Volatile Free Fatty Acids Research Articles

Deciphering pathogens inactivation mechanism during anaerobic co-digestion of food waste and waste activated sludge: The role of pH

Anaerobic co-digestion of waste activated sludge (WAS) with food waste (FW) represents a viable and effective approach to energy recovery and pollutant elimination. Little is known about the effect of alternating change of acid/alkaline conditions on pathogens inactivation within the co-digestion system of WAS and FW. In this study, pH changes during batch anaerobic co-digestion with WAS/FW ratios varying from 0.4 to 8, and their impact on pathogen inactivation were investigated using Escherichia coli as a model pathogen. At WAS/FW=1 and 2, serious acidification (pH<5.5) was effectively mitigated, and the pH transitioned from 6.4 during the acidogenesis stage to >8.5 during the methanogenesis stage. WAS/FW=1 achieved the highest methane yield (256.7 mL/g VS), which was 38.9 % greater than that in their mono-digestion scenario. A 2.37 − 3.87 log10 reduction in E. coli abundance occurred in all tested conditions during the acidogenesis stage, and a further reduction to an undetectable level was achieved only in WAS/FW reaching 1 subsequently. Low pH induced free volatile fatty acids dominated E. coli inactivation during the acidogenesis stage. During the methanogenesis stage, the combined action of OH− and the resultant free ammonia contributed to E. coli inactivation with free ammonia being more efficient in its action. This research contributes to the advancement of sustainable and secure management of organic wastes.

Dynamic microbial and metabolic changes during Apulian Caciocavallo cheesemaking and ripening produced according to a standardized protocol

The microbiota of a cheese play a critical role in influencing its sensory and physicochemical properties. In this study, traditional Apulian Caciocavallo cheeses coming from 4 different dairies in the same area and produced following standardized procedures were examined, as well as the different bulk milks and natural whey starter (NWS) cultures used. Moreover, considering the cheese wheels as the blocks of Caciocavallo cheeses as whole, these were characterized at different layers (i.e., core, under-rind, and rind) of the block using a multi-omics approach. In addition to physical-chemical characterization, culturomics, quantitative PCR, metagenomics, and metabolomics analysis were carried out after salting and throughout the ripening time (2 mo) to investigate major shifts in the succession of the microbiota and flavor development. Culture-dependent and 16S rRNA metataxonomics results clearly clustered samples based on microbiota biodiversity related to the production dairy plant as a result of the use of different NWS or the intrinsic conditions of each production site. At the beginning of the ripening, cheeses were dominated by Lactobacillus, and in 2 dairies (Art and SdC), Streptococcus genera were associated with the NWS. The analysis allowed us to show that although the diversity of identified genera did not change significantly between the rind, under-rind, and core fractions of the same samples, there was an evolution in the relative abundance and absolute quantification, modifying and differentiating profiles during ripening. The real-time PCR, also known as quantitative or qPCR, mainly differentiated the temporal adaptation of those species originating from bulk milks and those provided by NWS. The primary starters detected in NWS and cheeses contributed to the high relative concentration of 1-butanol, 2-butanol, 2-heptanol, 2-butanone, acetoin, delta-dodecalactone, hexanoic acid ethyl ester, octanoic acid ethyl ester, and volatile free fatty acids during ripening, whereas cheeses displaying low abundances of Streptococcus and Lactococcus (dairy Del) had a lower total concentration of acetoin compared with Art and SdC. However, the subdominant strains and nonstarter lactic acid bacteria present in cheeses are responsible for the production of secondary metabolites belonging to the chemical classes of ketones, alcohols, and organic acids, reaffirming the importance and relevance of autochthonous strains of each dairy plant although only considering a delimited production area.

Generation of High Concentration Free Volatile Fatty Acids from Continuous Anaerobic Digestion of Chicken Manure by Suppressing the Decomposition of Nitrogenous Components.

