The enzymatic activity was determined according to the methods recommended by IUPAC [18 ]. The activity of total cellulases and endoglucanases was determined by quantifying the reducing sugars from a filter paper (Whatman No.1) and with soluble cellulose (carboxymethylcellulose, CMC) respectively, as substrate. For total cellulase activity (FPase), 0.5 ml of the culture supernatant (crude enzyme extract) was incubated for 1 h at 50 °C with 1 ml of 0.05 M sodium citrate buffer (pH 4.8) containing 50 mg of filter paper Whatman No. 1 (1 x 6 cm strip). To determine the activity of endoglucanases (CMCase), 0.5 ml of the enzyme extract was incubated with 0.5 ml of 2% carboxymethylcellulose in 0.05 M sodium citrate buffer (pH 4.8) at 50 °C for 30 min. After incubation, the samples were centrifuged at 7000 rpm for 10 min and 4 °C, and an aliquot of 0.5 ml was taken. The free reducing sugars were quantified by the DNS method; controls of sole substrate, enzyme and a reference solution of glucose (1%) were also included. One cellulase/ endoglucanase unit (IU/ml) was defined as the amount of enzyme that produces 1 μmol equivalent of glucose per minute.
Whatman no 1
Manufactured by Cytiva
Sourced in United Kingdom, United States, Switzerland, Germany, Japan
Whatman No. 1 is a grade of filter paper commonly used in laboratory applications. It is a general-purpose filter paper designed for a wide range of filtration tasks, including gravity filtration, vacuum filtration, and other laboratory procedures requiring the separation of solids from liquids.
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Lab products found in correlation
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Whatman no 1 filter paper, by Cytiva (4 mentions)
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Uv vis spectrophotometer, by Mecasys (2 mentions)
Agilent chemstation, by Agilent Technologies (2 mentions)
Sugar samples, by Merck Group (2 mentions)
50 ml centrifuge tube, by Sarstedt (2 mentions)
Whatman no 1 paper, by Cytiva (2 mentions)
Freeze dryer, by IlShinBioBase (1 mentions)
Milli q apparatus, by Merck Group (1 mentions)
Acetonitrile, by Merck Group (1 mentions)
Optizen 2120 uv plus, by Mecasys (1 mentions)
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330 protocols using whatman no 1
1
Enzymatic Assay for Cellulases and Endoglucanases
2
Preparation of Medicinal Plant Decoctions
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All the plant materials A.
ceylanica (leaves), E.
quinquangulare (whole plant),
M. cerviana (whole plant),
S. parvifolia (leaves)
were prepared separately as decoctions according to the proportions followed
by Ayurvedic practitioners. The plant materials described above were washed
separately with tap water followed by distilled water and de-ionized water,
dried to achieve a constant weight. Each plant material was cut into small
pieces and ground to a fine powder using a clean kitchen blender. Powdered
samples (30 g) except S.
parvifolia were boiled with 800 ml of deionized water until the total volume reduced to 100 ml (1/8th of the original volume) using a beaker. Powdered
leaves of S. parvifolia(30 g) was refluxed with 800 ml of
deionized water to prepare the aqueous extract of the plant material. The
decoctions were sonicated and filtered through a cotton wool plug and then
using filter paper (Whatman No. 1). The filtrates were centrifuged at
2000 rpm for 10 min. The
supernatants were freeze dried. The freeze dried samples were weighed, and
stored at −20 °C in sterile tubes until further use. A
weight of 2.0 g of C.
sinensis (black tea powder) was added into
200 ml of boiling water and allowed to stand in a
closed beaker to prepare black tea infusion. The solution was allowed to
cool to room temperature, filtered through filter paper (Whatman No. 1) and
the filtrate was used for the experiments.
ceylanica (leaves), E.
quinquangulare (whole plant),
M. cerviana (whole plant),
S. parvifolia (leaves)
were prepared separately as decoctions according to the proportions followed
by Ayurvedic practitioners. The plant materials described above were washed
separately with tap water followed by distilled water and de-ionized water,
dried to achieve a constant weight. Each plant material was cut into small
pieces and ground to a fine powder using a clean kitchen blender. Powdered
samples (30 g) except S.
parvifolia were boiled with 800 ml of deionized water until the total volume reduced to 100 ml (1/8th of the original volume) using a beaker. Powdered
leaves of S. parvifolia(30 g) was refluxed with 800 ml of
deionized water to prepare the aqueous extract of the plant material. The
decoctions were sonicated and filtered through a cotton wool plug and then
using filter paper (Whatman No. 1). The filtrates were centrifuged at
2000 rpm for 10 min. The
supernatants were freeze dried. The freeze dried samples were weighed, and
stored at −20 °C in sterile tubes until further use. A
weight of 2.0 g of C.
sinensis (black tea powder) was added into
200 ml of boiling water and allowed to stand in a
closed beaker to prepare black tea infusion. The solution was allowed to
cool to room temperature, filtered through filter paper (Whatman No. 1) and
the filtrate was used for the experiments.
