Effect of sterilization on the physicochemical and sensory characteristics of “cau cau” (Peruvian creole food) of giant squid (Dosidicus gigas) in flexible pouches

Abstract

This study was elaborated on the “cau cau” (Peruvian creole food) of giant squid (SCC) in flexible pouches and treated at different sterilization times at 121 °C in a horizontal water immersion autoclave during 25 min (SCC1), 30 min (SCC2), and 35 min (SCC3), obtaining sterilization values (F₀) of 6, 9 and 13 min, and cooking values (C₀) of 57, 73 and 91 min, respectively. Proximate analysis, color analysis, instrumental texture profile analysis (TPA), commercial sterility test, and sensory evaluation were performed on each SCC. The products were commercially sterile and there was no significant difference (p > 0.05) in the physicochemical tests between SCC. On the other hand, there were significant differences (p < 0.05) in color and texture profile between the SCC ingredients. Regarding the sensory evaluation of each SCC, there were no marked differences, and they had a moderate acceptance among the panelists (n = 30).

Keywords:
Retort processing; F₀ value; Sterilizable flexible pouches; Convenience food; Texture Profile Analysis; Color analysis

Highligths

Sterilization treatments applied to “cau cau” (SCC) of giant squid affected the color and texture profile; however, there was no marked difference according to sensory evaluation

Sterilization treatments (in triplicate) for “cau cau” of giant squid indicated a maximum of 7.71% and 3.41% CV for F₀ and Co, respectively

The SCC1 (F₀ = 6.84 min) and SCC2 (F₀ = 9.36 min) can be stored up to 25 °C; SCC3 (F₀ = 13.83 min) is suitable for tropical areas (up to 40 °C)

1 Introduction

Sterilized foods correspond to convenience foods, which are ready-to-eat or convenient foods (Galizio & Diaz, 2020; Lee & Shin 2023). These types of convenience foods are experiencing increased consumption due to consumers' modern, fast-paced lifestyles (Kirse-Ozolina et al., 2019; Ditudompo et al., 2022; Wang et al., 2024), and were in greatest demand during the Coronavirus Disease (COVID) pandemic, with a forecast annual growth of 7.7% from 2022 to 2032 (Future Market Insights, 2022). Sterilized foods are popular in the International Market due to their simple transportation, ease of consumption, and long shelf life (Shan, 2015)

Sterilization is a heat treatment method used for food preservation, where food inside an airtight and resistant container is exposed to high temperatures for a certain time to destroy microorganisms responsible for food spoilage and extend its shelf life (Jyoti et al., 2024). Compared to conventional sterilized canning technology (which uses tinplate packaging), the use of flexible sterile or retort packaging has the advantage of affecting to a lesser extent the sensory and nutritional properties of the product, lower production costs, lower energy expenditure and reduced processing time (Majumdar et al., 2017a; Pal et al., 2019; Soni et al., 2020; Punia Bangar & Whiteside, 2024). The reason is that it requires less heat and time to reach commercial sterility (reducing from 30% to 50%) since the product in the flexible pouches has less thickness and a high ratio of surface area to volume (Durance & Collins, 1991; Topno et al., 2013; Nelluri et al., 2022). However, food sterilization in flexible packaging requires special autoclaves that use water immersion as a heating method (Jimenez et al., 2024), and a source of overpressure; since during sterilization an external pressure must be applied to the flexible pouches to resist the sterilization process (tinplate packaging resists due to its double sealing); this external pressure, called overpressure or backpressure, is higher than 1.05 kg/cm² or 15 psi (Costa-Viver & Suárez-Morales, 2015).

Sterilizable flexible pouches usually consist of i) polypropylene (inside, contact with food); ii) aluminum; iii) nylon; and iv) polyester (outer layer, where the printing goes) (Canadian Food Inspection Agency, 2020; Kontominas et al., 2021). In Peru, the hydrobiological products industry in sterilizable flexible pouches is still incipient, due to the lack of consumer adaptation to these products and the limited supply of product alternatives by companies.

The giant squid "Dosidicus gigas" is a cephalopod that is exported mainly as frozen raw material by Peru (2023). On the other hand, "cau cau" is one of the most representative dishes of Peruvian creole food which consists of a Peruvian spicy dish made from cow stomach (mondongo) or mutton cut into small pieces, accompanied by potatoes, carrots, peas, yellow peppers, garlic, and onion (PeruInfo, 2020). To add value and produce food with stew or convenience foods, sterilizing these products in flexible pouches represents an alternative for diversifying the offer to new potential consumers.

The sterilization value (F₀) initially developed by Bigelow in 1920 is a parameter to evaluate the effectiveness of heat treatment or safety, through the decrease logarithm of the microorganism Clostridium botulinum (Holdsworth & Simpson, 2016). Likewise, the heat application, apart from generating microbial destruction, also causes nutrient degradation, texture change enzymatic inactivation. These changes are represented by the cooking value (C₀), developed by Mansfield in 1962 (Holdsworth & Simpson, 2016; Ohlsson, 1980), through the decrease of vitamin B1 (thiamine).

