Triacylglycerol Profiles Research Articles

Novel approach for determination of milk fat adulteration with non-milk fat by RP-HPLC

The present work studied the method for quickly determining milk fat adulteration with non-milk fat. The approach was based on analyzing characteristic fingerprints of triacylglycerols profile in combination with chemometric techniques and a multidimensional vector model. Nineteen fat samples of bovine milk, 20 samples of cow cheese, and ten samples of cow butter were analyzed by reverse-phase high-performance liquid chromatography. The areas of 3 triacylglycerol peaks with equivalent carbon numbers of 35.6, 46.2, and 47.2 were used for diagnosing chemometrics and building the vector model. It was shown that the proposed approach could be used to quickly determine milk fat adulteration by non-milk fat with a concentration from 1.5 to 100%. Application of the vector model to 20 samples of cow cheese showed that most of them were made of authentic cow milk; however, the milk fat in 7 of these samples had been replaced with non-milk fat. For ten samples of cow butter, only three samples were produced from milk fat alone, while for other samples, the mass fraction of non-milk fat, expressed as palm oil, was determined to be in the range of 10–90%.

Detection of walnut oil adulterated with high-linoleic acid vegetable oils using triacylglycerol pseudotargeted method based on SFC-QTOF-MS

Authentication of walnut oil (WO) is challenging due to the adulteration of high-linoleic acid vegetable oils (HLOs) with similar fatty acid composition. To allow the discrimination of WO adulteration, a rapid, sensitive and stable scanning method based on supercritical fluid chromatography quadrupole time-of-flight mass spectrometry (SFC-QTOF-MS) was established to profile 59 potential triacylglycerol (TAGs) in HLOs samples within 10min. Limit of quantitation of the proposed method is 0.002µgmL-1 and the relative standard deviations range from 0.7% to 12.0%. TAGs profiles of WO samples from various varieties, geography origins, ripeness, and processing methods were used to construct orthogonal partial least squares-discriminant analysis (OPLS-DA) and OPLS models that were highly accurate in both qualitative and quantitative prediction at adulteration levels as low as 5% (w/w). This study advances the TAGs analysis to characterize vegetable oils and holds promise as an efficient method for oil authentication.

Lipidomic characteristics of three edible cold-pressed oils by LC/Q-TOF for simple quality and authenticity assurance.

Distinguishing oil samples from each other is challenging but it is crucial for ensuring food quality, and for detecting and preventing the possible adulteration of these products. Lipidomic profiling is believed to provide sufficient information to get fit-to-purpose confidence of oil identification as well as to deliver oil-specific lipid features which could be used as targets for routine authenticity testing of camelina, flax, and hemp oil in food control laboratories. Conducted di- and triacylglycerol profiling by LC/Q-TOFMS yielded successful differentiation of the oils. A marker panel consisting of 27 lipids (both DAGs and TAGs) useful for quality verification and authenticity assurance of the oils was established. Moreover, sunflower, rapeseed, and soybean oils were analysed as potential adulterants. We identified 6 lipid markers (DAGs 34:6, 35:2, 40:1, 40:2, 42:2, and TAG 63:1) which can be used for revealing the adulteration of camelina, hemp, and flax seed oils with these oils.

Study on the lipid and flavor compounds of Hibiscus manihot L. oil induced by different pressing method

Hibiscus manihot L. seed oil is an emerging high-value edible vegetable oil that also has a variety of important health functions, and concerns about its pressing method have great significance for commercial and consumer health. This study compared triacylglycerols (TAGs), diacylglycerol (DAG), free fatty acids (FFAs), and volatile flavor compounds for discriminating cold-pressed oil from hot-pressed oil. The profiles of TAGs, DAGs, FFAs, and flavor compounds were established by a lipidomic approach and electronic-nose (E-nose). Our results demonstrated that a slight decrease in TAG content and a significant increase in FFA and DAG contents were observed after roasting. The E-nose results showed that sulfides, terpenes, methane, compounds containing oxygen and nitrogen, alcohol and aromatic compounds greatly influenced the oil flavor. The PCA result of the E-nose data indicated that flavor compounds showed great promise for distinguishing the two samples. The quality indices and the oxidative stability of oil samples were detected to identify their nutritional value and edible quality. This study revealed that roasting greatly influenced the lipid and flavor substances of the Hibiscus manihot L. seed oil. Our findings could provide useful information on the impact of the pressing method for Hibiscus manihot L. seed oil.

