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Bovine milk contains a variety of endogenous peptides, partially formed by milk proteases that may exert diverse bioactive functions. Milk storage allows further protease activities altering the milk peptidome, while processing, e.g., heat treatment can trigger diverse chemical reactions, such as Maillard reactions and oxidations, leading to different posttranslational modifications (PTMs). The influence of processing on the native and modified peptidome was studied by analyzing peptides extracted from raw milk (RM), ultra-high temperature (UHT) milk, and powdered infant formula (IF) by nano reversed-phase liquid chromatography coupled online to electrospray ionization (ESI) tandem mass spectrometry. Only unmodified peptides proposed by two independent software tools were considered as identified. Thus, 801 identified peptides mainly originated from αS- and β-caseins, but also from milk fat globular membrane proteins, such as glycosylation-dependent cell adhesion molecule 1. RM and UHT milk showed comparable unmodified peptide profiles, whereas IF differed mainly due to a higher number of β-casein peptides. When 26 non-enzymatic posttranslational modifications (PTMs) were targeted in the milk peptidomes, 175 modified peptides were identified, i.e., mostly lactosylated and a few hexosylated or oxidized peptides. Most modified peptides originated from αS-caseins. The numbers of lactosylated peptides increased with harsher processing.

This study investigated the antimicrobial efficacy of bioactive plant compounds along with high pressure processing (HPP) against pathogens Bacillus cereus and Cronobacter sakazakii in infant formula and infant rice cereal. The influence of these applications on antimicrobial activity, shelf-life and sensory attributes of infant foods were examined. Trans-cinnamaldehyde (TC), (-)-Epigallocatechin gallate (EGCG) and [10]-Gingerol (GI) were incorporated (0.05%) in infant rice cereal reconstituted with infant formula. The cereal was inoculated with either B. cereus (ATCC 14579) or B. cereus spores (107-108 log CFU g-1). All the samples were stored at 7, 23 or 37°C for 0, 4, 8 and 24 h. TC showed the highest antimicrobial activity by inhibiting the B. cereus and its spores up to 2.72 and 3.8 log CFU g-1, respectively. HPP (600 MPa for 5 m), and TC (0.05-0.1%) along with Chitosan (CH) (1%), were applied to reconstituted powder infant formula which was inoculated with either 3 strains of C. sakazakii (ATCC 29544, ATCC 12868, and ATCC BAA 894) or 5 strains of B. cereus spore (ATCC 14579, ATCC 33018, ATCC 12826, ATCC 4342, and Difco Spores) cocktail (107-108 log CFU ml-1). All the samples were stored at 7, 23 or 45°C for 5-8 weeks. HPP and TC (0.1%) combination exhibited the highest inhibition (P < 0.05) by reducing the B. cereus spores 2.97 log CFU ml-1 after 7 d. C. sakazakii was fully inactivated by HPP, TC (0.05%) and C (1%) combination following 8 weeks of storage at 7 and 23°C and 2 weeks storage at 45°C. The combination of HPP and bioactive compounds exhibited additive antimicrobial effect. Gradual decrease (P < 0.05) in pH was observed in rice cereal and non-HPP formula samples due to the microbial growth and metabolic activity. Significant differences (P < 0.05) were found in color, aroma and general appearance of EGCG and GI applied cereal samples, while TC only did exhibit a cinnamon taste. In summary, the antimicrobial findings suggest that TC, EGCG, GI and CH could be incorporated in infant foods along with HPP as natural and safe alternatives to synthetic preservatives and thermal applications.

