Cold Saponification Research Articles

Pemberdayaan Masyarakat Pondok Pesantren (Santriwati) dalam Pembuatan Sabun Cuci Pakaian Berbahan Dasar Minyak Jelantah

Waste cooking oil/used cooking oil which is blackish brown is residue from the frying process that still contains fatty acids which have the potential to be used to make soap. At the Al Ihsan Puteri Islamic boarding school, Banjarmasin used cooking oil waste has not been managed well. This activity aims to educate the Islamic boarding school community (santriwati) and boarding school administrators about making washing soap from used cooking oil as a solution so that used cooking oil can be reused in other forms. Participants in the activity were 15 female students at the Al Ihsan Puteri Banjarmasin Islamic Boarding School. This activity consists of discussion (counseling) and direct practice. The chemistry knowledge education material explains the impact of used cooking oil on the body, while the soap-making training material explains the cold saponification method through demonstrations. The service results show that almost 100% of the students who initially did not know how to reuse used cooking oil in soap, now know how to reuse used cooking oil. Students also increase their knowledge and skills in reacting with materials in the saponification process.

Utilizing Used Cooking Oil and Organic Waste: A Sustainable Approach to Soap Production

This research examined the potential for utilizing waste materials generated during the production of dishes/meals and organic waste. Specifically, it evaluated the use of orange peel (OP), spent coffee grounds (SCG), and waste cooking oil in the production of soaps. For the purposes of this study, homemade soaps were made from used food oils using the cold saponification method using sodium hydroxide. During the soap preparation, spent coffee grounds and orange peel were added to the samples in increasing concentrations of 1%, 2.5%, and 5%. The quality of the individual types of homemade soaps was evaluated on the basis of physicochemical properties such as pH, moisture, total alkalinity, total fatty matter, malondialdehyde content, fat content, foaminess, and hardness. All soaps produced using the cooking oil met the ISO quality criteria and reveal a high TFM content, low moisture content, and also very good foam stability and satisfactory foaming stability. However, no relationship was observed between the use of OP and SCG in soap production and these parameters. However, according to the ABTS test, OP and SCG significantly contributed to the antioxidant properties of the soaps, while SCG-impregnated soaps performed slightly better in this respect. Soaps with SCG also had the highest levels of flavonoids. On the other hand, the fillers used for the soap formulation reduced their hardness. All soaps showed 100% solubility in water, thus confirming the biodegradability of the product. This study demonstrated the novel potential of incorporating waste products like orange peel, spent coffee grounds, and waste cooking oil into homemade soaps, highlighting their contributions to its antioxidant properties and water solubility while ensuring high quality standards.

Valorizing Olive Oil Mill Wastewater: Transforming Waste into Natural Soaps

In this research, the main objective is to find a solution to the problem of olive mill wastewater (OMWW). This solution involves the recovery of liquid waste produced by the crushing units, which contain a significant amount of oily residues despite their initial treatment. The concept is based on separating these discharges into aqueous and oil phases and using the latter to manufacture natural soaps. The liquid waste from the extraction of olive oil has a significant content of oily residues, characterized by a very high acidity (2.73%), exceeding the value of edible olive oil, as well as a high saponification index (186.2 mg KOH/g), making it an ideal source of fat for saponification. The results of the valorization of the OMWW in soap manufacturing reveal several significant elements. First, following the characterization of cold and hot products, a clear preference emerges in favor of soaps made using the cold process. In addition, the production yield is notable, with a rate of 94% for cold saponification and 89.9% for hot saponification, highlighting the efficiency of the process. Finally, the study highlights the importance of the quantities of fatty acids used in the formulation of soaps. An optimal formula is identified, comprising 40% oils recovered from the OMWW, 30% olive oil, 20% cocoa oil, and 10% castor oil, demonstrating the need for a precise balance to obtain quality soaps. This study solves the problem of the OMWW by reusing them to make natural soaps, thus reducing industrial waste. It also opens up new economic opportunities by creating a profitable and environmentally responsible production chain, promoting the transition to a circular economy.

