A screening method for the determination of ascorbic acid in fruit juices and soft drinks

A simple and rapid liquid chromatographic method based on a new stationary phase Teknokroma, Tr-010065 Mediterranea sea₁₈ (15 cm × 0.4 cm, id 3 μm), to determine ascorbic acid in beverages is reported. With the proposed method the samples were analysed by direct injection without a previous treatment. The total analysis time does not exceed 6 min. The method showed a good repeatability (RSD < 2%: n = 6) and an excellent sensitivity (LOD = 0.01 mg/l). Seventeen samples were analysed, including fruit juices, soft drinks and isotonic beverages. Ascorbic acid contents ranged from 6.6 to 840 mg/l. The ascorbic acid stability in some beverages during their shelf-life was also evaluated. Degradation of about 54% was observed in a tea drink.     

A simple and highly sensitive spectrophotometric determination of Cr(VI) in food samples by using 3,4-dihydroxybenzaldehydeisonicotinoylhydrazone (3,4-DHBINH)

A simple, rapid, sensitive and inexpensive method has been developed for the determination of trace amounts of chromium(VI) using 3,4-dihydroxybenzaldehydeisonicotinoylhydrazone (3,4-DHBINH). The metal ion gives a yellow coloured complex with 3,4-DHBINH in acetate buffer of pH 5.5 with 1:1 (metal:ligand) composition. The complex shows a maximum absorption at 400 nm. Beer’s law is obeyed in the range 0.5–7.7 ppm of Cr(VI). The molar absorptivity, Sandell’s sensitivity and detection limit were found to be 1.35 × 10⁴ L mol⁻¹ cm⁻¹, 0.0075 μg cm⁻² and 0.0045 μg mL⁻¹, respectively. The correlation co-efficient and regression co-efficient of the Cr(VI)–3,4-DHBINH complex were 0.99 and 0.12, respectively. Major cations and anions did not show any interference. The developed method has been successfully applied for the analysis of Cr(VI) in food samples (leafy vegetables), comparing the results simultaneously with those obtained using an Atomic Absorption Spectrophotometer, whereby the validity of the method has been tested.     

Identification and quantification of fungi and mycotoxins from Pu-erh tea

Pu-erh tea originates from the province of Yunnan in south-western China. As this tea is produced by so called Aspergillus post-fermentation the question arises which molds and mycotoxins may be found in this tea. In total 36 samples of Pu-erh tea were investigated for their content of filamentous fungi and the mycotoxins aflatoxins B₁, B₂, G₁, and G₂, fumonisins B₁, B₂, and B₃, and ochratoxin A. Fungi were isolated from all samples in a concentration of 1.0 × 10¹ to 2.6 × 10⁶ colony forming units (cfu)/g tea, all together 19 fungal genera and 31 species were identified. The most prevalent species were Aspergillus acidus and Aspergillus fumigatus, followed by Zygomycetes and Penicillium species. Aflatoxins and fumonisins were not found in the samples investigated, ochratoxin A was detected in 4 of 36 teas (11.1%).     

Quantitative analysis of acrylamide in tea by liquid chromatography coupled with electrospray ionization tandem mass spectrometry

An effective sample preparation procedure was optimized and a liquid chromatography–tandem mass spectrometry (LC–MS/MS) was developed for the quantitative analysis of acrylamide in tea. [¹³C₃]-acrylamide was used as internal standard. Acrylamide was extracted at 25 °C for 20 min by 10 ml water followed by 10 ml acetonitrile, and then 4 g of magnesium sulfate and 0.5 g of sodium chloride were added to the above mixture under stirring thoroughly. In order to increase the response of acrylamide, 9 ml acetonitrile layer was taken and concentrated to 0.5 ml. Solid-phase extraction with an Oasis MCX cartridge was carried out for clean-up. The limit of detection (LOD) and limit of quantification (LOQ) were 1 and 5 ng/ml, respectively. The recovery efficiency of the extraction procedure ranged between 74% and 79%. The levels of acrylamide in 30 tea samples were less than 100 ng/g. Black, oolong, white and yellow tea samples had quite low acrylamide contents (<20 ng/g). Higher acrylamide levels occurred in baked, roasted, and one sun-dried green tea samples (46–94 ng/g).     

