This research was supported by the Institute of International Education&#39;s Scholar Rescue Fund (IIE-SRF).

Separation of Aquafaba from Chickpeas
<ol>
	<li>Obtain cans of chick peas from local grocers and open with a manual can opener.</li>
	<li>Label cans from A to J.</li>
	<li>Separate chickpeas from aquafaba using a stainless steel meshed kitchen strainer and weigh the separated chickpeas and aquafaba.</li>
</ol>
Obtain a Representative Sample of Chickpeas and Aquafaba for Chemical Analysis.
<ol start="4">
	<li>Randomly select ten chickpeas after draining the aquafaba to determine moisture content.</li>
	<li>Place the selected chickpeas in a drying tin and heat at 100 &#177; 2 &#176;C for 16-18 h in a drying oven.</li>
	<li>After drying, grind the chickpeas to a powder for further use (e.g. protein and carbon content analysis).</li>
	<li>Freeze aquafaba and aquafaba foam from each source to -20 &#176;C and then dry in a freeze dryer. The dried aquafaba will be used for nitrogen and carbon content determination.</li>
</ol>
Aquafaba Functional Properties
<ol start="8">
	<li>Determine aquafaba viscosity at 25 &#176;C using a No. 2 shell cup.
	<ol>
		<li>Immerse the shell cup in the aquafaba.</li>
		<li>Record the time required to drain the cup11. A timer is started when the cup was lifted from the aquafaba and stopped when the stream leaving the cup stops.</li>
		<li>The aquafaba viscosity can be ascertained by a chart supplied with the cup that relates viscosity and drainage time.</li>
	</ol>
	</li>
	<li>Foaming capability
	<ol>
		<li>Blend the aquafaba solution (100 mL) in a stainless steel bowl with a hand mixer at speed setting of 10 for 2 min.</li>
		<li>Record the foam volume immediately after blending 100 mL of the aquafaba solution as Vf100 by placing the foam in a graduated measuring cup.</li>
		<li>Allow the foam to stand and dry over time to observe foam stability and obtain a sample of dried foam.</li>
	</ol>
	</li>
</ol>
Color Parameters of Chickpea Seed
<ol start="10">
	<li>Randomly select chickpea seed from each can for color determination.</li>
	<li>Calibrate the Hunter lab colorimeter using white, black and reference standards prior to measurements.</li>
	<li>Color coordinate values are determined by the CIE Lab system12, including the L (positive represents white and 0 represents dark), a (positive is red and negative is green), and b (positive is yellow and negative is blue) in triplicate with day light 65&#176;.</li>
</ol>
Protein and Carbon Contents
<ol start="13">
	<li>Determine protein and carbon contents by combustion using an elemental analyzer13. Protein content is estimated as nitrogen content multiplied by 6.2514.</li>
</ol>
Moisture Content
<ol start="14">
	<li>Determine seed and aquafaba moisture contents by heating samples at 100 &#177; 2 &#176;C for 16-18 h in a drying oven15.</li>
</ol>
NMR Spectrometry
<ol start="15">
	<li>Sample preparation
	<ol>
		<li>Prior to spectrometry, centrifuge aquafaba samples at 9200 &#215; g for 10 min. After centrifugation, pass the supernatant through a 0.45 &#181;m syringe filter (25 mm syringe filter with polytetrafluoroethylene (PTFE) membrane). Transfer the aquafaba sample (0.4 mL) into a NMR tube wuth 50 mg added deuterium oxide inside and subject the sample for NMR analysis.</li>
		<li>For the dried foam sample previously described, the sample (25 mg) is in deuterium oxide (500 mg) and the solution is subjected to NMR analysis.</li>
		<li>Place all samples for 13C-NMR and 1H-NMR in capped 5 mm NMR tubes. Add deuterium oxide and 3-(trimethylsilyl)propionic-2,2,3,3-d4 acid sodium salt (TMSP) to each NMR tube to provide a solvent lock signal and act as an internal standard, respectively.</li>
	</ol>
	</li>
	<li>NMR conditions
	<ol>
		<li>Acquire proton NMR spectra with a 500 MHz NMR or similar high field NMR spectrometer) with at least 16 scans per spectrum using a water suppression program such as the double pulse field gradient spin echo proton NMR (DPFGSE-NMR) pulse sequence16.</li>
		<li>Adjust the spectral shift to place the TMSP peak at 0 ppm then use the integration software to determine relative amounts of compounds present.</li>
	</ol>
	</li>
</ol>
Electrophoresis
NOTE: For this step, aquafaba that yielded the most stable foam (brand H) was selected. This brand did not contain salt.
<ol start="17">
	<li>Sample preparation
	<ol>
		<li>Introduce aquafaba to the upper reservoir of three centrifugal regenerated cellulose ultrafiltration devices each with different molecular weight cut offs (MWCOs) of 3, 10, or 50 kDa.</li>
		<li>Place the centrifugal filter units in a suitable centrifuge at 4 &#176;C and centrifuge at 3900 &#215; g for 2 h to obtain supernatant and retentate fractions. The supernatant fractions were used for 1H-NMR analysis of smaller molecules in the absence of higher molecular weight (MW) species. The 3 kDa membrane retentate fraction was used for electrophoretic separation as it was believed that this membrane would retain most proteins.</li>
		<li>Estimate the protein contents of both supernatant and retenate using a modified Bradford method using bovine serum albumin as a standard17.</li>
		<li>Place samples of supernatant and retentate fractions in Eppendorf tubes and subject these fractions to shaking at a frequency of 25 per s (10 min) in a suitable shaker18. Observe the shaken solution to determine if a foam has formed from shaking.</li>
		<li>Dissolve the retentate by adding 2.0 mL of distilled water to the ultrafiltration device for a second centrifugation treatment to achieve diafilteration. Subject the ultrafiltration device to a second centrifugation at 4 &#176;C and 3900 &#215; g for 2 h.</li>
