Calibration standard solutions were prepared with TBBPA in a 50% ethanol/water solution with final concentrations of 0, 2, 6, 11, 20, 23 mg/kg. When measuring using the spectrophoto meter, a 1.9 mL aliquot of the sample was placed in an acrylic cuvette (optical length 1 cm). After adding 0.1 mL of a 2% ferric iron coloring reagent (final concentrations 0.1%) to the cuvette, and was held for 2 min, the absorbance at 690 nm was measured. With a UV-Visible double-beam spectrophotometer (Carry 60, Agilent Technologies, Santa Clara, CA, USA). Prior to each measurement, zero calibratin was performed with a blank solution at 690 nm. The absorbances were measured with a UV-Visible double-beam spectrophotometer (Carry 60, Agilent Technologies, Santa Clara, CA, USA). As supplementary data, the reagent concentration and the reaction time were optimized with TBBPA at 23 mg/kg. Figures S1(a) and S1(b) (Supporting Information) show that 0.1% ferric coloring reagent is sufficient to complete the reaction in 2 min.
Carry 60
Manufactured by Agilent Technologies
Sourced in United States, Malaysia
The Carry 60 is a portable vacuum pump designed for laboratory use. It provides reliable vacuum performance in a compact and durable package.
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Lab products found in correlation
Lc 20 ad t, by Shimadzu (1 mentions)
2010 plus, by Shimadzu (1 mentions)
Emx spectrometer, by Bruker (1 mentions)
Empyrean diffractometer, by Malvern Panalytical (1 mentions)
Dsa100, by Krüss (1 mentions)
Jsm 7000f, by JEOL (1 mentions)
Uv vis spectrometer, by Agilent Technologies (1 mentions)
Dtu 1c, by TAITEC (1 mentions)
Qp2010 ultra, by Shimadzu (1 mentions)
15 protocols using carry 60
1
Spectrophotometric Determination of TBBPA
2
Proximate, Mineral, Amino Acid Analysis of Food
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Proximate analysis was done following official methods of analysis of AOAC International8 . The moisture and dry matter contents were determined using a hot air oven by drying the sample at 105 °C for 8 hours. Ash content was determined at 600 °C. Nitrogen content was estimated by Kjeldahl method (Kelplus Classic, DX VA, Pelican Equipment) and crude protein was calculated by multiplying nitrogen percentage by 6.25. Lipid was determined by the solvent extraction method by Soxtec system (SOCS Plus, SCS-6, Pelican Equipment) using diethyl ether (boiling point, 40–60 °C) as a solvent. The crude fiber was determined by digesting the moisture and fat-free sample successively with dilute (1.25%) acid and alkali using Fibre cap (Foss tecator, Sweden). Nitrogen free extract (NFE) was determined as per the following formula
Fatty acid, amino acid, and mineral analysis were carried out following AOAC8 . Calcium, magnesium, chromium, manganese, iron, chromium analysis was performed in Absorption Spectrophotometer (Model 280 FS- Make Agilent Technology). Phosphorus content was estimated using UV-Vis Spectrophotometer (Model - Agilent Carry-60). Amino acids were analyzed using HPLC (Model - Shimadzu LC-20 AD/T)). Fatty acids were analyzed through gas chromatography (Model - Shimadzu 2010 Plus).
Fatty acid, amino acid, and mineral analysis were carried out following AOAC8 . Calcium, magnesium, chromium, manganese, iron, chromium analysis was performed in Absorption Spectrophotometer (Model 280 FS- Make Agilent Technology). Phosphorus content was estimated using UV-Vis Spectrophotometer (Model - Agilent Carry-60). Amino acids were analyzed using HPLC (Model - Shimadzu LC-20 AD/T)). Fatty acids were analyzed through gas chromatography (Model - Shimadzu 2010 Plus).
3
Cellulase Activity Measurement Protocol
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The cellulase activity was measured in accordance with the methodology of Deng et al. [38 (link)] with minor modifications. After incubating the supernatant at 37 °C for one hour, 0.1 mL of the extract was added to a cellulase activity test mixture that consisted of 0.4 mL of 1% (w/v) carboxy methylcellulose and 0.5 mL of 0.1 M sodium acetate buffer with a pH of 5.0. The mixture was heated for 5 min at 100 °C and then cooled to room temperature. The enzyme activity was reported as U·mg−1 protein after the absorbance was read at 540 nm on UV-VIS spectrophotometer (Carry 60, Agilent, USA). One unit of cellulase activity was defined as the amount of the enzyme that catalyzed the formation of one µ-mole reducing groups per hour per mg protein.
4
Multimodal Characterization of Novel Material
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X-ray diffraction experiments were carried out using an Empyrean diffractometer (Panalytical) equipped with a copper anode. The CuKα radiation was selected by a monochromator and the measurements were carried out at room temperature using an accelerating voltage and a current of 40 kV and 30 mA, respectively. We used the standard formula for the hexagonal crystal system26 to obtain the lattice constants a and c from X-ray measurements. The TEM study (SAED and HRTEM) was performed using a JEOL 2010 electron microscope, operating at 200 kV. EPR measurements were performed at a 9.4 GHz frequency using an X-band Bruker EMX spectrometer equipped with a continuous nitrogen flow cryostat. The spectrometer was operated at a modulation frequency of 100 kHz and a microwave power of 20 mW. The studied sample was sealed directly into glass tubes (with a mass of approximately 10 mg). The concentration of the paramagnetic species was evaluated based on the use of a reference sample of CuSO4 × 5H2O with calibrated content of the unpaired spins from Cu2+. Magnetic properties were measured using the SQUID S700× magnetometer from 5 K to 300 K. The UV-vis absorption spectra were recorded using a UV-vis spectrometer (Agilent technologies Carry 60) with a 150 W Xe light source.