Os ácidos graxos polinsaturados atuam no organismo humano em diversos processos fisiológicos e metabólicos, além de serem importantes na nutrição infantil. A quantificação dos ácidos graxos polinsaturados, devido à presença de vários sítios reativos na molécula, deve envolver processos de extração da gordura em condições amenas. Este trabalho teve como objetivos a comparação dos métodos analíticos para determinação de lipídios totais e ácidos graxos polinsaturados (ácido linoléico, ácido &#945;-linolênico, ácido araquidônico e ácido docosahexaenóico) em fórmula infantil, a quantificação dos ácidos graxos polinsaturados nas fórmulas infantis comerciais e o acompanhamento da estabilidade destes ácidos graxos neste alimento. Foram analisadas 15 amostras, sendo uma amostra da Nacional Institute of Standards and Techonology (NIST 1849) e 14 fórmulas infantis comercializadas no Estado de São Paulo. Os métodos analíticos comparados para extração de lipídios foram: gravimétricos (Bligh Dyer, Roese Gottlieb e hidrólise ácida AOAC 963.15) e por cálculo (AOAC 996.06 e método direto adaptado de Golay et al. (2006)). Para a preparação dos ésteres metílicos de ácidos graxos utilizaram-se metodologias descritas pela IUPAC, Hartman e Lago e método direto adaptado de Golay et al. (2006). Compararam-se diferentes padrões interno, sendo estes dos ésteres metílicos de ácidos graxos 13:0, 21:0 e 23:0 e fatores de resposta para quantificação dos ácidos graxos polinsaturados em relação aos ácidos graxos 16:0, 18:0 e 23:0. O estudo de estabilidade durou 8 meses, e as análises foram realizadas nos meses de março, abril, julho e outubro de 2008. Os melhores resultados, para gordura total e para ácidos graxos polinsaturados, foram obtidos pelo método oficial (Roese Gottilieb). Quanto ao cálculo dos ácidos graxos polinsaturados, o uso do padrão interno 23:0 e o fator de resposta de correção teórico em relação ao 23:0 revelam resultados mais satisfatórios. Das fórmulas infantis comerciais analisadas, 85,7% apresentaram pelo menos um analito em desacordo com a informação nutricional e 100% com relação à legislação brasileira e o Codex Alimentarius. Em relação à estabilidade dos ácidos graxos polinsaturados, apenas três fórmulas infantis não apresentaram diferença estatisticamente significativa (p>0,05) nos teores de ácido graxos no período de 8 meses. / Long chain polyunsaturated fatty acids are involved in several physiological and metabolic process of human organism. The quantification of polyunsaturated fatty acid, must involve the fat extraction in mild conditions, due the reactive sites in the molecule. This work had as objective the comparison of the analytical methods for determination of polyunsaturated fatty acid (linoleic acid, &#945;-linolenic acid, arachidonic acid and docosahexaenoic acid) and lipids in infant formula, quantification of polyunsaturated fatty acids in commercial infant formulas and polyunsaturated fatty acids stability evaluation in these foods. A 15 samples of infant formulas were analyzed, these one from NIST and 14 commercial infant formulas. The analytical methods to lipids extraction were: Bligh and Dyer, Roese Gottlieb, acid hydrolyze AOAC 963.15, acid hydrolyze AOAC 996.06 and direct method adapted from Golay et al. (2006). To prepared esters metilics fatty acids utilized the methods: IUPAC, Hartman and Lago and direct method. The quantization of polyunsaturated fatty acid was realized with different standards internal, like fatty acid methyl ester 13:0, 21:0 and 23:0, and flame ionization detector response factors of correction in relation fatty acids 16:0, 18:0 and 23:0. The stability study during 8 months and analysis were analyzed in the months March, April, July and October. The best results, for lipids and polyunsaturated fatty acid, were obtained by the official method, Roese Gottlieb and metilation followed Hartman and Lago. In accordance with the present study the internal standard 23:0, and the theoretical correction factors with relation 23:0 showed satisfactory trueness and precision for calculation of polyunsaturated fatty acid. Among of commercial infant formula analyzed, 85.7% had at least one analyte in disagreement with the nutritional label facts and 100% with respect to Brazilian Legislation and the Codex Alimentarius. Regarding polyunsaturated fatty acids stability, three infant formulas showed no statistically significant difference (p > 0.05) in levels of these fatty acids during the period of analysis (8 months).

During this unique moment of feminist inquiry wherein breastfeeding has been a focal point of interdisciplinary research, little sociological scholarship has been presented which has centered on the various meanings that African American mothers, as a diverse group, attach to their experiences with breastfeeding and/or infant formula use. While patterns of behavior have been explored in a cross-racial context, most social science studies have not focused on how the choice between breastfeeding, using infant formula, or using a combination of the two has impacted (or has been shaped by) African American mothers’ constructs of self, motherhood/mothering, their birth experiences, and their sexuality. In order to understand the interplay of the decision-making process and these constructs, I conducted a qualitative study in which I participated in face-to-face interviews with a diverse group of thirty African-American mothers. They ranged in age from 18 years-old to 50-years-old. At the time of her interview, each mother had at least one child who was three-years-old or younger. Through our discussions, we explored how pre-pregnancy perceptions, lived experiences as a mother, familial influences, and the discourses surrounding motherhood within an African-American context affected the perceptions and experiences that the mothers in the study had with their infant feeding practice(s). Findings suggest that while African Americans mothers know that “breast is best,” that knowledge is not the only reason for their decisions. The first step in understanding why African-American mothers choose the feeding method(s) that they choose is embracing the reality that choosing is an ongoing and dynamic process which is often informed by what she does versus “is supposed to do” versus how she is portrayed weighed with the consequences of her choice(s) for herself and her family. Further, African American mothers are in the active process of negotiating an evolving definition of themselves within this post-Civil Rights, Affirmative Action context wherein choices appear abundant, but the choosing always comes with a price.

