Meat Science 202 (2023) 109217

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Meat Science
journal homepage: www.elsevier.com/locate/meatsci

Effects of the use of raw or cooked chickpeas and the sausage cooking time
on the quality of a lamb-meat, olive-oil emulsion-type sausage
S.A. Kasaiyan a, I. Caro b, *, D.D. Ramos c, B.K. Salvá d, A. Carhuallanqui c, M. Dehnavi a, J. Mateo a
a
Departamento de Higiene y Tecnología de los Alimentos, Universidad de León, Campus Vegazana s/n, 24007 León, Spain
b
Facultad de Medicina, Universidad de Valladolid, Avenida Ramón y Cajal 7, 47005 Valladolid, Spain
c
Laboratorio de Salud Pública y Salud Ambiental, Facultad de Medicina Veterinaria, Universidad Nacional Mayor de San Marcos, Av. Circunvalación Cuadra 28, San
Borja, Lima, Peru
d
Facultad de Ciencias de los Alimentos, Universidad Le Cordon Bleu, Av. General Salaverry, Magdalena del Mar, Lima 3180, Peru

A R T I C L E I N F O A B S T R A C T

Keywords: Reformulation of cooked sausages using high-protein plant-based food such as chickpea as meat extenders and
Meat extender vegetable oils to replace animal fat can be a suitable approach to promote the consumption of smaller portions of
Pulse meat. The pre-processing of chickpea and the sausage cooking intensity can potentially affect the quality of
Cooked sausage
reformulated sausages. In this study, an emulsion-type sausage made with lamb meat, chickpea and olive oil was
Sustainability
prepared in triplicate following three different formulations containing the same targeted levels of protein
(8.9%), lipids (21.5%), and starch (2.9%): control sausage (CON; control, without chickpea), and raw (RCP) and
cooked chickpea (CCP) sausages (both with 7% chickpea). Sausages were cooked at 85 ◦ C for two heating times
(40 min or 80 min) and were analysed for weight loss, emulsion stability, colour, texture, lipid oxidation and
volatile composition. Compared to CON sausages, the use of raw chickpea reduced the elasticity and significantly
increased lipid oxidation during the sausage-making process resulting in major changes in the volatile compo­
sition. The use of previously cooked chickpea, however, resulted in the sausages having greater cooking loss,
hardness and chewiness than CON sausages, while there was no difference in lipid oxidation, and differences in
volatile compounds were scarce. The reformulation with cooked chickpea could provide a sausage with more
similarity to the CON sausage. The extended heating time of 80 min at 85 ◦ C did not significantly affect the
quality traits in either CON or reformulated sausages except for a higher cooking loss.

1. Introduction promote the consumption of smaller portions of meat (De Boer, Schösler,
& Aiking, 2014). Meat products reformulated in this manner must not
Meat and meat products, rich in high-quality protein, vitamins B12, only be as nutritious as conventional products, but also have sensory
iron, zinc and selenium, are important sources of nutrients in the human characteristics comparable to conventional products to avoid rejection
diet (Godfray et al., 2018). However, meat production can be associated by potential consumers (Hoek et al., 2011; Mateo et al., 2021).
with large environmental and climate footprints (Eckl, Bierbroek, van’t Grain legumes (pulses), in addition to cereals and pseudocereals,
Veer, & Geleijnse, 2021) and, when meat intake levels are high, or meat oilseeds and mushrooms, as well as protein extracts obtained from them,
is consumed on a long-term basis, dietary meat may be associated with have been the suggested plant-based ingredients to replace meat in
adverse health effects, such as colorectal cancer (Godfray et al., 2018; reformulated meat products (Asgar, Fazilah, Huda, Bhat, & Karim,
Richi et al., 2015). In a context of increasing rate of demand for meat and 2017). Pulses have a high nutritional value (Pintado & Delgado-Pando,
meat products and considering the above concerns, reformulation of 2020) and their incorporation into the daily diet has been shown to have
meat products by partially replacing meat with protein-rich plant foods advantages and physiological effects in the control and prevention of
or replacing animal fat with vegetable oils could be alternatives of various diseases prevalent in many countries around the world (Mudryj,
choice for the meat industry. These approaches fall under the concept of Yu, & Aukema, 2014). Among vegetable oils, olive oil has been proposed
hybrid meat products (Grasso & Jaworska, 2021) and would aim to as a candidate oil to partially or totally replace animal fat in cooked

* Corresponding author.
E-mail address: [email protected] (I. Caro).

https://doi.org/10.1016/j.meatsci.2023.109217
Received 7 February 2023; Received in revised form 30 April 2023; Accepted 3 May 2023
Available online 5 May 2023
0309-1740/© 2023 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-
nc-nd/4.0/).
S.A. Kasaiyan et al. Meat Science 202 (2023) 109217

emulsion-type sausages because of its beneficial nutritional properties activity, which in short-heating treatments require temperatures be­
and favourable sensory and functional properties (Shin et al., 2022; tween 70 ◦ C and 90 ◦ C for denaturation (He et al., 2014; Shi et al., 2020;
Shin, Lee, Lee, Jo, & Choe, 2020). Liburdi et al., 2021).
The effect of the incorporation of pulse flours in meat products on The aim of this research was to determine the effects of the use of
their quality has been studied for several decades and there is currently a chickpea as a partial meat substitute, the pre-treatment of chickpeas
renewed interest in research on this topic (Grasso & Jaworska, 2021; (raw vs. cooked) and the intensity of sausage cooking on the quality
Hill, 2022; Pintado & Delgado-Pando, 2020; Verma, Ledward, & Lawrie, characteristics of an emulsion-type cooked sausage made with lamb
1984a). However, most research has been carried out on fresh meat meat and olive oil, namely: yield, texture, oxidative stability and flavour
products, such as burgers, and less on cooked sausages. compounds.
The amount of pulse used in experimental reformulations has been
up to 50% of the meat batter, although recommended levels have 2. Materials and methods
typically been between 5% and 10%.
The use of pulses at limited quantities seems to produce minor sen­ 2.1. Sausage ingredients
sory differences compared to other meat extenders (Chigwedere,
Wanasundara, & Shand, 2021). However, pulses can significantly affect The following ingredients were used in the sausage formulation: leg
the flavour of meat products if used in high amounts. Pulses in refor­ lean meat (obtained from 4-month-old lambs, which were reared on a
mulated meat products have been associated with “bean” and “hay-like” local farm under same intensive conditions; legs were purchased two
off-flavours (Chigwedere et al., 2021), which can have a significant days after slaughter, frozen at − 18 ◦ C, stored for up to two months and
negative effect on consumer acceptance. The colour of meat products thawed for two days under refrigeration before use; meat composition
can also be affected by the effect of pigments provided by pulses and, in ranged 73–75% moisture, 19–21% protein, 3–4% fat), cream-coloured
the case of raw pulses, by an oxidative effect on myoglobin (Verma et al., chickpea (Cicer arietium, Pedrosillano variety; La Asturiana, Vidanes,
1984a). León, Spain; containing 41% starch, 18% protein, 14% fibre and 6% fat),
Among the variety of pulses used to reformulate meat products, 0.4%-acidity refined olive oil, tap water, food-grade potato starch, so­
chickpea (Cicer arietinum L) has been one of the most widely used (Mateo dium chloride, polyphosphates (Tai K-7, Pilarica, Paterna, Spain), and
et al., 2021). Chickpea has a high nutritional value, being a rich and powdered black pepper; all purchased from the local market.
cheap source of protein, with levels normally above 20%. The protein
quality of pulses in terms of essential amino acid ratio is worse than that 2.2. Experiment design
of meat due to a low content of sulphur amino acids and tryptophan;
however, it can be improved when combined with rice or egg (Farooq & Emulsion-type sausages made with lamb meat, chickpea and olive oil
Boye, 2011). Chickpea is also a rich source of fibre and a good source of were prepared following three different formulations (three treatments),
iron, zinc, magnesium, calcium, vitamin E and B9 (Kaur & Prasad, each with the same target levels of protein, lipid and starch (8.9, 21.5
2021). Moreover, due to the functionality of chickpea components, such and 2.9%, respectively): a control sausage (CON; without chickpea) and
as proteins, starch, polyphenols, or carotenoids, this pulse is in demand two sausages with part of the meat replaced by chickpea, one with raw
as a functional ingredient in a variety of food applications (Kaur & chickpea (RCP) and the other with cooked chickpea (CCP). These sau­
Prasad, 2021; Sofi, Muzaffar, Ashraf, & Gupta, 2020). The functional sages were prepared in triplicate (three batches, each prepared on a
properties of proteins and starch from chickpeas when used as meat different day) at the Food Processing Hall of the Faculty of Veterinary
extenders can contribute positively to the functional properties of meat Medicine, University of León (León, Spain). The sausages were cooked
proteins, such as water holding capacity (WHC), binding, gelation and following two heating times: conventional (72–74 ◦ C core temperature)
emulsification (Li et al., 2020; Vatansever, Tulbek, & Riaz, 2020; Wang and prolonged (82–84 ◦ C core temperature). Therefore, the experi­
et al., 2023). mental setup consisted of 3 treatments (formulations) x 2 cooking con­
The processing of pulses prior to use, i.e. grinding, shelling, soaking ditions. To evaluate the effects of treatment and cooking condition on
or cooking, can influence the impact of pulses on the quality of the meat sausage quality, the following laboratory analyses were performed the
product. Boiling pulses or roasting pulse flours prior to use have the day after cooking: weight losses, pH colour, texture, thiobarbituric
advantage of effectively removing the content of antinutritional factors reactive substances (TBARS) and volatile compounds. Two sausage
(ANFs), thus improving their digestibility (Bessada, Barreira, & Oliveira, pieces were prepared for each combination treatment-heating condition,
2019). Moreover, heating inactivates undesirable enzymes such as and the quality analyses were performed in duplicate for each sausage
oxidative enzymes, i.e. lipoxygenase (LOX; Shariati-Ievari et al., 2016; piece the day after cooking. Furthermore, to find out whether refor­
Shi, Mandal, Singh, & Pratap-Singh, 2020), or hydrolytic enzymes, i.e. mulation or heating conditions could affect the oxidation stability of the
α-amylase (Guldiken et al., 2022), thus preventing discoloration, lipids during storage, a portion of each sausage piece was covered with
off-flavours, or texture changes in reformulated meat products. On the cling film, stored for 12 days under refrigeration (4 ◦ C) and used for an
other hand, heating negatively affects the technological functionality of additional TBARS test.
pulse starch or protein (Aguilera, Esteban, Benitez, Molla, &
Martin-Cabrejas, 2009), which might result in poorer binding, gelling, 2.3. Sausage preparation and sampling
emulsion or stability performances.
Emulsion-type sausages, after stuffing, are heated in ovens or water The formulations for each of the three experimental treatments are
for pasteurisation, normally at 80 ◦ C to a core temperature near to 72 ◦ C. shown in Table 1. For the three batches of sausages, each experimental
The heating intensity is a relevant factor for gel structure, strength and treatment consisted of 2 kg of sausage mix. Each batch was made with
cooking yield in sausage making (Barbut, Gordon, & Smith, 1996; Glo­ leg meat from a different lamb. The main ingredients in CON sausage
rieux et al., 2019). In reformulated sausages containing raw pulses, formulation were meat, oil and starch (44.5, 20 and 2.9%, respectively).
conventional heating (72 ◦ C core temperature) may not be intense The formulation of the sausages with chickpeas, raw (RCP) or cooked
enough to obtain a complete gelatinisation of pulse starches, whose (CCP) sausages, was designed to have quantities of protein, lipids and
gelatinisation temperature ranges between 70 and 90 ◦ C (Farooq & starch equal to those expected in CON sausages. Considering the con­
Boye, 2011), which in turn could lead to sub-optimal texture and water centration of starch in chickpeas (41%), the amount of chickpea to be
retention. Higher than usual heating intensities, i.e. 80–90 ◦ C core used in the sausage formulation, in order to achieve 2.9% starch (2.9 g
temperature, may also be required to achieve a significant inactivation starch/100 g sausage batter), was 7% (7 g chickpea per 100 g batter).
of undesirable pulse bioactive compounds such as lectins or LOX This amount of chickpea contributed 1.3% protein to the sausage batter,