Detection of irradiated black tea (Camellia sinensis) and rooibos tea (Aspalathus linearis) by ESR spectroscopy

Detailed ESR investigation on irradiated black and rooibos tea was carried out in the dose range of 0.5–10 kGy. Unirradiated black and rooibos tea samples exhibit a weak, symmetric ESR (electron spin resonance) singlet centered at g = 2.0043 ± 0.0010 with peak-to-peak line widths (ΔH_pp) of 1.00 ± 0.05 and 0.64 ± 0.05 mT, respectively. Irradiation caused a significant increase in ESR signal intensity of both samples without any changes in spectral patterns and these increases were found to be explained by a quadratic and/or an exponentially varying functions. Variation of ESR signal intensity, for the irradiated samples, at room temperature (295 K) with time in a storage period of 39 days showed that free radical responsible from the ESR spectrum of black tea is more stable than that of the rooibos tea. However, variable temperature and annealing studies show that the free radical responsible from the ESR spectrum of rooibos tea, with activation energy of 46.0 ± 3.5 kJ mol⁻¹, is more resistant to the temperature than that of the black tea with activation energy of 33.8 ± 3.1 kJ mol⁻¹.     

Content of essential oil, terpenoids and polyphenols in commercial chamomile (Chamomilla recutita L. Rauschert) teas from different countries

Commercial samples of German chamomile (Chamomilla recutita (L.) Rauschert) tea (n = 13), packaged in different countries and purchased in Estonia either at food markets (9) or at retail pharmacies (4), were analysed for the essential oil and terpenoid content and constituents by GC-MS. Also the tea infusions were analysed for polyphenols spectrophotometrically and chromatographically by LC-DAD-MS/MS. The oils were obtained from chamomile flowers with yields of 0.10 - 0.61%. The existence of two types of chamomile tea, one rich in bisabolol oxides and the other in (−)-α-bisabolol, was established. The total content of polyphenols in gallic acid equivalents, estimated by the classical Folin-Ciocalteu method (TP_FC), was almost independent of the sample origin, but the total content of polyphenols (in chlorogenic acid equivalents) calculated on the basis of net areas under the chromatographic curves (AUC) of the infusions at 280 nm (TP_AUC) showed a significant variability as well as content of total flavonols (0.29-1.21%) or total phenolic acids (7.7-91.4 mg/200 ml). The major phenolic compounds in the chamomile infusions were chlorogenic acids, ferulic acid glycosides, dicaffeoyl quinic acids and apigenin glycosides. Based on the amounts of essential oil, terpenoids, total flavonols and major phenolic compounds, the quality of the commercial chamomile teas was very variable, and the chamomile teas available in pharmacies should be preferred for the medical purposes.     

Hexavalent chromium removal by Osage Orange

This study explores the sorption potential of Osage Orange (Maclura Pomifera) for the removal of Cr(VI) ion. The influence of contact time, solution pH, initial metal concentration, amount of biosorbent and ionic strength on the removal of Cr(VI) ion was studied. The biosorption of Cr(VI) with pulp and peel was investigated in a batch arrangement. The initial and equilibrium concentrations of Cr(VI) ions in aqueous phase were determined by spectrophotometry. The sorption process was pH and concentration dependent. The sorption of Cr(VI) ions increased with a decreasing pH until pH 2. The increase in initial Cr(VI) ions concentration in aqueous phase increased the sorption. The sorption data fitted well with the Langmuir sorption model within the concentration range studied. The observed maximum biosorption capacity by Langmuir sorption model at pH of 2 for M. Pomifera pulp was 0.92 mmol of Cr(VI)/g and for M. Pomifera peel was 0.55 mmol of Cr(VI)/g.     

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).     

