Browsing by Author "Stone, Martha, advisor"

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Open Access
Comparison and acceptabilty of gluten-free yeast breads made with quinoa flour
(Colorado State University. Libraries, 2014) Chase, Caraline Bianchetto, author; Stone, Martha, advisor; Bunning, Marisa, committee member; Timpson, William, committee member
Quinoa (Chenopodium quinoa Willd) is a plant that is native to South America and is grown in the Andean mountains. The quinoa plant is resistant to harsh weather conditions and drought. It is a gluten free (GF) grain and has significance in making a GF yeast bread for people who cannot safely consume gluten proteins due to celiac disease, gluten allergies, or other issues. Most GF yeast breads on the market are made with a large portion of white or brown rice flour, which is neutral in taste, easily digestible, but low in vitamins and nutrients compared to quinoa flour. Quinoa flour is more expensive than rice flour and can impart negative aftertastes. The objectives of this study were to develop GF yeast bread formulas incorporating quinoa flour for rice flour and potato starch at 0, 36, 72, or 100% and obtain sensory and instrumental data on the breads. Specific gravity was calculated on the batter before baking with significant (p<0.05) differences existing among all batters. The 100% GF quinoa yeast bread was (p<0.05) smaller in volume than the other breads. The GF bread made with 100% rice flour and potato starch was significantly (p<0.05) softer (less hardness or firmness values) than the GF breads containing quinoa flour, while the 100% GF quinoa flour bread was firmest and least tender. Crust and crumb color did not (p>0.05) differ among any of the breads. The 100% GF quinoa yeast bread had the lowest water activity (p<0.05). Sensory analysis showed that for tenderness, flavor, and overall acceptability the 100% GF quinoa yeast bread was liked less (p<0.05) compared to the other breads. Based on the instrumental and sensory data collected, both the 36 and 72% QF yeast breads are acceptable GF yeast bread options containing QF.
Open Access
Development and evaluation of a weight control program for obese preadolescent children
(Colorado State University. Libraries, 1992) Hammarlund, Virginia Anne, author; Anderson, Jennifer, advisor; Stone, Martha, advisor; Bowden, David, committee member; Bechtel, Peter, committee member; Fettman, Martin J., committee member
Obesity is a common nutritional problem of American children. Health problems associated with childhood obesity and its continuation into adulthood underscore the need for effective weight control treatment for obese preadolescent children. The purpose of this study was to develop an effective weight control program that incorporated a structured low fat diet, regular exercise, behavior modification, family involvement and a fun learning environment. No weight control treatment for obese preadolescent children has incorporated these factors. The program was designed to teach obese preadolescent children methods to reduce their daily fat intake to between 25 and 30 percent of their total daily caloric intake. Thirty-two obese preadolescent children were recruited from the Cheyenne, Wyoming, area with the assistance of local health care practitioners. Children were randomly assigned to the special intervention group or the standard care group. Children assigned to the special intervention group participated in a newly designed ten week weight control program for obese preadolescent children. Children in the special intervention group changed the nutrient quality but not the fat quantity of their diets. These children, as indicated on their program evaluations and confirmed by nutrient analysis increased their consumption of fruits, vegetables, dairy products and meat. They did not use low fat food substitutions that were emphasized in the program. Analysis of activity factors confirmed that children in the special intervention group had increased their level of physical activity at weeks 10 and 22. Children in the standard care group received the nutritional counseling usually provided by a registered dietitian at the local medical clinic. Children in the standard care group exhibited a significant improvement (P [less than or equal to] 0.05) in relative weight and BMI at weeks 10 and 22. Changes in dietary fat intake of children in the standard care group were associated with changes in their weight status indicators. The greater success of the standard care group was related to the level of family functioning and the type of individualized counseling provided as usual care. Results of this study suggest that obese preadolescent children from dysfunctional families could achieve a greater level of weight control success with individualized care than with a group based program.
