Classification and quantification of sucrose from sugar beet and sugarcane using optical spectroscopy and chemometrics.

Sucrose, obtained from either sugar beet or sugarcane, is one of the main ingredients used in the food industry. Due to the same molecular structure, chemical methods cannot distinguish sucrose from both sources. More practical and affordable methods would be valuable. Sucrose samples (cane and beet) were collected from nine countries, 25% (w/w) aqueous solutions were prepared and their absorbances recorded from 200 to 1380 nm. Spectral differences were observable in the ultraviolet-visible (UV-Vis) region from 200 to 600 nm due to impurities in sugar. Linear discriminant analysis (LDA), classification and regression trees, and soft independent modeling of class analogy were tested for the UV-Vis region. All methods showed high performance accuracies. LDA, after selection of five wavelengths, gave 100% correct classification with a simple interpretation. In addition, binary mixtures of the sugar samples were prepared for quantitative analysis by means of partial least squares regression and multiple linear regression (MLR). MLR with first derivative Savitzky-Golay were most acceptable with root mean square error of cross-validation, prediction, and the ratio of (standard error of) prediction to (standard) deviation values of 3.92%, 3.28%, and 9.46, respectively. Using UV-Vis spectra and chemometrics, the results show promise to distinguish between the two different sources of sucrose. An affordable and quick analysis method to differentiate between sugars, produced from either sugar beet or sugarcane, is suggested. This method does not involve complex chemical analysis or high-level experts and can be used in research or by industry to detect the source of the sugar which is important for some countries' agricultural policies.

Phenolic profiling and antioxidant evaluation of extracts from Southern African indigenous fruits byproducts.

The call to build climate-resilient food systems in Africa has revived interest in indigenous fruits, which, however, remain under-researched. In this study, the phenolic content and antioxidant profiles of seed and pulp of ethanolic extracts from eight Southern African indigenous fruits were evaluated using UPLC-ESI-Q-TOF/MS and four antioxidant assays. Total phenols and hydroxycinnamic acids were highest in Dovyalis caffra (Kei apple) seed (5084.5 and 3403.83 mg/kg). Flavonoids were most abundant in Colpoon compressum (Colpoon) seed (1127.23 mg/kg), while hydrolysable tannins were highest in Syzygium guineense (Water pear) seed (666.13 mg/kg). Proanthocyanidins were abundant in Harpephyllum caffrum (Wild plum) pulp while anthocyanins were highest in Olea africana (Wild olive) pulp. Hierarchical clustering heatmap analysis showed similar concentration and diversity in the composition of reported compounds. Syzygium guineense seed had the lowest DPPH values. ORAC values were highest in O. africana pulp while H. caffrum pulp had the highest FRAP values and lipoxygenase inhibition capacity. In conclusion, the study revealed a diverse profile of phenolics in indigenous fruits extracts, to which their bioactivity is attributed. Specifically, H. caffrum pulp and S. guineense seed have potential as natural sources of phenolic antioxidants for food application.

Biopreservative efficacy of grape (Vitis vinifera) and clementine mandarin orange (Citrus reticulata) by-product extracts in raw ground beef patties.

Beef patties were treated with 450 µg/g of extracts from grape (Vitis vinifera) seeds (GSE), pomace (GPE) or orange (Citrus reticulata) pomace (OPE) and compared to negative (no extract; CTR) and positive (sodium metabisulphite; SMB) controls for their effect on colour, lipid and protein oxidation and bacterial growth under simulated retail display conditions (4 °C) for 9 d, and sensory quality. Antioxidant activity and redness of beef patties increased in the order of CTR < OPE = GPE < GSE < SMB. The order of thiobarbituric acid reactive substances and carbonyl values were CTR > GPE = OPE > GSE > SBM, while that of bacterial counts were CTR > GSE = GPE > OPE > SMB. Retail display period had significant effect on all the shelf-life parameters. Overall, intensity of aroma, beef-like aroma and flavour in beef patties were highest in OPE. Results suggested that GSE and OPE could be commercially valorised as natural antioxidants and antibacterials in beef patties, respectively.

Assessing similarity analysis of chromatographic fingerprints of Cyclopia subternata extracts as potential screening tool for in vitro glucose utilisation.