Evaluation of the carotenoid and fat-soluble vitamin profile of industrial hemp inflorescence by liquid chromatography coupled to mass spectrometry and photodiode-array detection

Industrial hemp (Cannabis sativa L.) is a plant matrix whose use is recently spreading for pharmaceutical and nutraceutical purposes. Detailed characterization of hemp composition is needed for future research that further exploits the beneficial effects of hemp compounds on human health. Among minor constituents, carotenoids and fat-soluble vitamins have largely been neglected to date despite carrying out several biological activities and regulatory functions. In the present paper, 22 target carotenoids and fat-soluble vitamins were analyzed in the inflorescences of seven Italian industrial hemp varieties cultivated outdoor. The analytes were extracted by cold saponification to avoid artifacts and analyzed by high-performance liquid chromatography coupled with Selected reaction monitoring mass spectrometry. Phytoene, phytofluene, and all-trans-β-carotene were the most abundant in all analyzed samples (31–55 µg g−1, 11.6–29 µg g−1, and 7.3–53 µg g−1, respectively). Besides the target analytes, liquid chromatography coupled with photodiode-array detection allowed us to tentatively identify several other carotenoids based on their retention behavior and UV–vis spectra with the support of theoretical rules and data in the literature. To the best of our knowledge, this is the first comprehensive characterization of carotenoids and fat-soluble vitamins in industrial hemp inflorescence.

FORMULATION AND EVALUATION OF HERBAL SOAP TAKING DIFFERENT BIOACTIVE PLANTS BY COLD SAPONIFICATION METHOD

Objective: The final goal of this research is to develop and evaluate herbal bath soap taking various bioactive herbal plants extract with different ethinic and dermatological importance in ayurveda, namely Solanum lycopersicum, Sapindus mukorossi, Acacia nilotica, Citrus limon, Aloe barbadensis, Piper betle, Curcuma longa and Cocus nucifera.
 Methods: The extract of all plant materials were obtained through various extraction procedures suitable for them. The extract was then mixed with lye and fatty oil for preparing a soap using the cold saponification method.
 Results: The prepared soap was evaluated against marketed soap. The prepared soap was found to be good in appearance, color and odor. pH, % free alkali content, foamability, foam stability, moisture content and alcohol insoluble matter were found to be 9.6, 0.22, 15 cm, 14.5 cm, 3.26 and 17.25, respectively. The antioxidant activity and antibacterial studies were done, which signifies prepared soap to be a potent antioxidant and antibacterial source.
 Conclusion: Based on the study results it can be concluded that herbal soap can be formulated using cold process method, taking different parameters in consideration as that of skin condition and as that of herbal potentials and its activity. This sought of herbal formulation can bring a big difference in the field of herbal cosmetic as there are many alignment and related flaws in different polyherbal or chemical-based formulations which can be removed.

Evaluation of Antibacterial and Antifungal Activities of a Medicinal Soap Made from Zingiber officinale and Syzygium aromaticum Essential Oils

Aims: The objective of this study is to evaluate the antibacterial and sensory properties of a formulated medicinal soap.
 Study Design: Extraction of essential oils, purchase of vegetable oils and caustic soda, soap formulation, testing of its antibacterial and antifungal activities, evaluation of its acceptability.
 Place and Duration of Study: Research unit of biochemistry of medicinal plant, food and nutritional sciences, Department of Biochemistry, Faculty of Science, University of Dschang from October 2019 to July 2020.
 Methodology: Cold saponification method was used for the production of soap using the following ingredients: Palm kernel oil, olive oil, coconut oil, Palm oleic, caustic soda, essential oils from ginger roots and cloves buds. The formulated soap was used for the evaluation of its anti-bacterial and anti-fungal activities as well as its acceptability using the 9-points hedonic scale. The soap was tested on the following microorganisms: Pseudomonas aeruginosa PA01, Staphylococcus aureus 56, Staphylococcus aureus 18, Staphylococcus aureus ATCC 25923, Candida albican, Candida tropicalis.
 Results: Results showed that the produced soap has bactericidal activities against staphylococcus aureus 18 and staphylococcus aureus ATCC 25923 and a bacteriostatic activity against staphylococcus aureus 56. Its activity against fungi was very low. The results of the sensory analysis of this product showed that it can be accepted by consumer since the lowest score was 6.55 in a scale of 9.
 Conclusion: Results showed that the produced soap has bactericidal activities against staphylococcus aureus 18 and staphylococcus aureus ATCC 25923 and a bacteriostatic activity against staphylococcus aureus 56. It was accepted with a score of at least 6.55/9 by the participants.