Equivalent carbon number-based targeted odd-chain fatty acyl lipidomics reveals triacylglycerol profiling in clinical colon cancer

Odd-chain FAs (OCFAs) are present in very low level at nearly 1% of total FAs in human plasma, and thus, their functions were usually ignored. Recent epidemiological studies have shown that OCFAs are inversely associated with a variety of disease risks. However, the contribution of OCFAs incorporated into complex lipids remains elusive. Here, we developed a targeted odd-chain fatty acyl-containing lipidomics method based on equivalent carbon number and retention time prediction. The method displayed good reproducibility and robustness as shown by peak width at half height within 0.7 min and coefficient of variation under 20%. A total number of 776 lipid species with odd-chain fatty acyl residues could be detected in the ESI mode of reverse-phase LC-MS, of which 309 lipids were further validated using multiple reaction monitoring transitions. Using this method, we quantified odd-chain fatty acyl-containing lipidome in tissues from 12 colon cancer patients, revealing the remodeling of triacylglycerol. The dynamics of odd-chain fatty acyl lipids were further consolidated by the association with genomic and proteomic features of altered catabolism of branched-chain amino acids and triacylglycerol endogenous synthesis in colon cancer. This lipidomics approach will be applicable for screening of dysregulated odd-chain fatty acyl lipids, which enriches and improves the methods for diagnosis and prognosis evaluation of cancer using lipidomics.

Fatty acids, triacylglycerols, thermal properties, morphology and antioxidant activity of Canarium odontophyllum seed oil

Canarium odontophyllum Miq. (Dabai) is an underutilized fruit and its oil has the potential to broaden the availability of healthy edible oil. The objective of this study was to determine the physiochemical and thermal properties, fatty acid and triacylglycerol profiles, morphological behavior, and antioxidant capabilities of dabai seed oil (DSO) by using chromatographic and thermal techniques. The results showed that DSO has a low unsaturation values, high peroxide and refractive index values with a melting point at body temperature. DSO showed 56.47% saturated fatty acids, 40% unsaturated fatty acids and 29.88% of 1,3-dipalmitoyl-2-oleoyl-glycerol content. Some crystals showed larger than 30 µm in size and others smaller, including spherical, dot-covered spherulites between 0.5 and 5 µm in size. In addition, the amount of ferric reducing power and phenolic contents of DSO were 3.62±0.10 mMol of Fe2+/g and 21.73±0.98 mg GAE per 100 g of samples. The DSO mimics the composition of palm stearin and palm oil mid fraction and showed comparable physicochemical and thermal properties to that of commercial cocoa butter. The results showed that the DSO has a traceable amount of antioxidant activity and good potential to be developed as a healthy edible oil resource.

Lipidomics analysis facilitate insight into the molecular mechanisms of urate nephropathy in a gout model induced by combination of MSU crystals injection and high-fat diet feeding.

Renal injury is one of the most common clinical manifestations of patients with hyperuricaemia/gout. The precise pathophysiological mechanism(s) for the renal injury is still unknown. Furthermore, it is also unclear whether the clinical therapies (e.g., colchicine and febuxostat) could prevent its progression or not. Lipids are involved in almost all of important biological processes and play critical roles in maintaining the renal functions. Herein, shotgun lipidomics was performed for class-targeted lipid analysis of cellular lipidomes in renal tissue of a gouty model induced by combination of monosodium urate crystals injection and high-fat diet feeding with/without treatment with either colchicine or febuxostat. Serum uric acid (UA), proinflammatory cytokines (i.e., TNF-α and IL-6), xanthine oxidase activity, footpad swelling, and pain threshold were determined to evaluate the gouty severity. Renal histopathological changes, blood urea nitrogen, creatinine, and kidney index were used to reflect renal injury. Lipidomics analysis revealed that altered triacylglycerol (TAG) profile, impaired mitochondrial function resulted by decreased tetra 18:2 cardiolipin, reduced 4-hydroxyalkenal (HNE) species, and elevated lysophospholipids were already present in the kidneys at early stage of renal injury, probably contributing to its occurrence and development. In addition to significantly reduce the UA level and relief the gouty severity, treatment with either colchicine or febuxostat could restore HNE bioavailability, thereby delaying the progression of renal injury. However, both of them could not recover the altered TAG profile and the impaired mitochondrial function, indicating that treatment with either of them could not completely prevent the development of renal injury in the gouty model.