Free volatile fatty acids (free VFA) play a crucial role in the inactivation of pathogens during the anaerobic digestion of animal manure. However, the decomposition of nitrogenous components can release alkaline ammonium-N, which might increase the pH and reduce the concentration of free VFA. In this study, continuous anaerobic digestion of high-solid chicken manure was conducted for 150days. The results indicated the process stabilized at a pH of approximately 6.0, with total ammonia nitrogen (TAN) of around 7.0g/L. The resulting concentration of free VFA was only about 3.1g/L, which might not sufficiently effective for pathogen inactivation. On the 70th day, hydrogen chloride was added into the reactor to adjust the pH to 5.5. This led to a further decrease in pH to 4.3 and TAN to 2.3g/L. As a result, the concentration of free VFA significantly increased, reaching up to 12.6g/L. These findings support the potential for generating high levels of free VFA even for nitrogen-rich manure by implementing an appropriate process regulation.

Effects of milk types used in Antep cheese production on some cheese organoleptic quality parameters and brine composition during 5‐month ripening

Antep cheese is a local Turkish cheese characterized by scalding during production and ripened in brine. In this study, Antep cheeses were produced using mixtures of different milk types (cow, sheep, and goat milk) and ripened for 5 months. The composition, proteolytic ripening extension index (REI), free fatty acid (FFA) content, and volatile compounds of the cheeses and the variation of the brines were analyzed for the 5-month ripening period. Low proteolytic activity in cheese during ripening caused the cheeses to have low REI values (3.92%-7.57%), although it was observed that some parts of the water-soluble nitrogen fractions diffused into the brine, causing a lower REI. As a result of lipolysis during ripening, total FFA (TFFA) concentrations in all cheeses were increased, whereas the highest increases were detected in the concentrations of short-chain FFAs. The highest FFA concentrations were determined in cheese produced using goat milk, and the volatile FFA ratio in TFFA exceeded 10% in the third month of ripening. Although it was observed that the milk types used in the production had significant effects on the change of volatile compounds of the produced cheeses and their brines, the impact of the ripening time was more important. PRACTICAL APPLICATION: This study investigated Antep cheese made with different milk types. Volatile compounds and soluble nitrogen fractions were transferred to the brine by diffusion during ripening. The volatile profile of the cheese varied with milk type, but ripening time was the primary factor influencing volatile compounds. This suggests that the targeted organoleptic properties of the cheese are determined by ripening time and conditions. Additionally, changes in the brine's composition during the ripening process provide insight into how to manage the brine as waste.

Altering the Chain Length Specificity of a Lipase from Pleurotus citrinopileatus for the Application in Cheese Making.

In traditional cheese making, pregastric lipolytic enzymes of animal origin are used for the acceleration of ripening and the formation of spicy flavor compounds. Especially for cheese specialities, such as Pecorino, Provolone, or Feta, pregastric esterases (PGE) play an important role. A lipase from Pleurotus citrinopileatus could serve as a substitute for these animal-derived enzymes, thus offering vegetarian, kosher, and halal alternatives. However, the hydrolytic activity of this enzyme towards long-chain fatty acids is slightly too high, which may lead to off-flavors during long-term ripening. Therefore, an optimization via protein engineering (PE) was performed by changing the specificity towards medium-chain fatty acids. With a semi-rational design, possible mutants at eight different positions were created and analyzed in silico. Heterologous expression was performed for 24 predicted mutants, of which 18 caused a change in the hydrolysis profile. Three mutants (F91L, L302G, and L305A) were used in application tests to produce Feta-type brine cheese. The sensory analyses showed promising results for cheeses prepared with the L305A mutant, and SPME-GC-MS analysis of volatile free fatty acids supported these findings. Therefore, altering the chain length specificity via PE becomes a powerful tool for the replacement of PGEs in cheese making.