		<li>Mix the retentate (0.025 g) with 1 mL of 0.02 M Tris-HCl pH 7.4 or phosphate-buffered saline (PBS) pH 7.4 to dissolve protein.</li>
		<li>Centrifuge the mixture at 21000 x g for 1 min.</li>
		<li>Use the supernatant to determine protein content using the modified Bradford as previously mentioned.</li>
	</ol>
	</li>
	<li>Electrophoretic separation 
	NOTE: The 3 kDa MWCO membrane retentate (described above) is subjected to electrophoretic separation using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).
	<ol>
		<li>Prepare polyacrylamide gels using a 15% polyacrylamide resolving gel and a 5% polyacrylamide stacking gel.</li>
		<li>Apply a sample of 20 &#181;g protein to one lane of the gel and PageRuler Prestained Protein Ladder with a range of 10-170 kDa to a separate polyacrylamide gel lane.</li>
		<li>Subject the gels to an electrical current in a Mini-Protein Tetra Cell system as modified from Laemmli (1970)19. For electrophoresis operating conditions, gel staining, and destaining follow Ratanapariyanuch et al. (2012)20.</li>
	</ol>
	</li>
</ol>
Peptide Mass Fingerprinting
NOTE: Cut bands from the SDS-PAGE gel of 3 kDa retentate (MWs of approximately 8, 10, 13, 14, 15, 20, 22, 31, 37, 55, and 100 kDa) for trypsin digestion according to Ratanapariyanuch et al. (2012)20 and perform mass spectral analysis.
<ol start="19">
	<li>In-gel digestion
	<ol>
		<li>De-stain the bands twice by immersing in 100 &#181;L of 200 mM ammonium bicarbonate (NH4HCO3) in 50% acetonitrile (ACN) and incubate at 30 &#176;C for 20 min.</li>
		<li>After the completion of de-staining, treat the gel samples with ACN (100 &#181;L) for 10 min and then dry under vacuum, using a centrifugal vacuum evaporator, for 15 min at room temperature.</li>
		<li>Immerse the dried gel samples in 100 &#181;L of 10 mM dithiothreitol (DTT) in 100 mM NH4HCO3 solution and incubate at 56 &#176;C for 1 h.</li>
		<li>Remove excess reducing buffer and replace it with 100 &#181;L alkylating buffer [100 mM iodoacetamide in mass spectrometry (MS) grade water].</li>
		<li>Incubate the gel samples at room temperature in the dark for 30 min.</li>
		<li>Wash gel samples twice with 200 mM NH4HCO3 (100 &#181;L), shrink the gels by immersing them in ACN (100 &#181;L), re-swell the gels with 200 mM NH4HCO3 (100 &#181;L) and again shrink by treating with ACN (100 &#181;L).</li>
		<li>Drain the ACN and dry the gel samples under vacuum in a centrifugal vacuum evaporator for 15 min.</li>
		<li>Re-swell dried gel samples using 20 &#181;L trypsin buffer (50 ng/&#181;L trypsin in 1:1,200 mM NH4HCO3: Trypsin stock solution) followed by addition of 30 &#181;L of 200 mM NH4HCO3 to each sample.</li>
		<li>Incubate samples on a Thermomixer overnight at 30 &#176;C with shaking at 300 rpm.</li>
		<li>Stop the trypsin digestion reaction by adding 1/10 of the final volume (volume of mixture after step 8) 1% trifluoracetic acid and extract the tryptic peptides from gel samples using 100 &#181;L of 0.1% trifluoracetic acid in 60% ACN and dry under vacuum using a centrifugal vacuum evaporator.</li>
		<li>Store tryptic peptides in -80 &#176;C prior to mass spectrometric analysis.</li>
	</ol>
	</li>
	<li>Mass spectrometry
	<ol>
		<li>Reconstitute tryptic peptides by adding 12 &#181;L of MS grade water: ACN:formic acid (97:3:0.1, v/v) and then vortex for 1 to 2 min to dissolve peptides.</li>
		<li>Centrifuge the resulting solution at 18,000 g for 10 min at 4 &#176;C.</li>
		<li>Transfer solution aliquots (10 &#181;L) to MS vials for liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis on a quadrupole time-of-flight (QTOF) mass spectrometer equipped with a liquid chromatography instrument and a LC-MS interface.</li>
		<li>Perform chromatographic peptide separation using a high-capacity HPLC-Chip: consisting of a 360 nL enrichment column and a 75 &#181;m &#215; 150 mm analytical column, both packed with C18-A, 180 &#197;, 3 &#181;m stationary phase.</li>
		<li>Load samples onto the enrichment column with 0.1% formic acid in water at a flow rate of 2.0 &#181;L/min.</li>
		<li>Transfer samples from the enrichment column to the analytical column.</li>
		<li>Peptide separation conditions are: a linear gradient solvent system consisting of solvent A (0.1% formic acid in water) and solvent B (0.1% formic acid in ACN). The linear gradient begins with 3% solvent B that increases to 25% solvent B over 50 min. Subsequently solvent B increases from 25 to 90% over 10 min at a flow rate of 0.3 &#181;L/min.</li>
		<li>Acquire positive-ion electrospray mass spectral data using a capillary voltage set at 1900 V, the fragment ion set at 360 V, and the drying gas (nitrogen) set at 225 &#176;C with a flow rate of 12.0 L/min.</li>
		<li>Collect spectral results over a mass range of 250-1700 (mass/charge; m/z) at a scan rate of 8 spectra/s. Collect MS/MS data over a range of 50-1700 m/z and a set isolation width of 1.3 atomic mass units. Select the top 20 most intense precursor ions for each MS scan for tandem MS with a 0.25 min active exclusion.</li>
	</ol>
	</li>
	<li>Protein identification
	<ol>
		<li>Convert spectral data to a mass/charge data format using Agilent MassHunter Qualitative Analysis Software.</li>
		<li>Process data against the UniProt Cicer arietinum (chickpea) database, using SpectrumMill as the database search engine.</li>
		<li>Include search parameters such as a fragment mass error of 50 ppm, a parent mass error of 20 ppm, trypsin cleavage specificity, and carbamidomethyl as a fixed modification of cysteine.</li>
	</ol>
	</li>
</ol>