Performance of Liquid Eversa on Fatty Acid Ethyl Esters Production by Simultaneous Esterification/Transesterification of Low-to-High Acidity Feedstocks

Liquid Eversa was evaluated in hydrolysis of acylglycerols from soybean oil deodorizer distillate (SODD), as well as simultaneous esterification/transesterification of SODD with low-to-high free fatty acids (FFAs) content using ethanol as acyl acceptor. Hydrolysis of SODD at mild temperature (37 °C) and without pH control (water:SODD mass ratio of 4:1) increased its FFAs content from 17.2 wt.% to 72.5 wt.% after 48 h reaction. A cold saponification of SODD allowed a saponification phase (SODD-SP) to be recovered with 93 wt.% saponification index and 2.25 wt.% FFAs content, which was used to find the experimental conditions for simultaneous esterification/transesterification reactions by experimental design. Temperature of 35 °C, enzyme concentration of 8.36 wt.%, and molar ratio of 3.64:1 (ethanol:SODD-SP) were found as the best conditions for fatty acid ethyl esters (FAEEs) production from SODD-SP (86.56 wt.% ester yield after 23 h reaction). Under the same reaction conditions, crude SODD (17.2 wt.% FFAs) and hydrolyzed SODD (72.5 wt.% FFAs) yielded products containing around 80 wt.% FAEEs. Caustic treatment could increase the ester content to around 90 wt.% and reduce the FFAs content to less than 1 wt.%. Our results show the good performance of liquid Eversa in aqueous (hydrolysis reactions) and organic (esterification/transesterification reactions) media.

Formulation and Antifungal Activities of Syzygium Aromaticum Essential Oil Contained in Natural Bath Soap

Syzygium aromaticum essential oil (clove oil) has a promising potential as an antifungal. However, the potential has never been applied in bath soaps. This study aimed to produce an antifungal natural soap using clove oil as the additive. Natural bath soap containing clove oil was made through a cold saponification reaction of vegetable oils, sodium hydroxide, and various contents of clove oil (1, 2, and 3% w/w) as an antifungal agent. The soap quality was tested based on Indonesian National Standards (INS), including water content, total fat, insoluble matters in ethanol, free alkali, and unsaponified fatty matter. With a Kirby-Bauer diffusion method, the researchers carried out an antifungal activity est of clove oil soap on Candida albicans and Trichophyton mentagrophytes. All samples in the quality test of clove oil natural bath soap have met the standard. The antifungal activities of all tested clove oil soap performed growth inhibition of Candida albicans yeast and Trichophyton mentagrophytes mould in all treatments. However, there is no relationship between the clove oil concentration and antifungal activities in the range of clove oil content in this study. The formulation of clove oil in natural bath soap executed excellent antifungal activities against both yeast and mould. Due to the potential of clove oil as an antifungal agent, clove oil in bath soap is efficient to be used as natural cosmetics and medicinal soap in the future.

Differential Thermal Isomerization: Its Role in the Analysis of Vitamin D3 in Foods.