Open Access
Effect of farm to fork operations on bioactive compounds in white-fleshed and color-fleshed potatoes
(Colorado State University. Libraries, 2015) Amer, Fauzi Saleh Massoud, author; Vanamala, Jairam, advisor; Stone, Martha, advisor; Reddivari, Lavanya, committee member; Holm, David, committee member
The potato, Solanum tuberosum L., is one of the most commonly consumed food crops worldwide, and is the leading vegetable crop in the United States with 69% of per capita consumption as processed potatoes. In addition to micro- and macro-nutrients, color-fleshed potatoes are one of the richest plant sources for health promoting components such as resistant starch, polyphenols, and carotenoids. In contrast, potatoes are well known to contain naturally occurring glycoalkaloids (GA; α-chaconine and α-solanine) and processing-induced acrylamide (AL). Potatoes can be stored up to one year before being processed/consumed and the effect of genotype, storage (4°C or 10°C; 3 or 6 months) and processing (baking and frying) on both toxic and health beneficial compounds remains unknown. We hypothesized that cultivar, storage and processing alters bioactive content in potato tuber and potato products. To test this hypothesis, raw, baked, and chipped of white-, yellow-, red-, and purple-fleshed potatoes from initial (fresh) and stored tubers were evaluated for AL/vitamin C and GAs using Ultra Performance Liquid Chromatography (UPLC) and High Performance Liquid Chromatography-Diode Array Detector (HPLC-DAD/UPLC-DAD), respectively. Total phenolic content (Folin - Ciocalteu reducing), anthocyanin content (pH differential method), antioxidant activity (DPPH and ABTS assay) were also determined. Raw potatoes were analyzed for reducing sugars (glucose and fructose) using a spectrophotometer. Sensory attributes (9-point hedonic scale) of baked and potato chips were assessed using untrained consumer panelists (n= 94 – 114). The content of GA/AL increased with storage, dependent on cultivar. Reducing sugar content in raw potatoes increased with storage, thereby, AL content in potato chips positively correlated with reducing sugars. Purple-fleshed potatoes had higher (p ≤ 0.05) total phenolic content, anthocyanin content, and antioxidant activity than red-fleshed potatoes and white-fleshed potatoes. The interaction effect of storage time and temperature on total phenolic content, anthocyanin content, and antioxidant activity was genotype-dependent. Baking led to a significant (p ≤ 0.05) increase in total phenolic content, anthocyanin content, and antioxidant activity; whereas chipping led to significant losses in total phenolic content, anthocyanin content, and antioxidant activity. However, red- and purple-fleshed potatoes could serve as potential sources of non-nutrient health-benefiting compounds in the human diet even after storage and processing. Vitamin C content in potato tubers and processed potatoes was genotype-dependent. Vitamin C content rapidly declined with storage after six months of storage irrespective of storage temperature (4°C or 10°C). Chipping and frying resulted in significantly reduced vitamin C levels compared to baked potatoes and unprocessed potatoes among all tested cultivars. An increase in GA and AL content, bioactive compounds, and antioxidant activity found with storage was cultivar dependent. However, vitamin C decreased with storage. Thus, it is critical to measure GA and AL content not only in the fresh tubers but also in the final potato products such as baked and chipped potatoes. It is critical to adjust food systems processes to consistently deliver lower GA and AL content, while retaining the beneficial bioactives, vitamin C, and sensory attributes of the final potato products. Adjusting farm-to-fork operations to retain the health-benefiting compounds in food crops while reducing natural and process-induced toxicants will aid in countering growing epidemic of chronic diseases globally.