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The authors have nothing to disclose.

In this research, we have found that chickpea aquafaba from different commercial sources produces foams that vary in both properties (volume and stability of foam) and chemical composition. There was a positive correlation between aquafaba viscosity and moisture content. Foam volume increase (Vf100) was not related to these parameters. Additives such as salt and disodium EDTA might suppress viscosity and foam stability as aquafaba from chickpea canned with these additives had lower viscosity and produced foams...

Increasingly vegetarian products are being developed that mimic the properties of meat, milk, and eggs. The functional properties of pulses are important in their current uses in food applications and their properties are being explored in the development of replacements for animal protein. For example, dairy alternatives sales were $8.80 Billion USD in 2015 and this market is growing rapidly. This market is projected to grow to $35.06 billion by 2024. Moreover, the upward trend in demand for plant-based milk substitutes is, in part, a result of consumer health concerns regarding cholesterol, antibiotics, and growth hormones often used in milk production1. Similarly, vegetable protein and hydrocolloid egg replacer markets are rapidly expanding and a compound annual growth rate of 5.8% is expected for these materials over the next 8 years with sales of $1.5 billion USD expected in 20262. A growing number of consumers prefer vegan protein sources, allergen reduced diets and reduced carbon footprint for food products. Demand for pulse-based products, especially from lentil, chickpea, and faba bean are steadily growing due to the high protein content, dietary fiber and low saturated fat content of pulses3. Pulses also contain phytochemicals with potentially beneficial biological activity4.
Commercial entities, scientists and private individuals have taken different approaches to communicate the quality properties of chickpea based egg and milk replacements. Gugger et al.5 produced a milk-like product from high starch grains including adzuki bean and chickpea. In their described methods the proponents attempted to show that their product is unique and different from &#34;aquafaba&#34;6. In another commercial approach elucidated by Tetrick et al.7 a plant-based egg substitute was developed. Their patent application describes methods of combining pulse flour with known thickeners that emulate the function of egg white in baked materials. Typical formulae include 80-90% pulse flour and 10-20% thickening additives.
Peer reviewed literature also indicates the functionality possible with chickpea and has demonstrated that albumin protein fractions obtained from kabuli and desi chickpea flour have good emulsification properties. They have also found a significant effect of chickpea source on the albumin yield and performance8.
After the initial internet report describing &#34;aquafaba&#34; by French chef Jo&#235;l Roessel, an open source movement is showing the utility of aquafaba as a replacement of egg white and dairy protein in many food applications. There are many highly viewed webpages and YouTube videos showing the incorporation of aquafaba in foods that emulate the qualities of ice cream, meringue, cheese, mayonnaise, scrambled eggs, and whipped cream. Most pioneers providing open source aquafaba applications (recipes) obtain their material by straining canned chickpeas and using the liquid in their recipes. These individuals are mostly not trained scientists. Video comments sections indicate that the respondents have copied the recipes and some have failed to replicate the successes of the aquafaba advocates.
All three approaches (corporate, scientific and open source) to developing egg and milk replacements have merit but are missing an important dimension. Applied scientists, basic scientists and individuals promulgating pulse-based products have incompletely characterized and standardized their input material. Standardization of a product for a specific use is a normal industrial practice. Chickpea cultivars have not been standardized for aquafaba quality and industrial canning practices are standardized to produce consistent chickpeas not aquafaba.
Based on studies of other commodities, it is predictable that both genotype and environment will contribute to pulse aquafaba quality. It is known that both genotype and environment affect kabuli chickpea canning properties9. Typically, genotypic effects are large between related species and smaller within members of a species. Variation in physical and chemical properties can be minimized through identity preservation that allows the selection of cultivars with desired properties. Environmental effects can also be large and are managed by quality evaluation and blending to standard performance in specific tests10.
There are many genetically distinct cultivars of chickpea in commercial production. For examples, the University of Saskatchewan Crop Development Centre, a major source of commercial chickpea germplasm, has released 23 chickpea cultivars since 1980 of which 6 are currently recommended for cultivation in Canada. While scientific manuscripts often describe the cultivar used in a study, the patents and internet pages that were surveyed did not indicate the cultivar used or the provenance of the chickpeas. The standardization of cultivars and handling could help users increase their success in using chickpea but this information is not available on canned chickpea products.
The objective of this research is to determine the aquafaba components that contribute foaming properties. Here,the rheological properties of aquafaba from commercial chickpea brands were compared and the chemical properties were studied by NMR, electrophoresis, and peptide mass fingerprinting. To our knowledge, this is the first research which describes the chemical composition and the functional properties of the aquafaba viscosifier components.