Due to the above advantages, processing in these flexible pouches has gained popularity in India (Majumdar et al., 2017b). Also, from the literature search conducted, research articles have been found primarily from Asia using flexible pouches with products of mackerel balls "Rastrelliger kanagurta" (Sarifudin et al., 2022), shrimp in masala - sauce from India (Puthanangadi Dasan et al., 2021), skipjack tuna "Katsuwonus pelamis" (Azhari et al., 2023) shrimp analog from lizardfish "Saurida tumbil" (Hema et al., 2021), anchoveta "Stolephorus commersonnii" (Bindu et al., 2010), where different values of F₀ (thermal death), C₀ (quality loss), texture profile, among others were evaluated.

Therefore, the objective of this study was to develop convenience food based on the mantle of giant squid (Dosidicus gigas) using flexible sterilizable bags and evaluate their changes in aspects of physicochemical and sensory properties to improve the quality and safety of the product.

2 Material and methods

2.1 Raw materials

In April 2022, 67 kg of fresh giant squid (SCC) mantle (Dosidicus gigas) (n = 5) in tube cut (LT mantle: 52.9 ± 0.52 cm and mantle thickness of 1.5 ± 0.18 cm) was acquired from Ventanilla Fishing Terminal (Callao, Peru) under refrigerated conditions, inside isothermal boxes and mixed with crushed ice (3 °C). It was transported within 30 min to the processing facilities of the CITEPesquero Callao of the Technological Institute of Production (TIP), in Peru. They were processed on the same day, and the quality characteristics are reported in the results of the proximate analysis.

In the TIP, the SCC mantles were cut longitudinally, obtaining an extended mantle layer, which was then cleaned by making cuts to remove remnants of organs (gills, feathers, stomach, and gonads). Afterwards, the SCC mantle was washed with cold water (3 °C).

Then, the SCC mantles were cut manually with a knife into strips (width of 2 cm), placed to cook by immersion in water at 95 °C for 30 min, and cooled by immersion in ice water (3°C). Finally, the SCC strips were cut every 2 cm. The product obtained was called SCC mantle cubes. The production yield was 48% S/S (giant squid cube/fresh giant squid mantle).

2.2 Elaboration on the “cau cau” of the giant squid

Cubes of SCC mantle and other ingredients (Table 1) were used for the preparation of the “cau cau” of SCC.

Table 1
Ingredients of “cau cau” of giant squid (SCC).

Ingredients	(%)*
Precooked giant squid cubes	35.7
Potato	11.9
Peas	4.0
Carrot	4.0
Vegetable oil	8.9
Yellow chili	5.3
Garlic	2.2
Onion	1.8
Salt	0.9
Xanthan gum	0.2
Ground cumin	0.2
Ground black pepper	0.1
Water	24.8

• (*) The amount is expressed in W/W (weight for weight).

All the vegetable ingredients were procured fresh and stored at room temperature. Before use, they were washed under running water. The potato and carrot were peeled and cut into cubes of approximately 1 cm³. Peas and garlic were peeled. Potato, carrot, and pea were blanched at 95 °C for 2, 3, and 2 min, respectively, and cooled. The yellow chili pepper (previously removed from its internal veins) was cut into thin strips, and the onion was cut into small cubes (0.5 cm³). Garlic, yellow chili pepper, and onion were pre-fried with oil for 10 minutes. Then, in an industrial blender (20 L capacity), the pre-fried ingredients were added, and the remaining oil was added along with salt, cumin, pepper, xanthan gum, and water. This mixture was blended for 15 minutes; this mixture was called “cau cau” sauce. After having all the ingredients prepared, 61 g of SCC mantle, 20 g of potato, 7 g of carrot, 7 g of pea, and 75 g of “cau cau” sauce (170 g net weight) were weighed on a scale. All the above was poured into a flexible sterilizable pouch 180 mm long and 130 mm wide, with a thickness of about 133 µm +/- 10%. The flexible pouch was internally composed of HB (high barrier) PET (12 µm), in the middle nylon (15 µm), and on the outside white CPP (cast polypropylene) (100 µm) (Dongwon Systems Corp., South Korea).

Subsequently, the flexible pouch was sealed at 60 mbar with a vacuum sealer (MULTIVAC C500, USA) and then cooled by ice water immersion (3 °C). A total of 750 packages of “cau cau” of SCC in flexible pouches were obtained.

2.3 Heat treatment “cau cau” of giant squid

The flexible pouches of “cau cau” of SCC were randomly divided into three equal parts (250 units each), which were sterilized at 121 °C with 2.3 kg/cm² in a 1.8 m³ water immersion horizontal autoclave (Kamigaki Works Co. Ltd., Japan) (Figure 1A). Each group was sterilized for 25 min (SCC1), 30 min (SCC2), and 35 min (SCC3), respectively, obtaining three treatments.

Figure 1
Horizontal water immersion autoclave (A). Arrangement of thermocouples in flexible pouches (B).