5
Quantification and Analysis of Antioxidant Enzymes
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Tissue samples from each group were accurately quantified, and tissue homogenization was prepared in KCl (0.15). Subsequently, the tissue samples were centrifuged for 15 min at 10000 g to get supernatants. The supernatants thus obtained stored in ice-cold water until further analysis. At the time of analysis, a fraction of the supernatant was taken and used to determine SOD, catalase, TBARs, PC, and GSH according to the previous method described elsewhere. The absorbance of the samples was taken through a UV-Visible spectrophotometer (Agilent Technologies, Carry 60) [55 (link), 56 (link)].
6
Encapsulation Efficiency Determination
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Two milliliters of the L-PC or CL-PC/α-TOC suspension was transferred precisely into a centrifugal ultrafiltration device with a molecular weight cutoff of 100 kDa and centrifuged at 3,000 rpm for 10 min. The content of procyanidins in the supernatant was determined by measuring the absorbance at 280 nm using a spectrophotometer (Carry 60, Agilent, Penang, Malaysia). The EE was calculated using equation (5) :
7
Spectrophotometric Sulfide Quantification
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The zinc precipitation method for sulphide determination was followed with slight modifications. Concisely, 500 µl of tissue supernatant was added to 400 µl of premixed zinc acetate (350 µl, 1% w/v) solution and 50 µl of sodium hydroxide (1.5 M) followed by centrifugation. The collected pellets were re-suspended with 160 µl of Milli-Q water and mixed with 40 µl of dye (20 µl of 20 mM NNDP in 7.2 M HCl and 20 µl of 30 mM FeCl3 in 1.2 M HCl). The mixture was incubated at 37 °C for 10 min and read absorbance at 670 nm using UV spectrophotometrically (Carry60, Agilent Technologies, CA 95051, USA) (Ang et al., 2012 ).
8
DPPH Radical Scavenging Activity of Procyanidins
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The DPPH free radical scavenging activity of non-encapsulated procyanidins was evaluated using a modified procedure [18] . Briefly, a 1 mL solution of procyanidins at different concentrations was mixed with 4 mL of 25 mg/L ethanol solution of DPPH. The mixture was mixed thoroughly and incubated at room temperature for 60 min. Then, the absorbance was read at 517 nm using a spectrophotometer (Carry 60, Agilent, Penang, Malaysia). The DPPH radical scavenging efficiency (%) of the samples was calculated by equation (2) :
where Ai is the absorbance of 1 mL of sample and 4 mL of DPPH, Aj is the absorbance of 1 mL of sample and 4 mL of ethanol, and A0 is the absorbance of 1 mL of ethanol and 4 mL of DPPH.
where Ai is the absorbance of 1 mL of sample and 4 mL of DPPH, Aj is the absorbance of 1 mL of sample and 4 mL of ethanol, and A0 is the absorbance of 1 mL of ethanol and 4 mL of DPPH.
9
Microalgal Growth in Varying Treated Wastewater
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Comparative growth profile of microalgal isolate ASK25 was studied employing varying concentrations of TWW (20, 40, 60, 80, and 100%) diluted with BG-11. The BG-11 growth medium was used as the control. Autoclaved TWW at various concentrations (20, 40, 60, 80 and 100%) was employed to study the growth of the microalgal isolate ASK25 and the initial pH for all the concentrations was adjusted to 6.8 ± 0.2 using 0.1 M NaOH and 0.1 M HCl. Homogenous algal suspension (10%, v/v) was used for inoculation having an optical density of 2.0 at 680 nm. The 4 L bubble columns reactors were used for microalgal isolate ASK25 containing 3 L of working volume employing different concentrations of TWW and BG-11(as depicted in Fig. 2 ) under a photon flux of approximately 50–60 μmol m−2 s−1, photoperiod of 16:8 h at 28 ± 2 °C. The growth of the microalgal isolate ASK25 was monitored during 24 h intervals by measuring optical density at 680 nm using UV–visible spectrophotometer (Agilent, Carry 60).![]()
Experiment set to grow algal isolate in different test media. (Control: standard BG11 medium, T1 (TWW: dH2O, 1:4); T2 (TWW: dH2O, 2:3); T3 (TWW: dH2O, 3:2); T4 (TWW: dH2O, 4:1) and T5 (TWW: dH2O, 5:0). Keys: 1: magnetic stirrer; 2: culture medium; 3: bubble column and 4: magnetic bar
10
Structural and Optical Characterization
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The morphology and structure of the products were characterized using a scanning electron microscope (JEOL, JSM-7000F). The optical properties were characterized using ultraviolet–visible spectroscopy (Agilent, Carry 60). The contact angle was measured on an optical contact angle instrument (KRUSS, DSA 100).
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