Similarity analysis of the phenolic fingerprints of a large number of aqueous extracts of Cyclopia subternata, obtained by high-performance liquid chromatography (HPLC), was evaluated as a potential tool to screen extracts for relative bioactivity. The assessment was based on the (dis)similarity of their fingerprints to that of a reference active extract of C. subternata, proven to enhance glucose uptake in vitro and in vivo. In vitro testing of extracts, selected as being most similar (n = 5; r ≥ 0.962) and most dissimilar (n = 5; r ≤ 0.688) to the reference active extract, showed that no clear pattern in terms of relative glucose uptake efficacy in C2C12 myocytes emerged, irrespective of the dose. Some of the most dissimilar extracts had higher glucose-lowering activity than the reference active extract. Principal component analysis revealed the major compounds responsible for the most variation within the chromatographic fingerprints, as mangiferin, isomangiferin, iriflophenone-3-C-ß-D-glucoside-4-O-ß-D-glucoside, iriflophenone-3-C-ß-D-glucoside, scolymoside, and phloretin-3',5'-di-C-ß-D-glucoside. Quantitative analysis of the selected extracts showed that the most dissimilar extracts contained the highest mangiferin and isomangiferin levels, whilst the most similar extracts had the highest scolymoside content. These compounds demonstrated similar glucose uptake efficacy in C2C12 myocytes. It can be concluded that (dis)similarity of chromatographic fingerprints of extracts of unknown activity to that of a proven bioactive extract does not necessarily translate to lower or higher bioactivity.

Chemometric analysis of chromatographic fingerprints shows potential of Cyclopia maculata (Andrews) Kies for production of standardized extracts with high xanthone content.

Cyclopia species are used for the production of honeybush tea and food ingredient extracts associated with many health benefits. A species-specific high-performance liquid chromatography (HPLC) method for Cyclopia maculata, developed and validated, allowed quantification of the major compounds in extracts from "unfermented" and fermented C. maculata. Two xanthones were tentatively identified for the first time in a Cyclopia species, whereas an additional four compounds were tentatively identified for the first time in C. maculata. "Fermentation" (oxidation) decreased the content of all compounds, with the exception of vicenin-2. Similarity analysis of the chromatographic fingerprints of unfermented C. maculata aqueous extracts showed extremely low variation (r ≥ 0.97) between samples. Some differences between wild-harvested and cultivated seedling plants were, however, demonstrated using principal component analysis. Quantitative data of selected compounds confirmed the low level of variation, making this Cyclopia species ideal for the production of standardized food ingredient extracts.

Enhancing aspalathin stability in rooibos (Aspalathus linearis) ready-to-drink iced teas during storage: the role of nano-emulsification and beverage ingredients, citric and ascorbic acids.

BACKGROUND: The effects of citric and ascorbic acids on the stability of aspalathin in rooibos (Aspalathus linearis) ready-to-drink (RTD) formulations containing fermented rooibos extract (FR), aspalathin-enriched green rooibos extract (GR) and aspalathin-enriched green rooibos extract ascorbic acid solubilisate (GR-solubilisate) were investigated during storage (12 weeks at 25 °C). RESULTS: Storage of iced tea formulations containing FR and GR extracts reduced their flavonoid content. The aspalathin content of FR iced tea without citric or ascorbic acid was reduced to undetectable levels by week 8 of storage. Addition of citric acid resulted in improved stability of aspalathin, but ascorbic acid did not impart additional stability. Iso-orientin and orientin were less affected than aspalathin, presumably owing to partial conversion of aspalathin to these flavones. Similar results were obtained for GR iced tea formulations. Improved stability of aspalathin was noted in iced tea containing GR-solubilisate with or without citric acid. Lower pH was shown to favour stability, especially for fermented rooibos iced teas. CONCLUSION: Citric and ascorbic acids contribute to the stability of rooibos flavonoids during storage. Differences in stability between formulations are not due to pH differences but may be related to the matrix.

Use of green rooibos (Aspalathus linearis) extract and water-soluble nanomicelles of green rooibos extract encapsulated with ascorbic acid for enhanced aspalathin content in ready-to-drink iced teas.