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Table 1 cling film, stored for 12 days under refrigeration (4 ◦ C) and used for an
Formulations of the control and reduced meat emulsion-type sausages. additional TBARS test.
CON RCP CCP
2.4. Batter composition and technological quality traits
Main ingredients (%)
Lamb meat 44.5 38 38
Chickpea (untreated)& – 7 7 For each batter sample, moisture, fat, protein and ash were deter­
Olive oil 20 20 20 mined according to the methods recommended by the AOAC (1999). All
Water$ 32.6 35 35 these analyses were performed in duplicate. The pH of the chickpea
Starch 2.9 – –
paste and the batters were measured in duplicate using a Basic 20
pH-meter with serial number 047021 (Crison Instruments, Barcelona,
Salts and spices (g/kg)
Spain) equipped with a penetration electrode (Hach Lange, Düsseldorf,
Sodium chloride 18 18 18
Phosphates 3 3 3 Germany). The pH meter was calibrated daily prior to use with two
Powdered black pepper 2 2 2 calibration buffers (pH 7.0 and pH 4.0), and the calibration and mea­
surements of samples were carried out with the buffers and samples at
CON: Control sausage, formulated without chickpea; RCP: sausage formulated
with soaked raw chickpeas; CCP: sausage formulated with cooked chickpeas.
room temperature (20 ± 2 ◦ C). The emulsion stability was determined,
&
The percentage refers to raw chickpea before pre-treatment (soaking or in duplicate, from a sample of 25 ± 1 g of raw batter which was placed
soaking and boiling). into a Falcon tube and centrifuged for 1 min at 600 ×g using a 2–15
$
Included in the percentage are the added water and the water absorbed by Sigma centrifuge (Osterode am Harz, Germany), following the proced­
the chickpeas during soaking and eventually cooking. ure described by Hughes, Cofrades, and Troy (1997). The filled Falcon
tubes were heated in a water bath for 30 min at 70 ◦ C and centrifuged
which led to a reduction of 6.5 % units in the amount of meat, with 20% again for 5 min at 1200 ×g. The supernatants were poured into
protein, in the RCP and CCP sausages (from 44.5% to 38%) – this meant pre-weighed crucibles and dried overnight at 100 ◦ C, and the pelleted
that 15% of the animal protein was replaced by vegetable protein, i.e. samples were extracted from the tubes and weighed. In the analysis the
CON sausage would contain 8.9% animal protein and chickpea sausages dry extracts from the supernatants were considered to be fat. The per­
7.6% animal protein. The formulations of the RCP and CCP sausages centages of fluid released from the batter after cooking and centrifuga­
complies with the permissible limits for plant-origin ingredients laid tion and the amount of dry extract in the fluid (fat) was calculated as a
down in the Meat Products Quality Standard Regulation (Spain, 2014): percentage of the initial batter.
no stablished limit for edible oils (quantum satis), <3% plant-origin
protein, <10% starch. 2.5. Sausage technological quality traits
For the RCP treatment, the chickpeas were previously soaked in tap
water for 16 h at room temperature (21 ◦ C) following the recommen­ The day after cooking, using a portion of each cooked sausage, the
dations from the provider and for the CCP treatment they were soaked in water retention capacity of the cooked sausage was determined in
the same conditions and then traditionally boiled in water for 3 h. Just duplicate following the method described by Hughes et al. (1997) with
before the mixing of the ingredients, the meat was minced using a meat slight modifications. Two grams of sausage, cut into fine strips, previ­
grinder (Mainca PM-12; Granollers, Spain) equipped with a 6-mm plate ously weighed using a precision balance (4 decimal places) were placed
size, and the soaked-raw or soaked-then-cooked chickpeas were grinded into 5-ml plastic centrifuge tubes containing a small amount of cotton, c.
with ice (1/1, w/w) in a Stephan UMC5 (Saint-Cannat, France) for 8 min a. 0.10 g, packed in the bottom. The tubes were then centrifuged at 9000
at 4 ◦ C and 2400 rpm to form a chickpea paste. A sample of the pulse xg for 10 min, at 4 ◦ C, (J2–21 centrifuge, Beckman Coulter, Barcelona,
paste was taken for pH and TBARS analyses. The sausage mix was then Spain). Afterwards, the strips of sausage were extracted from the tube
prepared in the Stephan UMC5 operating at 4 ◦ C, 2400 rpm and 50.7 × and weighed to determine the loss of water during centrifugation from
103 Pa pressure in two steps: i) minced meat, ice, NaCl, polyphosphate the weight difference.
and either starch (for CON) or the pretreated chickpeas (for RCP or CCP) Sausage colour was measured in duplicate on the cut surface of
were mixed for 4 min, and ii) olive oil was added and the mixing recently-cut 1-cm-thick sausage slices. Three different measurement
continued for another 4 min period. Afterwards, for each sausage points for each of two replicates were taken using a CM-700d portable
treatment, two portions of 70 g of the emulsified batter were sampled for spectrophotometer (Konica Minolta, Osaka, Japan) operating with D65
immediate analysis of pH, proximate composition and emulsion stabil­ illuminant D65, SCI mode, 8 mm aperture for measurement and a visual
ity, and the rest of the batter was stuffed into 34–36 mm-diameter beef angle of 10◦ . The values of the lightness (L*), redness (a*) and yellow­
casings from small intestine (Navaher, León, Spain) obtaining four ness (b*) were recorded.
similar, c.a., 0.5-kg sausage pieces per treatment. The texture profile analysis was analysed in triplicate on three 1-cm
The sausage pieces were weighed and then cooked suspended in a sized cubes each obtained from the inner part of the corresponding 1-cm
programmed oven at 85 ◦ C and 95% relative humidity for two different sausage slices. A texture analyzer TA.XT2i (Stable Micro Systems,
times. Two pieces were cooked for 40 min (conventional heating; Godalming, Surrey, UK) equipped with a 5-cm diameter cylindrical
72–74 ◦ C core temperature) and the other two for 80 min (prolonged probe running with a cross head speed of 1 mm/s and a compression
heating; 82–84 ◦ C core temperature). After cooking, the sausages were depth of 80% was used. Values for hardness, elasticity and cohesiveness
cooled in tap water for 10 min, drained at room temperature (21 ◦ C) for were recorded, and chewiness was calculated as the product of the three
30 min and weighed to calculate the cooking loss. Each (0.5-kg) sausage values (hardness × cohesiveness × springiness).
piece was then cut into three similar portions and each portion was
wrapped with polyethylene cling film and cooled in a cold chamber 2.6. Thiobarbituric acid-reactive substances (TBARS) and headspace
(4 ◦ C). The day after cooking, two portions of sausage were used for volatile composition in sausages
analysis. Prior to analysis, each of these two sausage portions was
further cut into two parts, one was used to assess water loss by centri­ The TBARS of the chickpea paste and the sausages were determined
fugation, pH, colour and texture, and the other was homogenised using a using the procedure described by Nam and Ahn (2003) with slight
domestic food processor (DP8108, 1000 W, Moulinex, Alençon, France) modification. A 2-g sample was homogenised with 20 mL distilled and
and used for analysis of chemical composition, TBARS and volatile deionised water using an IKA T-18 basic Ultra Turrax (Staufen, Ger­
compounds. The remaining (third) sausage portion was covered with many) working at 9500 rpm for 60 s. The homogenate was filtered
through a wire-mesh filter and then 1 mL of homogenate was mixed in a

