Polyphenolic composition and in vitro antioxidant activities of native‐ and tannase‐treated green tea extracts

The antioxidant activities of native‐ and tannase‐treated green tea extracts along with their major polyphenol components were investigated. The polyphenolic content and composition of the tea before and after tannase treatment were determined by liquid chromatography coupled with mass spectrometry (LC‐MS). Approximately 99% of the (?)‐epigallocatechin gallate (EGCG) and (?)‐epicatechin gallate (ECG) in green tea extract were converted by tannase to (?)‐epigallocatechin (EGC) and (?)‐epicatechin (EC), respectively, after 30 min. Biotransformed green tea exhibited a significantly higher DPPH˙ radical scavenging activities than native green tea (EC₅₀ value of 0.024 ± 0.001 and 0.044 ± 0.001 mg mL^?1, respectively). Kinetic parameters such as scavenging rate and stoichiometry were calculated. The rate of DPPH˙ radical scavenging activities for tannase‐treated green tea extract was shown to be higher than native green tea extract.     

Effect of chestnut tannin extract (Castanea sativa Miller) on the proliferation of Cladosporium cladosporioides on sheep cheese rind during the ripening

Strains belonging to the genus Cladosporium can cause black spots on the surface of sheep cheese, making it impossible to sell. Two water solutions of chestnut tannin extract (i) 200 g L⁻¹ (CHE200) and (ii) 400 g L⁻¹ (CHE400), and the chestnut tannin extract powder (CHEP) were tested in a cheese making trial in which 60 cheese units were allotted to 5 experimental groups (each of 12 cheeses: C1, control 1 without any treatment; C2, control 2 treated with a silver ion solution; and cheeses LCHE200, LCHE400, and LCHEP, treated with CHE200, CHE400, and CHEP, respectively). The cheeses were ripened in a room polluted with Cladosporium cladosporioides with the aim to create conditions for the proliferation of this fungus on the cheeses. The results indicated that chestnut tannin extract at a concentration of 200 g L⁻¹ is capable of completely inhibiting C. cladosporioides proliferation, avoiding spoilage of the sheep cheese.     

Ethanolic guava leaf extract with different chlorophyll removal processes: Antioxidant properties and its preventive effect on lipid oxidation in Pacific white shrimp

Sedimentation process (GLE-S) or chloroform (GLE-Ch) were used for dechlorophyllisation of ethanolic guava leaf extract (EGLE). EGLE without chlorophyll removal (GLE-C) was served as control. Among all the extracts, GLE-S had the lowest chlorophyll a, chlorophyll b, and total chlorophyll contents (P < 0.05). Extraction yield was highest in GLE-C, followed by GLE-S and GLE-Ch, respectively. Total phenolic content and antioxidant properties were highest in GLE-S (P < 0.05). When Pacific white shrimp (PWS) was pre-treated with GLE-S at varying doses (0.5% and 1%), lipid oxidation products were monitored as compared to the control (CON) and those pre-treated with 1.25% sodium metabisulphite (SMS-1.25) during refrigerated storage (4°C) for 12 days. Lower lipid oxidation was recorded for PWS pre-treated with GLE-S, especially at 1%. Therefore, sedimentation process, a green method, showed promising efficacy for producing EGLE free of green colour with improved antioxidant properties. The extract effectively reduced lipid oxidation in PWS during refrigerated storage.     

Ethanolic Noni (Morinda citrifolia L.) leaf extract dechlorophyllised using sedimentation process: Antioxidant,antibacterial properties and efficacy in extending the shelf-life of striped catfish slices

Antioxidant and antimicrobial activities of ethanolic noni leaf extract (ENLE) without and with chlorophyll removal by sedimentation method were comparatively investigated. Total chlorophyll content was reduced by 82% in the top fraction (CR-ENLE) collected after 24 h at 4 °C as compared to that of ENLE. Antioxidant and antimicrobial activities were lower in the bottom fraction rich in chlorophyll (Chlo-ENLE) than others (P < 0.05). Based on the microbiological limit, the shelf-life of striped catfish slices pre-treated with 400 mg kg⁻¹ C-R-ENLE was extended to 9 days as compared to the 3 days recorded for the control (without pre-treatment). Slices treated with CR-ENLE had lower lipid oxidation than those treated with ENLE during refrigerated storage (P < 0.05). The sedimentation process was therefore a potential green method for producing ENLE having improved antioxidant and antimicrobial activities without green colour. It can be used as a natural additive for shelf-life extension of fish slices.     