For nutritional purposes, the measurement of vitamin D3 (defined as the sum of vitamin D3 and previtamin D3) is required to obtain an accurate and reliable estimate of its content in foods. An often neglected aspect in the development of methods for the analysis of vitamin D3 is accounting for any potential analytical bias in the results associated with differential thermal isomerization between previtamin D and vitamin D. For LC-UV methods using a vitamin D2 internal standard, cold saponification, or direct lipid extraction techniques should be avoided, unless chromatographic separation of vitamin D2, vitamin D3, and their previtamin forms is achieved so that UV absorbance corrections can be made. For both LC-UV and LC-MS methods using calciferol internal standards, the simplest solution to avoid analytical bias due to the presence of previtamin D is to utilize heating conditions (typically during saponification) such that previtamin D and vitamin D in the sample and the internal standard reach an equivalent equilibrium state prior to instrumental analysis. Only under such circumstances is the integration of previtamin D unnecessary to obtain accurate results for vitamin D3. A detailed discussion of the quantitation of vitamin D3 in food with concise recommendations for avoiding measurement bias as a consequence of differential thermal isomerization.

Physico-chemical, Gas Chromatography-Mass Spectrometry (GC-MS) Analysis and Cold Saponification of Wild Grapes (Lannea microcarpa Engl & K. Krause) Seed Oil

Aims: Physico-chemical, Gas Chromatography-Mass Spectrometry (GC-MS) and Cold saponification was carried out on Lannea microcarpa (Wild grape) seed oil with the aim of identifying the quality and quantity of the oil and its suitability in soap production.
 Study Design: Experimental and instrumental study was done to determine the physicochemical characteristics, fatty acids present in the seed oil and its suitability for soap production.
 Place and Duration of Study: The study was conducted at the Biochemistry Laboratory, Kebbi State University of Science and Technology, Aliero, Nigeria from May to June, 2014.
 Methodology: The hexane extract of the sample was obtained by complete extraction using Soxhlet extractor, physicochemical analysis was carried out. A gas chromatography coupled with mass spectroscopy detector (GC-MS) system was used for the qualitative fatty acid determination. Simple cold method saponification was used in producing the soap.
 Results: The powdered seed (50 g) yielded 59.21±0.01% of the oil. Results from the physicochemical analysis showed the seed oil to be dark purple in colour and partially soluble in water with the acid, iodine, saponification and peroxide values at 16± 0.01 mgKOH/g, 121.6±0.1 gI2/100 g, 231.25±0.02 mgKOH/g, 3.02±0.01 meq H2O2respectively. The relative density and refractive index of the oil are at 0.5983±0.0001 (g/cm3) and 1.43±0.01 respectively. Qualitative GC-MS revealed the following fatty acids; Decanoic acid, Palmitic acid, Stearic acid, Margaric acid, 1-octadecanoic acid, Oleic and Erucic acid. The soap produced from the seed oil has pH and Foam height, 10.18±0.01 and 105.1±0.1 (cm³) respectively.
 Conclusion: The present study on seeds of wild grape revealed that the oil is a viable natural product that can be utilized in thefood and cosmetic industry.

Analyzing Physicochemical Properties of Wild Grapes (Lannea Microcarpa) Seed Oil

The purpose of this study was to analyze physicochemical properties of wild grape (Lannea microcarpa) seed oil. Several characterizations were conducted, including a gas chromatography and mass spectrometry (GC-MS) and cold saponification. The analyses were also supported by measuring acid, iodine, saponification, and peroxide values. Other analyses were relative density and refractive index. Experimental results showed that the oil was dark purple with the composition of oil of 59%. The qualitative GC-MS revealed the oil contained several fatty acids, including decanoic acid, palmitic acid, stearic acid, margaric acid, 1-octadecanoic acid, oleic, and erucic acid. The soap produced from the seed oil has basic pH and relatively high foam value. When the high concentration of oil was used, the appearance of oil was very dark purple and slightly soluble in water. This is due to the fact that most of the oil compositions were non-polar structure. This result confirmed the potential use of the oil for soap and other cosmetic materials.

Large-scale profiling of carotenoids by using non aqueous reversed phase liquid chromatography – photodiode array detection – triple quadrupole linear ion trap mass spectrometry: Application to some varieties of sweet pepper (Capsicum annuum L.)