The temperature during sterilization of each treatment was recorded in triplicate. For this purpose, a compression fitting for probes (2.0 mm in diameter and 42 mm in length) was fitted into the geometric center of three containers for each treatment (E-Val Pro, GKJ31009C042, Denmark), which were inserted into two buckets of SCC from each flexible pouch. Then, the remaining ingredients and covering liquid were added, vacuum was generated, and the containers were hermetically sealed. A T-type thermocouple (SSA-TS, Denmark) was placed in each compression fitting for temperature recording during sterilization (Figure 1B). The three packages were placed at the coldest point of the autoclave, previously determined based on the heat distribution (data not shown). Likewise, the autoclave temperature was recorded in quadruplicate using 4 T-type thermocouples. The product and autoclave temperature were recorded every second in real time, using the thermal validation cable system (brand ELLAB, Val Suite Pro 6.0 16.0, Denmark). As an additional verification measure, the autoclave temperature was monitored using a calibrated digital thermometer (TEINCO brand, T-1556, Spain).

Once the SCC flexible pouches are inside the autoclave, the sterilization process consists of: i) heating 1.6 m³ of water to 125 °C of the upper chamber by indirect contact with saturated water steam and air injection to raise the chamber pressure (to 2.3 kg/cm²); ii) lowering hot water to the sterilization tank (where the flexible pouch SCC is placed); iii) sterilizing the SCC flexible pouches by direct contact with water at overpressure, and; iv) cooling the samples, for which the hot water is pumped to the upper chamber and stored until the next are located use. Immediately, water (1.6 m³) at room temperature enters and remains in contact with the SCC flexible pouches for about 10 min until it reaches 40 °C. Subsequently, the autoclave was opened, and the “cau cau” of the SCC flexible pouches was removed and immersed in ice water for 40 min. The flexible pouches were drained, dried, and quarantined for 15 days for further evaluation.

The F₀ value was determined using Formula 1 (Ling et al., 2015; Soleymani Serami et al., 2021; Kim et al., 2020)

$\mathrm{F}\mathrm{o}={\int }_0^t{10}^{\left(\frac{T-Tref}{Z}\right)}dt$ (1)

where: "Z" is 10 °C (thermal resistance coefficient), "T" is the recorded temperature considered as the slowest heating zone, and "t" is the time (min) of the lethal velocity.

For the sterilization value (F₀), 121.1 °C is commonly used as a reference (temperature Tref.) and indicates the cumulative thermal effect in the reduction of C. _{botulinum (with} a Z value of 10 °C). In that sense and knowing the decimal logarithmic reduction time for C. botulinum (D = 0.25 min), it is advisable to perform a 12-log cycle reduction to ensure a statistical probability of survival of less than 1 spore in 1 billion (10¹²), known as 12D; therefore, a heat treatment at 121.1 °C for 3 min was performed to achieve 12 decimal log reductions in the population C. botulinum (Hall, 1997); however, the F₀ ^{value of 3 min} comes from another concept (Holdsworth, 2004). For sterilized canned goods, it is considered to achieve at least an F₀ value of 3 min (Bratt, 2013; Gopal et al., 1998), however, in the canning industry an F₀ value of 6 to _{14 min is obtained to} provide an additional margin of safety and to compensate for some possible inaccuracies in temperature measurement during sterilization (Hall, 1997).

In this sense, this study was designed to obtain F₀ values of 6, 9, and 13 _{min for} canned SCC in flexible pouches. To know the decrease in quality and degree of texture, among other ^{sensory attributes} generated by the heat treatment, the cooking value ₍C₀⁾ was determined based on Formula 2 (Ling et al., 2015):

$\mathrm{C}\mathrm{o}={\int }_0^t{10}^{\left(\frac{T-Tref}{Z}\right)}dt$ (2)

where: "Z" is 33.1 °C and "t" is the time (min) of the quality loss rate. Generally, the reference temperature at the cooking value (C₀) is 100 °C, which corresponds to the loss of quality during a thermal process equivalent to cooking at 100 °C (Lund, 1977). This is typically assessed using the degradation of thiamine as a reference, with a Z-value of 33.1 °C (Z equal to 33.1 °C). (Ohlsson, 1980; Ding et al., 2022)

2.4 Proximate analysis

Samples of: a) raw material: raw and skinless SCC mantle (MSS), and; b) finished product: three treatments of sterilized SCC flexible pouches, whose contents were previously drained for 3 minutes, were analyzed for their proximate chemical composition using the following methods: i) moisture determination by oven drying at 100 °C at constant weight (AOAC 952.08A, 2019) (Association of Official Analytical Chemists, 2019d); ii) fat determination by chemical hydrolysis method and subsequent extraction with diethyl ether (AOAC 948.15, 2019) (Association of Official Analytical Chemists, 2019c); iii) protein determination by Kjeldahl method with digestion with sulfuric acid and subsequent titration by neutralization with NaOH (0.1 N) (AOAC 940.25, 2019) (Association of Official Analytical Chemists, 2019b); iv) ash determination by calcination in a muffle at 550 °C (AOAC 938.08, 2019) (Association of Official Analytical Chemists, 2019a) and; v) carbohydrate determination by difference calculation.