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			<td height="21" style="height:21px;width:219px;">Stoppering Tray Dryer</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:219px;">Mixer&#160;</td>
			<td style="width:217px;"></td>
			<td style="width:100px;"></td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:219px;">Stainless steel hand mixer&#160;</td>
			<td style="width:217px;">Loblaws</td>
			<td style="width:100px;">PC2200MR</td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:219px;">Viscosity Measurement&#160;</td>
			<td style="width:217px;"></td>
			<td style="width:100px;"></td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:219px;">Shell cup No. 2&#160;</td>
			<td style="width:217px;">Norcross Corp.</td>
			<td style="width:100px;"></td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:219px;">Color Measurement&#160;</td>
			<td style="width:217px;"></td>
			<td style="width:100px;"></td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:219px;">Colorflex HunterLab spectrophotometer&#160;</td>
			<td style="width:217px;">Hunter Associates Laboratory Inc.</td>
			<td style="width:100px;"></td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:219px;">Protein and Carbon Contents&#160;</td>
			<td style="width:217px;"></td>
			<td style="width:100px;"></td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:219px;">Elemental analyzer&#160;&#160;</td>
			<td style="width:217px;">LECO Corp.</td>
			<td style="width:100px;">CN628</td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:219px;">NMR Spectrometry</td>
			<td style="width:217px;"></td>
			<td style="width:100px;"></td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:219px;">Spectrafuge 24D&#160;&#160;</td>
			<td style="width:217px;">Labnet International Inc.</td>
			<td style="width:100px;"></td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:219px;">Syringe filters&#160;</td>
			<td style="width:217px;">VWR International</td>
			<td style="width:100px;">CA28145-497</td>
			<td style="width:267px;">25 mm, with 0.45 &#181;m PTFE membrane</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:219px;">Deuterium oxide&#160;</td>
			<td style="width:217px;">Cambridge Isotope Laboratories Inc.</td>
			<td style="width:100px;">7789-20-0</td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:219px;">3-(trimethylsilyl)propionic-2,2,3,3-d4 acid sodium salt</td>
			<td style="width:217px;">Sigma-Aldrich</td>
			<td style="width:100px;">169913-1G</td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:219px;">Bruker Avance 500 MHz NMR spectrometer&#160;</td>
			<td style="width:217px;">Bruker BioSpin</td>
			<td style="width:100px;"></td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:219px;">TopSpin 3.2 software&#160;</td>
			<td style="width:217px;">Bruker BioSpin GmbH</td>
			<td style="width:100px;"></td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:219px;">Electrophoresis&#160;</td>
			<td style="width:217px;"></td>
			<td style="width:100px;"></td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:219px;">Regenerated cellulose membrane&#160;</td>
			<td style="width:217px;">Millipore Corp.</td>
			<td style="width:100px;"></td>
			<td style="width:267px;">3, 10, 50 kDa (MWCO)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:219px;">Centrifugal filter unit&#160;</td>
			<td style="width:217px;">Millipore Corp.</td>
			<td style="width:100px;"></td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:219px;">Benchtop centrifuge&#160;</td>
			<td style="width:217px;">Allegra X-22R, Beckman Coulter Canada Inc.</td>
			<td style="width:100px;"></td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:219px;">Mixer Mill MM 300&#160; bead mill&#160;</td>
			<td style="width:217px;">F. Kurt Retsch GmbH &#38; Co. KG</td>
			<td style="width:100px;"></td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:219px;">Eppendorf centrifuge 5417C</td>
			<td style="width:217px;">Eppendorf</td>
			<td style="width:100px;"></td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:219px;">Phosphate buffered saline, pH 7.4</td>
			<td style="width:217px;">Sigma-Aldrich</td>
			<td style="width:100px;">P3813-10PAK</td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:219px;">Tris-HCl buffer pH 7.4&#160;</td>
			<td style="width:217px;">Sigma-Aldrich</td>
			<td style="width:100px;">T6789-10PAK</td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="29">
			<td height="29" style="height:29px;width:219px;">PageRuler Prestained Protein Ladder&#160;</td>
			<td style="width:217px;">Fisher Scientific</td>
			<td style="width:100px;"></td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:219px;">Mini-Protein Tetra Cell system</td>
			<td style="width:217px;">BioRad</td>
			<td style="width:100px;"></td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:219px;">Peptide Mass Fingerprinting</td>
			<td style="width:217px;"></td>
			<td style="width:100px;"></td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:219px;">Thermo-Savant SpeedVac</td>
			<td style="width:217px;">BioSurplus</td>
			<td style="width:100px;"></td>
			<td style="width:267px;">Centrifugal vacuum evaporator&#160;</td>
		</tr>
		<tr height="46">
			<td height="46" style="height:46px;width:219px;">Trypsin buffer&#160;</td>
			<td style="width:217px;"></td>
			<td style="width:100px;"></td>
			<td style="width:267px;">20 &#181;L trypsin in 1 mM hydrochloric acid and 200 mM NH4HCO3</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:219px;">Iodoacetamide</td>
			<td style="width:217px;">Sigma-Aldrich</td>
			<td style="width:100px;">I1149-5 g</td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:219px;">Trifluoroacetic acid&#160;</td>
			<td style="width:217px;">Fluka</td>
			<td style="width:100px;">BB360P050</td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:219px;">Acetonitrile</td>
			<td style="width:217px;">Fisher Scientific&#160;</td>
			<td style="width:100px;">L14734</td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:219px;">Formic acid&#160;</td>
			<td style="width:217px;">Sigma-Aldrich</td>
			<td style="width:100px;">33015-500mL</td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:219px;">Mass spectrometry vial&#160;</td>
			<td style="width:217px;">Agilent Technologies Canada Ltd.</td>
			<td style="width:100px;"></td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:219px;">Agilent 6550 iFunnel quadrupole time-of-flight mass spectrometer&#160;</td>
			<td style="width:217px;">Agilent Technologies Canada Ltd.</td>
			<td style="width:100px;"></td>
			<td style="width:267px;">Agilent 1260 series LC instrument and Agilent Chip Cube LC-MS interface</td>
		</tr>
		<tr height="64">
			<td height="64" style="height:64px;width:219px;">HPLC-Chip II: G4240-62030 Polaris-HR-Chip_3C18&#160;</td>
			<td style="width:217px;"></td>
			<td style="width:100px;"></td>
			<td style="width:267px;">360 nL enrichment column and 75 &#181;m &#215; 150 mm analytical column, both packed with Polaris C18-A, 180&#197;, 3 &#181;m stationary phase.&#160;</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:219px;">Agilent MassHunter Qualitative Analysis Software</td>
			<td style="width:217px;">Agilent Technologies Canada Ltd.</td>
			<td style="width:100px;"></td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:219px;">SpectrumMill data extractors</td>
			<td style="width:217px;">Agilent Technologies Canada Ltd.</td>
			<td style="width:100px;"></td>
			<td style="width:267px;"></td>
		</tr>
	</tbody>
</table>