Als Schlüsselenzym im Renin-Angiotensin System übernimmt das Angiotensin Converting Enzyme (ACE) eine wichtige Rolle bei der Blutdruckregulierung. ACE spaltet das biologisch inaktive Angiotensin I zum vasokonstriktorisch wirksamen Angiotensin II, was zu einem Anstieg des Blutdruckes führt. Tier- und Humanstudien zeigten, dass die Aufnahme bekannter, aus dem Proteinabbau stammender ACE-Inhibitoren eine Absenkung des Blutdruckes bewirkte. In der Lebensmittelindustrie finden Hydrolysate von Milchproteinen, im speziellen von Molkenproteinen, Einsatz in hypoallergenen (HA) Säuglingsnahrungen. Obwohl das Phänomen einer ACE-Inhibierung durch HA-Nahrungen in vitro in der Literatur bereits Erwähnung fand, existieren bislang keine Angaben zu einer potentiellen Wirkung in vivo. In der vorliegenden Arbeit konnte für kommerzielle hypoallergene (HA) Säuglingsnahrung eine sehr starke ACE-Hemmung in vitro zeigen (IC50-Werte zwischen 20 und 104 mg Protein/liter), welche die für fermentierte Sauermilchprodukte dokumentierte Wirkung bei weitem überstieg.] Mittels RP-HPLC und ESI-TOF-MS konnte neben zahlreichen bekannten Peptiden mit ACE-hemmendem Effekt erstmals das aus der Primärsequenz von -Lactalbumin freigesetzte Dipeptid Ile-Trp in den HA-Nahrungen identifiziert und quantifiziert werden. Ile-Trp ist der bislang potenteste in Lebensmitteln nachgewiesene ACE-Inhibitor (IC50 = 0,7µM). HA-Nahrungen zeigten auch ex vivo im Zellsystem (HUVECs) einen stark ACE-hemmenden Effekt. Aus diesem Grunde wurde ein möglicher Einfluss der HA-Nahrungen auf den Blutdruck spontan hypertensiver Ratten untersucht. Hierfür wurden die Tiere im Rahmen einer Fütterungsstudie über 14 Wochen mit standardisiertem Futter, welchem HA-Nahrung (Gruppe 1), konventionelle Säuglingsnahrung (Gruppe 2) bzw. der bekannte ACE-Inhibitor Captopril (Gruppe 3) zugesetzt war, gefüttert. Eine vierte Gruppe mit Standardfutter diente als Kontrolle. Der Blutdruck wurde am wachen Tier nichtinvasiv mittels tail-cuff-Methode gemessen. Der systolische Blutdruck sank bei Verabreichung der HA-Nahrung nach 7 Wochen signifikant um 21 ± 8 mmHg ab im Vergleich zur Kontrollgruppe bzw. den mit konventioneller Säuglingsnahrung gefütterten Tieren. Captopril führte zur einer Blutdrucksenkung um 30 ± 7 mmHg.