The pH value in the “cau cau” sauce (before sterilization) was 6.31. The pH was determined with a pH meter (Mettler Toledo, model Seven Compact S220, Spain), where the electrode was immersed directly in the sauce. With this pH value, the “cau cau” product of the SCC is considered a low-acid food (pH > 4.6) (U.S. Food and Drug Administration, 2010). Therefore, this product must be sterilized to ensure its safety.

2.5 Color analysis

The color was measured instrumentally using a colorimeter (Konica Minolta, CM-5, Japan) on the CIELAB color scale, where: i) L*: lightness (0 to 100); ii) a*: from red (+) to green (-), and iii) b*: from yellow (+) to blue (-). At room temperature, the pre-cooked ingredients are listed as follows: giant squid; potato; carrot; pea; and the “cau cau” sauce (n = 5). These ingredients in their respective three treatments (SCC1, SCC2, and SCC3 (n = 15)) were poured directly into a well and measured in the colorimeter.

2.6 TPA texture test analysis

At ambient temperature, the pre-cooked ingredients (giant squid, potato, carrot, and pea (n = 6)) and their respective three treatments (n = 18) were characterized in terms of their texture profile using a texturometer (Brookfield, model CT3-1500, USA). A TA39 cylindrical probe was used, and each sample was subjected to a double compression to 30% deformation (stress normal), and a probe velocity of 0.5 mm/s with a waiting time of 5s between compressions. The texture parameters evaluated were as follows: hardness (N); adhesiveness (mJ); fracturability (N); cohesiveness; elasticity (mm); and chewiness (mJ).

2.7 Commercial sterility test

The commercial sterility test was performed on the three treatments (SCC1, SCC2, and SCC3), where four flexible pouches of each treatment were incubated at 37 °C for 14 days, using different culture media (for mesophilic and thermophilic aerobic microorganisms), according to the Food and Agriculture Organization (FAO) methodology.

2.8 Sensory evaluation

The sensory evaluation was a preference test (Stone et al., 2021; Anzaldúa, 1994; Świąder & Marczewska, 2021), conducted with the participation of 30 panelists (15 males and 15 females), all over 18 years of age, who were selected based on their regular consumption of traditional cau cau, approximately twice a month. The flexible pouches of “cau cau” of the SCC of the three treatments were tempered by immersing them in a kettle at 90 °C for 5 min. Subsequently, the contents of one package (undrained) of each treatment were divided into two white porcelain dishes and presented to two panelists for sensory. Each evaluation panelist received a dish of each treatment (Figure 2).

Figure 2
“cau cau” of giant squid SCC1 (A), SCC2 (B) and SCC3 (C).

For the registry of the sensory evaluation, each panelist received (via cell phone) a survey entitled: "Evaluation of product attributes of “cau cau” of giant squid” using Google Forms. This survey included the identification of the panelists (name, gender, and age) and the instructions for the sensory evaluation, which was also verbally induced.

The instructions for the sensory evaluation were:

Observe and try the product in front of you and answer the following statements.

Mark with an "X" in the corresponding statement. There were five statements:

1. In my opinion, the color of the product is:

2. In my opinion, the smell of the product is:

3. In my opinion, the texture of the giant squid is:

4. In my opinion, the taste of the product is:

5. In my opinion, the overall appearance of the product is:

Each statement used a 9-point rating scale (indicated by a circle), where each number corresponds to the following rating:

1 : Dislike it extremely, 2 : Dislike it very much, 3 : Dislike it moderately, 4 : Dislike it slightly, 5 : Neither like nor dislike it, 6 : Like it slightly, 7 : Like it moderately, 8 : Like it very much, and 9 : Like it extremely.

2.9 Statistical analysis

Statistical analysis of the results was performed with the SPSS program (version 25, IBM Corp., States). Normal distribution was evaluated with the Shapiro-Wilk test (n < 30), Levene's test, and Welch's one-factor Analysis of Variance (ANOVA), a means comparison and the mean obtained will be evaluated with the Games-Howell test. A significance level of p = 0.05 was used. Texture and sensory evaluation results had a non-parametric distribution and were compared using the Kruskal-Wallis test (p = 0.05).

3 Results and discussions

3.1 Proximate analysis

Table 2 shows the tests performed on the raw skinless SCC mantle (MSS) and on the three treatments “cau cau” of SCC (SCC1, SCC2, and SCC3). Proximate analysis results (moisture, fat, protein, carbohydrate, and Total Volatile Basic Nitrogen - TVBN) of MSS were like reports from other studies (Albrecht-Ruiz et al., 2017), where it is reflected that SCC meat is lean, with low-fat content.

Table 2
Proximate analysis of raw material and finished product samples.