Impact of composition and aeration on the migration of TAGs in sandwich type confectionery products

A sandwich type confectionery product is made with a soft filling and a chocolate coating. The fats used for these two parts are generally different to provide specific organoleptic sensations. Thus, their compositions, in terms of the triacylglycerols (TAGs) profile, are different. Depending on the ambient temperature conditions, the chemical potential gradient at the interface for TAGs, and the microstructures in the bulk of two parts, the migration of TAGs is influenced. We have studied the impact of different filling recipes on the migration of specific TAGs from the filling to the coating and vice versa with a newly developed tool, referred to as lipstick method. Also, the influence of the micro-aeration of the filling on the transfer process is evaluated. Furthermore, Fick’s law of diffusion-based model is developed. The migration of TAGs as predicted by the model is compared to the experimental measurements, and limitations of the model are discussed. This approach can be used to tune the recipes of coating and filling to enhance the shelf-life stability of such products while delivering on specific liking attributes of taste.

Comparison and enrichment of sn-2 palmitoyl triacylglycerols (OPO/OPL) in fish oil for its potential application as human milk fat substitutes

Triacylglycerols (TAG) are differences in fatty acid distributions between infant formula and human milk. In this study, fish oil (Tilapia, Golden pompano, Tiger grouper, and Basa) showed the potential as the source of human milk fat substitutes by comparing TAG profiles with infant formula and human milk. The total lipids and TAG of fish were concentrated in the by-products of fish (head and viscera) and contained high levels of palmitic acid, oleic acid, and linoleic acid. Compared with infant formula, fish oil was closer to human milk in sn-2 fatty acid distribution, and sn-2 palmitic acid level in fish oil exceeded 52 % of total palmitic acid, Golden pompano head was the highest (64.46 %). Further research showed that the content of sn-2 palmitoyl TAG (OPO and OPL dominated) increased from 157.16 mg/g TAG to 305.18 mg/g TAG by isopropanol enrichment (solid–liquid ratio: 1:4, temperature: −12 °C, time: 4 h).

Ion Mobility–Mass Spectrometry Approach for the Comparison of Sheep and Goat Milk Lipidomes

In this work, we report an analytical procedure to investigate the lipid compositions of sheep and goat milk. This approach is based on an ion mobility–high-resolution mass spectrometric method to facilitate the identification of complex lipid species and their regiochemistry. A common triacylglycerol profile was observed for sheep and goat milk samples, while a higher abundance of medium-chain fatty acids was observed at the sn-2 position for sheep milk. Furthermore, differences can be also observed in the levels of saturated and unsaturated fatty acids at the sn-2 position. In terms of lipid classes, goat milk showed higher levels of triacylglycerols, phosphatidylinositols and ether-linked phosphatidylethanolamines, while sheep milk showed higher levels of free fatty acids, lysophosphatidylethanolamines, lysophosphocholines and non-hydroxy fatty acid-dihydrosphingosine ceramides when compared with goat milk.

Impact of Conservation Processes on the Lipid Profile and Immunological Factors IL-10 and TGF-β1 in Whey Separated from Discarded Human Milk

Reuse made by the separation of whey can reduce the waste of human milk. However, the authors are not aware of the literature on treatments in human whey, made available by pasteurization holder, freeze-drying, spray drying, and high hydrostatic pressure. In this context, the present study applied treatments to human whey and evaluated their lipid and immunological quality. Among the results, a different formation in the triacylglycerol profile was evidenced after the application of spray drying and high hydrostatic pressure, while pasteurization and freezedrying presented ion intensity close to the control human whey. In addition, pasteurization proved to be adequate for lipid nutritional quality and transformation factor-β1 (TGF-β1) concentration, while an increase in interleukin-10 (IL-10) levels was promoted, between 73 and 80%, after freeze-drying, spray drying, and high hydrostatic pressure. Through the principal component analysis, it is noteworthy that the processes presented divergences in terms of the effects caused, with similarity only between pasteurization and freeze-drying in the composition of fatty acids. However, it was observed that all processes were able to maintain the nutrients. Nevertheless, it is relevant to consider individual characteristics presented and the interest in the desired quality, which can be promising as a complementary product to infant feeding.