Heat-induced changes in aspalathin, iso-orientin, and orientin content of ready-to-drink (RTD) green rooibos iced tea formulations were investigated. An organic-solvent-based aspalathin-enriched extract prepared from green rooibos was used "as-is" or encapsulated with ascorbic acid in a water-soluble nanomicelle-based carrier system. The common iced tea ingredients, ascorbic acid, and/or citric acid were added to the iced tea containing green rooibos extract. Only citric acid was added to the iced tea containing the nanomicelles. Heat treatments consisted of pasteurization (93 °C/5 min and 93 °C/30 min), normal-temperature sterilization (NTS; 121 °C/15 min), and high-temperature sterilization (HTS; 135 °C/4 min). Pasteurization had little or no effect on the flavonoid content. NTS and HTS induced significant losses in the flavonoids. The addition of citric and ascorbic acids improved the stability of the flavonoids, but encapsulation of green rooibos extract with ascorbic acid in nanomicelles did not offer additional stability. The only benefit of using the water-soluble nanomicelles was the improved clarity of the RTD product. Iso-orientin and orientin contents were substantially less affected than aspalathin by the heat treatments, partially because of conversion of aspalathin to these flavones, which countered losses. 5-Hydroxymethylfurfural (HMF), a known dehydration product of hexoses under mild acidic conditions and also a degradation product of ascorbic acid, was observed in formulations containing citric and/or ascorbic acids.

In vitro hepatic biotransformation of aspalathin and nothofagin, dihydrochalcones of rooibos (Aspalathus linearis), and assessment of metabolite antioxidant activity.

Aspalathin (2',3,4,4',6'-pentahydroxy-3'-C-beta-d-glucopyranosyldihydrochalcone) is the major flavonoid present in the South African herbal tea rooibos. In vitro metabolism of aspalathin and a structural analogue nothofagin, lacking the A ring catechol group, was investigated by monitoring the formation of glucuronyl and sulfate conjugates using Aroclor 1254 induced and uninduced rat liver microsomal and cytosolic subcellular fractions. Following glucuronidation of both aspalathin and nothofagin, HPLC-DAD, LC-MS, and LC-MS/MS analyses indicated the presence of two metabolites: one major and one minor. Only one aspalathin metabolite was obtained after sulfation, while no metabolites were observed for nothofagin. Two likely sites of conjugation for aspalathin are 4-OH or 3-OH on the A-ring. For nothofagin, the 4-OH (A-ring) and 6'-OH (B-ring) seem to be involved. The glucuronyl conjugates of aspalathin lack any radical scavenging properties in online postcolumn DPPH radical and ABTS radical cation assays. Deconjugation assays utilizing glucuronidase and sulfatase resulted in the disappearance of the metabolites, with the concomitant formation of the unconjugated form in the case of the glucuronidated product. The balance between conjugated and unconjugated forms of aspalathin could have important implications regarding its role in affecting oxidative status in intra- and extracellular environments in vivo.

Effect of pre-drying treatments and storage on color and phenolic composition of green honeybush (Cyclopia subternata) herbal tea.

The effect of various pre-drying treatments and storage temperatures on the color (L*, a*, b*, chroma, and hue angle) and phenolic composition of green Cyclopia subternata was investigated. Pre-drying treatments, which included comminution+drying (T2), steaming (ca. 90-93 degrees C/60 s)+comminution + drying (T3), and comminution+steaming+drying (T4), had a detrimental effect (p<0.05) on the color parameters relative to the control (T1) (drying of intact leaves). All drying took place at 40 degrees C and 30% relative humidity (RH). Of the pre-drying treatments (T2, T3, and T4), the best retention of the green leaf color was observed for T3 (p<0.05). T2 reduced the SS and TP contents of the leaves (p<0.05) as well as the content of individual phenolic compounds, including mangiferin, isomangiferin, and eriocitrin. Scolymoside did not follow the same trend as the other compounds, with the intact leaves having the lowest content. It is postulated that oxidation of eriocitrin (eriodictyol-7-O-rutinoside) to scolymoside (luteolin-7-O-rutinoside) occurred with comminution and steaming. The phenolic composition of samples, subjected to T1 and T3 and a 6 month storage period at 0 and 30 degrees C, remained stable. Storage at 30 degrees C resulted in detrimental color changes, leading to the loss of the green leaf color irrespective of the pre-drying treatment.

Effect of heat on aspalathin, iso-orientin, and orientin contents and color of fermented rooibos (Aspalathus linearis) iced tea.