Analysis of carotenoids is very complex and demanding in terms of both separation and detection. In this article, an analytical strategy relying on high-performance liquid chromatography-photodiode array detection-tandem mass spectrometry (HPLC-PAD-MS/MS) is presented for the large-scale screening of these phytochemicals. Separation was realized by means of Non-Aqueous Reversed Phase (NARP) chromatography on a triacontyl stationary phase kept at subambient temperature, using a mobile phase compatible with atmospheric pressure chemical ionization (APCI). The standards of 14 analytes were used to optimize the method and to predict the chromatographic behaviour of untargeted carotenoids. MS and MS/MS data, obtained during Information Dependent Acquisition (IDA) experiments, were utilized to set up a sensitive HPLC- selected reaction monitoring (SRM) method. Relative abundance between SRM ion currents (ion ratio) allowed the MS distinction of structural isomers (for example, bicyclic, monocyclic and acyclic isomers), while the identification of geometrical isomers was based on Qratio and fine structure, as calculated from UV–vis spectra. The comparison of LC-PAD- SRM chromatograms, acquired after applying two different extraction procedures (matrix solid-phase dispersion and overnight cold saponification), allowed verifying that sweet peppers are a good source of xanthophylls, prevalently occurring as esterified forms. The overall strategy could identify more than 40 carotenoids in some sweet pepper varieties (Jolly, horn and sweet chili pepper) available on the Italian and European food market.

The Effects of Cold Saponification on the Unsaponified Fatty Acid Composition and Sensory Perception of Commercial Natural Herbal Soaps.

Saponification is the process in which triglycerides are combined with a strong base to form fatty acid metal salts during the soap-making process. The distribution of unsaturated and saturated fatty acid determines the hardness, aroma, cleansing, lather, and moisturizing abilities of soaps. Plant extracts, such as rosemary, vegetable, and essential oils are frequently added to soaps to enhance quality and sensory appeal. Three natural soaps were formulated using cold saponification to produce a base or control bar (BB), hibiscus rosehip bar (H), and a forest grove bar (FG). Rosemary extract (R) or essential oil (A) blends were added as additives to each formulation prior to curing to evaluate the effects of natural plant additives on the lipid composition and sensory characteristics of these natural herbal soaps. A total of seven natural soaps, three without additives (BB, H, FG) and four with additives (BBR, HA, FGR, FGA), were manufactured and studied. The majority (86–99%) of the polyunsaturated fatty acids (5.0–7.0 µg/mg) remained unsaponified in the manufactured natural soaps regardless of feedstock used. Principal component analysis (PCA) analyses showed the unsaponifiable fatty acids were different in the hibiscus bar compared to the other bars. There was a very strong correlation between the content of unsaponified C18:3n3 and C18:1n9 in all natural soaps. These results indicate that unsaponified fatty acids are important contributors to the quality and overall sensory perception and preference of natural herbal soaps following manufacturing by cold saponification.

Non-aqueous reversed-phase liquid-chromatography of tocopherols and tocotrienols and their mass spectrometric quantification in pecan nuts

An easy and effective analytical method was developed for the simultaneous quantification of four tocopherols (Ts) and four tocotrienols (T3s) in three pecan nut cultivars (Stuart, Sioux, and Pawnee). The analytes were separated on a C30 column kept at 15°C, under isocratic non-aqueous reversed-phase (NARP) conditions, in only 18min and detected by atmospheric pressure chemical ionization–tandem mass spectrometry (APCI-MS/MS). The HPLC-APCI-MS/MS method was validated, according to the main FDA guidelines, and then applied for the characterization of the real samples. Analytes were extracted by cold saponification with recoveries greater than 87%. The limits of detection (LOD) and limits of quantification (LOQ) were in the range of 0.3–10μg/100mg and 1–30μg/100mg, respectively. Compared to other nuts, vitamin E composition of pecan nuts (Carya illinoinensis) has only partially been elucidated. Results have evidenced the prevalence of γ-forms for both Ts and T3s and clear quantitative differences of the identified vitamers among the studied cultivars. The richest variety in vitamin E was Sioux with a total content of about∼32mg/100g wet weight, followed by Stuart (∼16mg/100g) and Pawnee (∼9mg/100g).