Physicochemical and antioxidative properties of ultrasound-assisted extraction of mahua (Madhuca longifolia) seed oil in comparison with conventional Soxhlet and mechanical extractions.

Ultrasound-assisted solvent extraction (UAE) was applied to extract underutilized Madhuca longifolia seed oil. The effect of extraction time, temperature, solvent type, solvent/sample ratio, and amplitude on the oil yield and recovery were investigated. Approximately 56.97% of oil yield and 99.54% of oil recovery were attained using mild conditions of 35min, 35°C, 40% amplitude, isopropanol to acetone (1:1), and solvent to sample (20mL/g). UAE oil yield and recovery were comparable with Soxhlet extraction (SXE) whilst mechanical pressing (ME) yielded<50% of UAE recovery. UAE does not affect the fatty acids composition (46% C18:1; 22% C16:0; 21% C18:0, 10% C18:2), and triacylglycerol profile (23% POO, 17% POS, 16% SOO, and 14% POP). Interestingly, UAE extracted oil conferred remarkably (P<0.05) higher antioxidant capacity (IC50 of DPPH 106.60mg/mL and ABTS 39.80mg/mL) than SXE (IC50 of DPPH 810.40mg/mL and ABTS 757.43mg/mL) or ME (IC50 of DPPH 622.38mg/mL and ABTS 392.87mg/mL).

First report on quality and purity assessment of sweet almond oil in Brazilian body oils by gas chromatography and mass spectrometry.

Sweet almond oil is a raw material with high-added value used in different products. Then, the aim of this study is to evaluate the quality and purity of 10 body oils based on sweet almond oils currently available in the Brazilian market. Fatty acid composition and triacylglycerol (TAG) profile were determined by gas chromatography with flame ionization detector (GC-FID) and atmospheric solids analysis probe mass spectrometry (ASAP-MS), respectively. The authenticity of samples was assessed using an analytical curve equation. Soybean oil was chosen as the adulterant because it is the cheapest vegetable oil commercialized in Brazil. Hierarchical clustering analysis (HCA) in conjunction with ASAP-MS classified product samples according to the type of vegetable oil (soybean and sweet almond oils). The addition of soybean oil (8.79% to 99.70%) was confirmed in samples. However, only two samples stated in their label the presence of soybean oil as an ingredient. These findings highlight the need for better oversight by regulatory bodies to ensure that consumers acquire high quality and authentic products based on equally high quality and purity of sweet almond oils.

Milk fat depression and plasma lipids in dairy cows and goats

This study examines the effects of diets supplemented with various lipids selected to induce divergent milk fat content responses (including a milk fat depression) between dairy cows and goats on plasma lipid composition. The objective was to better understand the mechanisms behind the regulation of milk fat secretion in these two ruminant species. Twelve Holstein cows and 12 Alpine goats were fed a basal diet not supplemented (CTL) or supplemented with corn oil plus wheat starch (COS, 5% DM intake (DMI)), marine algae powder of Schizochytrium sp. (MAP, 1.5% DMI), or hydrogenated palm oil (HPO, 3% DMI), in a replicated 4 × 4 Latin square design, during 28 days. On day 27, blood samples were collected for lipid analysis. Plasma lipid classes were quantified by high-performance thin-layer chromatography, with triacylglycerol (TAG) and free fatty acid (FFA) fractions analysed for FA composition by GLC. Plasma molecular species of TAG and ceramides were determined by HPLC–high-resolution MS and by liquid chromatography–triple quadrupole, respectively. Irrespective of diet, plasma total lipid content was higher in cows than goats (+61%), and TAG concentration was higher in goats than cows (+157%). In cows, conversely to goats, COS increased the trans-10 C18:1 proportion in the free FA (+248%) and the TAG (+195%) fractions. In cows and goats, MAP induced increases in cholesterol esters, cholesterol and phospholipids compared to CTL and changes in the plasma free FA and FA of TAG profiles. In both ruminant species, the concentrations of the lipid fractions were unchanged by HPO compared to CTL. Our results point to species specificities and different diet effects in plasma concentrations and compositions of lipid fractions in cows and goats. These new data highlight how diets, that induce large variations in milk fat secretions, affect the plasma lipid classes available for milk fat synthesis.