Testing	Mantle of skinless giant squid (MSS)*	“Cau cau” of giant squid 1 (SCC1)*	“Cau cau” of giant squid 2 (SCC2)*	“Cau cau” of giant squid 3 (SCC3)*
Moisture (g/100 g)	85.7a ± 0.71	77.8b ± 0.10	78.0b ± 0.23	78.1b ± 0.25
Fat (g/100 g)	0.8a ± 0.10	6.6b ± 0.36	6.2b ± 0.16	6.6b ± 0.04
Protein (g/100 g)	12.4a ± 0.51	8.5b ± 0.59	9.1b± 0.45	9.14b ± 0.07
Ash (g/100 g)	1.0a ± 0.04	1.3b ± 0.01	1.3b ± 0.01	1.3b ± 0.01
Carbohydrate (g/100 g)	0.1a ± 0.06	5.7b ± 0.31	5.4b ± 0.83	4.9b ± 0.26
Total volatile basic nitrogen (TVBN) mg/100 g)	65.9a ± 3.80	73.8b ± 0.15	73.6b ± 0.30	73.6b ± 0.17
Commercial sterility	Not applicable	Commercially Sterile	Commercially Sterile	Commercially Sterile

• (*) Different letters in superscript in the same row indicate significant differences (p < 0.05) between samples.

According to Table 2, the proximate analysis showed no significant difference (p > 0.05) between treatments SCC1, SCC2, and SCC3. The significant increase in fat and carbohydrates in SCC1, SCC2, and SCC3 were due to the use of ingredients such as oil and potato, among others.

The TVBN content in MSS was 65.9 mg/100 g and showed a slight increase in SCC1, SCC2, and SCC3 (ranging from 73.6 to 73.8 mg/100 g). It should be noted that, unlike in fish, the TVBN content is not considered an indicator of freshness in SCC and does not have an established maximum limit (Peru, 2016). The components that make up the TVBN (mainly ammonium, chloride, among others) are related to physiological and metabolic functions of the species (Maza et al., 2007), serving as an osmoregulatory role that allows control of body, density swimming performance, and buoyancy (Lida et al., 1992)

3.2 Commercial sterility

The three treatments (SCC1, SCC2, and SCC3) were commercially sterile (see Table 2), showing no observations after incubation for 14 days at 37 °C.

3.3 Heat treatment

The sterilization value (F₀) and cooking value (C₀) of the three treatments (SCC1, SCC2, and SCC3) are shown in Table 3. In the treatments performed, there was a rise time of about 4 min (known as CUT: "come-up time"), i.e., time required from the beginning of heating to reach the regime temperature (121.1 °C), which is reflected in Figure 3.

Table 3
F₀ and C₀ of sterilized products.

	SCC1 (25 min)			SCC2 (30 min)			SCC3 (35 min)
	F0 (min)	C0 (min)	C0/F0	F0 (min)	C0 (min)	C0/F0	F0 (min)	C0 (min)	C0/F0
n1	7.65	57.38	7.5	10.90	73.71	6.8	15.14	91.82	6.1
n2	7.20	57.17	7.9	10.42	71.79	6.9	14.06	88.40	6.3
n3	6.84	55.38	8.1	9.36	68.87	7.4	13.83	88.85	6.4
Average	7.23	56.64	7.8	10.23	71.46	7.0	14.34	89.69	6.3
Standard Deviation	0.41	1.10	0.3	0.79	2.44	0.3	0.70	1.86	0.2
% CV	5.61	1.94	3.9	7.71	3.41	4.5	4.88	2.07	2.9
C0/F0 per treatment	8.4			7.9			6.6

• %CV: %Coefficient of Variation.

Figure 3
Heat treatment for SCC1 (A), SCC2 (B), and SCC3 (C), showing ^autoclave temperature ( ), product core temperature ( ), F₀ ( ), and _firing value _{( ).}

It should be noted that, among the three measurements taken for each treatment, ^{the lowest} value and the highest C₀ value (highlighted in bold) were selected to evaluate the minimum degree of safety of the SCC due to microbial destruction and the maximum impact on sensory characteristics and quality of the sterilized products, respectively.

According to Table 3, treatment 1 (SCC1) had an F₀ value of 6.84 min and C₀ of 57.38 min; treatment 2 (SCC2) had a value F₀ 9.36 min and C₀ of 73.71 min and treatment 3 (SCC3) had an F₀ value of 13.83 min and C₀ of 91.82 min. Likewise, according to the coefficient of variation (%CV), F₀ exhibited greater variability (from 4.88% to 7.71%) compared to C₀ (from 1.94% to 3.41%); however, it is not possible to compare the variability between different measurements; however, both parameters (F₀ and C₀) of each treatment can be considered to have acceptable variability. The _{variability margin} depends on _{the type} of experiment or parameter being evaluated, such as, for example: 8% for crop evaluation; 12% for fertilization; and 15% for pesticide evaluation, among others (Gordón-Mendoza & Camargo-Buitrago, 2015)