Determination of target fat-soluble micronutrients in rainbow trout's muscle and liver tissues by liquid chromatography with diode array-tandem mass spectrometry detection.

This paper describes an analytical approach, based on LC-diode array detector-MS/MS (LC-DAD-MS/MS), for characterizing the fat-soluble micronutrient fraction in rainbow trout (Oncorhynchus mykiss). Two different procedures were applied to isolate the analytes from liver and muscle tissue: overnight cold saponification to hydrolyze bound forms and to simplify the analysis; matrix solid-phase dispersion to avoid artifacts and to maintain unaltered the naturally occurring forms. Analytes were separated on a C30 analytical column by using a nonaqueous reversed mobile phase compatible with the atmospheric pressure chemical ionization. Compared to other works, the most relevant advantage of the here illustrated method is the large amount of information obtained with few analytical steps: nine fat-soluble vitamins (3,4-dehydroretinol, retinol, cholecalciferol, ergocalciferol, α-tocopherol, γ-tocopherol, δ-tocopherol, phylloquinone, and menaquinone-4) and eight carotenoids (all-trans-lutein, all-trans-astaxanthin, all-trans-zeaxanthin, all-trans-β-cryptoxanthin, all-trans-canthaxanthin, all-trans-ζ-carotene, all-trans-β-carotene, and all-trans-γ-carotene) were quantified after the method validation, while other untargeted carotenoids were tentatively identified by exploiting the identification power of the LC-DAD-MS/MS hyphenation.

Liquid chromatography–tandem mass spectrometry method for the determination of vitamin K homologues in human milk after overnight cold saponification

Human milk is the only source of vitamin K for exclusively breastfed neonates. This vitamin is crucial both for blood coagulation (vitamin K1) and for the normal neurological and skeletal development of the foetus and newborn (vitamin K2). Since vitamin K is ubiquitous in foods, deficiency is not common in adults, but plasma levels and hepatic storage are very low at birth. In light of the importance of this valuable micronutrient, a non-invasive method for verifying that exclusively breastfed infants are receiving an adequate supply of the vitamin is clearly a topic of great significance. In spite of this, the determination of the several vitamin K homologues in human milk has still not been completely elucidated. This paper presents an HPLC–MS/MS method for the simultaneous determination of phylloquinone, menaquinone-4 (MK-4), and menaquinone-7 (MK-7) in human milk after a simple and effective isolation procedure. Overnight cold saponification and extraction of the analytes with hexane provided yields above 75%. This procedure, combined with high performance liquid chromatography–tandem mass spectrometry (HPLC–MS/MS), made it possible to achieve limits of detection (LODs) below 0.8ng/mL. After a complete validation study, the method was applied to measure vitamin K congeners in several human milk samples; results showed vitamin K1 concentrations comparable with those reported in the literature. In addition, this is the first study performed for the determination of MK-4 and MK-7 in the maternal milk of Italian women.

Analysis of phytosterols and phytostanols in enriched dairy products by Fast gas chromatography with mass spectrometry.

A Fast gas chromatography and mass spectrometry method for plant sterols/stanols analysis was developed, using a short capillary gas chromatography column (10 m × 0.1 mm internal diameter × 0.1 μm film thickness) coated with 5% diphenyl-polysiloxane. A silylated mixture of the main plant sterols/stanols standards (β-sitosterol, campesterol, stigmasterol, campestanol, sitostanol) was well separated in 1.5 min, with a good peak resolution (>1.4, determined on a critical chromatographic peak pair (β-sitosterol and sitostanol)), repeatability (<13%), and sensitivity (<0.017 ng/mL). The suitability of this Fast chromatography method was tested on plant sterols/stanols-enriched dairy products (yogurt and milk), which were subjected to lipid extraction, cold saponification, and silylation prior to injection. The analytical performance (sensitivity < 0.256 ng/mL and repeatability < 10.36%) and significant reduction of the analysis time and consumables demonstrate that Fast gas chromatography-mass spectrometry method could be also employed for the plant sterols/stanols analysis in functional dairy products.