Open Access
Effect of genotype, storage and processing on the polyphenolic content, composition, in vitro anti-cancer activity and sensory attributes of colored-flesh potatoes
(Colorado State University. Libraries, 2012) Madiwale, Gaurav, author; Vanamala, Jairam, advisor; Stone, Martha, advisor; Reddivari, Lavanya, committee member; Bunning, Marisa, committee member; Holm, David, committee member
The potato (Solanum tuberosum L.) is the world's third largest food crop with per capita consumption of ~126 lbs. annually in the US. The 2010 US Potato Board Report revealed that over the past ten years, while consumption of traditional potatoes (mashed, baked, fried, steamed, boiled and french fries) declined, specialty/colored potato consumption increased by 17%, possibly due to their putative health benefits. Specialty/colored potatoes, which are rich in anti-proliferative and pro-apoptotic anthocyanins and/or carotenoids, can be an attractive "delivery system" for these bioactive compounds in humans. However, colored potatoes can undergo 3-6 months of storage before processing/consumption and the effect of storage and processing on their anti-cancer properties remains unknown. We hypothesized that potatoes retain polyphenolic content (TP), antioxidant activity (AA) and chemopreventive properties against early stage (HCT-116) and advanced stage (HT-29) human colon cancer cells even after 3 months of storage and processing (baking and chipping). To test this hypothesis, we utilized white-, yellow- and purple-fleshed potato clones and tested their phenolic (Folin-Ciocalteu) and anthocyanin (pH-differential) content, antioxidant activity (ABTS and DPPH), metabolite profile (UPLC-MS), anti-cancer properties (cell proliferation via cell counting and BrdU assays, and apoptosis via Caspase-Glo 3/7 assay), and sensory attributes (9-point hedonic scale). Purple-fleshed potatoes had higher phenolic content and antioxidant activity as compared to their white- and yellow-fleshed counterparts. The antioxidant activity of all clones increased with storage; however, an increase in total phenolic content was observed only in purple-fleshed clones. Baking caused minimal losses while chipping reduced the phenolic and anthocyanin content, and antioxidant activity of the potatoes. With storage, total phenolic and anthocyanin content, and antioxidant activity increased in baked samples while in the chipped samples, they remained constant. Principal component analysis of approximately 1600 peaks obtained by UPLC-MS analysis revealed that storage caused a shift in the metabolite profiles of potato clones. In general, ethanol extracts of uncooked, baked and chipped samples suppressed proliferation and elevated apoptosis (p < 0.05) in human colon cancer cell lines (HCT-116, HT-29). However, chipped samples did not have any effect on HT-29 cell lines. Anti-proliferative and pro-apoptotic properties of baked potatoes were similar to that of raw potatoes while chipping caused a significant reduction in the biological activity. Storage generally negatively affected the anti-cancer properties of the potato extracts. Sensory analysis revealed comparable acceptance of purple-fleshed baked and chipped potatoes when compared with traditional cultivars. Consumers were willing to pay a premium for colored-flesh potatoes if they were educated on their potential health benefits. Phenolic content and antioxidant activity of purple-fleshed potatoes, after baking, were comparable with those of anthocyanin-rich berries. Hence, locally grown purple-fleshed potatoes can be a healthier choice as they possess greater levels of bioactive compounds and in vitro anti-cancer properties even after processing as compared to their white- and yellow-fleshed counterparts.
Open Access
Effects of cooking methods on antioxidant properties, quality attributes, and sensory characteristics of selected leafy greens
(Colorado State University. Libraries, 2014) St. Onge, Matthew Mark, author; Bunning, Marisa, advisor; Stone, Martha, advisor; Newman, Steven, committee member
Kale, arugula, spinach, and other types of leafy vegetables are rich sources of vitamins and minerals and well-suited to growing in Colorado's climate. This project focused on the chemical, quality, and sensory assessment of a selection of 6 specialty leafy greens (arugula, cherokee lettuce, mache, pac choi, red kale, and spinach). Total phenolic (TP) content, radical scavenging capacity (1,1-diphenyl-2-picrylhydrazyl, DPPH), color measurement, instrumental texture characteristics, and sensory attributes of raw versus cooked (boiled, microwaved, or steamed) samples of the select taxa were analyzed and evaluated. All 6 varieties were cultivated in a greenhouse using organic planting medium. Samples of the greens were freeze-dried as raw or post-heat treatment, then underwent an extraction procedure, and were analyzed for total phenolics compared to gallic acid standards and radical scavenging using DPPH compared to trolox standards. Fresh and cooked comparisons were tested for color differences using a HunterLab ColorFlex spectrophotometer and changes in texture utilizing a TA-XT2 texture analyzer. Consumer (n=50; n=51) sensory analysis was administered using a 9-point hedonic scale. Fresh, uncooked mache had higher (p<0.05) total phenolics and radical scavenging ability than all other cultivars. Cooked samples revealed that spinach values for each test did not differ (p>0.05) from fresh samples and microwaved and steamed samples of red kale contained more total phenolics than fresh, but less (p<0.05) was observed in boiled samples. Other significant findings included reductions (p<0.05) in lightness of cooked samples and cooked spinach samples after all heating treatments were observed to be more tender than fresh samples (p<0.05). This research helps to fill information gaps which exist in leafy greens research. Many studies focus on one cultivar using one testing method, but little research has been conducted on these types of greens using several analytical testing methods to obtain comparable data.