The aim of this study was to observe biofilm formation by Enterobacter sakazakii (E. Sakazakii) from different clinical, dairy and environmental origins on three infant feeding tubes made of different materials. Infant formula milk was selected as the medium for E. sakazakii growth. Seventeen isolates from different origins were retrieved and tested for purity, using a plating method and biochemical tests to eliminate the non E. sakazakii strains from this study. A method to rapidly and accurately detect viable cells of E. sakazakii on infant feeding tube surfaces using of the BacTrac® 4000 microbiological growth analyser was developed. The sources of errors such as from cleaning, operation and handling procedures were assessed prior to experimental runs. The strength of biofilm formation by different isolates of E. sakazakii on plastic surfaces was scrutinised using a microtiter plate assay. The results from the microtitre plate assay were based on the absorbance at 550 nm of crystal violet stained films and showed that all the clinical isolates were able to attach and form strong biofilms on the plate. Some environmental isolates formed strong or weak biofilms and some did not produce biofilm at all. However, dairy isolates formed both strong and weak biofilms in the microtitre plate when incubated in 10% reconstituted infant formula milk. The further studies were to quantify biofilm formation by three isolates of different origin on three different materials of infant feeding tubes using a batch system. Tubing pieces were incubated with infant formula milk inoculated with E. sakazakii cells at approximately 8 log CFU mL-1 and the biofilm formation was assessed at three time intervals: 4, 12 and 24 hours. Biofilm formation on the tubing by clinical isolates was also observed using epifluorescence microscopy and the scanning electron microscope. E. sakazakii from clinical, dairy and environmental isolates were able to form biofilm on three different materials of infant feeding tubes. The results showed that the initial attachment at 4 h on silicone tubing was low compared with the other two tubes. The scanning electron micrographs showed the surface characteristics of each tubing and the biofilm formation by E. sakazakii clinical isolates after 4, 12 and 24 hours. Silicone tubing appeared to be the best choice for premature babies that need feeding using feeding tubes, as it was slow to become colonised compared with the PVC and polyurethane tubing.

Thesis (MSc)--University of Stellenbosch, 2007. / ENGLISH ABSTRACT: The World Health Organisation (WHO) estimates that at least 75% of infants receive infant formula milk (IFM) either entirely or in conjunction with breast milk during the first four months after birth. The presence of the emerging pathogen Enterobacter sakazakii in IFM has been associated with rare but fatal cases of neonatal infections and deaths. There is thus a need for accurate methods for the rapid detection of E. sakazakii in foods. At present, the methods used to detect and identify this micro-organism are inadequate, controversial and contradictory. The aim of this study was to determine the most suitable method for E. sakazakii detection after evaluation of the currently available methods. A further aim was to optimise a polymerase chain reaction (PCR) method for the detection of only viable E. sakazakii cells utilising the DNA-intercalating dyes ethidium monoazide (EMA) and propidium monoazide (PMA). The Food and Drug Administration (FDA) method for E. sakazakii detection was utilised to select 50 isolates from IFM and 14 from the environment, regardless of colony appearance. These isolates were identified by sequencing a 1.5 kilobase (kb) fragment of the 16S ribosomal DNA (rDNA) and by using the National Centre for Biotechnological Information (NCBI) database to confirm the closet known relatives. Seven of the 50 (14%) IFM isolates and six of the 14 (43%) environmental isolates were identified as E. sakazakii. The methods that were evaluated for accuracy in detecting and identifying these E. sakazakii isolates included yellow pigment production on tryptone soy agar (TSA), chromogenic Druggan-Forsythe-Iversen (DFI) and Enterobacter sakazakii (ES) agars and PCR using six different species-specific primer pairs described in the literature. The suitability of the FDA method was lowered by the low sensitivity, specificity and accuracy (87%, 71% and 