The role of oxidation-reduction potential as an early warning indicator, and a microbial instability mechanism in a pilot-scale anaerobic mesophilic digestion of chicken manure

To identify an early warning indicator for the anaerobic digestion of chicken manure and reveal a microbial mechanism of instability, a bench-scale digester with a working volume of 55 L was utilized by raising the organic loading rates (OLRs). Three characteristics were identified and posited as early warning indicators, namely: one-way changes, a longer early response time, and sudden variations. The core early warning indicator set was constructed, which included the oxidation-reduction potential (ORP), individual volatile fatty acids, and CH4/CO2. Considering the convenience, economic nature, and reliability of industrial biogas plants, the ORP was recommended as a final early warning indicator with an inhibition threshold of > −540 mV. The initial instability was caused by the accumulation of ammonia, which suppressed methanogenic activity, although it stimulated the growth of Methanosarcina. The accumulation of free volatile fatty acids, which inhibited both the growth and methanogenic activity of Methanosarcina, was responsible for the final collapse.

Ultrasonication retains more milk fat globule membrane proteins compared to equivalent shear-homogenization

Ultrasonication, like common shear homogenization, can reduce the milk fat globule size and may change the milk fat globule membrane (MFGM). This work compared the effect of ultrasonication to equivalent shear homogenization on MFGM proteins and lipid-derived volatile components. Results showed that treating milk with ultrasound at 35 kJ/L would realize a similar size distribution of the milk fat globules as shear-homogenization at 20 MPa. Proteomics analysis revealed that in total 192 MFGM proteins were identified and quantified and a number of these proteins were lost after both treatments; however, more MFGM proteins remained after ultrasonication than after shear-homogenization. SDS-PAGE results showed that milk plasma proteins, and especially caseins, were absorbed on the milk fat globules after both treatments. In addition, the amount of the volatile free fatty acids increased after both treatments.Industrial relevance: Ultrasonication, as an innovative food processing technology, in comparison to traditional homogenization, was shown to equally efficiently decrease the MFG size, but lead to less damage to native MFGM proteins, which may be due to its longer homogenization time window. These results increased knowledge on the biochemical changes of milk fat globules after their size reduction and showed that ultrasonication could be used as a novel approach to improve dairy product quality.

Impact of Functional Stirred Low Fat Yoghurt Supplemented With Spirulina platensis Powder on Some Quality Characteristics and Therapeutic Effects In Vivo تأثير تدعيم يوغورت مقلب وظيفي منخفض الدهن بمسحوق Spirulina platensis على بعض خصائص الجودة والتأثيرات العلاجية في کائن حي

Yoghurt is one of the most consumed healthy and nutritious foods worldwide. Nutrition-related diseases are regularly found in developed or underdeveloped countries, wherein enriched dairy products can dramatically reduce the risk of these diseases. Spirulina platensis powder (SPP) is considered a good source of bioactive compounds exhibiting high antioxidant properties. In the present research, physicochemical and nutritional attributes of yoghurt supplemented with 0.5, 1, and 2% of SPP were investigated. The results showed that the addition of SPP decreased coagulation time, syneresis, and pH values. The supplementation with SPP increased yoghurt acidity as lactic acid, total solids content, total protein, fat, and total volatile free fatty acids. Stirred yoghurt treatments containing 0.5% of SPP displayed higher sensory acceptability than other treatments. Furthermore, in vivo therapeutic effects of the produced yoghurt were evaluated. The results revealed that SPP-supplemented yoghurt provided a novel technological approach to manufacture a functional food exhibiting prevention effects to cardiovascular diseases and improved liver, kidney, and heart functions.