Typical Characterization of Commercial Camellia Oil Products Using Different Processing Techniques: Triacylglycerol Profile, Bioactive Compounds, Oxidative Stability, Antioxidant Activity and Volatile Compounds.

The processing technique is one of the key factors affecting the quality of camellia oil. In this study, camellia oils were obtained using four different processing techniques (cold-pressed, roast-pressed, fresh-pressed, and refined), and their triacylglycerols (TAGs) profile, bioactive compound (tocopherols, sterols, squalene, and polyphenols) level, oxidative stability, and volatile compounds were analyzed and compared. To further identify characteristic components in four camellia oil products, the TAG profile was analyzed using UPLC-QTOF-MSE. Five characteristic markers were identified, including OOO (m/z 902.8151), POL (m/z 874.7850), SOO (m/z 904.8296), PPL (m/z 848.7693), PPS (m/z 852.7987). Regarding the bioactive compound level and antioxidant capacity, the fresh-pressed technique provided higher α-tocopherols (143.15 mg/kg), β-sitosterol (93.20 mg/kg), squalene (102.08 mg/kg), and polyphenols (35.38 mg/kg) and showed stronger overall oxidation stability and antioxidant capacity. Moreover, a total of 65 volatile compounds were detected and identified in four camellia oil products, namely esters (23), aldehydes (19), acids (8), hydrocarbons (3), ketones (3), and others (9), among which pressed oil was dominated by aldehydes, acid, and esters, while refined oil had few aroma components. This study provided a comprehensive comparative perspective for revealing the significant influence of the processing technique on the camellia oil quality and its significance for producing camellia oil of high quality and with high nutritional value.

Changes in Chemical Composition of Flaxseed Oil during Thermal-Induced Oxidation and Resultant Effect on DSC Thermal Properties.

To investigate the changes in chemical composition of flaxseed oil during thermal-induced oxidation and the resultant effect on thermal properties, samples with different oxidation levels were obtained by being heated at 180 °C for two hours and four hours. The oxidation degree was evaluated using peroxide value (PV), extinction coefficient at 232 nm and 268 nm (K232 and K268), and total polar compounds (TPC). Using chromatography, the fatty acid profile and triacylglycerol (TAG) profile were examined. Differential scanning calorimetry (DSC) was used to determine the crystallization and melting profiles. Thermal-induced oxidation of flaxseed oil led to a significant increase (p < 0.05) in PV, K232, K268, and TPC, but the relative content of linolenic acid (Ln) and LnLnLn reduced dramatically (p < 0.05). TPC derived from lipid degradation affected both crystallization and melting profiles. Statistical correlations showed that the onset temperature (Ton) of the crystallization curve was highly correlated with K232, TPC, and the relative content of LnLnLn (p < 0.05), whereas the offset temperature (Toff) of the melting curve was highly correlated with the relative content of most fatty acids (p < 0.05). This finding provides a new way of rapid evaluation of oxidation level and changes of chemical composition for flaxseed oils using DSC.

Effects of pasteurization and high-pressure processing on the fatty acids, triacylglycerol profile, Dornic acidity, and macronutrients in mature human milk

The effects of high-pressure processing (HPP; 300, 400, 500 and 600 MPa for 5 min) compared to Holder pasteurization treatments on the macronutrients, acidity in Dornic, total coliforms, fatty acids (FA) composition, and the triacylglycerols (TAG) profile of mature human milk (HM) were evaluated. The results showed that both processes eliminated the microorganisms present, and the concentration of macronutrients and Dornic acidity did not show significant differences in processes. A total of 34 FAs were identified by gas chromatography with flame ionization detector, with palmitic and oleic acids having the highest concentrations. Regarding the TAG profile determined by direct infusion by electrospray ionization mass spectrometry, TAG PLO (P: palmitic acid; L: linoleic acid; and O: oleic acid) was the one with the highest estimated concentration (6.52–7.27%). The evaluation of hierarchical clustering analysis results suggests that processing with lower pressures was more beneficial for HM due to the similarity between the intensities of the TAGs identified in each sample and the control HM sample. Besides, the results obtained on the evaluated components suggested that HPP could be a promising alternative to HoP applied in human milk banks since it maintained the characteristics of HM and is faster than HoP.

