Research project

I have a research project that I need help with. I have done mostly half of the research. I only needs help with:oResults: You can start with some of your own observations if you have any. Primarily you will be using data that I provide you with which is fictional but should be realistic to an actual experiment. Summarize your findings in focused, brief statements. You should include something from both objective and subjective testing oCharts and Graphics: These will be produced by putting your findings and results into charts or graphs. You should include something from both objective and subjective testing. oFood Science Component: Food chemistry NOT NUTRITON science. Use lecture material, journal articles you found, textbooks. Cite all references clearly, even my notes. oConclusions: Summarize your conclusions with succinct, focused statements. Did you answer the research question? oLimitations: Every experiment has definite limitations. Some limitations are obvious and apply to all the teams. Do not state you used fake data. Pretend that the data is real. oImplications: What does your work show that food scientists, consumers, food industry would benefit from? Maybe more needs to be done before conclusions are so obvious. I have attached multiple of articles to help with the food science component (only this part requires some sources).NOTE: I have attached this article to take some idea about what’s needed. (Using Egg Replacers in A Custard Style Pumpkin Pie)I will also send the research paper that needs to be completed once its assigned.


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Eur Food Res Technol (2018) 244:97–104
Application of pulses cooking water as functional ingredients:
the foaming and gelling abilities
Sophie E. Stantiall1 · Kylie J. Dale1 · Faith S. Calizo1 · Luca Serventi1
Received: 1 May 2017 / Revised: 9 June 2017 / Accepted: 24 June 2017 / Published online: 5 July 2017
© Springer-Verlag GmbH Germany 2017
Abstract Recent interest in pulses has resulted in the
application of their cooking water (PCW) as replacement
for egg white in meringues. Nonetheless, scientific understanding of their chemical composition, physicochemical
properties and effect on food quality is lacking. This study
analysed the PCW of haricot beans, garbanzo chickpeas,
whole green lentils and split yellow peas, determining their
composition and effect on meringues’ quality. The PCW
tested contained different amounts of sugar, soluble and
insoluble fibre, protein, ash and saponins. All PCW exerted
foaming ability (39–97%), directly correlated to their
protein content and lower than that of egg white (400%).
Moreover, gelling ability was observed, resulting in the
development of egg-like meringues, with their hardness
inversely correlated to the content of insoluble fibre. Whole
green lentils expressed the highest foaming ability while
garbanzo chickpeas resulted in the strongest gelling ability.
Sensory analysis of meringues depicted low acceptance for
the taste of meringues made with the PCW of haricot beans
and whole green lentils. On the contrary, high acceptance
was determined for garbanzo chickpeas and split yellow
peas, similarly to that of the egg white.
Keywords Fibre · Functional ingredients · Protein ·
Pulses · Texture
* Luca Serventi
Department of Wine, Food and Molecular Biosciences,
Faculty of Agriculture and Life Sciences, Lincoln University,
RFH Building, Lincoln, PO Box 85054, Christchurch 7647,
New Zealand
Foaming and gelling ingredients are often used in food to
provide desired textural quality. Foaming agents are commonly used in angel cake, meringue and mousse to produce
food with light texture [1]. A foam is a two-phase system
of gas dispersed within a liquid phase, formed through
whipping or other means of agitation where gas bubbles
are entrapped within the foam by two different films [2].
The foaming ability of a solution is influenced by surface
activity, surface tension of the water–air interface, the filmforming properties of the foaming agent and the ability to
rapidly absorb on the air–liquid interface [1]. Proteins are
a major component of foam formation as they are amphiphilic, consisting of a hydrophilic end attracted to the water
phase and a hydrophobic end attracted to the air phase [3].
Egg white is a common foaming agent due to its high albumin content, providing stable foams in bakery and confectionary foods [1].
Gelling agents used in food applications are predominately hydrocolloids [4] that form a gel by dissolving in a
liquid phase as a colloid mixture, thus developing a weak
cohesive internal structure [5]. Several studies described
the gelling ability of soluble fibre components such as cellulose and pectin, which are common hydrocolloids used
as gelling agents [4, 5]. Their ability to disperse into minute particles when in aqueous environments provides these
hydrocolloids with the capacity to gel [6].
Recent interest in plant-based foods has focused the
attention of research into alternative foaming agents to
replace egg white, such as chickpea cooking water [7].