3
S.A. Kasaiyan et al. Meat Science 202 (2023) 109217

screw-capped tube with 50 μL of 7.2% butylated hydroxytoluene etha­ Table 2

nolic solution and 1 mL of 20 mM thiobarbituric acid/15% trichloro­ Composition and emulsion stability of reformulated emulsion-type sausage
acetic acid. The mixture was vortexed, heated in a water bath at 80 ◦ C batters.
for 20 min, cooled in cold water for 10 min, centrifuged at 4000 ×g for CON RCP CCP P-level
20 min and filtered through a 0.45 μm hydrophilic polytetrafluoro­ Composition (%) and pH
ethylene 13 mm syringe filter (Membrane Solutions, LLC, Auburn, WA, Moisture 66.23 ± 0.28a 63.41 ± 0.12c 64.67 ± 0.24b <0.001
USA). The absorbance was then read at 531 nm. Quantification was Lipids 18.38 ± 0.26 18.92 ± 0.40 19.45 ± 0.49 0.19
performed on the basis of the absorbance of 1,1,3,3-tetra-ethoxypropane Protein 9.29 ± 0.06 9.31 ± 0.06 9.08 ± 0.02 0.09
Ash 2.87 ± 0.08 3.17 ± 0.13 2.91 ± 0.10 0.09
standard solutions. #
Starch + fibre 3.23 ± 0.16b 5.20 ± 0.42a 3.89 ± 0.60ab 0.049
The volatile composition in the sausage headspace was analysed as pH 6.28 ± 0.03b 6.27 ± 0.04b 6.35 ± 0.07a 0.010
previously described by Carballo et al. (2020) with slight modifications.
Briefly, volatile compounds were extracted from 20 mL vials containing
Emulsion stability (%)&
3 g of homogenised sausage previously incubated at 45 ◦ C for 20 min, Total fluid losses 0.66 ± 0.09 0.40 ± 0.08 0.76 ± 0.11 0.11
using a 75 μm Carboxen/polydimethylsiloxane 1-cm-coated fused-silica Oil losses 0.62 ± 0.09 0.36 ± 0.07 0.71 ± 0.10 0.10
fibre (Supelco, Bellefonte, PA, USA) for 40 min at 45 ◦ C. Afterwards,
Results are reported as means (average of triplicate batches and two samples per
volatile compounds were desorbed in the injection port of a GC 7890A batch; n = 6) ± standard errors of the mean.
chromatograph (Agilent System, Zwingen, Switzerland) for 2 min at CON: Control sausage, formulated without chickpea; RCP: sausage formulated
260 ◦ C in splitless mode, and separated with a 60 m × 0.25 mm sized, with soaked raw chickpeas; CCP: sausage formulated with cooked chickpeas.
0.25 mm film thickness DB-5MS column (Agilent System) using helium ab
Means within the same row without a common superscript are significantly
(1 mL/min) as the carrier gas, with oven programmed at 35 ◦ C (1 min), different (P < 0.05; Tukey post-hoc test).
#
35 ◦ C to 50 ◦ C (10 ◦ C/min), 50 ◦ C to 200 ◦ C (4 ◦ C/min), 200 ◦ C to 250 ◦ C Obtained by difference: 100 – (% moisture + % fat + % protein + % ash).
(50 ◦ C/min) and 250 ◦ C (11 min). Detection (MS 5975C detector, Agi­
&
Losses due to cooking and centrifuging the batter in Falcon tubes.
lent System) was carried out in electron impact mode at 70 eV with the
spectrometer transfer line and ion source temperatures of 240 ◦ C and The value of pH was slightly but significantly higher in CCP batter,
260 ◦ C, respectively, scanning from 40 m/z to 350 m/z. Identification suggesting that cooking of chickpeas increased pH value. Accordingly,
was performed by comparing the peak’s mass spectrum with those in the we found that the pH values of raw and cooked chickpea pastes were
NIST/EPA/NIH-98 mass spectral database, and comparing the linear respectively 6.40 ± 0.03 and 6.91 ± 0.03. The increase in pH of chick­
retention indexes calculated from a series of n-alkanes with those re­ peas due to cooking could be at least partially explained by the leaching
ported in the literature. The compound levels were expressed as area of soluble acidic compounds in the cooking water, as chickpea cooking
units x 106. water is known to be slightly acidic (Stantiall, Dale, Calizo, & Serventi,
2018).
2.7. Statistical analysis Water and oil losses in the batters (emulsion stability features) due to
cooking and centrifugation were minimal for all the formulations and
The general linear model analysis of variance (ANOVA) was per­ were not affected by the replacement of meat for raw or cooked chick­
formed using SPSS Statistics software (version 26; IBM, Somers, NY, peas (Table 2). In contrast, Dzudie, Scher, and Hardy (2002), Thushan-
USA). For the sausage batter before cooking, treatment (formulation) Sanjeewaa, Wanasundara, Pietrasika, and Shanda (2010) and Tahma­
was used as a fixed factor. For the sausages, treatment, cooking time and sebi, Labbafi, Emam-Djomeh, & Yarmand, (2016), all using raw pulse
interaction treatment × cooking time were used as fixed factors. In flours, reported that reformulation with pulses at levels of 4% to 11%
addition, replication consisting of the data from the analysis of two increased emulsion stability and WHC of cooked sausages. Discrepancies
portions of sausages for each treatment-cooking time combination could be attributed to differences in the experimental conditions be­
within each production batch was included as a random factor. When tween this study and those studies that can potentially affect the
the ANOVA proved to be significant (P < 0.05) for treatment or the emulsion stability, such as the type and amount of pulse used or the
treatment x cooking-time interaction, it was followed by the Tukey test percentage of meat replacement, the type of meat, fat, water, etc.
to perform pair-wise comparisons, considering statistical significance
for P < 0.05. 3.2. Sausage technological quality traits