Oxidation in HiOx-packaged pork Longissimus muscle predisposes myofibrillar and sarcoplasmic proteins to N-nitrosamine formation in nitrite-curing solution

The effect of meat protein in situ oxidation on the formation of N-nitrosodiethylamine (NDEA) was investigated. Fresh minced pork was untreated (Con) or treated with 700 mg/kg α-tocopherol (Toc) or 300 mg/kg tea polyphenols (PPE), packaged in a HiOx atmosphere (78.8% O₂, 18.8% CO₂, 2.4% N₂), then stored at 2 ± 1 °C for up to 10 days. Crude myofibrillar (MP) or sarcoplasmic (SP) protein (20 mg/mL) extracted from stored meat was reacted with 43 μM sodium nitrite at 80 °C for 1 h. Lipid oxidation was totally inhibited in PPE pork but increased in Con and Toc samples after 10 days. There was significant protein oxidation (losses of sulfhydryls, formation of protein carbonyls) in both MP and SP in all samples during storage. However, the Con group suffered more extensive protein oxidation than Toc and PPE and produced more NDEA (P < 0.05), indicating that protein oxidation promoted nitrosation.     

Assessment of plasma concentrations of (−)-epigallocatechin gallate in mice following administration of a dose reflecting consumption of a standard green tea beverage

The plasma exposure of (−)-epigallocatechin gallate (EGCg) is typically assessed following administration of EGCg at doses equivalent to the consumption of at least 10 cups of green tea in one sitting. This study determines the plasma concentrations of EGCg in mice following administration of a dose reflecting typical consumption of one standard green tea beverage. Swiss Outbred mice were orally administered 0.76 mg/kg EGCg, and using a validated HPLC method, the C_max of un-conjugated and total EGCg was determined to be 31.5 ± 3.3 and 34.3 ± 2.0 nM, respectively (mean ± s.d., n = 3–5). The area under the plasma concentration versus time curve for un-conjugated and total EGCg was 114.3 ± 4.1 and 116.4 ± 4.1 nM·h, respectively (mean ± s.d., n = 3–5). To minimise potential ex vivo plasma degradation, a novel stabilizing solution of 20 mM ascorbic acid (AA) and 13 mM tris[2-carboxyethyl]phosphine hydrochloride (TCEP) was employed. Comparative evaluation of the efficacy of the 20 mM AA and 13 mM TCEP stabilizing solution to the commonly used stabilizing solution of 114 mM AA and 0.13 mM Na₂EDTA, indicated that the AA/TCEP solution provided significantly greater (p < 0.05) protection to EGCg than the AA/Na₂EDTA stabilizing solution. Overall, this study demonstrates that plasma concentrations of EGCg are in the low nM range following oral administration to mice at a dose reflecting the consumption of a standard green tea beverage. In addition, a novel stabilizing solution has been identified which may be useful in stabilizing plasma samples obtained from pharmacokinetic studies.     

Variation in antioxidant potential and total polyphenol content of fresh and fully-fermented Sri Lankan tea

Tea polyphenols possess antioxidant properties and have been shown to have a protective effect against several degenerative diseases. The study aimed to determine the amounts of polyphenols and antioxidant properties for teas grown in Sri Lanka, over a period of 10 months. Water extracts of freeze-dried fresh (unfermented) and fully-fermented tea leaves were made for a structured set of samples (fermented and unfermented teas from six plantations; teas representing two harvesting seasons from four plantations) collected from the main tea growing regions in Sri Lanka. Total phenolic content (TPC), the ferric reducing antioxidant power (FRAP) and the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging activity were determined for each sample. The results highlight significant (P < 0.05) variations in antioxidant activity across the six plantations. FRAP and DPPH for both fermented and unfermented teas from the four highland plantations showed a significant (P < 0.05) interaction between season and plantation. A similar interaction between season and plantation was observed for total phenolics in unfermented teas from the four highland plantations. The variability of the total phenolics for fermented teas, however, was independent of seasonal variations. A significant correlation (r = 0.5, P < 0.05) was observed between FRAP and total phenolics.     

Chromium content in selected convenience and fast foods in Poland

The chromium content in selected convenience and fast foods was determined. Samples were wet digested with HNO₃ (69%) in a microwave digestion system. Chromium was determined by graphite furnace atomic absorption spectroscopy (GF-AAS). The chromium content in convenience food ranged on average from 2.22 to 18.2 μg/100 g, in fast food from 3.76 to 28.6 μg/100 g, and in instant food from 0.34 to 4.75 μg/100 g.     

