparation of standard solutions, dilution, and physiological salt solutions (Tyrode’s and Kreb’s solutions). Physiological salt solutions and dilutions of standard drugs were freshly made on the day of the experiment by the method of Naz et al. (2016 ).

For the experiments conducted in this study, various antibiotics, media, and reagents were utilized. The antibiotic disks were obtained from MAST (Mast Co., UK), while the powders of oxacillin and penicillin were acquired from Sigma-Aldrich (St. Louis, MO, USA). Media and agar supplies were obtained from Merck (Merck Co., USA). Specifically, mannitol salt agar, blood agar, Mueller-Hinton agar, Mueller-Hinton broth (MHB), sodium chloride (NaCl), and magnesium chloride (MgCl2) were sourced from Merck (Merck Co., USA). Chemical reagents including crystal violet, glucose, and agarose were purchased from Sigma-Aldrich (St. Louis, MO, USA). In addition, 96-well sterile polystyrene, non-treated microplates, available in both flat-bottom and round-bottom variants, were obtained from NEST (Biotechnology Co., Ltd., China), and Jet Biofil (Guangzhou, China).

The active ingredient of Tamsulosin hydrochloride (TMS) was purchased from Tolid Daru Pharmaceutical Co (Tehran, Iran). The citric acid (CA), Ethylenediamine (EDA), Rhodamine B (RhB), l-Cysteine (l-Cys), l-leucine (l-Lue), Creatinin, Sucrose, Glucose, Iron (ɪɪɪ) chloride, Aluminum chloride, Calcium chloride, copper (II) sulfate, Magnesium chloride, Sodium chloride, Potassium chloride, Zinc sulfate, Acetic acid, Phosphoric acid, Boric acid, and Methanol (Me-OH) with the analytical grade were purchased from Merck Co. (Darmstadt, Germany). Deionized (DI) water with a resistivity of more than 18 MΩ has been used in the whole of the experiments.