3. Results and discussion The weight loss of the sausages during cooking and the technological
quality traits of the cooked sausages after 24 h of refrigerated storage are
3.1. Meat batter composition and technological quality shown in Table 3. The reformulation showed significant differences in
cooking losses, which were mainly due to evaporation of moisture from
The use of chickpeas in sausage formulation resulted in a sausage the sausage surface (P < 0.001). In contrast, no significant difference in
batter composition with a lower percentage of moisture. (Table 2). This centrifugation losses was observed (P = 0.09). The CCP sausages showed
was expected because the difference in the water added to both RCP and a weight loss of 3 to 7% units higher than CON or RCP sausages. How­
CCP batters and that added to the CON batter (2.4 g of water/100 g ever, the partial substitution of meat by raw chickpea (RCP sausages)
batter; Table 1) was lower than the moisture provided from the addi­ showed no effect on cooking losses compared to CON sausages. The
tional amount of meat in the CON batter (6.5 g of meat/100 g batter; evaporation of water (cooking losses) from the sausage, when sausage
Table 1; contributing 4.8 g of water/100 g batter). The RCP batter diameter and cooking conditions are the same, depends on the migration
contained a higher concentration of the sum starch + fibre than CON of water inside the sausage and thus on the composition of the sausage
formulation. The concentration of starch + fibre in CON batter was and its WHC. Hence, the WHC of the functional components of raw
consistent with and similar to the amount of starch used, and the sig­ chickpeas, such as protein, starch, fibre, etc., would be comparable to
nificant difference between RCP and CON sausages would be attributed that of the amount of meat replaced.
to the fibre content in chickpeas. The intermediate concentration of The lower and higher cooking losses for, respectively, RCP and CCP
starch + fibre showed by CCP batter compared to RCP and CON batter sausages suggest a heat-induced decrease in WHC of proteins and starch
could be explained by partial losses in carbohydrates (fibre included) of cooked chickpeas compared to those of raw chickpeas. The addition of
from the chickpeas during cooking (Liu & Serventi, 2020; Ma et al., native (non-denatured) chickpea protein to myofibrillar protein sus­
2011). pensions increases water retention during thermal gelation of composite

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Table 3
Weight loss due to cooking and technological properties of reformulated emulsion-type sausages cooked for conventional (conv) or prolonged (prolong) time.
Formulations P-level

CON-conv CON-prolong RCP-conv RCP-prolong CCP-conv CCP-prolong F t F×t

Weight losses (%)

Cooking 4.26 ± 0.44d 6.84 ± 0.44bc 6.23 ± 0.41bcd 6.67 ± 0.89c 9.08 ± 0.25b 13.39 ± 0.58a <0.001 <0.001 0.005
Centrifugation 15.7 ± 1.2 14.4 ± 0.7 16.1 ± 0.7 17.8 ± 1.4 19.1 ± 1.9 16.6 ± 1.2 0.092 0.50 0.22
Colour
L* 75.9 ± 0.6 76.3 ± 0.6 77.0 ± 0.6 76.8 ± 0.7 77.8 ± 0.6 77.6 ± 0.6 0.055 1.00 0.091
a* 1.00 ± 0.09b 1.19 ± 0.11b 1.43 ± 0.13ab 1.75 ± 0.13a 1.01 ± 0.11b 1.31 ± 0.11ab 0.005 0.018 0.25
b* 15.7 ± 0.2c 15.4 ± 0.2c 15.7 ± 0.2c 15.7 ± 0.3bc 16.7 ± 0.2ab 16.8 ± 0.2a 0.002 0.88 0.70

Texture
Hardness (N) 26.2 ± 0.90b 24.0 ± 0.6b 24.6 ± 1.71b 25.5 ± 1.3b 28.7 ± 1.1ab 31.0 ± 1.2a 0.038 0.39 0.22
Cohesiveness 0.252 ± 0.010de 0.243 ± 0.090e 0.279 ± 0.006cd 0.273 ± 0.032bc 0.317 ± 0.003a 0.299 ± 0.006ab 0.001 0.16 0.14
Elasticity 0.428 ± 0.008a 0.399 ± 0.036ab 0.349 ± 0.024b 0.346 ± 0.015b 0.389 ± 0.050ab 0.365 ± 0.008b 0.011 0.17 0.09
Chewiness (N) 27.4 ± 0.9abc 24.9 ± 0.6c 25.5 ± 1.8bc 25.8 ± 1.3bc 30.7 ± 0.7ab 32.0 ± 1.1a 0.013 0.98 0.85

Results are reported as means (average of triplicate batches and two samples per batch; n = 6) ± standard error of the mean.
CON: Control sausage, formulated without chickpea; RCP: sausage formulated with soaked raw chickpeas; CCP: sausage formulated with cooked chickpeas; conv:
conventional cooking time; prolong: prolonged cooking time; F: formulation; t: cooking time.
abcd
Means within the same row without a common superscript are significantly different (P < 0.05; Tukey post-hoc test).

gels (Li et al., 2021; Wang et al., 2023). On the other hand, protein the higher moisture losses and thus higher protein concentration in the
denaturation in cooked chickpeas results in reduced protein function­ more intensely cooked sausages.
ality, i.e. reduced ability to interact with water molecules (Aguilera Reformulation had a significant effect on all textural characteristics
et al., 2009; Ma et al., 2011). Furthermore, in the RCP (and CON) sau­ of the sausages (Table 3). Compared to CON sausages, RCP and CCP
sages, unlike the CCP sausages, the gelatinisation of the starch took sausages showed respectively similar or greater hardness and chewiness.
place during the cooking of the sausage which might have also Results suggest that the chickpea components involved in the heat-
contributed to the higher water retention. induced gelation in the sausages, such as proteins, mucins, starch and
Cooking time exerted a significant effect on cooking losses, finding complex carbohydrates, would have compensated (for RCP) or
more losses for the prolonged heating (P < 0.001; Table 3). Comparing enhanced (for CCP) the bonding strength effect of the replaced meat
within each treatment, significant differences in cooking losses were proteins. Moreover, regardless how was the processing of chickpeas, the
observed between for CON and CCP sausages. In agreement, higher replacement of meat by chickpea paste tended to significantly increase
cooking losses are expected as sausage core temperature increases the sausage-gel cohesiveness and decrease its elasticity. In previous
(Glorieux et al., 2019; Shin et al., 2022). Unexpectedly, this effect was studies using raw pulse flours as meat extenders in emulsion-type
not found in RCP sausages. The different pattern observed in these cooked sausages, the effects of pulses on hardness and elasticity have
sausages, i.e. the lack of effect of heating time on moisture losses, could shown partial discrepancy, which might be explained by differences in
be related to the gelatinisation dynamics of raw chickpea starch during the experiment conditions. Dzudie et al. (2002), using bean meal at
the sausage cooking period. Singh, Singh-Sandhu, and Kaur (2004) re­ levels up to 10.0% to replace the same amount of beef, thus reducing the
ported temperature ranges for starch gelatinisation of different chickpea moisture level in the sausage with the addition of pulse flour, observed a
varieties between 62 and 74 ◦ C. Therefore, in RCP sausages the gelati­ decrease in hardness. In contrast, Tahmasebi, Labbafi, Emam-Djomeh,
nisation of raw chickpea starch may not have ended with the conven­ and Yarmand (2016) found that replacing beef with pigeon pea flour
tional heating and the ongoing gelatinisation during the extended period (up to 16.5%) combined with maize flour increased the toughness of the
of the prolonged heating may have had a slowing effect on moisture sausages. Neither study, in contrast to our results, found a significant
migration inside the sausage and this, in turn, may have decreased effect of meat substitution with pulse flour on elasticity. However, as
moisture evaporation. regards to cohesiveness, according to our results, the use of pulse flour
The colour of the CON sausage, whose values are shown in Table 3, increased cohesiveness in the reformulated sausages (Dzudie et al.,
seemed to be considerably different from that of cream-coloured 2002; Tahmasebi et al., 2016).
chickpeas, which according to Güzel and Sayar (2012) cream-coloured Comparing the CCP and RCP sausages separately, the CCP sausages
chickpeas showed L*, a* and b* values of 53–57, 7–8, and 17–23, for showed higher hardness, cohesiveness, and chewiness than the RCP
L*, a* and b*, respectively. Thus, the reformulation of sausages with sausages, despite the expected reduction of protein gelation function­
chickpea should have an effect on sausage colour, as was the case in our ality in the pre-cooked chickpea (Ma et al., 2011). This increasing effect
study. However, the specific effects of adding chickpea on the sausage could be related to the higher dry matter and protein contents in CCP
colour are difficult to predict and explain since several complex factors sausages resulting from the greater weight (moisture) loss during
are involved, such as the levels of chickpea and meat pigments in cooking (Table 2). According to Petridis, Vlazakis, Tzivanos, Derlikis,
sausage batter, pigment changes during sausage processing, sausage and Ritsoulis (2010), a higher protein content in frankfurters tends to
microstructure affecting light scattering, etc. Our results showed that the produce a harder and more cohesive gel network. Regarding cooking
difference in colour between CON and chickpea-containing sausages time, however, the prolonged heating, although it resulted in signifi­
depended on the chickpea pre-treatment. Compared to CON sausages, cantly higher moisture loss in the CON and CCP sausages, did not in­
the use of raw chickpea increased a* and that of cooked chickpea crease the texture values in any of the treatments. The protein gelation
increased b*. In agreement, Sanjeewaa, Wanasundara, Pietrasika, & was probably fully completed with the conventional heating (Barbut
Shanda (2010) found the use of raw chickpea flour (5%) in the formu­ et al., 1996), and the excess moisture losses due to prolonged heating
lation of a low-fat sausage to increase b*, whereas a* and L* were not were not high enough to significantly change the texture values.
affected. Cooking time affected the a* value (P = 0.018); however, this The differences found in the texture profile between the sausages
effect could not be evidenced when comparing between heating times studied would probably have changed their sensory texture perception.
within formulations. The effect of heating time on a* might be related to The decreasing effect on elasticity found in chickpea reformulated

