Di potassium hydrogen phosphate

Incubation was done in refrigerated shaker incubator of model Innova 4230, New Brunswick Scientific Co., Inc., (NJ, USA). Sample analysis was done by using a chiral column purchased from Phenomenex (USA) (Lux Cellulose-4; 250 × 4.6 mm; 5 µ particle size) and an Ultra Fast Liquid Chromatograph of Shimadzu (Kyoto, Japan) equipped with binary pump (LC 20AD), UV/Visible detector (LC 20A) and Rheodyne injector port. Lab solutions software was employed for the HPLC analysis.
Racemic Carvedilol was kindly gifted by Symed labs limited, Medak (India). (S)-(−)-Carvedilol and (R)-(+)-Carvedilol were purchased from the Toronto Pharmaceuticals (Canada). Calcium chloride, sodium alginate, sodium chloride and di-potassium hydrogen phosphate were purchased from Hi Media Laboratories Pvt. Ltd., Mumbai (India). Solvents (n-heptane, isopropanol and methanol) of HPLC grade were purchased from Sigma Aldrich Chemicals Pvt. Ltd., Maharashtra (India). Laboratory grade ethyl acetate was purchased from Finar limited, Ahmadabad, Gujarat (India).

The porcine pancreatic lipase (PPL) 5 (Type II, 100–500 U/mg protein using olive oil) was purchased from Sigma Aldrich Co. India. For enzyme activity analysis via the olive oil emulsion method, chemicals were obtained from Hi-Media Laboratories: dipotassium hydrogen Phosphate and Potassium dihydrogen phosphate (preparation of pH 7 buffer), gum arabic, pure olive oil, and Sodium hydroxide. Chromatographically pure monoolein and oleic acid were purchased from Sigma—Aldrich (Shanghai-china). Bradford reagent was prepared using Coomassie brilliant blue, ethanol, phosphoric acid (85% pure), and glycerol. Biosynthetic Iron (II) oxide nanoparticles were used as a support for immobilization which was prepared using leaf extract and 0.01 M FeCl₃. The leaf from Bauhinia tomentosa plant was used in the present study complies with institutional, national, and international guidelines and legislation. Permission to take leaf samples were obtained.

Luria–Bertani (LB) broth, agar, catechol, veratryl alcohol, magnesium sulfate, zinc sulfate, copper sulfate, ammonium sulfate, silver chloride, mercuric chloride, calcium chloride, sodium chloride, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, silver nitrate, sodium nitrate, potassium nitrate, ammonium nitrate and sodium thiosulfate, sulfanilamide, N-(1-naphthyl)ethylenediamine dihydrochloride (NEDD), 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), Diethyl aminoethyl (DEAE)-cellulose and Biogel P-150 were procured from Himedia (Mumbai, India). Coomassie brilliant blue R-250, hydrogen peroxide and n-propanol were obtained from Loba Chemie (Mumbai, India), SD Fine (Mumbai, India) and Qualigens (Mumbai, India), respectively. Ammonium sulphate, NADH, L-DOPA, guaiacol, hydroxyquinolone and tartaric acid were supplied by SRL Pvt Ltd (Mumbai, India).