Acetylacetone

Analytical grade chemicals and Milli Q water were used for the entire analyses. Ascorbic acid, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), dimethyl sulfoxide (DMSO), 1,1-diphenyl-2-picrylhydrazyl (DPPH), ethylenediaminetetraacetic acid (EDTA), β-nicotinamide adenine dinucleotide (NADH), 2,4,6-tris(2-pyridyl)-s-triazine (TPTZ), nitroblue tetrazolium (NBT), phenazine methosulfate (PMS), trichloroacetic acid (TCA), sodium nitroprusside and (N-(1-naphthyl)ethylenediamine dihydrochloride  (NED or Griess reagent) were supplied by Sigma-Aldrich Chemical Co (Bangalore, India). Aluminum chloride, ethanol, methanol, ferric chloride, Folin-Ciocalteu reagent, potassium chloride, sodium acetate, sodium carbonate, sodium hydroxide, sodium chloride, disodium hydrogen phosphate (anhydrous), potassium dihydrogen phosphate, potassium acetate, gallic acid, ferrous ammonium sulfate, ammonium acetate, glacial acetic acid and acetyl acetone were requisitioned from Merck (Mumbai, India).

Chloroform, toluene, 2-propanol, and acetyl-acetone were purchased from Merck (Germany). They were all in analytical grade with purity higher than 99.9%, 99.9%, 99.8%, and 99%, respectively. Tetra-n-butyl titanate with purity higher than 98% was supplied from Fluka (Switzerland). Granular activated carbon (with a mean diameter of 1.5-2 mm) with highest purity available (extra pure) was also obtained from Merck (Germany).

The C. militaris hyphae were collected from 6-day-old cultures on PDA plates, and their weight was recorded. Then, 1.2 mL of 50% EtOH was added to each tube, and the suspension was incubated in an ultrasonic bath with sonication at 70 °C for 15 min. One milliliter of the supernatant was then transferred to new tubes after centrifugation at 14,000 rpm (Eppendorf K-5418R). A total of 1.2 mL of a 10 mM sodium periodate solution (Merck, cat# 7790-28-5) was added to the suspension, and the mixture was shaken for 30 s. Then, 1.2 mL of a 0.2 M acetylacetone (Merck, cat# 123-54–6) solution was added to the former solution and kept in a water bath at 70 °C for 10 min. The sample absorbance was measured with a UV–Vis–NIR spectrophotometer (Cary 5000, version 3.00, Agilent, Scan Version 6.2.0.1588), and the glycerol concentrations were estimated using the formula Y = 0.0055*Ab-0.0012. [Y = glycerol concentration; Ab = absorbance at 413 nm. The formula was build based on a twofold serious dilution of glycerol (> / = 99% purification, Merck, cat # 56-81-5). The glycerol content (microgram glycerol/mg hyphal weight) was calculated by dividing the total glycerol content of each sample by its corresponding hyphal weight.

Precisa LS 220A SCS (Dietikon, Switzerland) analytical balance was used for weighing. GC-FID analysis was performed with Shimadzu GC-2010 Plus (Kyoto, Japon) equipped with Shimadzu AOC-5000 Plus (Kyoto, Japon) autosampler and Restek Rtx-Wax capillary column (30 m × 0.32 mm ID, 0.25 μm film thickness) (Pennsylvania, United States). For absorption measurements of the validation method, Shimadzu UV-1800 (Kyoto, Japon) spectrophotometer was used.
Formaldehyde standard, o-(2,3,4,5,6-Pentafluorobenzyl)hydroxylamine hydrochloride as derivatization reagent, 1-bromo-4-fluorobenzene (BFB) as internal standard were purchased from Sigma-Aldrich Chemie GmbH (Steinheim, Germany). Sodium chloride, potassium chloride, calcium chloride dihydrate, acetylacetone, acetic acid (glacial) were provided from Merck KGaA (Darmstadt, Germany).

Undoped BiVO₄ photoanodes were fabricated using a modified metal-organic deposition method, as previously described.^{20 ,26 (link)} Briefly, the precursor was prepared by mixing 0.2 M bismuth nitrate pentahydrate dissolved in acetic acid (99%, BDH) and 0.2 M vanadyl acetylacetonate in acetylacetone (Merck) for 1 hour at room temperature. The BiVO₄ photoanodes were deposited using the precursor on FTO substrates (TEC 15, Hartford Glass Co.) by the spin-coating method (1000 rpm, 20 s). After spin-coating each layer, the substrates were calcined at 450 °C for 15 min. The total number of layers was 15 for each BiVO₄ photoanode and the films were calcined at 450 °C for 5 hours.

Ferric hydroxide oxide (hydrated, 30–50 mesh), oleic acid (90%), oleylamine (90%), n-docosane (99%), ammonia (30%), tetraethylorthosilicate (TEOS, 99%), (3-chloropropyl)-triethoxysilane (95%), (3-iodopropyl)trimethoxysilane (95%), sodium iodide, potassium tertbutoxide (tBuOK), tert-butanol (tBuOH), europium(III) chloride hexahydrate (99.9%, trace metals basis), terbium(III) chloride hexahydrate (99.9%, trace metals basis), acetylacetone, pentane, diethyl ether, cyclohexane, acetone, and ethanol were purchased from Merck. Triton X-100 and 1-hexanol (99%) were purchased from Alfa Aesar (Haverhill, MA, USA).