3
Mycelia, Grains, and Basidiocarps Extraction
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Freeze dried mycelia, fermented wheat grains, and air dried basidiocarps (both wild and cultivated) were blended, soaked in 95% ethanol at 1: 100 (w/v) on 150 rpm shaker and incubated in room temperature (25 ± 2 °C) for seven days43 . The aliquots were then filtered through filter paper Whatman No.1 and evaporated to dryness using Eyela vacuum rotary evaporator at 40 °C and 173 Hpa. Hot aqueous extracts were prepared by boiling freeze dried samples (100 g/L) at 100 °C for two hours43 . The aliquots were filtered through filter paper Whatman No.1 and the filtrate was freeze dried.
4
Endogenous Hormone Extraction and Quantification
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Endogenous hormones (IAA, GA3, ABA, JA, SA) were extracted from the terminal buds of plants and determined by Knegt and Bruinsma [88 (link)] method as follows:
Five g of fresh leaves was collected and put overnight at 4 °C in dark in 80% redistilled cold MeOH (10 mL MeOH per g FW). Filtering of the extract by filter study (Whatman No.1) and then washed with 5 mL of 80% MeOH. The extract was then evaporated by rotary evaporator into aqueous phase. The residue, i.e., aqueous phase, was dissolved by 5 mL phosphate buffer (0.1 M, pH 8) and held at −18 °C for 24 h. The defrosted extract was centrifuged at 4 °C at a rate of 12,000× g, polyvinylpyrrolidone (PVP) was applied and filtered through Whatman No.1, then the filtrate was washed by 7 mL phosphate buffer (0.1 M, pH 8) and the total volume complete up to 30 mL. Partition extract was carried out against diethyl ether (1:2 volume) two times. Organic phase was discarded each time. Partition extract was performed two times against diethyl ether (1:2). Through time, organic phase was discarded. The aqueous phase pH was changed to pH 2.5 using 5-N HCl and partitioned twice with diethyl ether and discarded the aqueous phase. The organic phase was evaporated to dry film and then dissolved with 5 mL H2O (0.1 N acetic acid) and pH with 1 N acetic acid was changed again to 2.5.
Five g of fresh leaves was collected and put overnight at 4 °C in dark in 80% redistilled cold MeOH (10 mL MeOH per g FW). Filtering of the extract by filter study (Whatman No.1) and then washed with 5 mL of 80% MeOH. The extract was then evaporated by rotary evaporator into aqueous phase. The residue, i.e., aqueous phase, was dissolved by 5 mL phosphate buffer (0.1 M, pH 8) and held at −18 °C for 24 h. The defrosted extract was centrifuged at 4 °C at a rate of 12,000× g, polyvinylpyrrolidone (PVP) was applied and filtered through Whatman No.1, then the filtrate was washed by 7 mL phosphate buffer (0.1 M, pH 8) and the total volume complete up to 30 mL. Partition extract was carried out against diethyl ether (1:2 volume) two times. Organic phase was discarded each time. Partition extract was performed two times against diethyl ether (1:2). Through time, organic phase was discarded. The aqueous phase pH was changed to pH 2.5 using 5-N HCl and partitioned twice with diethyl ether and discarded the aqueous phase. The organic phase was evaporated to dry film and then dissolved with 5 mL H2O (0.1 N acetic acid) and pH with 1 N acetic acid was changed again to 2.5.
5
Carotenoid Extraction from Samples
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Extraction of carotenoids was performed according to the method previously described by Silva da Rocha et al. 29) . The ground sample (2 g) was added to 25 mL of acetone. The mixture was shaken by vortex for 10 min and filtered using filter paper (Whatman No. 1) and passed through a separation funnel. The filtrate was fractionated with 20 mL of petroleum ether and washed with 100 mL of distilled water to remove the acetone. These steps were repeated twice. Whatman No. 1 covered with anhydrous sodium sulfate (5 g) for removing residual water was used to filter the petroleum ether layer. The volume of the extracts was adjusted to 25 mL using petroleum ether. Then, absorbance was measured at 450 nm.