Open Access
Effects of dark malts, dry hopping, and filtration on xanthohumol content and bioactivity of American India Pale Ales
(Colorado State University. Libraries, 2011) Henley, Twila Jean, author; Stone, Martha, advisor; Vanamala, Jairam, advisor; Bunning, Marisa Lee, 1954-, committee member; Avens, John S., committee member; Reddivari, Lavanya, committee member; Stonaker, Frank H., committee member
Xanthohumol (XN), a prenylated chalcone found in hops (Humulus lupulus L.) has been shown to possess a wide spectrum of beneficial properties including anti-oxidant, anti-proliferative, pro-apoptotic, anti-inflammatory, anti-bacterial, anti-viral, and anti-malarial activities. Efforts have been made to increase the amount of XN in beers by modifying certain brewing ingredients and procedures. However, the effects of modifications such as addition of dark malts, dry hopping, and DE filtration on XN content and the biological activity of American India Pale Ales (IPAs) are not known. In this study, different brands of IPAs with and without addition of dark/roasted malts, dry hopping, and filtration and one non IPA as a standard were analyzed for XN, isoxanthohumol, total phenolic content, and antioxidant capacity. Isolated beer compounds and selected whole beer matrixes were used to determine the synergistic effect of beer compounds on proliferation and apoptosis of HCT 116 p53 +/+ colon cancer cells. No XN was found in the standard, and the XN content in IPAs ranged from 0.00 to 12.69 mg/L. A heavily dry hopped IPA made with increased amounts of dark malt contained higher amounts of XN compared to other IPAs. The use of dark malts was protective against the removal of XN and other phenolics after diatomaceous earth (DE) filtration and dry hopping increased XN content in beer. Whole beer matrixes with greater levels of XN suppressed proliferation and elevated apoptosis in colon cancer cells compared with isolated XN and/or IX, indicating that the biological effect of XN can be bolstered in the presence of other beer compounds.