The cooking water from different pulses such as beans,
chickpeas, lentils and peas might contain varying levels
of carbohydrates, protein and saponins that may provide
foaming and gelling abilities. After soaking and cooking
of chickpeas and kidney beans, a considerable decrease
in sugars was observed [8]. This would indicate that the
portion of soluble carbohydrates lost was dispersed into
the cooking water. These findings are in agreement with
another study [9] showing that a significant portion of
reducing sugars, sucrose, raffinose, stachyose and verbascose was lost through diffusion into the chickpea cooking
water and also in the germination process through hydrolysis. A recent study discovered that high concentrations
water-soluble polysaccharides from chickpea flour had the
ability to from stable foams [10]. Cooking was also shown
to decrease protein and ash content of chickpeas [11] thus
indicating that small fractions of protein might leach from
pulses to their cooking water. Also, phytochemicals contained in legumes and known as saponins are able to emulsify due to their chemical structure [12]. A previous review
[13] described how the amphipathic nature of these glycosides allows for the incorporation of water and air, causing
saponins to foam.
We hypothesised that carbohydrates, protein and saponins may impart foaming and gelling abilities to pulses
cooking water (PCW). Therefore, our objectives were to
assess the composition of the PCW, their physicochemical properties and effect on food quality. Meringues were
chosen as food matrix to study foaming and gelling abilities and to determine consumer acceptance of the PCW as
functional ingredients.
Materials and methods
Haricot beans (Sun Valley Foods, Auckland, New Zealand), garbanzo chickpeas (Kelley Bean Co, Scottsbluff,
NE, USA), whole green lentils (McKenzie’s, Victoria, Australia), split yellow peas (Cates, Ashburton, New Zealand)
and eggs (Weedons large sized eggs, New Zealand) were
tested for this study. Eggs were kept refrigerated at 4 °C and
the albumen (egg white) separated from the yolk by hand.
The egg white used was determined to have a moisture content of 88.4 ± 0.2 g/100 g. Soaking in a 1:3.3 weight ratio
(dry pulse:water) was performed for 16 h to achieve tenderness [14]. Then, soaked pulses were drained. Cooking
in boiling water (weight ratio 1:1.75 dry pulse:water) was
carried until tenderness of the cooked pulse [11]: this step
required 90 min with the exception of whole green lentils
(60 min). Cooking time was taken from the start of heating. Once boiled, the PCW were allowed to cool to room
temperature (25 °C); this process required about 120 min
and yielded approximately 0.6 g PCW/g pulse. Cooled
PCW were drained from the cooked pulses and stored at
Eur Food Res Technol (2018) 244:97–104
-18 °C, then thawed overnight at room temperature prior
to the analyses.
Meringue recipe
Each batch consisted of 160 g of foaming agent (PCW or
egg white), 115 g of icing sugar (Pams, New Zealand) and
of 115 g caster sugar (Pams, New Zealand). The foaming
agent was whipped for 3 min with a Delta Food Equipment
mixer, model number 500A (New Zealand), at a medium
speed (level 3). The caster and icing sugars were then added
to the mixture and whipped for a further 13 min on high
(level 5). Then, doses of 25 g were weighed and placed on
a baking tray. Meringue mixtures were baked at 100 °C for
75 min in a Moffat Ltd oven, model E32 M (Rolleston, IN,
Proximate composition and total saponin quantification
of PCW
Dry matter content was determined by oven drying at
105 °C overnight as by the AACC method 44-15A [15].
Samples were freeze-dried prior to proximate analysis and
oven dried (65 °C, overnight) prior to saponin analysis.
The extraction of high molecular weight (HMW) and low
molecular weight (LMW) water-soluble carbohydrates followed a method developed from Pollock and Jones [16],
while their quantification followed a method developed by
Jermyn [17]. Briefly, 25 mg of samples were weighed in
a 2 ml tube, vortexed with 1 ml of 80% aqueous ethanol
and shaken for 30 min at 65 °C. Samples were then centrifuged for 15 min at 13,000 rpm to separate the supernatant. This process was performed twice. For quantification,
12 µl of each LMW extract was pipetted into a microwell
plate using a single channel pipette and 188 µl of water
were added using a multi-channel pipette. Next, 40 µl of
the diluted extract were pipetted to a new 96 microwell
plate in triplicate along with each of the standard solutions
in triplicate. Finally, 200 µl of the anthrone reagent (in sulfuric acid) were added to each well (multi-channel) and
mixed; solutions were then incubated in an oven for 25 min
at 65 °C. Samples were analysed using a microplate reader
after being shaken for 5 s and at absorbance measured at
620 nm. For extraction of HMW the protocol above was
slightly modified: the 1 ml of ethanol was replaced with
1 ml of water. After centrifugation, the supernatant was
extracted into a new 2 ml Eppendorf tube. The supernatant samples were stored at -20 °C until quantification. For
quantification, sucrose and inulin standards were prepared,
respectively. The same method was used for HMW as
LMW except for 40 µl of the original extract used instead
of the 12 µl used for LMW analysis.