74%, respectively) of using yellow pigment production for E. sakazakii identification. DFI and ES agars were shown to be sensitive, specific and accurate (100%, 98% and 98%, respectively) for the detection of E. sakazakii. The specificity of the PCR amplifications was found to vary between 8% and 92%, with Esakf and Esakr being the most accurate of the primer pairs evaluated. The current FDA method for E. sakazakii detection requires revision in the light of the availability of more sensitive, specific and accurate detection methods. Based on the results obtained in this study, a new method is proposed for the detection of E. sakazakii in food and environmental samples. This proposed method replaces the culturing steps on violet red bile glucose agar (VRBGA) and TSA with culturing on chromogenic DFI or ES agar. For identification and confirmation of presumptive E. sakazakii isolates, the oxidase test, yellow pigment production and API biochemical profiling is replaced by DNA sequencing and/or species-specific PCR with the most accurate primer pair (Esakf and Esakr). The amendments to the current FDA method will reduce the time to detect E. sakazakii from approximately 7 days to 4 days and should prove to be more sensitive, specific and accurate for E. sakazakii detection. In this study, a novel PCR-based method was developed which was shown to be capable of discriminating between viable and dead E. sakazakii cells. This was achieved utilising the irreversible binding of bacterial DNA to photo-activated PMA or EMA in order to prevent PCR amplification from the dead cells. At concentrations of 50 and 100 μg.ml-1, PMA completely inhibited PCR amplification from dead cells, while causing no significant inhibition of the PCR amplification from viable cells. EMA was equally effective in preventing PCR amplification from dead cells, however, it also inhibited PCR amplification from viable cells. PMA-PCR in particular, will be useful for assessing the efficacy of processing techniques, as well as for monitoring the resistance, survival strategies and stress responses of E. sakazakii. This will be an important step in the efforts to eliminate E. sakazakii from food and food production environments. / AFRIKAANSE OPSOMMING: Die Wêreld Gesondheidsorganisasie (WGO) beraam dat ten minste 75% van alle babas net baba formule melk (BFM) of BFM in kombinasie met moedersmelk in die eerste vier maande na geboorte kry. Die teenwoordigheid van die voortkomende patogeen Enterobacter sakazakii in BFM is al geassosieer met skaars maar noodlottige gevalle van neonatale infeksies en sterftes. Akkurate metodes word dus benodig vir die vinnige deteksie van E. sakazakii in voedsel. Die metodes wat huidiglik gebruik word vir die deteksie en identifikasie van hierdie mikroörganisme is onvoldoende, kontroversieël en teenstrydig. Die doel van hierdie studie was om die beste metode vir die deteksie van E. sakazakii te bepaal, na 'n evaluasie van die metodes wat huidiglik beskikbaar is. 'n Verdere doel was om 'n polimerase ketting reaksie (PKR) metode vir die deteksie van slegs lewensvatbare E. sakazakii selle te optimiseer deur gebruik te maak van die DNSbindende kleurstowwe, etidium mono-asied (EMA) en propidium mono-asied (PMA). Die Voedsel en Medisyne Administrasie (VMA) se metode vir E. sakazakii deteksie is gebruik om, ongeag van die kolonie kleur, 50 isolate vanuit BFM en 14 isolate vanuit die omgewing te kies. Hierdie isolate is geïdentifiseer deur die DNS volgorde van 'n 1.5 kilo-basis (kb) fragment van die 16S ribosomale DNS (rDNS) te bepaal en die Nationale Sentrum vir Biotegnologiese Informasie (NSBI) databasis te gebruik om die mees verwante spesie te bevestig. Sewe van die 50 (14%) BFM isolate en ses van die 14 (43%) omgewings isolate is geïdentifiseer as E. sakazakii. Die metodes wat geëvalueer is in terme van akkuraatheid vir deteksie en identifikasie van hierdie E. sakazakii isolate het PKR met ses verskillende spesie-spesifieke peiler pare soos beskryf in die literatuur, geel-pigment produksie op triptoon soja agar (TSA) en chromogeniese Druggan-Forsythe-Iversen (DFI) en Enterobacter sakazakii (ES) agars ingesluit. Die geskiktheid van die VMA metode is verlaag deur die lae sensitiwiteit, spesifisiteit en akkuraatheid (87%, 71% en 74% onderskeidelik) van geel pigment produksie vir E. sakazakii identifikasie. Chromogeniese DFI en ES agars was sensitief, spesifiek en akkuraat (100%, 98% en 98% onderskeidelik) vir die identifikasie van E. sakazakii. Die spesifisiteit van die PKR produkte het gewissel tussen 8% en 92%, en Esakf en Esakr is as die akkuraatste geëvalueerde peiler paar geidentifiseer. Die huidige VMA metode vir E. sakazakii deteksie vereis hersiening aangesien meer sensitiewe, spesifieke en akkurate deteksiemetodes voortdurend beskikbaar word. 