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The Effects of Commercial Freezing on Vitamin Concentrations in Spinach (Spinacia Oleracea)

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The Effects of Commercial Freezing on Vitamin Concentrations in Spinach (Spinacia Oleracea) JULS Review Article The Effects of Commercial Freezing on Vitamin Concentrations in Spinach (Spinacia oleracea) Konrad Samsel1,2,* and Abizar Meghani2,3,4 ABSTRACT Commercial food processing has had a significant impact on reducing food spoilage and increasing accessibility to nutrient-dense vegetables. The commercial freezing process, in particular, has given producers the ability to store vegetables with minimized risk of microbial and enzymatic spoilage. Despite the effectiveness of freezing as a preservation method, there is evidence that pre-freezing procedures and prolonged storage can reduce the concentration of vitamins present within certain vegetables. Spinach, one of the most widely produced and consumed vegetables, is particularly susceptible to nutrient loss during the commercial freezing process due to its large surface area and high mineral content. This review summarizes the known effects of the freezing process on hydrophilic and lipophilic vitamins including vitamin C, thiamin, riboflavin, β-carotene, and α-tocopherol. There are two key mechanisms that lead to decreased vitamin concentrations, with the first being attributed to pre-freezing processes including washing and blanching which favours the leaching of hydrophilic vitamins. The second mechanism of vitamin loss is attributed to residual enzymatic activity during storage, where the degree of residual activity can be partially attributed to differences in blanching protocols and freezing practices. Understanding the mechanisms and extent of vitamin loss that the commercial freezing process imparts on leafy green vegetables can help inform future research on improved food processing methods that minimize nutrient loss. Implementing procedures that maintain nutrient retention in frozen vegetables has the potential to assist individuals in achieving their recommended daily intakes of micronutrients. Keywords: Nutrients, Frozen Storage, Spinach, Vitamins, Ice Nucleation Affiliations *Please direct correspondence to: 1Department of Nutritional Sciences, University of Toronto, Konrad Samsel Canada Email: [email protected] 2Department of Ecology and Evolutionary Biology, University of Toronto, Canada Published online on July 23, 2021 3Department of Physiology, University of Toronto, Canada 4Department of Physics, University of Toronto, Canada 1 The Journal of Undergraduate Life Sciences • Volume 15 • Issue 1 • July 2021 | https://doi.org/10.33137/juls.v15i1.37032 Review Articles The Effects of Commercial Freezing on Vitamin Concentrations in Spinach INTRODUCTION their recommended daily allowance of vitamin A12, identifying how vitamin concentrations decrease during The commercial freezing process has allowed commercial freezing can help inform future research on consumers to store foods for extended periods of improved processing and storage methods that help time with minimal food spoilage1. Storing foods at individuals reach their nutritional requirements. This sub-zero temperatures reduces enzyme activity, review will evaluate evidence related to the effects of prevents microbial growth, and is one of the most the commercial freezing process on vitamin availability effective preservation methods with regard to in spinach. mitigating nutrient loss during prolonged storage2. Freezing as a preservation method has allowed year- BACKGROUND INFORMATION round consumption of nutritious vegetables including spinach, which is known for its high amounts of vitamin Nutritional Composition of Spinach C, thiamin, riboflavin, β-carotene, and minerals such Spinach (Spinacia oleracea) is a commonly consumed as iron and magnesium3. commodity and is ingested and cultivated in many regions throughout the world13,14. There are numerous Research suggests that pre-freezing processes and commercially available cultivars of spinach14,15, each prolonged storage of spinach can result in considerable bred for beneficial qualities such as yield, organoleptic losses of vitamin C, thiamin, riboflavin, and β-carotene, properties, or resistance to various biotic and abiotic and minor losses of α-tocopherol4–8. The observed large conditions13,16. With regard to the nutritional content of losses of water-soluble vitamins are mostly attributed fresh spinach, studies have found that variation can to pre-freezing processes, namely blanching, which be influenced by several intrinsic and extrinsic factors results in the leaching of vitamins and other soluble including growing season, environmental conditions, components of the food4,6. In this context, leaching can and cultivar16–20. The representative values for fresh be defined as the loss or extraction of nutrients