The screening of early warning indicators and microbial community of chicken manure thermophilic digestion at high organic loading rate

Anaerobic digestion of chicken manure (CM) with high nitrogen content is easily suffered inhibition of ammonia, especially under thermophilic and high organic loading rate (OLR) condition. To screen the early warning indicators for instability, thermophilic digestion of CM was conducted in a bench-scale reactor (working volume 55 L) under OLR from 1.0 to 6.0 g VS/(L·d). Microbial community analysis was used to reveal the mechanism of instability at high OLR. The system collapsed at an OLR of 6.0 g VS/(L·d). The initial ammonia inhibition evoked serious acidification, subsequently, the dual suppression of free volatile fatty acids and free ammonia caused the final collapse. The most likely mechanism of iso-butyrate degradation was propionate pathway. The ratio of n-butyrate to iso-butyrate was screened as an early warning indicator with the threshold of above 0.45. The high OLR of 6.0 g VS/(L·d) broke the balance of syntrophic oxidation bacteria (norank_o__MBA03, Gelria, and norank_o__D8A-2), hydrogenotrophic methanogens (Methanothermobacter) and fermentative and acid-producing bacteria (Ruminiclostridium_1, Bacteroides, Defluviitalea, norank_o__Clostridia, and norank_o__M55-D21). The non-functional acid tolerant bacteria (Defluviitoga, norank_f__Family_XI, Corynebacterrium_1, Caldicoprobacter, Lactobacillus, and Erysipelothrix) became the dominant after the system collapsed.

Volatilome and Bioaccessible Phenolics Profiles in Lab-Scale Fermented Bee Pollen.

Bee-collected pollen (BCP) is currently receiving increasing attention as a dietary supplement for humans. In order to increase the accessibility of nutrients for intestinal absorption, several biotechnological solutions have been proposed for BCP processing, with fermentation as one of the most attractive. The present study used an integrated metabolomic approach to investigate how the use of starter cultures may affect the volatilome and the profile of bioaccessible phenolics of fermented BCP. BCP fermented with selected microbial starters (Started-BCP) was compared to spontaneously fermented BCP (Unstarted-BCP) and to unprocessed raw BCP (Raw-BCP). Fermentation significantly increased the amount of volatile compounds (VOC) in both Unstarted- and Started-BCP, as well as modifying the relative proportions among the chemical groups. Volatile free fatty acids were the predominant VOC in Unstarted-BCP. Started-BCP was differentiated by the highest levels of esters and alcohols, although volatile free fatty acids were always prevailing. The profile of the VOC was dependent on the type of fermentation, which was attributable to the selected Apilactobacillus kunkeei and Hanseniaspora uvarum strains used as starters, or to the variety of yeasts and bacteria naturally associated to the BCP. Started-BCP and, to a lesser extent, Unstarted-BCP resulted in increased bioaccessible phenolics, which included microbial derivatives of phenolic acids metabolism.

Antioxidant Properties and the Preservative Effect of Whole Radish Extract on Quality of Deccan Mahseer (Tor khudree) Steaks during Chilled Storage

ABSTRACT The effect of dip treatment with solvent-free whole radish extract (WRE) on quality characteristics of Deccan mahseer steaks was evaluated during the chilled storage. In-vitro antioxidant activities of the extract were evaluated based on total phenolic and flavonoid contents, DPPH radical scavenging activity, and metal chelating ability prior to the dip treatment. The quality parameters, including total volatile base nitrogen (TVB-N), trimethylamine nitrogen (TMA-N), and free fatty acid (FFA), were significantly (p < .05) reduced in WRE treated steaks compared to the control (CT) steaks. WRE treatment of fish steaks significantly controlled the lipid oxidation parameters of thiobarbituric acid reactive substances (TBARS) and peroxide value (PV). Total plate count (TPC) of CT sample was higher than the treated sample and remained within the limit of 7 log10 cfu/g at the end of storage. As per the sensory analysis, shelf life of Deccan mahseer was determined to be 15 days for WRE treated steaks and 9 days for control steaks during chill storage. Based on the results, it can be concluded that WRE treatment can limit the progress of fish spoilage bacteria in fish steaks and can be used as a substitute for synthetic additives.