The phenolic quality of commercial South African fermented rooibos iced teas in terms of aspalathin, iso-orientin, and orientin contents in comparison to a "cup of tea" was shown to be inferior. The role of the different manufacturing stages of powdered extract used in iced tea formulation and, more specifically, the impact of pasteurization and sterilization on the color and phenolic content of the beverage, were assessed as potential causes of its inferior phenolic quality. Aspalathin and its corresponding flavones, iso-orientin and orientin, were found to be present at all stages of the powdered extract production process. Spray-drying did not significantly (P ≥ 0.05) alter the aspalathin, iso-orientin, or orientin content of concentrates. Simulated normal-temperature sterilization (NTS at 121 °C/15 min) and high-temperature sterilization (HTS at 135 °C/4 min), but not necessarily pasteurization (93 °C/30 min), significantly (P < 0.05) reduced the aspalathin, iso-orientin, and orientin contents of different iced tea formulations. Heat-induced losses of iso-orientin and orientin were lower than those for aspalathin. Conversion of aspalathin to the flavones is implicated. The addition of ascorbic acid and/or citric acid to the base iced tea formulation containing only rooibos extract and sugar proved to be beneficial, especially for the retention of aspalathin. Browning, that is, absorbance at 420 nm, was significantly (P < 0.05) increased in the base formulation. In the case of the formulations also containing ascorbic acid and/or citric acid, absorbance remained unchanged or decreased when subjected to NTS and HTS treatments. This was attributed to removal of brown polymers from solution as the pH values of these formulations were lower than that of the base formulation.

Effect of species variation and processing on phenolic composition and in vitro antioxidant activity of aqueous extracts of Cyclopia spp. (Honeybush Tea).

The in vitro antioxidant activity of aqueous extracts prepared from four Cyclopia spp. (unfermented and fermented) was assessed using radical (ABTS *+) scavenging, ferric ion reduction, and inhibition of Fe2+-induced microsomal lipid peroxidation as criteria. Aqueous extracts of unfermented and fermented Aspalathus linearis (rooibos) and Camellia sinensis teas (green, oolong, and black) were included as reference samples. Qualitative and quantitative differences in phenolic composition were demonstrated for the Cyclopia spp. The xanthone glycoside, a.k.a. mangiferin, was the major monomeric polyphenol present in the Cyclopia extracts, with both unfermented and fermented C. genistoides extracts containing the highest quantities. Fermentation resulted in a significant reduction in extract yields and their total polyphenolic and individual polyphenol contents. Unfermented plant material should preferentially be used for preparation of extracts, as fermentation significantly ( P < 0.05) lowered antioxidant activity of all species, except in the case of C. genistoides, where the ability to inhibit lipid peroxidation was not affected. Unfermented plant material also retained the highest concentration of mangiferin. Overall, extracts of unfermented Cyclopia were either of similar or lower antioxidant activity as compared to the other teas. However, the presence of high levels of mangiferin merits the use of Cyclopia spp. and, in particular, C. genistoides, as an alternative herbal tea and potential dietary supplement.

Evaluation of spectrophotometric methods for screening of green rooibos (Aspalathus linearis) and green honeybush (Cyclopia genistoides) extracts for high levels of Bio-active compounds.

The potential of UV spectrophotometry and an aluminium chloride (AlCl(3)) colorimetric method to determine the dihydrochalcone (DHC) and mangiferin contents of green rooibos and honeybush (C. genistoides) extracts, respectively, was investigated. The DHC content of rooibos water extracts, determined using UV spectroscopy, correlated with the sum of the aspalathin and nothofagin contents as quantified using HPLC (r = 0.98). A correlation coefficient of 0.91 was obtained when correlating the mangiferin content of C. genistoides methanol extracts, determined by the AlCl(3) colorimetric method, with the results obtained by HPLC. Using the linear equations from the correlations it was possible to predict the DHC and mangiferin contents of extracts from the respective spectrophotometric measurements to a reasonable accuracy as an alternative to HPLC. The total polyphenol (TP) content of rooibos water extracts can also be determined using UV spectrophotometry and aspalathin as a standard (r = 0.99) as an alternative to the Folin-Ciocalteau method. The TP content of rooibos extracts correlated (r = 0.99) with its total antioxidant activity (TAA) as determined with the ABTS radical cation scavenging assay, but the TP content of C. genistoides water extracts is not a good indication of their TAA (r = 0.27). The aspalathin content of rooibos extracts correlated with their TAA (r = 0.96), but the mangiferin content of honeybush water extracts only gave a moderate correlation with their TAA (r = 0.75).