5
S.A. Kasaiyan et al. Meat Science 202 (2023) 109217

sausages could be considered negative considering that a high elasticity (Karolkowski, Guichard, Briand, & Salles, 2021; Shariati-Ievari et al.,
is a key desired characteristic of frankfurters (Petridis et al., 2010). The 2016; Xu, Zhao, Gu, Rao, & Chen, 2020). The TBARS initially present in
relationship between instrumental texture and sensory data in refor­ the raw chickpea-water (1:1; w/w) paste, used in the formulation, would
mulated sausages, however, is not always easy to predict (Hayes, Ste­ have partially contributed to the TBARS levels of the cooked sausages,
panyan, Allen, O’Grady, & Kerry, 2011). Thus, a sensory analysis would since before being mixed with the meat, the TBARS level of the paste was
be needed to fully understand the effect of the observed changes in 1.16 ± 0.12 mg malondyaldehyde/kg of paste (mean ± standard devi­
texture profile parameters on sausage acceptance. In addition, further ation; n = 3). However, most of the TBARS increase (lipid oxidation) in
research should be conducted to evaluate the combined effect of using RCP sausages would have occurred during the mixing, stuffing and
chickpeas with texture-enhancing ingredients such as selected proteins initial period of the sausage heating, until the heat was able to inhibit
or hydrocolloids, thus improving elasticity. the enzymatic activity. Regarding LOX heat stability, Verma et al.
(1984b) observed that heating raw chickpea flour at 60 ◦ C for 1 h
inactivated its oxidation effect on meat batters.
3.3. Lipid oxidation and headspace volatile composition in sausages In an ad hoc supplementary experiment, we found that neither
ascorbic acid (0.5 g/kg) not potassium metabisulphite (0.45 g/kg), two
Raw pulses seem to produce lipid oxidation when they are mixed common additives used in the meat industry, were able to significantly
with other food ingredients, meat included, to prepare reformulated reduce neither the discolouration (decrease in a* value) of the RCP
food products, thus generating secondary lipid oxidation products batter, nor the lipid oxidation (TBARS levels) in the RCP sausage (Sup­
(Krause et al., 2022; Shariati-Ievari et al., 2016; Verma, Ledward, & plementary Table S1). In a previous study, Verma et al. (1984b) reported
Lawrie, 1984b). Accordingly, in our study, the use of raw chickpeas that an antioxidant containing α-tocopherol at levels of 1% of fat content
significantly increased TBARS levels in RCP sausage (Table 4). In rela­ was able to retard but not prevent the lipid oxidation related to the use
tion to the lipid oxidation in RCP sausages and in accordance with the of raw chickpea in a sausage, although it did not inhibit discoloration.
findings of Verma et al. (1984b), during mixing we observed that the The TBARS levels in the sausages within each treatment did not
RCP batter was also discoloured (myoglobin was oxidised) compared to change significantly between conventional and prolonged heating,
the CON and CCP batters (Supplementary Fig. S1). which indicate that lipoxygenase activity on lipid oxidation was
The TBARS increase in RCP sausages would have been mainly the completely inactivated by the conventional heating. No significant
result of enzymatic activity by the LOX contained in raw pulses

Table 4
Thiobarbituric-acid reactive substances (TBARS) and headspace volatile compounds& of the reformulated emulsion-type sausages cooked for conventional (conv) or
prolonged (prolong) time.
Formulations P-level

CON-conv CON-prolong RCP-conv RCP-prolong CCP-conv CCP-prolong F t F×t

TBARS 0.60 ± 0.13b 0.43 ± 0.09b 2.58 ± 0.05a 2.17 ± 0.08a 0.57 ± 0.08b 0.45 ± 0.05b <0.001 0.49 0.095

Aldehydes
Ethanal 4.4 ± 0.4b 6.2 ± 0.5b 71.0 ± 9.3a 50.0 ± 7.0a 4.8 ± 1.1a 3.4 ± 1.2b <0.001 0.061 0.005
Methylbutanal ND 0.8 ± 0.78 3.9 ± 2.5 2.2 ± 1.4 2.9 ± 1.9 4.1 ± 2.7 0.052 0.90 0.401
Pentanal 2.6 ± 1.6b 6.1 ± 2.2b 262.6 ± 17.9a 183.1 ± 17.3a 28.2 ± 8.4b 26.1 ± 9.5b 0.002 0.22 0.69
Hexanal ND ND 7693.2 ± 766.2a 5971.1 ± 359.6a 234.3 ± 97.7b 125.1 ± 96.4b – 0.003 –

Alcohols
Ethanol 26.0 ± 15.6b 31.1 ± 18.6b 452.6 ± 98.2a 469.9 ± 101.2a 84.2 ± 53.0b 7.3 ± 3.9b 0.003 0.71 0.28
Methylbutanol ND ND 41.1 ± 25.6 21.3 ± 8.7 ND ND – 0.10 –
Pentanol ND ND 60.2 ± 6.3 42.8 ± 3.4 45.6 ± 16.6 ND – 0.054 –
Hexanol ND ND 53.9 ± 21.5 38.4 ± 24.8 ND ND – 0.21 –

Hydrocarbons
Pentane 236.2 ± 15.9b 359.7 ± 18.4b 1471.4 ± 160.9a 1360.8 ± 104.8a 339.2 ± 68.1b 477.9 ± 27.1b 0.012 0.60 0.21
Hexane 6.8 ± 0.8b 11.3 ± 1.5ab 10.3 ± 1.4ab 7.2 ± 1.0b 26.5 ± 9.7a 14.2 ± 1.5ab 0.011 0.027 0.030
Heptane 10.0 ± 1.8c 15.4 ± 2.6c 63.5 ± 7.8a 50.1 ± 3.7ab 18.2 ± 4.0c 40.2 ± 6.8b 0.008 0.071 0.077
Octane 83.7 ± 13.8c 147.1 ± 17.5bc 248.3 ± 6.7a 190.3 ± 12.9ab 120.6 ± 38.3bc 257.4 ± 26.5a 0.007 0.088 0.016
2-Octene ND ND 10.6 ± 1.0 8.0 ± 0.9 ND ND – 0.20 –

Terpene compounds
α-Pinene 907.7 ± 41.6ab 909.3 ± 29.2ab 827.0 ± 65.9bc 659.9 ± 23.4c 896.7 ± 21.1ab 1030.1 ± 69.7a 0.004 0.14 0.013
Camphene 73.7 ± 6.6abc 84.2 ± 4.2ab 59.7 ± 7.0bc 43.7 ± 2.6c 93.2 ± 8.3a 96.8 ± 11.9a 0.018 0.94 0.39
β-Pinene 836.2 ± 33.9 657.9 ± 132.9 785.0 ± 58.7 638.7 ± 15.1 753.6 ± 20.8 870.2 ± 51.3 0.50 0.34 0.16
Myrcene 67.4 ± 3.1 71.4 ± 4.4 86.3 ± 6.9 80.4 ± 3.8 71.4 ± 6.0 67.6 ± 8.8 0.16 0.76 0.040
δ-3-Carene 3735.7 ± 149.8 3914.1 ± 136.7 4199.9 ± 102.1 3742.8 ± 77.8 3899.7 ± 64.8 4229.0 ± 175.5 0.19 0.68 0.038
o-Cymene 59.4 ± 3.4 61.2 ± 4.6 58.9 ± 3.0 49.7 ± 1.1 57.3 ± 2.6 67.7 ± 5.8 0.074 0.33 0.089
p-Cymene 223.8 ± 104.3 255.5 ± 17.1 250.7 ± 18.5 230.4 ± 6.6 245.3 ± 5.2 253.2 ± 8.1 0.62 0.48 0.067
Limonene 3118.0 ± 144.3bc 3395.9 ± 149.6abc 3652.0 ± 130.7ab 2961.6 ± 5.8c 3384.3 ± 72.4abc 3764.0 ± 254.9a 0.11 0.86 0.022
β-Caryophyillene 54.2 ± 2.2ab 53.5 ± 3.2ab 59.3 ± 2.6a 54.9 ± 3.0b 35.7 ± 9.6b 50.3 ± 2.6ab 0.056 0.43 0.015