Evaluation of hot and cold extraction of bioactive compounds in teas

This study evaluated the bioactive compounds of different types of tea by comparing hot and cold infusions. A multivariate data analysis was carried out, where the principal components analysis (PCA) and hierarchical cluster analysis (HCA) were used. Phenolic compounds varied between 267.27–2896.00 mg GAE L^?1 and 215.10–7351.33 mg GAE L^?1 for hot (80 °C) and cold extraction (20–25 °C), respectively. In the case of their antioxidant activity, results with DPPH were 43.10‐73.67% for hot extraction and 46.80‐77.13% for cold extraction. The average values for the ABTS˙⁺ method ranged between 2535.43 and 33 300.17 μmol TE L^?1 and between 1110.34 and 38 300.67 μmol TE L^?1, respectively, for hot and cold extraction. Different compounds were identified by liquid chromatography in the samples evaluated, where caffeine presented the higher concentrations in the teas. Samples of green and black tea (hot extraction) and white tea (cold extraction) showed bacteriostatic activity for S. aureus and E. coli. No extract had any bactericide activity. The current study revealed that cold infusion was more efficient in the extraction of bioactive compounds.     

Efficacy of sodium chlorite as an inhibitor of enzymatic browning in apple slices

Sodium chlorite (SC) is an effective sanitizer for inhibiting microbial growth. This investigation was conducted to determine the efficacy of SC as a browning control agent for use on fresh-cut apple slices, applied alone, or in conjunction with organic acids. Additionally, the authors compared the efficacy of SC to that of acidified sodium chlorite (ASC) and to several other salts and examined the effect of pH and several different organic acids on efficacy of SC. The fresh-cut apple slices were dipped in treatment solutions for 1 min, then drained and placed in plastic containers at 20 °C for 24 h, and finally stored in polyethylene bags at 5 °C for 2 weeks. Color was measured periodically during storage. Lightness (L) values for all treated and control samples measured at 4 h, 24 h, and 2 weeks of storage were compared to L value for untreated samples measured immediately after cutting. Percent decrease in L-values was calculated for each sample at each time interval. Apple slices treated in ASC or SC solution had a significantly smaller decrease in L value indicating less browning than those treated in citric acid or water control at 4 h (P < 0.01), and with the exception of 1 g L⁻¹ ASC and 0.1 g L⁻¹ SC, all other ASC and SC treated slices still had significantly less browning than those for the water control (P < 0.01) at 24 h. After 2 weeks of storage, only SC (0.5–1.0 g L⁻¹), sodium bisulfite (0.5 g L⁻¹) and calcium l-ascorbate (10 g L⁻¹) continued to inhibit browning. Treatment with 0.5 g L⁻¹ SC and pH adjusted in the range from 3.9 to 6.2 using citric acid (CA) reduced browning more effectively than 0.5 g L⁻¹ SC without pH adjustment. Two organic acids, salicylic acid and cinnamic acid, when added to SC solution, were found to achieve even better inhibition of browning than CA at the same pH value.     

Synthesis and application of a new functionalized resin for use in an on-line,solid phase extraction system for the determination of trace elements in waters and reference cereal materials by flame atomic absorption spectrometry

The synthesis and characterization of the resin Amberlite XAD-4 functionalized with 2,6-pyridinedicarboxaldehyde and its application in an on-line system for the preconcentration of cadmium, cobalt, copper, lead and manganese prior to determination using flame atomic absorption spectrometry (FAAS) is proposed. Metal ions retained on the modified resin were eluted using 1.0 mol L⁻¹ HNO₃ solution and aspirated directly to the nebulizer–burner system of a FAAS instrument using a flow injection system. Detection limits (3σ) were determined to be 0.13 μg L⁻¹ for Cd, 0.29 μg L⁻¹ for Cu, 0.23 μg L⁻¹ for Mn, 0.58 μg L⁻¹ for Co and 2.19 μg L⁻¹ for Pb using a 10 mL of water sample loading volume. The limits of detection would be 100 times higher with units of μg kg⁻¹ for the solid samples in which their dilution ratios as (volume/weight) were 100. Enrichment factors ranged from 23.6 to 28.9 (for Co and Mn, respectively). The proposed method was successfully applied to determination of the analytes in natural water samples and certified reference materials.     