composition and properties of aquafaba: water recovered from commercially canned chickpeas

Chickpea and other pulses are commonly sold as canned products packed in a thick solution or a brine. This solution has recently been shown to produce stable foams and emulsions, and can act as a thickener. Recently interest in this product has been enhanced through the internet where it is proposed that this solution, now called aquafaba by a growing community, can be used a replacement for egg and milk protein. As aquafaba is both new and being developed by an internet based community little is known of its composition or properties. Aquafaba was recovered from 10 commercial canned chickpea products and correlations among aquafaba composition, density, viscosity and foaming properties were investigated. Proton NMR was used to characterize aquafaba composition before and after ultrafiltration through membranes with different molecular weight cut offs (MWCOs of 3, 10, or 50 kDa). A protocol for electrophoresis, and peptide mass fingerprinting is also presented. Those methods provided valuable information regarding components responsible for aquafaba functional properties. This information will allow the development of practices to produce standard commercial aquafaba products and may help consumers select products of superior or consistent utility.

Each can of chick peas is labeled to indicate the ingredients added during canning. Ingredients included water, chick peas, salt, and disodium ethylenediamine tetraacetic acid (EDTA). In addition, two cans were labeled as &#34;may contain calcium chloride&#34;. Three distinct lining colours were observed; white, clear yellow and metallic (Table 1).
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	<tbody>
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			<td>Brand c...

Watch this Scientific Journal Video about Composition and Properties of Aquafaba: Water Recovered from Commercially Canned Chickpeas at JoVE.com

Composition and Properties of Aquafaba: Water Recovered from Commercially Canned Chickpeas

Aquafaba is a viscous juice from canned chickpea that, when stirred vigorously, produces a relatively stable white froth or foam. The primary research goal is to identify the components of aquafaba that contribute viscosifying/thickening properties using nuclear magnetic resonance (NMR), ultrafiltration, electrophoresis, and peptide mass fingerprinting.

Chickpea and other pulses are commonly sold as canned products packed in a thick solution or a brine. This solution has recently been shown to produce ...

composition-properties-aquafaba-water-recovered-from-commercially

Research

JoVE Journal

Environment

33.2K Views.  University of Saskatchewan. Aquafaba is a viscous juice from canned chickpea that, when stirred vigorously, produces a relatively stable white froth or foam. The primary research goal is to identify the components of aquafaba that contribute viscosifying/thickening properties using nuclear magnetic resonance (NMR), ultrafiltration, electrophoresis, and peptide mass fingerprinting. 