Production of enzyme-modified cheese (EMC) with ripened white cheese flavour: II- effects of lipases

Enzyme-modified cheeses (EMCs) have been used uniquely to enhance the cheese flavour in processed foods. In this paper, effects of lipolytic enzymes during EMC production were investigated. EMC with ripened white cheese flavour was produced by a two-stage process in which proteolysis is followed by lipolysis. The results of proteolysis as the first stage were discussed in our previous report (Bas et al., 2019). In the present paper, four different commercial lipolytic enzymes with 3 different incubation times were applied to reach the target ripened white cheese flavour and free fatty acid (FFA) profiles, volatile compounds and sensory properties were investigated and the relationships between these properties were evaluated. Results showed that a balance in FFA profile is important. Samples contained nearly 19% butanoic acid in total volatiles and in the range of 10.7–12.3% volatile FFA in total FFA were desirable. During lipolysis, 17 new volatiles were formed, and most of the acids and esters among them are compounds commonly found in ripened white cheese. Moreover, absence of n-aldehydes was desirable for ripened white cheese flavour and lipolysed samples did not contain n-aldehydes. Briefly, lipolysis is critical in the formation of ripened white cheese flavour and two different enzyme-incubation time combinations (24 h with Piccantase® A and 48 h with Lipomod™ 801MDP) were suggested to use in the production of EMC with ripened white cheese flavour.

No difference in inhibition among free acids of acetate, propionate and butyrate on hydrogenotrophic methanogen of Methanobacterium formicicum

Free volatile fatty acids such as free acetic acid (FAA) and free butyrate acid (FBA) are true inhibitors of hydrogenotrophic methanogens (HM) in mixed culture. However, their inhibitory effects on pure culture of HM remain unclear. In this study, a typical HM of Methanobacterium formicicum demonstrated no difference in toxicity conferred by FAA, free propionate acid (FPA), or FBA in regard to the specific methanogenic activity (SMA) based on the C50% (0.19, 0.17, and 0.23 g/L, respectively) and recoverable concentration values (0.97, 0.69, and 0.61 g/L, respectively). These results were within the same order of magnitude. The concentrations of FAA, FBA, and FPA all correlated well with the SMA values according to the inhibition model. Additionally, changes in the activity of the electron transport system also agreed well with the trend in the SMA variation. Together, the results of this study provide a benchmark to control methanogenesis during industrial applications.

Microbiological and Physicochemical Quality of Carp sausage Enriched With Propolis Natural Extract during Chilled Storage

ABSTRACTThis study aimed to determine the effect of propolis extract on quality of liquid smoked carp (Cyprinus carpio) sausage stored at 2°C for 9 weeks. Carp sausages were separated into three groups: control group, without propolis extract; second group, with 1% (w/w) propolis extract added; and third group, with 2% (w/w) propolis extract added. Propolis extract added carp sausages showed lower total volatile basic nitrogen (TVB-N) value, peroxide value (PV), and free fatty acid (FFA) content compared to control during the storage period. During 9 weeks of storage, propolis extract added carp sausages showed significantly lower total aerobic mesophilic bacteria (TAMB) and psychrophilic bacteria count (PBC) than those of control samples (p < .05). Yeast and mold were not detected in all fish sausages throughout the storage period.

Effects of using whey and maltodextrin in white cheese powder production on free fatty acid content, nonenzymatic browning and oxidation degree during storage

Use of maltodextrin and whey improves the physical properties of spray-dried cheese powder and, as cheese replacers, they decrease the raw material costs. However, it is necessary to evaluate their effects on the chemical quality and sensory properties of the product. Three powders, control (CON), whey-added cheese powder (WACP), and maltodextrin-added cheese powder (MACP) were produced to determine free fatty acid (FFA) content, degree of nonenzymatic browning, oxidation, and sensory (flavour) changes during 12 months' storage. In the emulsion preparation and drying process, total volatile FFAs decreased up to 9% and 53.5%, respectively, with higher decrease in CON than in WACP or MACP. Although whey increased oxidation degree of powder, oxidised flavour could not be perceived until the ninth month of storage. Maltodextrin slightly decreased cheese flavour and overall impression scores. There was no difference detected in the powders for scorched flavour.