Medium-chain triglyceride production in Nannochloropsis via a fatty acid chain length discriminating mechanism.

Depending on their fatty acid (FA) chain length, triacylglycerols (TAGs) have distinct applications; thus, a feedstock with a genetically designed chain length is desirable to maximize process efficiency and product versatility. Here, ex vivo, in vitro, and in vivo profiling of the large set of type-2 diacylglycerol acyltransferases (NoDGAT2s) in the industrial oleaginous microalga Nannochloropsis oceanica revealed two endoplasmic reticulum-localized enzymes that can assemble medium-chain FAs (MCFAs) with 8-12 carbons into TAGs. Specifically, NoDGAT2D serves as a generalist that assembles C8-C18 FAs into TAG, whereas NoDGAT2H is a specialist that incorporates only MCFAs into TAG. Based on such specialization, stacking of NoDGAT2D with MCFA- or diacylglycerol-supplying enzymes or regulators, including rationally engineering Cuphea palustris acyl carrier protein thioesterase, Cocos nucifera lysophosphatidic acid acyltransferase, and Arabidopsis thaliana WRINKLED1, elevated the medium-chain triacylglycerol (MCT) share in total TAG 66-fold and MCT productivity 64.8-fold at the peak phase of oil production. Such functional specialization of NoDGAT2s in the chain length of substrates and products reveals a dimension of control in the cellular TAG profile, which can be exploited for producing designer oils in microalgae.

Triacylglycerol fingerprint of sow milks during different lactation stages and from different breeds.

Sow milk fats not only provide energy but also essential nutrients for piglets. Thus, feeding strategies must be aligned with fat composition, especially triacylglycerols (TAGs) and their isomers. The triacylglycerol (TAG) profiles of sow milk fats from five typical breeds (Landrace × Large White, Landrace, Large White, Duroc, Pietrain) and two lactation stages (colostrum and milk) were systematically studied. A total of 45 major TAG species were identified using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. The most abundant TAG was oleic acid-palmitic acid-linoleic acid (O-P-L) (13.92% and 12.03% in colostrum and milk, respectively), which was not significantly different in colostrum among all breeds. TAG composition of sow milk was affected mainly by the lactation stage rather than sow breed. Furthermore, TAG compositions of sow milk fats were similar to those of human milk fats, but significant differences were observed between commercial piglet formulas and sow milk. Therefore, the results will contribute to the optimization of piglet formulas to improve the growth and wellness of piglets, as well as potentially providing a basis for food usage as a new source of nutrients for human infants in future.

Comprehensive Composition of Flavor Precursors in Kopi Luwak and Jacu Exotic Green Bioprocessed Coffees

Exotic coffees may be defined as extravagant and unique coffees, primarily due to their production mode, including unusual bioprocessing or fermentation conditions associated with superior sensorial characteristics. The aim of the present study was to investigate the influence of bioprocessing and of growing conditions on flavor precursors of Jacu and Kopi Luwak exotic green coffees, respectively. Moreover, this is the first study to perform a detailed chemical analysis of these exotic coffees. Thirteen green Coffea arabica bean samples were obtained, five from Espírito Santo state, Brazil, and eight Kopi Luwak from different regions of Indonesia. Samples were analyzed regarding their proximate composition, chlorogenic acids (CGA), sucrose, alkaloids, triacylglycerols (TAG), diacylglycerols, free fatty acids, sterols, diterpenes and tocopherols. Scanning electron micrography confirmed bioprocessing of Jacu and Kopi Luwak coffee samples. Bioprocessing by the Jacu bird caused reductions of 69 and 28% in caffeine and CGA contents, respectively. The TAG profile of Jacu coffee was modified. TAG containing two saturated fatty acids were preferably hydrolyzed in detriment to those containing two unsaturated fatty acids. Other coffee components were not affected by the bird's digestion of the beans. Kopi Luwak coffee samples had a chemical composition in accordance with reported ranges for non-bioprocessed green C. arabica samples, except for caffeine (0.48 g/100 g) and CGA (5.09 g/100 g), which were found in low amounts. Crop year rather than location or post-harvest processing discriminated Kopi Luwak coffee samples, suggesting that weather conditions would be the most crucial aspect for their chemical composition, especially in terms of total lipids, ashes, total CGA, sucrose and proteins.