Video: Composition and Properties of Aquafaba: Water Recovered from Commercially Canned Chickpeas

Laboratory-determined Phosphorus Flux from Lake Sediments as a Measure of Internal Phosphorus Loading

Eutrophication is a water quality issue in lakes worldwide, and there is a critical need to identify and control nutrient sources. Internal phosphorus (P) loading from lake sediments can account for a substantial portion of the total P load in eutrophic, and some mesotrophic, lakes. Laboratory determination of P release rates from sediment cores is one approach for determining the role of internal P loading and guiding management decisions. Two principal alternatives to experimental determination of sediment P release exist for estimating internal load: in situ measurements of changes in hypolimnetic P over time and P mass balance. The experimental approach using laboratory-based sediment incubations to quantify internal P load is a direct method, making it a valuable tool for lake management and restoration.
Laboratory incubations of sediment cores can help determine the relative importance of internal vs. external P loads, as well as be used to answer a variety of lake management and research questions. We illustrate the use of sediment core incubations to assess the effectiveness of an aluminum sulfate (alum) treatment for reducing sediment P release. Other research questions that can be investigated using this approach include the effects of sediment resuspension and bioturbation on P release.
The approach also has limitations. Assumptions must be made with respect to: extrapolating results from sediment cores to the entire lake; deciding over what time periods to measure nutrient release; and addressing possible core tube artifacts. A comprehensive dissolved oxygen monitoring strategy to assess temporal and spatial redox status in the lake provides greater confidence in annual P loads estimated from sediment core incubations.

Lake eutrophication is a water quality issue worldwide, making the need to identify and control nutrient sources critical. Laboratory determination of phosphorus release rates from sediment cores is a valuable approach for determining the role of internal phosphorus loading and guiding management decisions.

Eutrophication is a water quality issue in lakes worldwide, and there is a critical need to identify and control nutrient sources. Internal phosphorus (P) ...

EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. I. Collection of Virus Samples

EPA Method 1615 was developed with a goal of providing a standard method for measuring enteroviruses and noroviruses in environmental and drinking waters. The standardized sampling component of the method concentrates viruses that may be present in water by passage of a minimum specified volume of water through an electropositive cartridge filter. The minimum specified volumes for surface and finished/ground water are 300 L and 1,500 L, respectively. A major method limitation is the tendency for the filters to clog before meeting the sample volume requirement. Studies using two different, but equivalent, cartridge filter options showed that filter clogging was a problem with 10% of the samples with one of the filter types compared to 6% with the other filter type. Clogging tends to increase with turbidity, but cannot be predicted based on turbidity measurements only. From a cost standpoint one of the filter options is preferable over the other, but the water quality and experience with the water system to be sampled should be taken into consideration in making filter selections.

EPA Method 1615 uses an electropositive filter to concentrate enteroviruses and noroviruses in environmental and drinking waters. This manuscript describes the procedure for collecting samples for Method 1615 analyses.

EPA Method 1615 was developed with a goal of providing a standard method for measuring enteroviruses and noroviruses in environmental and drinking waters. ...

EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. Part III. Virus Detection by RT-qPCR

EPA Method 1615 measures enteroviruses and noroviruses present in environmental and drinking waters. This method was developed with the goal of having a standardized method for use in multiple analytical laboratories during monitoring period 3 of the Unregulated Contaminant Monitoring Rule. Herein we present the protocol for extraction of viral ribonucleic acid (RNA) from water sample concentrates and for quantitatively measuring enterovirus and norovirus concentrations using reverse transcription-quantitative PCR (RT-qPCR). Virus concentrations for the molecular assay are calculated in terms of genomic copies of viral RNA per liter based upon a standard curve. The method uses a number of quality controls to increase data quality and to reduce interlaboratory and intralaboratory variation. The method has been evaluated by examining virus recovery from ground and reagent grade waters seeded with poliovirus type 3 and murine norovirus as a surrogate for human noroviruses. Mean poliovirus recoveries were 20% in groundwaters and 44% in reagent grade water. Mean murine norovirus recoveries with the RT-qPCR assay were 30% in groundwaters and 4% in reagent grade water.

Here we present a procedure to quantify enterovirus and norovirus in environmental and drinking waters using reverse transcription-quantitative PCR. Mean virus recovery from groundwater with this standardized procedure from EPA Method 1615 was 20% for poliovirus and 30% for murine norovirus.

EPA Method 1615 measures enteroviruses and noroviruses present in environmental and drinking waters. This method was developed with the goal of having a ...

Protocol for Measuring the Thermal Properties of a Supercooled Synthetic Sand-water-gas-methane Hydrate Sample

Methane hydrates (MHs) are present in large amounts in the ocean floor and permafrost regions. Methane and hydrogen hydrates are being studied as future energy resources and energy storage media. To develop a method for gas production from natural MH-bearing sediments and hydrate-based technologies, it is imperative to understand the thermal properties of gas hydrates.
The thermal properties' measurements of samples comprising sand, water, methane, and MH are difficult because the melting heat of MH may affect the measurements. To solve this problem, we performed thermal properties' measurements at supercooled conditions during MH formation. The measurement protocol, calculation method of the saturation change, and tips for thermal constants' analysis of the sample using transient plane source techniques are described here.
The effect of the formation heat of MH on measurement is very small because the gas hydrate formation rate is very slow. This measurement method can be applied to the thermal properties of the gas hydrate-water-guest gas system, which contains hydrogen, CO2, and ozone hydrates, because the characteristic low formation rate of gas hydrate is not unique to MH. The key point of this method is the low rate of phase transition of the target material. Hence, this method may be applied to other materials having low phase-transition rates.

We present a protocol for measuring the thermal properties of synthetic hydrate-bearing sediment samples comprising sand, water, methane, and methane hydrate.

Methane hydrates (MHs) are present in large amounts in the ocean floor and permafrost regions. Methane and hydrogen hydrates are being studied as future ...