Antioxidant, Some Flavor Components, Microbiological and Microstructure Characteristics of Corn Milk Yoghurt

Corn milk is considered as a new innovation especially in the making of yoghurt based products. The present study was conducted to study antioxidants components such as (phenols compounds and DPPH scavenging activity), total volatile free fatty acids, microbiological and microstructure characteristics in yoghurt samples produced from two different ratios of buffalos and corn milk (70:30 coded A and 60:40 coded B) respectively. Results indicated that the increasing ratio of corn milk led to increase the content of phenolic components and DPPH scavenging activity. Total viable content decreased in samples (70:30 and 60:40). Streptococci and lactobacilli count decreased by the increase of corn milk in produced samples. Results also show that the trend of yeasts content was the opposite ones. Election microscopy examination revealed that adding corn milk to yoghurt had more smoothly and systematically distributed casein with a bit coarse structure less porosity in the network of casein. From these results could be concluded that adding 40% corn milk during processing of yoghurt is increasing the antioxidant activity and improves the microstructure as well.

Phytohormones and free volatile fatty acids from cyanobacterial biomass wet extract (BWE) elicit plant growth promotion

The present study highlights the importance of cyanobacteria in plant growth promotion and screened the effect of biomass wet extract (BWE 1% and BWE 10%) of twenty cyanobacterial strains on the morphometric parameters of Pisum sativum L. seedlings after germination. Cyanobacterial BWE treatment significantly increased the radicle, plumule and total seedling length along with percentage of response than the control treatment with water. At BWE 1% treatment, Scytonema bohneri MBDU 104 showed 73.3% of the total seedling response, whereas Dolichospermum spiroides MBDU 903 showed increased total seedling length of 5.69cm among other strains tested. All the tested strains varied among themselves at different parameters. To identify the most potent isolate, all the tested morphometric parameters were evaluated using a multi – criteria decision analysis (MCDA) of PROMETHEE-GAIA software. The isolate Scytonema bohneri MBDU 104 was selected as the potent plant growth promoting cyanobacteria and its BWE was purified using preparative – HPLC and analysed by LC-MS/MS for identifying possible plant growth elicitors. Phytohormones like indole-3 acetic acid (IAA), indole-3 butyric acid (IBA) and cinnamic acid along with free volatile fatty acids were identified and implicated for plant growth promotion by cucumber seed germination bioassay.

The microbiome of NLR knockout animals reveals insights into susceptibility to experimental colitis.

Abstract Upon sensing particular pathogen associated molecular patterns (PAMPs), inflammasome-forming NOD-like receptors (NLRs) initiate pyroptosis and the cleavage of pro IL-18 and pro IL-1β to their active forms. Consequently, the absence of an NLR or its malfunction in the gut has been shown to result in dysbiosis. There is significant evidence linking dysbiosis with chronic inflammation in the gut and colon cancer. Prior work in our laboratory has revealed that mice lacking NLR inflammasomes, specifically Nlrp1−/− mice, are highly susceptible to experimental colitis and colitis-associated cancer. Here, we extend these findings by evaluating the microbiome of mice lacking either Asc or Nlrp1. 16s rRNA sequencing revealed 6 unique genera prevalent in Nlrp1−/− mice compared to WT and 9 genera in Asc−/− mice compared to WT. Remarkably, the microbiome composition in both knockout animals was more pathogenic, including such genera as Helicobacter, Mucispirillum, and Clostridium. The abundance of volatile free fatty acids was assessed to investigate functional metabolic differences due to the uniqueness of the microbiome in these animals, and interestingly the abundance of acetate, butyrate, and proprionate were distinctly different in Nlrp1−/− and Asc−/− mice compared to WT. Crucially, our analyses were conducted prior to any experimental colitis model. The differences in the microbiomes of these mice have the potential to resolve conflicting data in the field regarding inflammasome deficient mice and subsequent predisposition to or protection from colitis and cancer. Future work will establish which NLR is responsible for sensing the specific bacteria present in the microbiome of the inflammasome-deficient mice at rest.