Agar films containing green tea extract and probiotic bacteria for extending fish shelf-life

A bioactive film composed of agar, incorporating green tea extract and probiotic strains (Lactobacillus paracasei L26 and Bifidobacterium lactis B94) was applied on hake fillets in order to evaluate the effect of the films during 15 days of storage. Hake was previously inoculated with Shewanella putrefaciens and Photobacterium phosphoreum (10³–10⁴ CFU/g) to simulate a spoilage process. The green tea and/or probiotic film provoked a reduction, particularly of H₂S-producing bacteria counts and total viable bacteria throughout the storage period. The probiotic strains added to the film could pass to the fish producing an increment of lactic acid bacterial counts, even in the presence of green tea extract. The effect of the films also caused a decrease in the indexes of fish quality (total volatile basic nitrogen (TVB-N), trimethylamine nitrogen (TMA-N) and pH). The total viable counts, H₂S-producing microorganisms and TVB-N were maintained below the limits of acceptability during 15 days for the fillet covered with the green tea + probiotic film, compared to the rest of the samples. Films with green tea and probiotic were able to extend shelf-life of hake at least for a week and increase the beneficial lactic acid bacteria in fish.     

Characterization of anthocyanins in Kenyan teas: Extraction and identification

Characterization and quantification of anthocyanins in selected tea cultivars processed into black (aerated) and green (unaerated) tea products was carried out in this study. The anthocyanins were extracted from tea products processed from a number of newly bred purple leaf coloured Kenyan tea cultivars (Camellia sinensis) using acidified methanol/HCl (99:1 v/v). Extracted anthocyanins were purified by C₁₈ solid phase extraction (SPE) catridges and characterised by HPLC-UV-Visible. They were identified according to their HPLC retention times, elution order and comparison with authentic standards that were available. Total monomeric anthocyanins were determined by the pH-differential method. Although the tea cultivars gave different yields of anthocyanins, the unaerated (green) teas had significantly (p ? 0.05) higher anthocyanin content than the aerated (black) teas. This was attributed to the degradation of anthocyanins by polyphenol oxidase products (catechin O-quinones) formed during the auto-oxidation (fermentation) process of black tea manufacture. Of the six most common natural anthocyanidins, five were identified in the purified extracts from purple leaf coloured tea, in both aerated (black) and unaerated (green) teas namely; delphinidin, cyanidin, pelargonidin, peonidin and malvidin. The most predominant anthocyanidin was malvidin in both tea products. In addition, two anthocyanins namely, cyanidin-3-O-galactoside and cyanidin-3-O-glucoside were also identified. Tea catechins were also identified in the tea products derived from the purple coloured tea cultivars namely, epigallocatechin (EGC), catechin (+C), epicatechin (EC), epigallocatechin gallate (EGCG), and epicatechin gallate (ECG). Correlation between the total catechins versus the total anthocyanins and anthocyanin concentration in unaerated teas revealed significant negative correlations (r = −0.723^∗ and r = −0.743^∗∗, p ? 0.05 and p ? 0.01, respectively). However, in aerated (black) tea the correlations were insignificant (r = −0.182 and r = −0.241, p > 0.05).     

Retardation of quality changes of Pacific white shrimp by green tea extract treatment and modified atmosphere packaging during refrigerated storage

The effect of modified atmosphere packaging (MAP) on the quality changes of Pacific white shrimp (Litopenaeus vannamei) treated with or without green tea extract (1 g/L; GTE) in combination with or without ascorbic acid (0.05 g/L; AA) during refrigerated storage of 10 days was investigated. Shrimp without treatment stored under MAP had lowered psychrotrophic bacteria, enterobacteriaceae and H₂S-producing bacteria count (P < 0.05) but similar lactic acid bacteria count (P > 0.05), in comparison with shrimp stored in air (control). The coincidental lowered rate of increase in pH, total volatile base (TVB) content and thiobarbituric acid reactive substances (TBARS) were obtained in shrimp stored under MAP (P < 0.05). However, MAP slightly lowered melanosis formation and improved likeness score to some extent. When shrimp were treated with GTE and stored under MAP, the lower microbiological and chemical changes as well as the lowest melanosis formation were observed, compared to shrimp kept under MAP without treatment and the control (P < 0.05). GTE treatment in combination with MAP could retard chemical changes and melanosis formation, regardless of AA incorporation (P > 0.05). Nevertheless, GTE in combination with AA had higher inhibition on microbial growth and yielded the shrimp with higher likeness, compared with the other treatments (P < 0.05). Therefore, shrimp treated with GTE in combination with AA prior to MAP had the lowest losses in quality during refrigerated storage.     