Results are reported as means (average of triplicate batches and two samples per batch; n = 6) ± standard errors of the mean.
CON: Control sausage, formulated without chickpea; RCP: sausage formulated with soaked raw chickpeas; CCP: sausage formulated with cooked chickpeas; conv:
conventional cooking time; prolong: prolonged cooking time; F: formulaton; t: cooking time.
ND: not detected.
abc
Means within the same row without a common superscript are significantly different (P < 0.05; Tukey post-hoc test).
&
: TBARS expressed as mg of malondialdehyde/kg of sausage and volatile compounds as area units x 106.

6
S.A. Kasaiyan et al. Meat Science 202 (2023) 109217

changes in TBARS were observed during refrigerated aerobic storage in This finding could be attributed to an indirect effect of the different
any of the sausages (data shown in the supplementary material for concentration of volatile compounds in the sausages (matrix) and in
brevity; Table S2). The storage-related oxidative stability could be their headspace (in equilibrium with the sausage matrix) between for­
attributed to the protective effect of the olive oil used in the sausages mulations. The higher amounts of aldehydes, alcohols and hydrocarbons
against further lipid oxidation (Pintado et al., 2015; Shin et al., 2022). in RCP sausages compared to CON and CCP sausages could thus have
The profile of volatile compounds in the headspace of sausages is resulted in a sausage-headspace equilibrium with lower relative pro­
shown in Table 4. The compounds detected were aldehydes, alcohols, portions of α-pinene and camphene in the headspace. Heating time did
aliphatic straight-chain hydrocarbons and terpenoids. The majority of not affect the level of terpenes in sausage headspace; however, a sig­
the aldehydes, alcohols and hydrocarbons, i.e. straight-medium-chain nificant interaction formulation × time was detected for α-pinene,
(C5-C8) compounds, would have been formed from lipid breakdown myrcene, o-cymene, limonene and β-caryophyllene. In RCP sausages,
(Resconi, Escudero, & Campo, 2013; Shahidi & Oh, 2020). Terpenoids their levels, and especially that of limonene and β-caryophyllene, tended
would have originated from the black pepper (Dosoky, Satyal, Barata, da to decrease with the longer heating time, while in CON and CCP sau­
Silva, & Setzer, 2019; Musenga et al., 2007) used in the formulation. sages this tendency was not observed. We have no other reason to
Formulation had a significant effect on the composition of volatile explain this observation, apart from a matrix effect, i.e. an increased
compounds, with the greatest effect found when comparing the RCP retention of these volatile compounds by the RCP sausage matrix after
sausages with the other two sausages. RCP sausages showed the highest prolonged heating.
concentration of most of the lipid-derived compounds detected, i.e.
pentanal, hexanal, pentane, octane and octene. This finding, which is 4. Conclusions
consistent with the higher levels of TBARS found in RCP sausages,
suggests that lipid oxidation mediated by raw chickpeas during the The effects of the reformulation of a lamb meat and olive oil emul­
preparation of RCP sausages was primarily responsible for the formation sion type sausage using 7% chickpea as a partial replacer of meat and
of these volatile compounds. In agreement, Shariati-Ievari et al. (2016) total replacer of potato starch on the sausage quality depended on the
associated the use of raw chickpea flour in burgers with increased values pre-treatment of the chickpeas. The performance of raw chickpea on
of medium-chain alcohols, aldehydes and aliphatic hydrocarbons pro­ emulsion stability, sausage yield, and sausage hardness and chewiness
duced through LOX activity. Krause et al. (2022) also reported the for­ was comparable to that of the meat and potato starch replaced. How­
mation of a range of lipid-derived volatile compounds attributed to ever, raw chickpea caused an increased lipid oxidation during the
oxidative enzymatic activity when raw pulses were used in reformulated sausage making process, leading to the formation of volatile compounds
baked cakes. with a potential effect on off-flavour development. Cooked chickpea
RCP-sausage headspace also showed the highest levels of ethanal and reduced cooking yield and, possibly due to the increased moisture loss,
ethanol. They could have been formed by microbial fermentation during led to an increase in sausage hardness, cohesiveness and chewiness.
the mixing and filling of the batter and the initial part of the cooking However, cooked chickpea had no effect on the oxidation of the sausage
(Krause et al., 2022). These two compounds have also been detected in and scarcely affected the volatile compound profile. Both raw and
raw chickpea seeds and flours (Karolkowski et al., 2021; Krause et al., cooked chickpeas reduced the elasticity and slightly changed the colour
2022) where, as suggested by Karolkowski et al. (2021), they might of the cooked sausages, with raw chickpeas increasing a*, and cooked
originate via amino acid degradation. This pathway could also be a chickpeas increasing b*. Further research is needed to assess the effects
source of methylbutanol and methylbutanal, which, taken together, of the volatile compounds associated with raw chickpea use on sausage
were more abundant in sausages containing chickpeas than in CON consumer acceptance or to explore approaches to reduce the oxidation.
sausages. No apparent advantage was gained by increasing the heating time at
The increased lipid-derived volatile compounds, ethanal and 85 ◦ C from 40 min to 80 min. On the contrary, prolonged heating may
ethanol, and methylbutanol in the RCP sausage headspace suggests a result in a lower yield of sausages with cooked chickpeas. Reformulation
more intense flavour in RCP sausages than in the other. Those com­ of sausage with cooked chickpea could be recommended. However,
pounds could probably impart the RCP sausages with a beany flavour. additional research is needed to improve the reformulation in order to
Shariati-Ievari et al. (2016) linked the use of raw chickpea flour (6%) in reduce moisture loss during cooking and to increase the elasticity of the
burgers and the subsequent increase of volatile compounds derived from sausage.
lipid oxidation to a less desirable aroma, and Krause et al. (2022), in
cakes, to the appearance of green-bean flavours. In contrast, Thushan- CRediT authorship contribution statement
Sanjeewaa et al. (2010), using flour from raw chickpea (up to 5%) in a
low-fat cooked sausage, reported sensory scores of flavour intensity and S.A. Kasaiyan: Methodology, Data curation, Investigation. I. Caro:
desirability of chickpea-containing sausages similar to those of the Visualization, Writing – review & editing. D.D. Ramos: Funding
control sausages, and low scores for beany flavour. Further sensory acquisition, Conceptualization, Project administration. B.K. Salvá:
analysis is therefore necessary to determine the possible effect, negative Conceptualization, Methodology. A. Carhuallanqui: Project adminis­
or otherwise, of the use of raw chickpeas (and also cooked chickpeas) on tration, Investigation. M. Dehnavi: Investigation, Methodology. J.
the flavour of the sausages in this study. Mateo: Supervision, Conceptualization, Methodology, Formal analysis,
Regarding the effect of heating time on the content of aldehydes, Data curation, Visualization.
alcohol and hydrocarbons in the headspace of the sausages, significant
effects were only observed for the levels of hexanal and hexane, with Declaration of Competing Interest
lower values of these compounds being found in the most intensively
heated sausages. These effects could be related to increased evaporation The authors declare that they have no known competing financial
of both compounds during cooking due to the longer heating time or, in interests or personal relationships that could have appeared to influence
the case of hexanal, to possible chemical reactions with other compo­ the work reported in this paper.
nents of the sausage (Pignoli, Bou, Rodríguez-Estrada, & Decker, 2009).
Formulation showed effects on the levels of terpenes, presumably Data availability
from pepper, in the headspace of the sausage. Although all sausages
were prepared with the same amount of pepper, lower amounts of the Data will be made available on request.
two volatile terpenes that eluted most rapidly in the chromatographic
analysis, i.e. α-pinene and camphene, were found for RCP sausages.