Screening for Endocrine Activity in Water Using Commercially-available In Vitro Transactivation Bioassays

In vitro transactivation bioassays have shown promise as water quality monitoring tools, however their adoption and widespread application has been hindered partly due to a lack of standardized methods and availability of robust, user-friendly technology. In this study, commercially available, division-arrested cell lines were employed to quantitatively screen for endocrine activity of chemicals present in water samples of interest to environmental quality professionals. A single, standardized protocol that included comprehensive quality assurance/quality control (QA/QC) checks was developed for Estrogen and Glucocorticoid Receptor activity (ER and GR, respectively) using a cell-based Fluorescence Resonance Energy Transfer (FRET) assay. Samples of treated municipal wastewater effluent and surface water from freshwater systems in California (USA), were extracted using solid phase extraction and analyzed for endocrine activity using the standardized protocol. Background and dose-response for endpoint-specific reference chemicals met QA/QC guidelines deemed necessary for reliable measurement. The bioassay screening response for surface water samples was largely not detectable. In contrast, effluent samples from secondary treatment plants had the highest measurable activity, with estimated bioassay equivalent concentrations (BEQs) up to 392 ng dexamethasone/L for GR and 17 ng 17&#946;-estradiol/L for ER. The bioassay response for a tertiary effluent sample was lower than that measured for secondary effluents, indicating a lower residual of endocrine active chemicals after advanced treatment. This protocol showed that in vitro transactivation bioassays that utilize commercially available, division-arrested cell &#34;kits&#34;, can be adapted to screen for endocrine activity in water.

Screening for Endocrine Activity in Water Using Commercially-available In Vitro Transactivation Bioassays

A protocol to screen for endocrine activity in organic extracts of water samples, including treated wastewater effluent and surface (receiving) water, was adapted using commercially available division-arrested ("freeze and thaw") in vitro transactivation bioassays.

In vitro transactivation bioassays have shown promise as water quality monitoring tools, however their adoption and widespread application has been ...

Extraction of Organochlorine Pesticides from Plastic Pellets and Plastic Type Analysis

Plastic resin pellets, categorized as microplastics (&#8804;5 mm in diameter), are small granules that can be unintentionally released to the environment during manufacturing and transport. Because of their environmental persistence, they are widely distributed in the oceans and on beaches all over the world. They can act as a vector of potentially toxic organic compounds (e.g., polychlorinated biphenyls) and might consequently negatively affect marine organisms. Their possible impacts along the food chain are not yet well understood. In order to assess the hazards associated with the occurrence of plastic pellets in the marine environment, it is necessary to develop methodologies that allow for rapid determination of associated organic contaminant levels. The present protocol describes the different steps required for sampling resin pellets, analyzing adsorbed organochlorine pesticides (OCPs) and identifying the plastic type. The focus is on the extraction of OCPs from plastic pellets by means of a pressurized fluid extractor (PFE) and on the polymer chemical analysis applying Fourier Transform-InfraRed (FT-IR) spectroscopy. The developed methodology focuses on 11 OCPs and related compounds, including dichlorodiphenyltrichloroethane (DDT) and its two main metabolites, lindane and two production isomers, as well as the two biologically active isomers of technical endosulfan. This protocol constitutes a simple and rapid alternative to existing methodology for evaluating the concentration of organic contaminants adsorbed on plastic pieces.

Microplastics act as vector of potentially toxic organic contaminants with unpredictable effects. This protocol describes an alternative methodology for assessing the levels of organochlorine pesticides adsorbed on plastic pellets and identifying the polymer chemical structure. The focus is on pressurized fluid extraction and attenuated total reflectance Fourier transform infrared spectroscopy.

Plastic resin pellets, categorized as microplastics (&#8804;5 mm in diameter), are small granules that can be unintentionally released to the environment ...

Capturing Flow-weighted Water and Suspended Particulates from Agricultural Canals During Drainage Events

The purpose of this study is to describe the methods used to capture flow-weighted water and suspended particulates from farm canals during drainage discharge events. Farm canals can be enriched by nutrients such as phosphorus (P) that are susceptible to transport. Phosphorus in the form of suspended particulates can significantly contribute to the overall P loads in drainage water. A settling tank experiment was conducted to capture suspended particulates during discrete drainage events. Farm canal discharge water was collected in a series of two 200 L settling tanks over the entire duration of the drainage event, so as to represent a composite subsample of the water being discharged. Imhoff settling cones are ultimately used to settle out the suspended particulates. This is achieved by siphoning water from the settling tanks via the cones. The particulates are then collected for physico-chemical analyses.

Nutrients present in particulate form can contribute significantly to the overall loads in agricultural drainage waters. This study describes a novel method to capture flow-weighted water and suspended particulates from farm canal drainage over the entire duration of the drainage event.

The purpose of this study is to describe the methods used to capture flow-weighted water and suspended particulates from farm canals during drainage ...