Дослідження ефективності використання бактеріальних препаратів на якісні характеристики делікатесних виробів

Досліджено упродовж визрівання фізико-хімічні, біохімічні та мікробіологічні характеристики делікатесного продукту з м’яса птиці, виготовленого зі застосуванням бактеріального препарату та нетрадиційних інгредієнтів рецептури. У статті показано, що корисна мікрофлора бактеріального препарату, який містить композицію бактеріальних штамів, позитивно впливає на сировину – філе курчат-бройлерів, в якому відбувається зміна не лише фізико-хімічних, а головне – біохімічних показників. Перетворення вільних амінокислот і летких жирних кислот не тільки характеризує біологічну цінність продукту, а насамперед, обумовлює його смак і аромат. Доказом, є більш високий (до 30%) вміст вільних амінокислот як в якісному, так і в кількісному складі, порівняно з контролем. Показано, що в продукті з бактеріальним препаратом на кінцеву добу ферментування вміст летких жирних кислот на 11,6% був вищим, ніж у контролі. Ефективність використання у продукті бактеріального препарату підтверджена і результатами мікробіологічних досліджень, за яких дослідний продукт був готовий на 3 доби раніше, ніж контроль. Отже, доведено, що використання бактеріального препарату змінює інтенсивність та спрямованість процесу визрівання продукту, надає та формує специфічні смако-ароматичні властивості та забезпечує продукту гарантовану якість.

Vacuum-assisted headspace-solid phase microextraction for determining volatile free fatty acids and phenols. Investigations on the effect of pressure on competitive adsorption phenomena in a multicomponent system

This work proposes a new vacuum headspace solid-phase microextraction (Vac-HSSPME) method combined to gas chromatography-flame ionization detection for the determination of free fatty acids (FFAs) and phenols. All target analytes of the multicomponent solution were volatiles but their low Henry's Law constants rendered them amenable to Vac-HSSPME. The ability of a new and easy to construct Vac-HSSPME sampler to maintain low-pressure conditions for extended sampling times was concurrently demonstrated. Vac-HSSPME and regular HSSPME methods were independently optimized and the results were compared at all times. The performances of four commercial SPME fibers and two polymeric ionic liquid (PIL)-based SPME fibers were evaluated and the best overall results were obtained with the adsorbent-type CAR/PDMS fiber. For the concentrations used here, competitive displacement became more intense for the smaller and more volatile analytes of the multi-component solution when lowering the sampling pressure. The extraction time profiles showed that Vac-HSSPME had a dramatic positive effect on extraction kinetics. The local maxima of adsorbed analytes recorded with Vac-HSSPME occurred faster, but were always lower than that with regular HSSPME due to the faster analyte-loading from the multicomponent solution. Increasing the sampling temperature during Vac-HSSPME reduced the extraction efficiency of smaller analytes due to the enhancement in water molecule collisions with the fiber. This effect was not recorded for the larger phenolic compounds. Based on the optimum values selected, Vac-HSSPME required a shorter extraction time and milder sampling conditions than regular HSSPME: 20 min and 35 °C for Vac-HSSPME versus 40 min and 45 °C for regular HSSPME. The performance of the optimized Vac-HSSPME and regular HSSPME procedures were assessed and Vac-HSSPME method proved to be more sensitive, with lower limits of detection (from 0.14 to 13 μg L−1), and better intra-day precision (relative standard deviations values < 10% at the lowest spiked level) than regular HSSPME for almost all target analytes. The proposed Vac-HSSPME method was successfully applied to quantify FFAs and phenols in milk and milk derivatives samples.