7
S.A. Kasaiyan et al. Meat Science 202 (2023) 109217

Acknowledgments processing, and nutrition (pp. 307–329). Hoboken, NJ: Wiley-Blacwell. https://doi.
org/10.1002/9781119776802.ch12.
Hoek, A., Luning, P. A., Weijzen, P., Engels, W., Kok, F. J., & de Graaf, C. (2011).
This work was supported financially by the Vicerrectorado de Replacement of meat by meat substitutes. A survey on person and product-related
Investigación y Posgrado, Universidad Nacional Mayor de San Marcos, factors in consumer acceptance. Appetite, 56, 662–673. https://doi.org/10.1016/j.
Project A2008001m. appet.2011.02.001
Hughes, E., Cofrades, S., & Troy, D. J. (1997). Effects of fat level, oat fibre and
carrageenan on frankfurters formulated with 5, 12 and 30% fat. Meat Science, 45(3),
Appendix A. Supplementary data 273–281. https://doi.org/10.1016/S0309-1740(96)00109-X
Karolkowski, A., Guichard, E., Briand, L., & Salles, C. (2021). Volatile compounds in
pulses: A review. Foods, 10. https://doi.org/10.3390/foods10123140. Article 3140.
Supplementary data to this article can be found online at https://doi. Kaur, R., & Prasad, K. (2021). Technological, processing and nutritional aspects of
org/10.1016/j.meatsci.2023.109217. chickpea (Cicer arietinum) - a review. Trends in Food Science & Technology, 109,
448–463. https://doi.org/10.1016/j.tifs.2021.01.044
Krause, S., Keller, S., Hashemi, A., Decharles, N., Bonazzi, C., & Rega, B. (2022). From
References flours to cakes: Reactivity potential of pulse ingredients to generate volatile
compounds impacting the quality of processed foods. Food Chemistry, 371. https://
Aguilera, Y., Esteban, R. M., Benitez, V., Molla, E., & Martin-Cabrejas, M. A. (2009). doi.org/10.1016/j.foodchem.2021.131379. Article 131379.
Starch, functional properties, and microstructural characteristics in chickpea and Li, J., Chen, Y., Dong, X., Li, K., Wang, Y., Wang, Y., Du, M., Zhang, J., & Bai, Y. (2021).
lentil as affected by thermal processing. Journal of Agricultural and Food Chemistry, 57 Effect of chickpea (Cicer arietinum L.) protein isolate on the heat-induced gelation
(22). https://doi.org/10.1021/jf902042r, 10,682-10,688. properties of pork myofibrillar protein. Journal of the Science of Food and Agriculture,
AOAC. (1999). Meat and meat products. In P. Cunniff (Ed.), Official Methods of Analysis of 101, 2108–2116. https://doi.org/10.1002/jsfa.10833
the AOAC International, 16th ed. Combining tradition and innovation, Gaithersburg, Li, K., Fu, L., Zhao, Y. Y., Xue, S. W., Wang, P., Xu, X., & Bai, Y. H. (2020). Use of high-
Maryland, USA (pp. 1–15). Vol. II. intensity ultrasound to improve emulsifying properties of chicken myofibrillar
Asgar, M. A., Fazilah, A., Huda, N., Bhat, R., & Karim, A. A. (2017). Nonmeat protein protein and enhance the rheological properties and stability of the emulsion. Food
alternatives as meat extenders and eat analogs. Comprehensive Reviews in Food Science Hydrocolloids, 98. https://doi.org/10.1016/j.foodhyd.2019.105275. Article 105275.
and Food Safety, 9, 513–529. https://doi.org/10.1111/j.1541-4337.2010.00124.x Liburdi, K., Esti, M., Petroselli, V., Mendler-Drienyovszki, N., Radicetti, E., &
Barbut, S., Gordon, A., & Smith, A. (1996). Effect of cooking temperature on the Mancinelli, R. (2021). Catalytic properties of lipoxygenase extracted from different
microstructure of meat batters prepared with salt and phosphate. LWT - Food Science varieties of Pisum sativum and Lens culinaris. Journal of Food Biochemistry, 45. https://
and Technology, 29, 475–480. https://doi.org/10.1006/fstl.1996.0073 doi.org/10.1111/jfbc.13617. Article e13617.
Bessada, M. F., Barreira, J. C. M., & Oliveira, M. B. P. P. (2019). Pulses and food security: Liu, S., & Serventi, L. (2020). Introduction: Wastewater generation. In L. Serventi (Ed.),
Dietary protein, digestibility, bioactive and functional properties. Trends in Food Upcycling legume water: From wastewater to food ingredients (pp. 13–25). Cham,
Science & Technology, 93, 53–68. https://doi.org/10.1016/j.tifs.2019.08.022 Swizerland: Springer Nature. https://doi.org/10.1007/978-3-030-42468-8.
Carballo, D. E., Andrés, S., Giráldez, F. J., Khanjari, A., Caro, I., Llamazares, D., … Ma, S., Boye, J. I., Simpson, B. K., Prasher, S. O., Monpetit, D., & Malcolmson, L. (2011).
Mateo, J. (2020). The effects of storage and hop extract on aroma and flavour Thermal processing effects on the functional properties and microstructure of lentil,
compounds in Balkan-style sausages packed under a CO2-containing anaerobic chickpea, and pea flours. Food Research International, 44, 2534–2544. https://doi.
atmosphere. Heliyon, 6. https://doi.org/10.1016/j.heliyon.2020.e05251. Article org/10.1016/j.foodres.2010.12.017
e05251. Mateo, J., Caro, I., Kasiayan, S., Salvá, B. K., Carhuallanqui, A., & Ramos, D. D. (2021).
Chigwedere, C. M., Wanasundara, J. P. D., & Shand, P. (2021). Sensory descriptors for Potential of pulses flours as partial meat replacers in heat-treated emulsion-type
pulses and pulse-derived ingredients: Toward a standardized lexicon and sensory meat sausages. Frontiers in Animal Science. https://doi.org/10.3389/
wheel. Comprehensive Reviews in Food Science and Food Safety, 21, 999–1023. https:// fanim.2021.693086. Article 2:693086.
doi.org/10.1111/1541-4337.12893 Mudryj, A. N., Yu, N., & Aukema, H. M. (2014). Nutritional and health benefits of pulses.
De Boer, J., Schösler, H., & Aiking, H. (2014). “Meatless days” or “less but better”? Applied Physiology, Nutrition, and Metabolism, 39(11), 1197–1204. https://doi.org/
Exploring strategies to adapt Western meat consumption to health and sustainability 10.1139/apnm-2013-0557
challenges. Appetite, 76, 120–128. https://doi.org/10.1016/j.appet.2014.02.002 Musenga, A., Mandrioli, R., Ferranti, A., Dórazio, G., Fanali, S., & Raggi, M. A. (2007).
Dosoky, N. S., Satyal, P., Barata, L. M., da Silva, J. K. R., & Setzer, N. W. (2019). Volatiles Analysis of aromatic and terpenic constituents of pepper extracts by capillary
of black pepper fruits (Piper nigrum L.). Molecules, 24(3), 4244. https://doi.org/ electrochromatography. Journal of Separation Science, 30(4), 612–619. https://doi.
10.3390/molecules24234244 org/10.1002/jssc.200600456
Dzudie, T., Scher, J., & Hardy, J. (2002). Common bean flour as an extender in beef Nam, K. C., & Ahn, D. U. (2003). Effects of ascorbic acid and antioxidants on the color of
sausages. Journal of Food Engineering, 52, 143–147. https://doi.org/10.1016/S0260- irradiated ground beef. Journal of Food Science, 68(5), 1686–1690. https://doi.org/
8774(01)00096-6 10.1111/j.1365-2621.2003.tb12314.x
Eckl, M. R., Bierbroek, S., van’t Veer, P., & Geleijnse, J. M. (2021). Replacement of meat Petridis, D., Vlazakis, E. L., Tzivanos, I., Derlikis, E. M., & Ritsoulis, C. (2010). Effects of
with non-meat protein sources: A review of the drivers and inhibitors in developed selected ingredients, and fat content on the sensory and mechanical properties of
countries. Nutrients, 13. https://doi.org/10.3390/nu13103602. Article 3602. frankfurter-type sausages. Journal of Texture Studies, 41(6), 880–898. https://doi.
Farooq, Z., & Boye, J. I. (2011). Novel food and industrial applications of pulse flours and org/10.1111/j.1745-4603.2010.00260.x
fractions. In B. K. Tiwari, A. Gowen, & B. McKenna (Eds.), Pulse foods processing, Pignoli, G., Bou, R., Rodríguez-Estrada, M. T., & Decker, E. A. (2009). Suitability of
quality and nutraceutical applications (pp. 283–324). Oxford: Academic Press. https:// saturated aldehydes as lipid oxidation markers in washed Turkey meat. Meat Science,
doi.org/10.1016/B978-0-1238-2018-1.00007-0. 83, 412–416. https://doi.org/10.1016/j.meatsci.2009.06.019
Glorieux, S., Steen, L., Van de Walle, D., Dewettinck, K., Foubert, I., & Fraeye, I. (2019). Pintado, T., & Delgado-Pando, G. (2020). Towards more sustainable meat products:
Effect of meat type, animal fat type, and cooking temperature on microstructural and Extenders as a way of reducing meat content. Foods, 9(8). https://doi.org/10.3390/
macroscopic properties of cooked sausages. Food and Bioprocess Technology, 12, foods9081044. Article 1044.
16–26. https://doi.org/10.1007/s11947-018-2190-6 Pintado, T., Herrero, A. M., Ruiz-Capillas, C., Triki, M., Carmona, P., & Jiménez-
Godfray, H. C. J., Aveyard, P., Garnett, T., Hall, J. H., Key, T. J., Lorimer, J., … Colmenero, F. (2015). Effects of emulsion gels containing bioactive compounds on
Jebb, S. A. (2018). Meat consumption, health, and the environment. Food Security, sensorial, technological, and structural properties of frankfurters. Food Science and
361. https://doi.org/10.1126/science.aam5324. Article eaam5324. Technology International, 22(2), 132–145. https://doi.org/10.1177/
Grasso, S., & Jaworska, S. (2021). Part meat and part plant: Are hybrid meat products fad 1082013215577033
or future? Foods, 9(12). https://doi.org/10.3390/foods9121888. Article 1888. Resconi, V., Escudero, A., & Campo, M. (2013). The development of aromas in ruminant
Guldiken, B., Konieczny, D., Franczy, A., Satiro, V., Pickard, M., Wang, N., … meat. Molecules, 18(6), 6748–6781. https://doi.org/10.3390/molecules18066748
Nickerson, M. T. (2022). Impacts of infrared heating and tempering on the chemical Richi, E. B., Baumer, B., Conrad, B., Darioli, R., Schmid, A., & Keller, U. (2015). Health
composition, morphological, functional properties of navy bean and chickpea fours. risks associated with meat consumption: A review of epidemiological studies.
European Food Research and Technology, 248, 767–781. https://doi.org/10.1007/ International Journal for Vitamin and Nutrition Research, 85(1), 70–78.
s00217-021-03918-4 Sanjeewaa, T. W. G., Wanasundara, J. P. D., Pietrasika, Z., & Shanda, P. J. (2010).
Güzel, D., & Sayar, S. (2012). Effect of cooking methods on selected physicochemical and Characterization of chickpea (Cicer arietinum L.) flours and application in low-fat
nutritional properties of barlotto bean, chickpea, faba bean, and white kidney bean. pork bologna as a model system. Food Res. Int., 43, 617–623. https://doi.org/
Journal of Food Science and Technology, 49, 89–95. https://doi.org/10.1007/s13197- 10.1016/j.foodres.2009.07.024
011-0260-0 Shahidi, F., & Oh, W. Y. (2020). Lipid derived flavor and off flavor of traditional and
Hayes, J. E., Stepanyan, V., Allen, P., O’Grady, M. N., & Kerry, J. P. (2011). Evaluation of functional foods: An overview. Journal of Food Bioactives., 10, 20–31. https://doi.
the effects of selected plant-derived nutraceuticals on the quality and shelf-life org/10.31665/JFB.2020.10224
stability of raw and cooked pork sausages. LWT - Food Science and Technology, 44, Shariati-Ievari, S., Ryland, D., Edel, A., Nicholson, T., Suh, M., & Aliani, M. (2016).
164–172. https://doi.org/10.1016/j.lwt.2010.05.020 Sensory and physicochemical studies of thermally micronized chickpea (Cicer
He, S., Shi, J., Ma, Y., Xue, S. J., Zhang, H., & Zaho, S. (2014). Kinetics for the thermal arietinum) and green lentil (Lens culinaris) flours as binders in low-fat beef burgers.
stability of lectin from black turtle bean. Journal of Food Engineering, 142, 132–137. Journal of Food Science, 81(5), S1230–S1242. https://doi.org/10.1111/1750-
https://doi.org/10.1016/j.jfoodeng.2014.06.008 3841.13273
Hill, H. (2022). Utilization of dry beans and other pulses as ingredients in diverse food Shi, Y., Mandal, R., Singh, A., & Pratap-Singh, A. P. (2020). Legume lipoxygenase:
products. In M. Siddiq, & M. A. Uebersax (Eds.), Dry beans and pulses production, Strategies for application in food industry. Legume Science, 2. https://doi.org/
10.1002/leg3.44. Article 2:e44.