Open Access
Effects of time and temperature during melanging on the volatile profile of dark chocolate
(Colorado State University. Libraries, 2019) Clark, Caitlin, author; Stone, Martha, advisor; Miller, Jeffrey, committee member; DiVerdi, Joseph, committee member
Small-batch chocolate-makers use distinct equipment and process steps from that of large-scale industry leaders. Only one of these steps is unique to small-batch chocolate-making and is known as melanging—a refining process that takes place in a stone wet-grinder. Despite being essential to flavor and overall quality development, it has never been studied, and all data on its application to chocolate are anecdotal. This research evaluated the melanging step at three different temperatures in order to determine, via Head Space Solid Phase Micro Extraction (SPME) coupled with GC-MS, which volatile flavor-active compounds are formed or lost during the melanging process and how the time and temperature affect the volatile composition of the chocolate mass. Univariate statistical analysis on the 36 samples (3 temperatures, 4 time points, and 3 technical replicates) was performed using analysis of variance (ANOVA) and multivariate statistical analysis was performed using principal component analysis (PCA) and orthogonal partial least squares (OPLS). These analyses found that time, rather than temperature, accounted for most of the variation among samples. Some trends among chemical classes were identified, and certain indicator compounds were highlighted for their particular relevance to the system. Pyrazines displayed the strongest pattern of any annotated chemical class. As a rule, these compounds started with moderate to high abundance at 0 hours and decreased to extremely low levels by 24 hours. However, most classes of chemical compounds analyzed in this study did not follow strong patterns of behavior predicted by chemical class. Particle size and pH were also measured for each sample. Majority particle size was found to be below 25 microns generally at all time points beyond 8 hours, and pH for all samples had a mean of 5.25 with a standard deviation of 0.177. Analysis showed significant temperature-dependent p-values for several compounds, but significant time-dependent p-values were apparent for a greater number of compounds. For compounds which showed dependency on both time and temperature, the time-dependent p-value was much smaller in most cases. Both PCA and OPLS analyses suggested the same trends. It was determined first, that melanging is a viable method to carry out successful chocolate refining. The final flavor of chocolate made in a melanger is far more dependent on time spent in the melanger than on the temperature of the chocolate system. In addition, classes of chemical compounds did not behave according to unified patterns of behavior in melanging systems (with the exception of nitrogen heterocycles, which decreased in relative abundance). Finally, literature previously published on conching may be successfully extrapolated to melanging.
Open Access
Influence of roasted barley on quality of beer
(Colorado State University. Libraries, 2013) Davis, Dave, author; Stone, Martha, advisor; Avens, Jack, committee member; DeVoe, Dale, committee member
The research examined the influence of roasted barley content of beers on the quality of the resulting product. Recent researchers have indicated that moderate consumption can be a source of beer specific antioxidants, help reduce the risk of cardiovascular disease, and lower the occurrence of certain types of cancers. Beers were made with two-row malted and six-row unmalted roasted barley. Brews contained varying levels of the unmalted six-row roasted barley with the balance of the grain bill composed of two-row malted barley. They were analyzed for total phenolic content, sensory properties, and the physical properties specific gravity, color, and calculated alcohol. Four test groups were prepared with four replications of each test group for a total of 16 brews. No significant differences (P > 0.05) were found between the test brews for total phenolics. Total phenolics ranged from 314.77 to 451.72 micromoles / 100 milliliters for the test brews. Using Kuskal-Wallis one-way analysis of variance, a significant difference was found (χ2 =14.328, p = 0.00249) which demonstrated an increase in beer color as the percentage of roasted barley increased for the beer treatments. A significant difference was found in the organoleptic properties of the finished beers for total score, but not in individual categories. Perception of the bitter and burnt characteristics imparted by roasted barley appeared to increase as the percentage of roasted barley increased. This was not necessarily viewed as a detriment to the beer, but as an increase in complexity that added to the overall flavor and balance of the beer. There was a linear progression (P > 0.05) in the scores for aroma and appearance as the percentage of roasted barley increased. No significant differences (P > 0.05) were found in original or final specific gravity or within calculated alcohol values.