6
Eucalyptus Extract Preparation and Dilution
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To evaluate the effect of a water-soluble extract of the Eucalyptus genus, fresh leaves of three Eucalyptus species were collected and washed thoroughly with tap water followed by distilled water to remove dust. After blotting dry with filter paper (Whatman No. 1) 250 g of fresh leaves was ground in 500 ml of distilled water in a household blender for 10 minutes. The extract was then filtered through two layers of Whatman No. 1 filter paper and residue was washed thoroughly 2 to 3 times and made it up to 1 liter by the addition of distilled water, thus the stock solution of 100% aqueous extract was prepared. The stock solution was then stored in a refrigerator in a dark place in conical flasks until required. The extract was further diluted to 10, 25, 50, and 75 % extract concentration.
7
Determination of Crude Fiber Content
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The crude fibre was determined using the AOAC (2012). Two grams (W3) of sample was dissolved in 200 mL of 1.25% (v/v) sulphuric acid in a conical flask and was placed on a hot plate and boiled for 30 min. The content was filtered using filter paper (Whatman No.1) and the residue on the filter paper was washed with 70 mL distilled water. The washed residue was transferred back into the flask and about 200 mL 1.25% (w/v) NaOH is added and boiled for 30 min. The content was filtered as described earlier and the residue obtained was washed with distilled water and then filtered again using filter paper (Whatman No.1). The residue is then transferred to an ashing dish and dried at 130°C for 2 h, cooled in a desiccator and weighed (W1). This was then ashed at 550°C inside the muffle furnace chamber (Carbolite AAF1100, United Kingdom) for 30 min, cooled and reweighed (W2). The ash obtained was subtracted from the residue and the difference expressed as percentage of the starting material as shown in equation below;
Where, Cf = Crude fibre (%) W1 = Mass of crucible with the dried residue (g) W2 = Mass of crucible with the ash (g) W3 = Mass of sample(g)
Where, Cf = Crude fibre (%) W1 = Mass of crucible with the dried residue (g) W2 = Mass of crucible with the ash (g) W3 = Mass of sample(g)
8
Plant Growth Regulator Extraction and Quantification
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Extractions and purifications were performed according to Battal and Tileklioğlu [17] with little modifications. The frozen samples (2-2.5 g) were powdered in liquid nitrogen. The cold methanol (80%) was added to powdered samples and homogenized by using a tissueblender (Jencons). Then, they were stored at 4 °C for 24 h in the dark and filtered through a filter paper (Whatman No.1). The supernatants were again filtered through a PTFE filter (0.45 µm) and concentrated at 35 °C by evaporation. The samples were re-dissolved in 0.1 M KH2PO4 buffer (pH 8.00) and centrifuged at 10.000 for 1 h at 4 °C. The supernatants were mixed 1 g polyvinyl polypyrrolidone (PVPP, Sigma Chemical Co, Dorset, UK) and filtered through Whatman No.1 [21] . The filtrates were passed through Sep-Pak C18 (Waters Hichrom Ltd., Berkshire, UK) cartridges. Cartridge adsorbing plant growth regulator were collected eluated with 80% methanol and the extracts were collected in vials. Then, the samples were analyzed to detect gibberellic acid (GA), zeatin (ZA), indole acetic acid, abscisic acid (ABA) levels [18, 19] .
9
Tailoring P(VDF-TrFE) Membranes for Microfluidics
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Several processing techniques and post-treatment protocols were employed to produce P(VDF-TrFE) membranes with tailored morphological and physicochemical properties suitable for microfluidic device development 14 (link) . NIPS technique was used to produce P(VDF-TrFE) membranes using ethanol as a weak non-solvent in order to tune the liquid-liquid demixing process to produce samples with large spherical microstructures, similar to commercial Hi-Flow Plus substrates from Millipore 24 (link) . On the other hand, TIPS technique was used to produce porebased microstructures as an alternative morphology 24 (link) . Further, ES has been used to obtain randomly oriented P(VDF-TrFE) fibre membranes, mimicking the morphology of commercial Whatman TM no.1 25 (link) , and oriented electrospun membranes, in order to study the effect of fibre orientation 15 (link) .
As the hydrophobic nature of P(VDF-TrFE) membranes hinders their use as microfluidic substrates, the as-processed membranes were submitted to a post-treatment with oxygen plasma and its effect on the physicochemical properties of the membranes was evaluated.
As the hydrophobic nature of P(VDF-TrFE) membranes hinders their use as microfluidic substrates, the as-processed membranes were submitted to a post-treatment with oxygen plasma and its effect on the physicochemical properties of the membranes was evaluated.
10
Phytochemical Extraction and Evaluation
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Exactly 5.0 g of the dried sample was ground to a fine powder and extracted with 50 mL of distilled water for 120 minutes at room temperature of 29 C. The dried sample was ground to a fine powder (15.0 g) and extracted with 150 mL of distilled water at 50 C, 70 C, and 90 C. The samples were filtered on Whatman TM No 1. The filtrate was used for the assessment of phenolic contents, ferric reducing power, and radical scavenging capacities.
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