At the time of writing this article, no reference was found indicating ^{the maximum} variability (expressed as %CV) for values F₀ (repetitions) in the _{canning sterilization process;} however, it is understood that it should be as low as possible due to the critical importance of ensuring product safety. Given this and after consulting the Food and Drug Administration (FDA) (e-mail sent on 20/08/2024, personal communication), they responded that there is no official document on the matter and that a %CV of 5% is universally accepted in the industry. The “cau cau” of SCC in this study can be considered as a non-homogeneous canned food (Institute for Thermal Processing Specialists, 2014) (consisting of solids and a significant amount of liquid). Additionally, it includes several ingredients (see Table 1) with greater variation in dimensions (length, width, and height) of the flexible pouches compared to homogeneous products (consisting mainly of solids and a reduced amount of liquid) that can be considered canned fish in tinplate packaging, as they usually contain fish, meat, salt, oil and/or water. Also, heat transfer in non-homogeneous canned foods (such as in this study) occurs primarily through convection and conduction, while for homogeneous canned foods, it occurs mainly by conduction (Chen & Ramaswamy, 2007). All these factors influence the variability of temperature _recording in the product during sterilization (affecting the variation of F₀), ^such variation is quite common and higher in non-homogeneous preserves (Institute for Thermal Processing Specialists, 2014), for example, the %CV of F₀ fluctuates from 34% to 60% in preserves in flexible, packaging while the %CV of F₀ is from 3% to 10% in canned foods in tinplate packaging ^(data not shown from industrial), scale records all this variability may not yet allow a consensus to establish a maximum %CV limit for F₀ ^{values in sterilization.}

On the other hand, based on Table 3, there is a decrease in the C₀/F₀ ratio: 7.8, 7.0, and 6.3 (average values for each treatment, underlined) as the time increases sterilization from 25 min, 30 min, and 35 min, respectively. Also, from the values previously chosen (in bold), the decrease in the ratio is observed in F₀ and C₀ C₀/F₀ (row bottom of Table 3) for each treatment: 8.4, 7.9, and 6.6 as the sterilization time increases.

From Table 3, the temperature record for each treatment of the chosen values for F₀ and C₀ (highlighted in bold) is shown in Figure 3.

In compliance with safety standards for canned foods, all treatments in this study ^{reached and exceeded the} recommended F₀ (F₀ greater than 3 min) (Bratt, 2013; Gopal et al., 1998). Regarding C₀, it is suggested to have a maximum range of 100 to 200 min, as a C₀ greater than this range is considered deteriorated quality (Awuah et al., 2007). The highest value C₀ obtained in the present study was 91.82 min.

Different factors influence the value F₀ in sterilization, such as type of _{packaging, type of} product and physicochemical composition, net weight, and sterilization temperature, among others. This study used 170 g of net weight of SCC caul ^{sterilized at 121.1 °C in} flexible pouches, obtaining an F₀ of 6.84 min with 25 min of _{sterilization time, such} sterilization time was close to those recorded by other studies, which sterilized at 121.1 °C in flexible pouches such as: i) 26.01 min (^weight 200 g ^net F₀ of 7 min) (Ali et al., 2005); ii) 27.54 min (150 g net weight, F₀ of 6 min) (Sreelakshmi et al., 2015); iii) 38.84 min (220 g net weight, F₀ of 6 min) _{(Majumdar et al., 2017a).}

In this sense, to reach an F₀ of 6.84 min according to the _{experimental conditions} mentioned in this study (flexible pouches), 25 min were required; however, the sterilization of products such as canned fish in tinplate of 170 g net weight requires about 43 to 50 min (data not shown from records of packaging made on an industrial scale). This shows that these flexible packages require about 58% of the sterilization process time compared to traditional tinplate packaging. In this regard, Ali et al. (2005) reported that the sterilization time for canned sardines in flexible packaging was 26.01 min and 33.16 min in tinplate, showing that approximately 78% of the time was required to achieve sterilization in flexible packaging compared to tinplate cans (maintaining the same net weight and temperature packaging sterilization).

3.4 Color analysis

Table 4 shows the color expressed in L*, a*, and b* of the giant squid, potato, carrot, pea, and “cau” sauce in the precooked state and after the sterilization process, obtaining SCC1, SCC2, and SCC3. All the precooked ingredients (giant squid, potato, carrot, pea, and cau sauce) had significant differences (p < 0.05) in L*, a*, and b* concerning the same ingredients present in SCC1, SCC2, and SCC3; these changes in color parameters (L*, a* and b*), for example, the decrease in lightness (L*) and increase in red color intensity (a*) (except in carrot), could be influenced by the migration of pigmentation from yellow chili pepper, onion and also by the effect of pre-frying of some ingredients that, due to the reduction of sugars, produced the Maillard reaction causing a moderate non-enzymatic browning (Bordin et al., 2013), being sensorially acceptable in these products.

Table 4
Color analysis of ingredients, “cau cau” sauce (precooked and with respective treatments) (n = 15).