from spinach provided by the United States Department the food by way of a fluid medium, namely water or of Agriculture show that it is a good source of vitamin condensed steam, that acts as a solvent. Prolonged C (28.1mg/100g), providing 31-37% of the DRI for storage at sub-zero temperatures has also been shown adults3,21. This vegetable also contains 0.078mg/100g to augment vitamin loss, with the proposed mechanism of thiamin, 0.189mg/100g of riboflavin, 469μg/100g of being residual activity of enzymes involved in vitamin vitamin A, 5626μg/100g of β-carotene, and 2.03mg/100g degradation9. This residual enzymatic activity can be of α-tocopherol3. A 100-gram serving is thus expected increased by the process of freeze concentration, where to provide 7% of the DRI for thiamin, 15-17% for enzymes and their substrates are concentrated within riboflavin, 52-67% for vitamin A, and 14% for vitamin a fluid medium present between intracellular ice crystal E in adults21. Determining these vitamin concentrations formations10. Decreases in vitamin concentrations in often involves homogenization of the spinach leaf frozen spinach may have an impact on public health, and addition of the homogenate to a solute specific with reduced vitamin levels affecting an individual’s to the vitamin being measured6,7,22. After the solution ability to reach their daily recommended intake (DRI) of containing the extract of the compound of interest is nutrients. With an estimated 47% of young Canadian filtered, the sample is analyzed via high pressure liquid adults having suboptimal or deficient levels of serum chromatography (HPLC)6,7,22,23. A protocol for ascorbic ascorbic acid11 and 29-46% of Canadians not meeting acid detection in spinach is detailed in Figure 1. Figure 1 Protocol for ascorbic acid detection in spinach. Measurement protocol adapted from Bouzari et. al.⁶ 2 The Journal of Undergraduate Life Sciences • Volume 15 • Issue 1 • July 2021 | https://doi.org/10.33137/juls.v15i1.37032 The Effects of Commercial Freezing on Vitamin Concentrations in Spinach Review Articles Figure 2 Impact of cooling rate on ice crystal formation and freeze concentration. Commercial Freezing Process of Spinach After blanching to reduce enzyme and microbial activity, The commercial freezing process includes several the product is then rapidly frozen at temperatures around pre-freezing steps to protect the quality and prevent -20°C to -40°C28. Lower temperatures will cause rapid spoilage of the final product. The most common pre- cooling of the product, resulting in increased numbers freezing processes are washing and steam-based of ice crystal initiation sites, known as nuclei29,30. This blanching. Blanching, which consists of a brief increased nucleation will limit the growth and final size application of heat to food, helps preserve product of the ice crystals29,30. One benefit of this finer crystal quality by decreasing enzyme activity and microbial size is that it can help preserve the texture and taste load24. Enzymes present in the cellular components of the spinach by minimizing freeze-concentration, can affect the colour, taste, and nutritional quality of the which occurs when solutes are aggregated due to vegetable following harvesting25. After the occurrence ice formation30. This physical process can increase of cellular damage from picking and handling , these substrate concentrations, increasing the kinetics of enzymes produce off-flavours, promote discolouring enzyme-mediated degradation reactions10. Preventing via enzymatic browning reactions, and promote vitamin the formation of large ice crystals also helps minimize degradation through enzyme induced oxidation25. The the likelihood of damage to cellular structures within temperature at which a product is blanched strongly the food. Damage to cellular structures affects the determines the degree of enzyme inactivation and nutritional and textural qualities of frozen food by subsequent residual enzymatic activity during storage. allowing the release of sequestered enzymes and by For example, studies examining the effects of blanching facilitating the loss of osmotic and turgor pressure28. on ascorbic acid oxidase (AAO) activity in vegetables This connection between cooling rate, nucleation, found that greater temperatures led to increased freeze concentration and crystal size is further depicted enzyme inactivation, helping to preserve vitamin C in Figure 2. content within the food over time26,27. While spinach blanching time and temperature may vary, a steam The frozen spinach is then stored at sub-zero blanch at 93.3°C for 3 minutes has been suggested to temperatures until it is defrosted or cooked. The be an effective pre-freezing treatment6. product often spends several months in commercial The Journal of Undergraduate Life Sciences • Volume 15 • Issue 1 • July 2021 | https://doi.org/10.33137/juls.v15i1.37032
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