Comparison of saponification methods for characterization of the nonsaponifiable fraction of virgin olive oil

Despite the interest of the nonsaponifiable fraction of olive oil (composed by sterols, tocopherols, fatty alcohols, hydrocarbons, phenols, pigments, and volatile compounds) there are no studies in the literature dealing with comparison of methods to isolate this fraction. Four saponification methods (viz. hot and cold saponification, and microwave‐ and ultrasound‐assisted saponification) have been compared by analysis of the resulting extracts using LC–MS in high‐resolution mode and data treatment with multivariate statistical tools. Principal component analysis and Venn diagrams were employed to compare qualitatively and/or semiquantitatively the metabolite profiles provided by the different methods. Hot saponification was the approach which provided the highest efficiency of this step in qualitative and quantitative terms. On the other hand, cold and microwave‐assisted saponification led to nonsaponifiable extracts with similar composition according to unsupervised analysis. These results revealed that microwave assistance does not promote chemical alteration of the nonsaponifiable fraction. Finally, ultrasound‐assisted saponification provided the extract with a more different composition.

Comparison of extraction methods for kahweol and cafestol analysis in roasted coffee

Kahweol and cafestol, diterpenes from the unsaponifiable fraction of coffee, present known effects on human health such as anticarcinogenic and hipercholesterolemic activities. There are discrepancies regarding the levels reported for these compounds in roasted coffee, probably due to the extraction processes. Therefore, four sample preparation methods were studied: direct hot saponification (DHS), direct cold saponification (DCS); and Bligh and Dyer (BD) or Soxhlet (SO) extraction followed by saponification. The levels of diterpenes and their dehydro derivatives obtained by high performance liquid chromatography with diode array and mass spectrometry detectors (HPLC-DAD-MS/MS) and the chromatographic profiles of roasted coffee, obtained by these four methods, were compared. DHS was more efficient for extraction, showing better separation of chromatographic peaks and levels of 930.2 (± 36.8), 113.2 (± 4.7), 568.6 (± 16.6) and 87.1 (± 3.7) mg 100 g-1 for kahweol, dehydrokahweol, cafestol and dehydrocafestol, respectively. The DHS extract presented a diterpene content (kahweol and cafestol) 15% superior to that of DCS and up to 88% superior than using SO and BD methods.

Comprehensive Profiling of Carotenoids and Fat-Soluble Vitamins in Milk from Different Animal Species by LC-DAD-MS/MS Hyphenation

This paper describes a novel and efficient analytical method to define the profile of fat-soluble micronutrients in milk from different animal species. Overnight cold saponification was optimized as a simultaneous extraction procedure. Analytes were separated by nonaqueous reversed-phase (NARP) chromatography: carotenoids on a C(30) column and fat-soluble vitamins on a tandem C(18) column system. Besides 12 target analytes for which standards are available (all-trans-lutein, all-trans-zeaxanthin, all-trans-β-cryptoxanthin, all-trans-β-carotene, all-trans-retinol, α-tocopherol, γ-tocopherol, δ-tocopherol, ergocalciferol, cholecalciferol, phylloquinone, and menaquinone-4), the DAD-MS combined detection allowed the provisional identification of other carotenoids on the basis of the expected retention times, the absorbance spectra, and the mass spectrometric data. Retinol and α-tocopherol were the most abundant fat-soluble micronutrients and the only ones found in donkey's milk along with γ-tocopherol. Ewe's milk also proved to be a good source of vitamin K vitamers. Bovine milk showed a large variety of carotenoids that were absent in milk samples from other species with the only exception of all-trans-lutein and all-trans-zeaxanthin.