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7 protocols using sodium acetate

1

Total Anthocyanin Content Determination

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TAC was evaluated by the pH differential method using two buffer systems: 25 mM potassium chloride (Sigma-Aldrich GmbH, Steinheim, Germany) solution (pH 1.0) and 0.4 M sodium acetate (Loba Chemie Pvt. Ltd., Mumbai, India) buffer (pH 4.5). PPEs were mixed with the corresponding buffers, and absorbances were determined simultaneously at 510 and 700 nm, respectively, after 15 min of incubation at 23°C [42 ]. A=A510A700 pH 1.0A510A700 pH 4.5.
TAC, expressed as mg of cyanidin-3-glucoside equivalents per 100 g of PPE (mg cy-3-glu/100 g), had been calculated as follows: TAC=A×MW×1000ε, where A: absorbance; MW: molecular weight (449.2 g/mol); and ε: molar absorptivity of cyanidin-3-glucoside (26,900 M−1·cm−1).
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2

Quantitative Analysis of Tetracycline Antibiotics

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Pure standard chlortetracycline hydrochloride (95%), tetracycline and oxytetracycline (98%) were obtained from Medchemexpress, USA. Methanol was obtained from Fisher Chemical, UK. Deionized water was obtained from a milli Q water purification system and was filtered through a 0.22 µm Millipore membrane filter (Milford, MA, USA). Acetonitrile was purchased from Sigma-Aldrich (France). Sodium acetate, sodium hydrogen phosphate, potassium dihydrogen orthophosphate, trifluoroacetic acid and ethylene diamintetraaceticacid were purchased from LOBA CHEMIE, Mumbai, India. Oxalic acid dihydrate was obtained from WINLAB, UK. Citric acid and tri-sodium citrate2-hydrate were purchased from Avonchem, UK.
Acetic acid was obtained from S.D.FineChem Ltd., India.
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3

Bacterial growth and polylactic acid production

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De Man, Rogosa, and Lactobacilli (MRS) agar and broth, and chitosan were purchased from HiMedia, Mumbai, India. Glucose, yeast extract, sodium acetate, dipotassium hydrogen phosphate (K2HPO4), potassium dihydrogen phosphate (KH2PO4), magnesium sulfate (MgSO4), manganese sulfate (MnSO4), ferrous sulfate (FeSO4), calcium carbonate (CaCO3), and antimony trioxide (Sb2O3) were procured from Loba Chemie. N-butanol was purchased from MERCK Life Science Pvt. Ltd., Bangalore, India. LA was purchased from Rankem laboratory reagent, Gurgaon, Haryana, India. Distilled water was used at each step. Commercial PLA was purchased from Balson Industries, Pune, India. All remaining chemicals and solvents used for analysis were of analytical grade depending on their utilizations.
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4

Phytochemical Analysis and Antioxidant Evaluation

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All the chemicals were of good grade and were purchased
from HiMedia
Leading BioScience Company. The chemicals and reagents, which are
used during the project work are as follows: sodium hydroxide, sodium
chloride, sodium acetate, methanol, petroleum ether, Folin–Ciocalteu
reagent, sodium hydroxide, sodium carbonate, sodium nitrite, and aluminum
chloride, which were obtained from Loba Chemie (Mumbai, India). Gallic
acid, quercetin, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) were obtained
from Sigma-Aldrich (St. Louis, Missouri, USA).
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5

Antioxidant and Enzyme Assays Protocol

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2,2-diphenyl-1-picryl-hydrazyl (DPPH), 2,4,6-Tripridyl-s-triazine (TPTZ), were purchased from Sigma-Aldrich, USA. supplied by Mumbai, India. Ascorbic acid, sodium acetate buffer (pH-3.6), glacial acidic acid, sodium acetate, were procured from Loba Chemie Pvt. Ltd., Mumbai, India. Glucose-6-phoshtate were dehydrogenase (G6PDH), β-nicotinamide adenine dinucleotide phosphate (β-NADP) were purchased from Sisco Research Laboratory Pvt., Mumbai, India. Serum glutamic oxaloacetic transaminase (SGOT) and serum glutamic pyruvic transaminase (SGPT) were obtained from ARKRAY Healthcare Pvt. Ltd., India. STZ was bought from HiMedia, Mumbai, India. All other chemicals used were of analytical grade.
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6

Analytical Characterization of BGR-34 Herbal Formulation

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HPTLC system (CAMAG, Muttenz, Switzerland), TLC Silica gel 60F254 (Merck KGaA, 64271 Darmstadt, Germany), and a Water’s
ACQUITY UPLC system (Waters Corp., MA, USA)were used for mass spectrometry
analysis. C18 column (ACQUITY UPLC BEH C18 1.7 μm, 2.1 ×
100 mm), Mass Lynx V4.1 (Waters, USA), and HPLC water were used. α-Amylase
(CAS RN 9000-90-2), α-glucosidase (CAS number: 9001-42-7), pNPG (CAS number: 2207-68-3), fast blue (CAS number: 14263-94-6), 2,2-diphenyl-1-picrylhydrazyl
(DPPH) (CAS number:1898-66-4), and ascorbic acid (CAS number:50-81-7)
were purchased from Sigma Aldrich Co., St Louis, USA. Folin Ciocalteu
reagent, sodium carbonate, aluminum chloride, sodium acetate, α-Amylase,
and glucosidase were procured from Loba Chemie Pvt. Ltd., Mumbai,
India. BGR-34 was procured as a gift sample from AIMIL pharmaceutical
India Ltd. All other solvents and chemicals used were of analytical
grade.
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7

Trifluoroacetic Acid Synthesis and Characterization

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Trifluoroacetic acid (TCA) was procured from Sisco Research Laboratories Pvt. Ltd, India. Sodium acetate was procured from Loba chemie, India. Other chemicals required in the study were procured from Sigma-Aldrich, Germany. All sample preparations and measurements were carried out using double distilled water (Cascada water purifier, UK) having conductance of 0.055 µS/ cm 2 .
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