Subacute oral toxicity of cocoa tea (Camellia ptilophylla) water extract in SD rats

Camellia ptilophylla, known as cocoa tea, is a natural decaffeinated tea plant which has been proved with potential bioactivities. This study was carried out to evaluate the safety of green cocoa tea water extract (CWE) by determining its potential toxicity after subacute administration in SD rats. In the subacute toxicity study, oral administration of CWE up to 800 mg kg^?1 body weight day^?1 for twenty‐eight consecutive days did not cause either mortality or toxicity in SD rats, regardless of gender. There were no adverse effects in body weight, food consumption, relative organ weight, haematological parameters, clinical chemistry, gross pathology and histopathology between treatment and control groups. Several parameters (e.g. neutrophils%, Ca²⁺, total Ca) in CWE‐treated rats with high dosage were significantly different from the control groups, but still in a safety level. A dose of 800 mg kg^?1 day^?1 was identified as the no observed adverse effect level (NOAEL) in this study.     

Effect of different extraction methods on the recovery of chlorogenic acids,caffeine and Maillard reaction products in coffee beans

Water and ethanolic extracts were obtained from green and roasted (3 different roast degrees) Arabica and Robusta coffee beans. Three types of water extracts were prepared from the examined, finely ground material through: (a) brewing with boiling water, (b) boiling in water, and (c) boiling in water under elevated pressure. All these extracts were lyophilized. Two types of ethanolic extracts were derived from the examined material through (a) extraction of the finely ground coffee beans and (b) extraction of the solid residue that remained after boiling the coffee beans in water under elevated pressure. These ethanolic extracts were dried. Both water and ethanolic extracts were analyzed for concentration of potential antioxidants such as chlorogenic acids and caffeine (by HPLC) and Maillard reaction products (measurements of absorbance at 420 nm). Concentration of chlorogenic acids in Robusta extracts varied between 0.4 and 36.0 g × 100 g⁻¹ dry extract weight (db.), while in Arabica extracts it ranged from 0.1 to 22.4 g × 100 g⁻¹ db. Extracts of dark roasted Arabica contained more chlorogenic acids than those of Robusta. Concentration of caffeine, which in green and roasted coffee beans is maintained at the similar level, tended to increase in Robusta extracts with the roast degree and temperature of extraction with water, while in case of Arabica extracts there was no noticeable tendency. Caffeine concentrations varied between 0.12 and 8.41 g × 100 g⁻¹ db. and between 0.03 and 6.53 g × 100 g⁻¹ db. in Robusta and Arabica extracts, respectively. Ethanolic extracts were characterized by relatively higher caffeine concentrations and lower contents of brown pigments and chlorogenic acids as compared to water extracts. The richest in antioxidants were extracts of green Robusta coffee beans derived through boiling in water under elevated pressure.     

In vitro antioxidant and anticancer activities of ethanolic extract of selenium-enriched green tea

Selenium-enriched green tea is now being increasingly produced in China and is well known as a bioactive beverage, due to its high content of active components. In this study, the antioxidant and anticancer activities of an ethanolic extract and an aqueous extract of Se-enriched green tea were investigated. The results indicated that the ethanolic extract possessed significantly higher antioxidant activity than the aqueous extract and the positive control α-tocopherol, by both α,α-diphenyl-β-picrylhydrazyl (DPPH) radical-scavenging and ferric thiocyanate (FTC) assays. The ethanolic extract inhibited the proliferation of human cervical adenocarcinoma HeLa cell and possessed a significantly higher antitumour activity than the aqueous extract and the positive control 5-fluorouracil (5-FU), in the dose range of 62.5–250 μg/ml. Moreover, the ethanolic extract could significantly inhibit the growth of lung carcinoma A549 and hepatoma HepG2 in a concentration-dependent manner, with IC₅₀ values of 278.6 μg/ml and 431.6 μg/ml, respectively. Selenium, tea polyphenols and polyphenols constituents, especially epigallocatechin gallate (EGCG), were significantly higher in the ethanolic extract than in the aqueous extract, which were possibly responsible for the higher antioxidant and antitumour activities of the ethanolic extract.     