Eur Food Res Technol (2018) 244:97–104
Protein content was determined by quantifying total
nitrogen. Freeze-dried samples were combusted in a Dumas
type elementar analyser (Elementar Rapid-N Exceed,
Hanu, Germany) according to the method 993.13 [18],
while fat content was determined by Soxhlet extraction in
hexane using a Soxhlet Extraction Unit E-816HE (Buchi,
Switzerland) by the method 991.36 [18]. Ash content was
determined via oven drying as by the method 930.22 [18].
Iodine test was performed to assess the presence of starch
by colour reaction to a triiodide anion (I3-) complex.
Insoluble fibre was determined by difference, subtracting
mean values of LMW and HMW carbohydrates, protein
and ash from the dry matter content.
Total saponin content of the PCW was determined
by colorimetric assay based on an optimized method by
Ncube and others [19]. Briefly, 0.1 g of PCW powder was
extracted in a centrifuge tube with 20 ml of 70% aqueous
ethanol (BioLab, Australia), vortexed for 30 s and then centrifuged (Heraeus Multifuge X1R, Thermo Scientific, MA,
USA) at 2000 rpm for 10 min. In a test tube, 500 µl of the
supernatant obtained from centrifugation was mixed with
500 µl vanillin (Sigma Aldrich, USA) in 8% ethanol solution and 5000 µl of 72% sulfuric acid (Fisher Scientific,
USA) in aqueous solution. The mixture was vortexed for
30 s and placed in a water bath at 65 °C for 10 min, then
cooled to room temperature (25 °C) in a cold water bath for
5 min and transferred into a 3 ml cuvette for spectrophotometric analysis. Absorbance at 544 nm was measured with
a spectrophotometer V-1200 (VWR, USA). Soyasaponin I,
95% pure (Sigma Aldrich, USA) was used as standard saponin to build a calibration curve. Total saponin content was
expressed in mg/g.
Physiochemical analysis of foaming agents
The four PCW were characterized for basic physicochemical properties such pH, density, viscosity and foaming ability, in comparison to egg white to evaluate their feasibility
as foaming agents. A Mettler Toledo pH metre (SevenEasy
pH, Switzerland) was used to measure the pH of the samples. Density was measured with a hydrometer (Peter Stevenson LTD, Scotland), while viscosity was determined
using the RM100 model viscometer (Lamy Rheology,
France). A 30 ml sample of foaming agent was analysed
at room temperature with measuring system 12 and shear
rate of 14.1 s-1. Finally, to measure the foaming ability
(FA) of the foaming agents, an optimized method based on
that proposed by Sathe and Salunke [20] was developed. A
50 ml sample of PCW or egg white was whipped for 7 min
in a Delta Food Equipment mixer, model number 500A, at
speed 5. The FA, expressed as a percent (%) was calculated
by the following equation, where Vf was the volume of the
final foam and Vi was the volume of the initial liquid:
FA(%) =
Vf – Vi
× 100
Characterisation of meringues
Moisture content of the meringues was analysed following
the AACC method 44-15A [15]. Meringues’ weight was
measured on an analytical scale. Height of the meringues
was measured with a calliper (1112-150, Insize Co. ltd,
Suzhou New District, China). Volume of the final products was measured via the AACC method number 10-05.01
[21]. A TA.XT. Texture Analyser (Stable Micro Systems,
Godalming, England) was used to record the hardness
and extensibility of the meringues upon single compression until breaking point. One whole meringue was placed
on the base for each measurement and the following settings were used: load cell 5 kg, aluminium probe P/25,
pre-test speed 1.0 mm/s, test speed 1.7 mm/s, post-test
speed 10.0 mm/s, auto trigger type of 5 g. Lastly, a CR-210
Chroma Metre colorimeter (Minolta, Japan) was used to
measure the colour of the meringues’ crust and expressed
on the CIE Lab scale.
Sensory analysis of meringues
Sensory analysis of meringues was conducted with
untrained panellists to gain information on consumer
acceptance of PCW as ingredients. 40 untrained panellists were recruited at Lincoln University (New Zealand).