Construction of a Low-cost Mobile Incubator for Field and Laboratory Use

Incubators are essential for a range of culture-based microbial methods, such as membrane filtration followed by cultivation for assessing drinking water quality. However, commercially available incubators are often costly, difficult to transport, not flexible in terms of volume, and/or poorly adapted to local field conditions where access to electricity is unreliable. The purpose of this study was to develop an adaptable, low-cost and transportable incubator that can be constructed using readily available components. The electronic core of the incubator was first developed. These components were then tested under a range of ambient temperature conditions (3.5 &#176;C - 39 &#176;C) using three types of incubator shells (polystyrene foam box, hard cooler box, and cardboard box covered with a survival blanket). The electronic core showed comparable performance to a standard laboratory incubator in terms of the time required to reach the set temperature, inner temperature stability and spatial dispersion, power consumption, and microbial growth. The incubator set-ups were also effective at moderate and low ambient temperatures (between 3.5 &#176;C and 27 &#176;C), and at high temperatures (39 &#176;C) when the incubator set temperature was higher. This incubator prototype is low-cost (&lt; 300 USD) and adaptable to a variety of materials and volumes. Its demountable structure makes it easy to transport. It can be used in both established laboratories with grid power or in remote settings powered by solar energy or a car battery. It is particularly useful as an equipment option for field laboratories in areas with limited access to resources for water quality monitoring.

This paper describes a method for building an adaptable, low-cost and transportable incubator for microbial testing of drinking water. Our design is based on widely available materials and can operate under a range of field conditions, while still offering the advantages of higher-end laboratory-based models.

Incubators are essential for a range of culture-based microbial methods, such as membrane filtration followed by cultivation for assessing drinking water ...

Composition and Distribution Analysis of Bioaerosols Under Different Environmental Conditions

Variable microorganisms in particulate matter (PM) under different environmental conditions may have significant impacts on human health. In this study, we described a protocol for multiple analyses of the biological compositions in environmental PM. Five experiments are presented: (1) PM number monitoring by using a laser particle counter; (2) PM collection by using a cyclonic aerosol sampler; (3) PM collection by using a high-volume air sampler with filters; (4) culturable microbes collection by the Andersen six-stage sampler; and (5) detection of biological composition of environmental PM by bacterial 16SrDNA and fungal ITS region sequencing. We selected hazy days and a livestock farm as two typical examples of the application in this protocol. In this study, these two sampling methods, cyclonic aerosol sampler and filter sampler, showed different sampling efficiency. The cyclonic aerosol sampler performed much better in terms of collecting bacteria, while these two methods showed the same efficiency in collecting fungi. Filter samplers can work under low temperature conditions while cyclonic aerosol samplers have a sampling limitation for temperature. A solid impacting sampler, such as an Andersen six-stage sampler, can be used to sample bioaerosols directly into the culture medium, which increases the survival rate of culturable microorganisms. However, this method mainly relies on culture while more than 99% of microbes cannot be cultured. DNA extracted from the culturable bacteria collected by the Andersen six-stage sampler and samples collected by cyclonic aerosol sampler and filter sampler were detected by bacterial 16S rDNA and fungal ITS region sequencing.All the methods above may have wide application in many fields of study, such as environmental monitoring and airborne pathogen detection. From these results, we can conclude that these methods can be used under different conditions and may help other researchers further explore the health impacts of environmental bioaerosols.

Understanding the biological composition of environmental particulate matter is important for the study of its significant impacts on human health and disease spread. Here, we used three types of bioaerosol sampling methods and a biological analysis of airborne microbes to better explore airborne microbial communities under different environmental conditions.

Variable microorganisms in particulate matter (PM) under different environmental conditions may have significant impacts on human health. In this study, ...

Xylem Water Distribution in Woody Plants Visualized with a Cryo-scanning Electron Microscope

A scanning electron microscope installed cryo-unit (cryo-SEM) allows specimen observation at subzero temperatures and has been used for exploring water distribution in plant tissues in combination with freeze fixation techniques using liquid nitrogen (LN2). For woody species, however, preparations for observing the xylem transverse-cut surface involve some difficulties due to the orientation of wood fibers. Additionally, higher tension in the water column in xylem conduits can occasionally cause artifactual changes in water distribution, especially during sample fixation and collection. In this study, we demonstrate an efficient procedure to observe the water distribution within the xylem of woody plants in situ&#160;by using a cryostat and cryo-SEM. At first, during sample collection, measuring the xylem water potential should determine whether high tension is present in the xylem conduits. When the xylem water potential is low (&#60; ca. &#8722;0.5 MPa), a tension relaxation procedure is needed to facilitate better preservation of the water status in xylem conduits during sample freeze fixation. Next, a watertight collar is attached around the tree stem and filled with LN2&#160;for freeze fixation of the water status of xylem. After harvesting, care should be taken to ensure that the sample is preserved frozen while completing the procedures of sample preparation for observation. A cryostat is employed to clearly expose the xylem transverse-cut surface. In cryo-SEM observations, time adjustment for freeze-etching is required to remove frost dust and accentuate the edge of the cell walls on the viewing surface. Our results demonstrate the applicability of cryo-SEM techniques for the observation of water distribution within xylem at cellular and subcellular levels. The combination of cryo-SEM with non-destructive in situ observation techniques will profoundly improve the exploration of woody plant water flow dynamics.

Observing the water distribution within the xylem provides significant information regarding water flow dynamics in woody plants. In this study, we demonstrate the practical approach to observe xylem water distribution in situ&#160;by using a cryostat and cryo-SEM, which eliminates artifactual changes in the water status during sample preparation.

A scanning electron microscope installed cryo-unit (cryo-SEM) allows specimen observation at subzero temperatures and has been used for exploring water ...

Composition and Properties of Aquafaba: Water Recovered from Commercially Canned Chickpeas

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Xylem Water Distribution in Woody Plants Visualized with a Cryo-scanning Electron Microscope