The influence of different water types and brewing durations on the colloidal properties of green tea infusion

Green tea infusions were prepared with three different water types: distilled water, commercial mineral water and CaCl₂ added distilled water (at 20 mg L^?1). The impact of different water type on the colloidal particle parameters, namely size, polydispersity index (PDI) and zeta potential, was examined within 24 h. Our results indicate that distilled water green tea infusion (DWT) brewed at 80 °C for 5 min contained the smallest particles (242.17 ± 11.78 nm) as examined by dynamic light scattering (DLS). Moreover, DWT showed smaller size increase during the 24 h storage than that of mineral water tea and CaCl₂ added water tea infusion. The morphology characteristics of green tea infusion particles were visualised by transmission electron microscope (TEM) which revealed that green tea infusion particles had heteromorphic shapes. Our results suggest that mineral composition of water impacts the colloidal size and stability of green tea infusion. Additionally, DLS and TEM could be useful tools to provide colloidal information which is important to understand quality characteristics of green tea infusion during processing and storage.     

Comparison of bioactive components in GABA tea and green tea produced in Taiwan

The aim of this study is to investigate the bioactive components of GABA (γ-aminobutyric acid) tea as compared with green tea produced in Taiwan. Using in total 56 tea samples (28 green tea and 28 GABA tea), moisture content, Hunter L, a and b values, phenolic compounds, amino acids including GABA, fatty acids and ascorbic acid were determined. The results showed that moisture, total free amino acids, crude fat, Hunter L value, total nitrogen, free fatty acids and reducing sugar did not differ significantly between GABA tea and green tea. However, GABA tea had higher Hunter a and b values, while green tea had higher total catechin and ascorbic acid contents (p < 0.05). Of major catechins, epicatechin and epigallocatechin gallate were found to be lower in GABA tea than in green tea. For free amino acids, GABA, alanine, ammonia, lysine, leucine and isoleucine were found to be significantly higher in GABA tea, while the glutamic acid, aspartic acid, and phenylalanine were higher in green tea (p < 0.05). Theanine, tryptophan, valine, threonine and methionine were not found to be different between the two kinds of tea.     

Daily green tea extract supplementation reduces prothrombotic and inflammatory states in dialysis patients

Cardiovascular disease (CVD) remains the most common cause for excess morbidity and mortality in patients with chronic kidney disease (CKD) and end stage renal disease (ESRD) under chronic dialysis. ESRD patients have increased oxidative stress and endothelial dysfunction alongside increased levels of inflammation related proteins, which has prompted the exploration of anti-inflammatory and antioxidant treatments to improve outcomes. As green tea is increasingly well recognized for its antioxidant properties, we probed the effect of consumption of 1 capsule daily of green tea as a commercially available, decaffeinated green tea capsule (1 g, catechin content 68 mg) for 6 months on fibrinogen and inflammation in dialysis patients. Chronic hemodialysis patients (N = 25) were recruited and fibrinogen, FDP-D-dimer, high sensitivity (hs) CRP and the mononuclear cell protein expression of p22^phox, were assessed before, i.e. baseline and after 6 months of ingestion of 1 green tea capsule per day. After 6 months of daily green tea capsule ingestion, dialysis patients showed reduced protein expression of p22^phox (p < 0.0001), reduced hsCPR (p = 0.032) and fibrinogen (p = 0.022) levels and increased FDP-D-dimer (p = 0.0019) compared to their values at baseline. These results document lower oxidative stress and inflammation with green tea capsule ingestion and suggest a likely positive impact of green tea treatment on the atherosclerotic process of ESRD patients under dialysis.     