Appearance was evaluated based on photos of the whole
meringues (Fig. 1), while taste, texture and overall preference were evaluated upon tasting. The test was conducted
in individual sensory booths and 9-point hedonic scales
were used for all ratings.
Statistical analysis
All measurements were taken in triplicates by analysing three different samples of PCW (liquid and dried) and
meringues. ­Microsoft® ­Excel® for Windows 2013 was used
for statistical analysis to process raw data in triplicate to
gain the mean, standard deviation and standard error. The
same software was used to calculate the following correlations: foaming ability with protein content of PCW;
meringues’ hardness with insoluble fibre content of PCW.
One-way analysis of variance (ANOVA) with Tukey test
was conducted through ­
Minitab®17. The ANOVA test
assumed equal variances for the analysis and a significance
level of a = 0.05.
Eur Food Res Technol (2018) 244:97–104
Fig. 1 ?Pictures of the
meringues: front view (a) and
top view (b)
Results and discussion
Determination of dry matter revealed significant differences (p < 0.001) in solid loss among the pulses studied (Table 1). Boiled garbanzo chickpeas released the highest amount of solid in the PCW (5.13/100 g) (Table 1). Similar values of dry matter were determined for the other pulses, with the exception of haricot beans, which only released 3.28/100 g of dry matter in the PCW (Table 1). Previous studies [8, 9, 11] reported significant losses of oligosaccharides, fibre and protein boiled legumes. The extent of the loss in the cooked legumes determined in those studies was comparable to the concentration of dry matter that we quantified in the PCW. Degradation of the external layer of chickpea and lentil seeds upon boiling was visible, possibly explained their high losses. Differently, the form of yellow peas (split) might have caused the high loss. Looking at specific components, a high amount of watersoluble carbohydrates (WSC) was found. Within WSC, the vast majority of them was determined to be LMW carbohydrates, likely sugars [17], with about 1/100 g content for all PCW, except for whole green lentils (0.54 ± 0.03/100 g, Table 1), in agreement with previous studies on cooking loss from pulses [8, 9, 11, 22]. Sugars likely consisted of sucrose, raffinose and stachyose as determined by others by analysis of raw and boiled pulses [9, 22]. The HMW carbohydrates were also identified, with particularly high quantity in the PCW of haricot beans (0.16 ± 0.20/100 g, Table 1). The HMW probably consisted of soluble fibre [17]. Total nitrogen determination by Dumas revealed a significant amount of protein in all PCW, as result of loss from the seeds upon boiling [11]. Estimated protein content ranged from 0.70/100 g of haricot beans to 1.51/100 g of whole green lentils (Table 1). The smaller size of whole green lentils and split yellow peas might explain their higher protein content due to superior solubilisation. No traces of fats were detected (Table 1). Previous studies on boiled chickpeas [9, 11] reported significant loss of small fractions of fats upon boiling. It is possible that in our case the loss was too small to be detected or that those fats degraded during processing. Interestingly, a high portion (almost half) of dry matter was not quantified by the methods described above and was attributed to water insoluble carbohydrates, with starch and insoluble fibre possibly representing it [23]. Iodine test resulted negative for all PCW, thus reasonably excluding the presence of starch. By difference, only insoluble fibre was the likely component of this fraction. Further research may elucidate whether the insoluble fibre fraction consisted of cellulose, hemicellulose, lignin or pectin. Previous studies on the effect of Table 1 ?Proximate composition and saponin content of the pulses cooking water (PCW) d Composition of the PCW Nutritional information Dry matter (g/100 g) LMW (g/100 g) HMW (g/100 g) Insoluble fibre (g/100 g, by difference) Protein (g/100 g) Fat (g/100 g) Ash (g/100 g) Saponins (mg/g) Haricot beans Garbanzo chickpeas Whole green lentils Split yellow peas 3.28 ± 0.5 0.73 ± 0.03c 0.16 ± 0.20a 0.93 ± 0.05d a 5.13 ± 0.02 1.20 ± 0.02a 0.04 ± 0.00c 2.37 ± 0.02a b 4.69 ± 0.02 0.54 ± 0.03d 0.07 ± 0.00b 2.09 ± 0.02b 4.41 ± 0.18c 1.02 ± 0.03b 0.09 ± 0.00b 1.63 ± 0.18c 0.70 ± 0.00d Purchase answer to see full attachment

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