Ingredients	Color Parameter (1)
	L*	a*	b*
Precooked squid	84.87a ± 0.51	-0.23a ± 0.26	10.68a ± 0.32
Squid in SCC1	72.09b ± 0.12	2.54b ± 0.00	18.97b ± 0.05
Squid in SCC2	71.76c ± 0.03	2.64c ± 0.03	19.08b ± 0.07
Squid in SCC3	69.69d ± 0.04	3.56c ± 0.03	18.79b ± 0.03
Precooked potato	72.50a ± 0.10	-1.58a ± 0.50	23.40a ± 1.44
Potato in SCC1	69.40b ± 0.01	1.49b ± 0.01	20.50b ± 0.18
Potato in SCC2	67.43c ± 0.13	2.16c ± 0.00	21.95a ± 0.03
Potato in SCC3	65.05d ± 0.04	2.92d ± 0.01	20.59b ± 0.03
Precooked carrot	52.46a ± 0.04	34.60a ± 0.47	38.12a ± 1.51
Carrot in SCC1	47.43b ± 0.01	24.82b ± 0.02	30.94b,c ± 0.02
Carrot in SCC2	46.74c ± 0.02	25.27c ± 0.04	30.87b ± 0.03
Carrot in SCC3	45.16d ± 0.03	23.49d ± 0.07	30.80c ± 0.16
Precooked peas	46.96a ± 0.24	-11.49a ± 0.03	20.46a ± 0.11
Peas in SCC1	51.13 b ± 0.08	1.14b ± 0.05	25.94b ± 0.06
Peas in SCC2	51.63 c ± 0.04	1.2c ± 0.04	24.02c ± 0.19
Peas in SCC3	50.34d ± 0.01	1.85d ± 0.00	24.37d ± 0.01
“Cau cau” sauce	73.35a ± 0.04	8.07a ± 0.20	50.96a ± 0.10
Sauce in SCC1	66.95b ± 0.04	8.35b ± 0.01	52.89b ± 0.02
Sauce in SCC2	65.29c ± 0.17	9.13c ± 0.01	51.68c ± 0.04
Sauce in SCC3	62.73d ± 0.01	9.75a ± 0.03	48.32d ± 0.03

• ⁽¹⁾
  Different letter in superscript indicates significant difference (p < 0.05) in the same column of each ingredient.

Likewise, comparing the three heat treatments, the longer the sterilization time (SCC3>SCC2>SCC1), the lower the lightness (L*) (except in pea), the higher the (a*) values (i.e., increase in red, except color intensity in carrot); there is no marked difference in color change in b* (except parameter in pea and sauce). At longer sterilization time, this study reported lower L*; Majumdar et al. (2017b) reported a decrease in L* in shrimp in curry sauce; Shah et al. (2017) showed that there was no variation in L* in meat and sauce of a traditional Indian dish. All the above indicate that the variation in color parameters is influenced not only by sterilization time but also by the composition of product ingredients, etc.

3.5 Texture profile analysis

The texture parameters (hardness, adhesiveness, fracturability, cohesiveness, springiness, and chewiness) that are part of the texture profile analysis and that simulate chewiness through 2 compression cycles in the present study are shown in Table 5.

Table 5
Texture profile analysis of precooked ingredients (n = 6) and with treatments (n = 18).

Samples	Texture parameters(1)
	Hardness 1 (N)	Adhesiveness (mJ)	Fracturability (N)	Hardness 2 (N)	Cohesiveness	Elasticity (mm)	Chewiness (mJ)
Precooked squid	0.57a	0.03a	0.66a	0.47a	0.79a	44.00a	21.51a
Squid in SCC1	3.08b	0.04a	3.08b	2.93b	0.87a,b	32.98b	79.85b
Squid in SCC2	2.68b,c	0.03a	2.68b,c	2.53b,c	0.82a	31.31b	70.89b
Squid in SCC3	2.03a,c	0.01a	2.03a,c	1.97a,c	0.93b	32.61b	61.72b
Precooked potato	3.20a	0.30a	2.27a	2.00a	0.25a	44.31a	37.21a
Potato in SCC1	1.79a	0.05b	1.78a	1.38a	0.33a	35.51b	23.51a
Potato in SCC2	1.70a	0.03b	1.70a	1.24a	0.38a	37.19b	24.25a
Potato in SCC3	1.55a	0.05b	1.55a	1.23a	0.36a	36.54b	20.50a
Precooked carrot	2.86a	1.13a	2.49a	1.80a	0.18a	19.60a	8.30a,b
Carrot in SCC1	1.06a,b	0.005b	0.43a,b	0.91a	0.53a	41.06a	19.94a
Carrot in SCC2	0.20c	0.03b	0.02b,c	0.16b	0.32a	41.61a	3.30b
Carrot in SCC3	0.29c	0.05b, a	0.02c	0.26b	0.25a	40.63a	2.14b
Precooked peas	2.40a	0.02a	1.64a	1.42a	0.53a	45.22a	42.78a
Peas in SCC1	1.88b	0.03a	1.88a	1.44a,b	0.51a	36.83b	29.23a,b
Peas in SCC2	1.12a,b	0.03a	0.57a,b	0.97a,b	0.55a	38.14b,c	24.32a,b
Peas in SCC3	0.42b	0.02a	0.27b	0.38b	0.63a	39.07a,c	10.86b

• ⁽¹⁾
  A different letter in superscript indicates a significant difference (p < 0.05) in the same column.