Rapid determination of soluble solids content and acidity of Black currant (Ribes nigrum L.) juice by mid-infrared spectroscopy performed in series

Black currant juice quality is a prime importance for growers and industrials. Such quality relies on the empirical and destructive measurements of soluble solids content and acidity. These measurements are time consuming and expensive. The serial analysis of the Black currant juice quality could allow the valuation of the product for the producer and saving of time for the manufacturer. Our objective was to develop predictive models of soluble solids content and acidity based on the mid-infrared spectroscopy performed in series.We showed that infrared spectroscopy allowed determining efficiently the quality of Black currant juices and that such technique could be performed in series. Soluble solids content were determined with a R²-value of 0.97 and an RMSECV of 1.14°Brix for a range of values comprised between 12.9°Brix and 16.42°Brix. Acidity content was determined with a R²-value of 0.96 and an RMSECV of 2.61 g L⁻¹ in a large range of values comprised between 14.87 g L⁻¹ and 36.27 g L⁻¹.These results allow asserting that the soluble solids content and the acidity of Black currant juice can be accurately determined by infrared spectroscopy. Furthermore, a large number of measures can be performed in series, what opens perspectives for a practical application.     

Preparation of galacto-oligosaccharides using membrane reactor

Galacto-oligosaccharides (GOS) have many biological functions. They are produced from lactose by a glycosyl transfer of one or more d-galactosyl units onto the d-galactose moiety of lactose catalysed by β-galactosidase. This reaction can proceed in batch or in continuous system. Membrane reactor with ultrafiltration ceramic membrane (150 kDa) was used and batch system as well. Lactose in phosphate buffer (197.9 g L⁻¹), recombined whey (197 g L⁻¹) and evaporated ultrafiltration permeate (200.7 g L⁻¹) were used as substrates. The used concentration of enzyme Maxilact LX 5000 was 6 U mL⁻¹. The highest concentration of GOS in buffer was reached after 30 min of batch reaction and it was 25.5 g L⁻¹, for recombined whey it was 31.1 g L⁻¹ and for ultrafiltration permeate it was 32.9 g L⁻¹. For continuous process was obtained 8.8 g L⁻¹ of GOS for buffer and 21.0 g L⁻¹ for recombined whey and 16.7 g L⁻¹ of GOS for ultrafiltration permeate.     

Microstructure characteristics of tea seed dietary fibre and its effect on cholesterol,glucose and nitrite ion adsorption capacities in vitro: a comparison study among different modifications

The effects of xylanase–cellulase hydrolysis, ultrasonic modification and enzymatic-ultrasonic treatment on the physicochemical properties, morphological structures and adsorption capacities in vitro of purified fibre (PF) from tea seed were determined. The results showed the ultrasonically treated fibre (UTF) had a higher water-holding capacity (60.15 g g⁻¹), oil-binding capacity (30.42 g g⁻¹), swelling capacity (29.93 mL g⁻¹), emulsification activity (381.36 m² g⁻¹) and emulsification stability (20.20) than PF, enzymatically hydrolysed fibre (EHF) and enzymatic-ultrasonic treated fibre (EUF). The structures of PF, UTF, EHF and EUF were characterised by scanning electron microscope, Laser particle size analyzer, X-ray diffraction, thermogravimetric analysis and Fourier transform infrared spectra. Furthermore, compared with PF, the adsorption capacity of UTF, EHF and EUF for cholesterol, glucose and nitrite ions during simulative gastrointestinal tract was improved to different degrees. This study can provide guidance for the comprehensive utilisation of byproduct of tea seed and designing novel functional dietary fibre.     

Cream formation and main chemical components of green tea infusions processed from different parts of new shoots

The formation of green tea cream and its chemical components were investigated. The green tea infusions were processed from different parts of new shoots: the bud and the first leaf, the second leaf and its implicative stem, the third leaf and its implicative stem, and the fourth leaf and its implicative stem, which were named as first part, second part, third part and fourth part, respectively. The results showed that the formation of tea cream and its settlement slowed down gradually from the first part to the fourth part, and that the amount of tea cream also decreased accordingly. The amount of tea cream was influenced remarkably by the chemical components in the green tea infusion. The main components of green tea cream were polyphenols (29.86 –78.66%), total sugar (14.47–27.62%) and caffeine (2.35–10.43%). Catechins (12.8–42.5%) were the main components of polyphenols which participated in tea cream formation. The main components in the catechins were found to be (−)-epigallocatechin (EGC), (−)-epigallocatechin gallate (EGCG), (−)-epicatechin (EC) and (−)-epicatechin gallate (ECG). Seven minerals were found in the tea cream, including Ca²⁺, K⁺, Mg²⁺, Mn²⁺, Na⁺, Zn²⁺ and Ni²⁺.