'n Nuwe metode, gebaseer op die resultate van hierdie studie, word voorgestel vir die deteksie van E. sakazakii in voedsel- en omgewingsmonsters. Die voorgestelde metode vervang die kwekingsstap op violet rooi gal glukose agar (VRGGA) en TSA deur kweking op chromogeniese DFI of ES agars. Verder word die oksidase toets, geel pigment produksie en API biochemiese profiele van vermoeidelike E. sakazakii isolate vervang deur DNS volgorde bepaling en/of spesie-spesifieke PKR met die mees spesifieke peiler paar (Esakf and Esakf) vir die identifikasie en bevestiging van E. sakazakii. Die voorgestelde wysigings van die VMA metode sal die tydsduur van E. sakazakii identifikasie van 7 dae na 4 dae verminder, en behoort ook meer sensitief, spesifiek en akkuraat te wees vir die deteksie van E. sakazakii. 'n Nuwe PKR-gebaseerde metode wat tussen lewensvatbare en dooie E. sakazakii selle kan onderskei is in hierdie studie ontwikkel. Dit is bereik deur die onomkeerbare binding van bakteriële DNS aan lig-geaktiveerde EMA of PMA om die PKR amplifisering van dooie selle te voorkom. Konsentrasies van 50 en 100 μg.ml-1 PMA het PKR amplifikasie heeltemal geïnhibeer, terwyl geen inhibisie van lewensvatbare selle bespeur kon word nie. EMA was ook suksesvol in die voorkoming van die PKR amplifikasie van dooie selle, alhoewel daar ook 'n mate van DNS inhibisie was tydens die amplifikasie van lewensvatbare selle. PMA-PKR kan ook van nut wees vir die assessering van die doeltreffendheid van prosesseringstegnieke, en ook vir die waarneming van die weerstandigheid, oorlewingsstrategieë en stresresponse van E. sakazakii. Dit sal 'n belangrike stap wees in pogings om E. sakazakii van voedsel en voedsel produksieomgewings te elimineer.

Os ácidos graxos polinsaturados atuam no organismo humano em diversos processos fisiológicos e metabólicos, além de serem importantes na nutrição infantil. A quantificação dos ácidos graxos polinsaturados, devido à presença de vários sítios reativos na molécula, deve envolver processos de extração da gordura em condições amenas. Este trabalho teve como objetivos a comparação dos métodos analíticos para determinação de lipídios totais e ácidos graxos polinsaturados (ácido linoléico, ácido &#945;-linolênico, ácido araquidônico e ácido docosahexaenóico) em fórmula infantil, a quantificação dos ácidos graxos polinsaturados nas fórmulas infantis comerciais e o acompanhamento da estabilidade destes ácidos graxos neste alimento. Foram analisadas 15 amostras, sendo uma amostra da Nacional Institute of Standards and Techonology (NIST 1849) e 14 fórmulas infantis comercializadas no Estado de São Paulo. Os métodos analíticos comparados para extração de lipídios foram: gravimétricos (Bligh Dyer, Roese Gottlieb e hidrólise ácida AOAC 963.15) e por cálculo (AOAC 996.06 e método direto adaptado de Golay et al. (2006)). Para a preparação dos ésteres metílicos de ácidos graxos utilizaram-se metodologias descritas pela IUPAC, Hartman e Lago e método direto adaptado de Golay et al. (2006). Compararam-se diferentes padrões interno, sendo estes dos ésteres metílicos de ácidos graxos 13:0, 21:0 e 23:0 e fatores de resposta para quantificação dos ácidos graxos polinsaturados em relação aos ácidos graxos 16:0, 18:0 e 23:0. O estudo de estabilidade durou 8 meses, e as análises foram realizadas nos meses de março, abril, julho e outubro de 2008. Os melhores resultados, para gordura total e para ácidos graxos polinsaturados, foram obtidos pelo método oficial (Roese Gottilieb). Quanto ao cálculo dos ácidos graxos polinsaturados, o uso do padrão interno 23:0 e o fator de resposta de correção teórico em relação ao 23:0 revelam resultados mais satisfatórios. Das fórmulas infantis comerciais analisadas, 85,7% apresentaram pelo menos um analito em desacordo com a informação nutricional e 100% com relação à legislação brasileira e o Codex Alimentarius. Em relação à estabilidade dos ácidos graxos polinsaturados, apenas três fórmulas infantis não apresentaram diferença estatisticamente significativa (p>0,05) nos teores de ácido graxos no