According to Table 5, concerning precooked SCC, hardness 1 and 2 first and second compression cycles) and fracturability increased significantly (p < 0.05) by about 4 to 6 times in the three heat treatments (SCC1, SCC2, and SCC3); likewise, but to a lesser extent, cohesiveness and chewiness also increased. The springiness of the SCC decreased (p < 0.05) after the heat treatments. However, the other precooked ingredients such as carrots and peas had a significant decrease (p < 0.05) in toughness (first and second cycles), adhesiveness, and fracturability with heat treatments (SCC1, SCC2, and SCC3) (except in the adhesiveness of the pea). This contrast of change in toughness behavior is because SCC flesh, formed mainly by myofibrillar and connective protein (collagen) (Torres-Arreola et al., 2008), has been thermally contracted, changing its volume and affecting its different bonds, causing a significant increase in its toughness (Grajales-Lagunes et al., 2007). On the other hand, potatoes, carrots, and peas, constituted mainly by carbohydrates and starch, among others, significantly decreased their hardness after sterilization. In addition, it could be noted that Greve et al. (1994) reported a similar result. Likewise, the fracturability of carrots and peas has decreased (p < 0.05) after thermal treatments. Potato, due to its high variation in fracturability in the precooked state, was statistically different (p > 0.05) after heat treatments.

Adhesiveness did not change (p > 0.05) in the precooked or heat-treated state in the SCC and peas, but there was a decrease in adhesiveness (p < 0.05) in the potato and carrot.

3.6 Sensory evaluation

Results of the sensory evaluation with the attributes: i) color; ii) odor; iii) texture; iv) flavor; and v) general appearance. The results are shown in Table 6.

Table 6
Sensory evaluation “cau cau” of giant squid (SCC) (n = 30).

Sensory attributes “cau cau” of giant squid *,1
Treatments	Color	Odor	Texture	Taste	General Appearance
SCC1	7.0a	7.0a	7.0a	7.0a,*	7.0a,*
SCC2	7.0a	6.0a,b	7.0a	7.0a	6.5a,b
SCC3	7.0a	6.5b	7.0a	7.0a	6.0b

• ^*
  All results were not normally distributed (except SCC1 for Taste and General Appearance). (_¹) Different letters in superscript in the same column indicate significant (difference p < 0.05) between the medians of the 3 treatments.

According to the results in Table 6, there were no significant differences (p > 0.05) in the attributes of color, texture, and flavor among the three treatments (SCC1, SCC2, and SCC3), whose result had a value of 7.0, indicating that the 30 panelists moderately liked the “cau cau” of SCC. On the other hand, there were significant differences (p < 0.05) in odor and general appearance among the three treatments of the SCC, where in both attributes, the treatment SCC1 had the highest score, with a value of 7.0, indicating that the “cau cau” of SCC in flexible pouches sterilizable was moderately liked by the participating panelists. In general, no marked differences were found in the sensory evaluation product of these products sterilized at different sterilization times, similar results were reported by Sarifudin et al. (2022).

In this regard and not finding marked sensory differences in these three heat treatments applied to the “cau cau” of the SCC in flexible pouches, the products SCC1 ⁽F₀ ^{= 6.84 min)} and SCC2 (F₀ = 9.36 min) can be intended to normal storage conditions, where temperature is less than 25 °C (Heinz & Hautzinger, 2007).

This heat treatment design has the objective of inactivating the spores _{C. botulinum with} F₀from 3 min to 121.1 °C (Hall, 1997); while the SCC3 product with a higher F₀ value (13.83 min), can be intended for commercialization in tropical areas where the _{storage temperature} may exceed 25 °C (up to 40 °C) (Heinz & Hautzinger, 2007), such as jungle areas or even in military incursions. Sterilization targeted for products in tropical areas require a value F₀ of 12 to 15 min (Heinz & Hautzinger, 2007), necessary to significantly reduce spores of Geobacillus stearothermophilus, which have been reported as a contaminating agent in these products exposed to such environmental temperature conditions (Rigaux et al., 2014; Nnko et al., 2001). Spores of G. stearothermophilus are up to 20 times more resistant than spores of C. botulinum (Iciek et al., 2008).

4 Conclusions

The canned “cau cau” of (Peruvian creole food) of giant squid (SCC) in flexible pouches was sterilized at 121.1 °C for 25 min, 30 min, and 35 min, obtaining values F₀ for 6, 9, and 13 min, respectively; where there were some significant differences (p < 0.05) in color analysis, instrumental texture profile, and sensory evaluation among the three treatments. In general, and according to the sensory evaluation, the three treatments of “cau cau” of SCC had a moderate acceptance preference among the panelists. This research proposes that SCC sterilized at 121 °C in a horizontal water immersion autoclave during 35 minutes can be marketed in tropical areas (temperatures ranging from 25 °C to 40 °C), while treated during 25 and 30 minutes can be marketed under normal storage conditions (temperatures below 25 °C). As a result, diversification of SCC consumption is appropriate for these sterilized products.

Data Availability Statement

All data generated or analyzed in this study are included in this published article.

• Cite as:
  Bustinza-Hilari, J. V., Cuba-Mayo, F. E., & Aldoradin-Puza, E. (2025). Effect of sterilization on the physicochemical and sensory characteristics of “cau cau” (Peruvian creole food) of giant squid (Dosidicus gigas) in flexible pouches. Brazilian Journal of Food Technology, 28, e2025059. https://doi.org/10.1590/1981-6723.05925
• Funding:
  None.

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Edited by

• Associate Editor:
  Rogério Souza de Jesus.

Publication Dates

• Publication in this collection
  23 Jan 2026
• Date of issue
  2025

History

• Received
  07 July 2025
• Accepted
  23 Oct 2025