Open Access
Physicochemical and sensory quality of chiffon cake prepared with rebaudioside-A and erythritol as replacement for sucrose
(Colorado State University. Libraries, 2012) Lothrop, Robert S., author; Stone, Martha, advisor; Avens, John, committee member; Hyatt, Doreene, committee member; Miller, Jeffrey, committee member
Rebaudioside-A has been used for many years throughout the world as a non-nutritive sweetener in many different food systems (Goyal et al. 2010), however, up until 2008, it was not an approved food sweetener in the United States (FDA 2008). Prior to this approval, stevia extracts were found in herbal and health food stores throughout the United States since the 1970's (Carakostas et al. 2008). Since the passage of the Dietary Supplement Health and Education Act (DSHEA) in 1994, stevia extracts were legally sold as "dietary supplements" in the United States marketplace (Carakostas et al. 2008). However, under the DSHEA, these products were not permitted to be marketed with any packaging or advertising language suggesting that they be used as a sweetener (Carakostas et al. 2008). Since 2008, Truvia™ brand sweetener produced and marketed by Cargill (2010) has been available on the consumer market in the United States. This sweetener is a mixture of rebaudioside-A and erythritol that is made for a consumer to replace sucrose in applications at a 1:1 ratio (Cargill 2010). The sweetening characteristics of rebaudioside-A appear to be approximately 300 times that of sucrose (Lin and Lee 2005) and digestion, if any, occurs in the large intestine by gastrointestinal microflora, lending a minimal caloric intake to the user. Truvia™ and other blends of rebaudioside-A and erythritol can be used as a natural non-nutritive sugar alternative and may prove to be an effective and acceptable replacement to sucrose in baked systems such as chiffon cake (Cargill 2010). Rebaudioside-A is an leaf extract of the Stevia rebaudiana plant (Cargill 2010) and erythritol is a four-carbon polyol widely found in nature in such food as melons, grapes, pears, seaweeds, fungi as well as naturally occurring in fermented food products (Moon et al. 2010). The purpose of this research was to determine the physicochemical and sensory effects of replacing a mixture of rebaudioside-A and erythritol for sucrose at varying levels (0, 25, 50, 75 and 100%) in chiffon cake. Analytical testing of specific gravity, texture, volume, water activity, moisture, color and differential scanning calorimetry was conducted on the five cake formulations in addition to nutritional analysis and a consumer sensory evaluation (n = 40). Five treatments were prepared using four replications in a randomized complete block one factor design. ANOVA was used to determine significant differences. If so, differences among means were examined using Tukey's honestly significant difference. There were no differences in instrumental color of crust or crumb among any of the replacement levels with the exception that crumb color of both the control and 25% reduced sucrose samples were "more yellow" than the 100% reduced sucrose cakes (p < 0.05). Water activity ranged from 0.86 to 0.91 and some statistical differences were found among treatments. However, there were no differences in percent moisture. Texture analysis showed the 25 and 50% reduced sucrose cakes were more tender than the 100% reduced sucrose cake (p < 0.05). Volume and specific gravity tests had no differences among treatments. Differential scanning calorimetry (DSC) results showed that as the replacement level of sucrose with a mixture of rebaudioside-A and erythritol increased, both the onset and peak temperatures decreased. These results corresponded with DSC testing conducted by Lim and others (1992) on wheat starch, sucrose and water interactions. Enthalpy was consistent for the control, 25 and 50% reduced sucrose samples and increased for both the 75 and 100% samples; however, differences were small and would likely not be detectable by consumers. A nine-point hedonic scale ranging from "like extremely" to "dislike extremely" was used to measure overall liking of color of crumb, tenderness, sweetness, aftertaste and overall acceptability. Panelists were asked to rank preference of the five samples from one to five with one being the most preferred. Sensory panelists "liked" all of the chiffon cake samples and the highest "overall acceptability" scores were for cakes prepared with 25 and 50% reduction in sucrose (p < 0.05). These two samples also ranked highest in preference (p < 0.05). Additionally panelists "liked" the tenderness of the 25 and 50% reduced sucrose cakes more than the other samples (p < 0.05). For sweetness and aftertaste, panelists "liked" the 25, 50 and 75% reduced sucrose cakes (p < 0.05). Chiffon cake prepared with 50% sucrose and 50% rebaudioside-A and erythritol resulted in a product with high overall consumer acceptability and 20% fewer calories than one formulated with 100% sucrose.