8
S.A. Kasaiyan et al. Meat Science 202 (2023) 109217

Shin, D. J., Lee, H. J., Lee, D., Jo, C., & Choe, J. (2020). Fat replacement in chicken formulation optimization, emulsion stability and textural characterization. LWT
sausages manufactured with broiler and old laying hens by different vegetable oils. Food Science and Technology, 68, 76–84. https://doi.org/10.1016/j.lwt.2015.12.011
Poultry Science, 99, 2811–2818. https://doi.org/10.1016/j.psj.2020.01.008 Thushan-Sanjeewaa, T. W. G., Wanasundara, J. P. D., Pietrasika, Z., & Shanda, P. J.
Shin, D. J., Yim, D. G., Kwon, J. A., Kim, S. S., Lee, H. J., & Jo, C. (2022). Effect of cutting (2010). Characterization of chickpea (Cicer arietinum L.) flours and application in
time and cooking temperature on physicochemical properties of chicken breast meat low-fat pork bologna as a model system. Food Research International, 43, 617–623.
emulsion sausage with olive oil. Poutry Science, 101. https://doi.org/10.1016/j. https://doi.org/10.1016/j.foodres.2009.07.024
psj.2021.101554. Article 10155. Vatansever, S., Tulbek, M. C., & Riaz, M. N. (2020). Low- and high-moisture extrusion of
Singh, N., Singh-Sandhu, S., & Kaur, M. (2004). Characterization of starches separated pulse proteins as plant-based meat ingredients: A review. Cereal Foods World, 65(4).
from Indian chickpea (Cicer arietinum L.) cultivars. Journal of Food Engineering, 63, https://doi.org/10.1094/CFW-65-4-0038. Article CFW-65-40038.
441–449. https://doi.org/10.1016/j.jfoodeng.2003.09.003 Verma, M. M., Ledward, D. A., & Lawrie, R. A. (1984a). Utilization of chickpea flour in
Sofi, S. A., Muzaffar, K., Ashraf, S., & Gupta, I. (2020). Chickpea. In A. Manickavsagan, & sausages. Meat Science, 11, 109–121.
P. Thirunathan (Eds.), Pulses: Processing and product development (pp. 55–76). Cham, Verma, M. M., Ledward, D. A., & Lawrie, R. A. (1984b). Lipid oxidation and
Swizerland: Springer Nature. https://doi.org/10.1007/978-3-030-41376-7. metmyoglobin formation in sausages containing chickpea flour. Meat Science, 11,
Spain. (2014). Real Decreto 474/2014, de 13 de junio, por el que se aprueba la norma de 171–189.
calidad de derivados cárnicos. BOE no. 147 (pp. 46058–46078). Boletín Oficial del Wang, Y., Yuan, J., Li, K., Chen, X., Wang, Y., & Bai, Y. (2023). Evaluation of chickpea
Estado: Madrid. protein isolate as a partial replacement for phosphate in pork meat batters: Techno-
Stantiall, S. E., Dale, K. J., Calizo, F. S., & Serventi, L. (2018). Application of pulses functional properties and molecular characteristic modifications. Food Chemistry,
cooking water as functional ingredients: The foaming and gelling abilities. European 404. https://doi.org/10.1016/j.foodchem.2022.134585. Article 134585.
Food Research and Technology, 244(1), 97–104. https://doi.org/10.1007/s00217- Xu, M., Zhao, J., Gu, Z., Rao, J., & Chen, B. (2020). Changes in odor characteristics of
017-2943-x pulse protein isolates from germinated chickpea, lentil, and yellow pea: Role of
Tahmasebi, M., Labbafi, M., Emam-Djomeh, Z., & Yarmand, M. S. (2016). Manufacturing lipoxygenase and free radicals. Food Chemistry, 314. https://doi.org/10.1016/j.
the novel sausages with reduced quantity of meat and fat: The product development, foodchem.2020.126184. Article 126184.