período de 8 meses. / Long chain polyunsaturated fatty acids are involved in several physiological and metabolic process of human organism. The quantification of polyunsaturated fatty acid, must involve the fat extraction in mild conditions, due the reactive sites in the molecule. This work had as objective the comparison of the analytical methods for determination of polyunsaturated fatty acid (linoleic acid, &#945;-linolenic acid, arachidonic acid and docosahexaenoic acid) and lipids in infant formula, quantification of polyunsaturated fatty acids in commercial infant formulas and polyunsaturated fatty acids stability evaluation in these foods. A 15 samples of infant formulas were analyzed, these one from NIST and 14 commercial infant formulas. The analytical methods to lipids extraction were: Bligh and Dyer, Roese Gottlieb, acid hydrolyze AOAC 963.15, acid hydrolyze AOAC 996.06 and direct method adapted from Golay et al. (2006). To prepared esters metilics fatty acids utilized the methods: IUPAC, Hartman and Lago and direct method. The quantization of polyunsaturated fatty acid was realized with different standards internal, like fatty acid methyl ester 13:0, 21:0 and 23:0, and flame ionization detector response factors of correction in relation fatty acids 16:0, 18:0 and 23:0. The stability study during 8 months and analysis were analyzed in the months March, April, July and October. The best results, for lipids and polyunsaturated fatty acid, were obtained by the official method, Roese Gottlieb and metilation followed Hartman and Lago. In accordance with the present study the internal standard 23:0, and the theoretical correction factors with relation 23:0 showed satisfactory trueness and precision for calculation of polyunsaturated fatty acid. Among of commercial infant formula analyzed, 85.7% had at least one analyte in disagreement with the nutritional label facts and 100% with respect to Brazilian Legislation and the Codex Alimentarius. Regarding polyunsaturated fatty acids stability, three infant formulas showed no statistically significant difference (p > 0.05) in levels of these fatty acids during the period of analysis (8 months).

Als Schlüsselenzym im Renin-Angiotensin System übernimmt das Angiotensin Converting Enzyme (ACE) eine wichtige Rolle bei der Blutdruckregulierung. ACE spaltet das biologisch inaktive Angiotensin I zum vasokonstriktorisch wirksamen Angiotensin II, was zu einem Anstieg des Blutdruckes führt. Tier- und Humanstudien zeigten, dass die Aufnahme bekannter, aus dem Proteinabbau stammender ACE-Inhibitoren eine Absenkung des Blutdruckes bewirkte. In der Lebensmittelindustrie finden Hydrolysate von Milchproteinen, im speziellen von Molkenproteinen, Einsatz in hypoallergenen (HA) Säuglingsnahrungen. Obwohl das Phänomen einer ACE-Inhibierung durch HA-Nahrungen in vitro in der Literatur bereits Erwähnung fand, existieren bislang keine Angaben zu einer potentiellen Wirkung in vivo. In der vorliegenden Arbeit konnte für kommerzielle hypoallergene (HA) Säuglingsnahrung eine sehr starke ACE-Hemmung in vitro zeigen (IC50-Werte zwischen 20 und 104 mg Protein/liter), welche die für fermentierte Sauermilchprodukte dokumentierte Wirkung bei weitem überstieg.] Mittels RP-HPLC und ESI-TOF-MS konnte neben zahlreichen bekannten Peptiden mit ACE-hemmendem Effekt erstmals das aus der Primärsequenz von -Lactalbumin freigesetzte Dipeptid Ile-Trp in den HA-Nahrungen identifiziert und quantifiziert werden. Ile-Trp ist der bislang potenteste in Lebensmitteln nachgewiesene ACE-Inhibitor (IC50 = 0,7µM). HA-Nahrungen zeigten auch ex vivo im Zellsystem (HUVECs) einen stark ACE-hemmenden Effekt. Aus diesem Grunde wurde ein möglicher Einfluss der HA-Nahrungen auf den Blutdruck spontan hypertensiver Ratten untersucht. Hierfür wurden die Tiere im Rahmen einer Fütterungsstudie über 14 Wochen mit standardisiertem Futter, welchem HA-Nahrung (Gruppe 1), konventionelle Säuglingsnahrung (Gruppe 2) bzw. der bekannte ACE-Inhibitor Captopril (Gruppe 3) zugesetzt war, gefüttert. Eine vierte Gruppe mit Standardfutter diente als Kontrolle. Der Blutdruck wurde am wachen Tier nichtinvasiv mittels tail-cuff-Methode gemessen. Der systolische Blutdruck sank bei Verabreichung der HA-Nahrung nach 7 Wochen signifikant um 21 ± 8 mmHg ab im Vergleich zur Kontrollgruppe bzw. den mit konventioneller Säuglingsnahrung gefütterten Tieren. Captopril führte zur einer Blutdrucksenkung um 30 ± 7 mmHg.