Open Access
Sensory quality of cheddar cheese made with bulk starter and direct to vat starter culture
(Colorado State University. Libraries, 2024) Musetti, James, author; Stone, Martha, advisor; Bunning, Marisa, committee member; Narayanan Nair, Mahesh, committee member; Miller, Jeff, committee member; Romero, Dennis, committee member
The production of cheese in the world consumes around 35% of the total milk production and has increased on average by 4% per year over the past 30 years (Fox et al., 2017). Cheddar consumption had modestly and steadily increased in the past several years in the United States and has increased 2.22% from 9.87 pounds per capita to 10.09 pounds per capita (USDA ERS Dairy Data, n.d.). Over the past several decades modernization of cheddar production in the United States has enabled producers to increase throughput with fewer resources resulting in more efficient production and consistent quality. This includes the common practice of standardizing cheese making procedures on a strict timing basis and using reliable and consistent rate and extent of acidification through culture selection and dosage. One such advancement was the development of defined starter cultures produced in a frozen or lyophilized state to be applied directly to the vat as a direct to vat inoculant (DVI) by the cheesemaker. Previously lactic acid bacterial cultures, defined or natural, were propagated by the cheesemaker prior to cheese production by a preceding fermentation of milk or whey and used to inoculate the milk for cheddar production. The current research investigated if any differences in cheddar cheese biochemical and sensory characteristics exist among cheeses made with bulk starter and DVI technologies. Cheeses were produced using bulk starter culture technology, DVI technology, and DVI technology with pre-acidification then ripened for 90 days. The rate and extent of acidification in the process was analyzed with cheese composition, extent of the catabolism of protein and fat during ripening, and sensory characteristics of the cheese analyzed. MANOVA model analysis reported that the treatments had a significant effect on the cheesemaking process (p=0.00381). Coagulation time was the only response found to be statistically significant (p=0.00081) from the process, biochemical, and sensory responses after mixed model analysis was completed. The make data or milk batch was found to have a significant effect on the cheese production process (p=0.00036), biochemistry (p=0.04391), and sensory characteristics (p=0.00002) of the cheeses. Therefore, it can be concluded that there was no difference in cheddar proteolysis, lipolysis, and sensory characteristics in cheeses manufactured with bulk starter and DVI, and there was no difference in cheddar proteolysis, lipolysis, and sensory characteristics in cheeses manufactured with bulk starter and DVI culture preparations with recipe adjustment for coagulation. The null hypotheses cannot be rejected.
Open Access
Species specific detection of adulteration of water buffalo milk with cow milk using high resolution melting analysis
(Colorado State University. Libraries, 2015) Musetti, James, author; Stone, Martha, advisor; Bunning, Marisa, advisor; Hyatt, Doreene, committee member
Economically motivated adulteration, a subset category of food fraud, was defined in 2009 by the Food & Drug Administration (FDA) as "the fraudulent, intentional substitution or addition of a substance in a product for the purpose of increasing the apparent value of the product or reducing the cost of its production," and can often encompass effects of public safety through the unknown addition of allergens, toxins, and hygienic risks (Wheatley & Spink, 2013). According to the FDA database which tracks scholarly records of adulterated foods, dairy products are the second most reported adulterated food products (Moore, Spink, & Lipp, 2012). Water buffalo milk represents the second most produced milk worldwide and higher value dairy raw products make it subject to economically motivated adulteration by means of dilution with cow milk (Bonfatti, Giantin, Rostellato, Dacasto, & Carnier, 2013). It is of interest to consumers, manufacturers and governing bodies to have a simple, fast, accurate, and sensitive method to detect adulteration in water buffalo milk. Several methods have been developed to detect species adulteration in dairy products including immunological, chromatography, and molecular methods (Mayer, 2005). In this investigation a real-time PCR assay was developed to specifically detect and relatively quantify cow milk adulteration in water buffalo milk utilizing high resolution melting analysis. A duplex real-time PCR reaction was performed targeting mitochondrial DNA in both species of interest and the resulting amplified products were analyzed after a melt cycle was performed. Results indicated that the assay was specific, efficient, and validated the use of real-time PCR for analysis. The use of high resolution melting analysis allowed for the discrimination of dilution of water buffalo milk with cow milk down to 0.1%. This study indicates the feasibility of real-time PCR and high resolution melting analysis in the detection of adulteration in dairy products.