Removal of Cr(VI) from aqueous solutions by modified walnut shells

Walnut shell (WNS) (Juglans regia) has been utilised as an adsorbent for the removal of Cr(VI) ions from aqueous solutions after treatment with citric acid. The modification reaction variables, such as citric acid level (5-10 g), reaction time (0-24 h), and temperature (110-130 °C) were studied in batch experiments. The rate of adsorption was studied under a variety of conditions, including initial Cr(VI) concentration (0.1-1.0 mM), amount of adsorbent (0.02-0.20 g), pH (2-9), temperature and contact time (10-240 min). Adsorption of Cr(VI) is in all cases pH-dependent showing a maximum at equilibrium pH values between 2.0 and 3.0 for citric acid modified walnut shell (CA-WNS). The applicability of the Langmuir, Freundlich and D-R adsorption isotherms has been tested for the equilibrium. Maximum adsorption capacities of CA-WNS and untreated WNS under experimental conditions were 0.596 and 0.154 mmol/g for Cr(VI) ions, respectively.     

Caffeine in Chiang Rai tea infusions: Effects of tea variety,type, leaf form,and infusion conditions

Caffeine in Chiang Rai tea infusions was found to be dependent on infusion conditions (water temperature and infusion time), and leaf form (non-ground or ground) but independent of tea variety and type. For non-ground leaf samples, the higher the water temperature and the longer the infusion time, the higher the caffeine concentrations in tea infusions. After infusing for longer than 15 min, the dissolution rate of caffeine became slower and the concentration was essentially constant. For ground leaves, the caffeine content was not influenced by infusion time. Caffeine concentrations in tea infusions from Camellia sinensis var. sinensis (26.8 ± 0.81 and 22.3 ± 5.55 mg/100 ml for ground and non-ground samples, respectively) were not significantly different from that of Camellia sinensis var. assamica (24.4 ± 0.66 and 20.3 ± 5.07 mg/100 ml for ground and non-ground samples, respectively). The difference in caffeine concentration between green tea (28.1 ± 8.19 mg/100 ml) and oolong tea (20.3 ± 1.52 mg/100 ml) was not statistically significant.     

Green tea: A promising anticancer agent for renal cell carcinoma

Renal cell carcinoma (RCC) is one of the most lethal amongst the urologic malignancies, comprising three percent of all human neoplasms, and its incidence appears to be rising. RCC is refractory to both chemotherapy and radiotherapy. Therefore, the discovery of new strategies for therapeutic intervention remains a priority. Green tea (Camellia sinensis) and tea polyphenols have been proposed to exert protective effects against several types of cancer, based on preclinical and clinical trial data; however, the anticarcinogenic activity of green tea towards RCC is unknown. In this study, a targeted metabolite analysis on a green tea leaves methanolic extract was performed by HPLC/DAD and the antiproliferative activity of the extract was assayed using human renal cancer cell lines A-498 and 769-P. The total phenolic content was very high (31.8% of methanolic extract), and the main compounds were flavan-3-ols (94.3% of the total phenolic content), and especially (−)-epigallocatechin-3-gallate (35.9% of the total phenolic content). In addition, two methylxanthines – theophylline and caffeine – were also present in the extract, caffeine being the most abundant. Green tea extract strongly inhibited the growth of both RCC cell lines in a concentration-dependent manner, with IC₅₀ values of 54 ± 10 and 129 ± 28 μg/ml for A-498 and 769-P cells, respectively. This is the first report showing that green tea is likely to be an effective anticancer agent for renal cell carcinoma.     

Determination of lead and cadmium in food samples by the coprecipitation method

In this study, the coprecipitation method developed using a combination of 2-mercaptobenzothiazole (MBT) as a chelating reagent and copper as coprecipitate carrier was used for the determination of trace lead and cadmium in various food samples by graphite furnace atomic absorption spectrometry (GFAAS). The method was applied for the determination of Pb(II) and Cd(II) in salami, sausage, chicken, anchovy, spinach, cabbage, onion, dill, parsley, lettuce, tea and rice samples. The matrix modifiers were added as 50 μg NH₄H₂PO₄ + 3 μg Mg(NO₃)₂ for both Pb(II) and Cd(II). The signals were measured as peak area. The concentrations of Pb(II) and Cd(II) in the food samples were found to be in the range of 6.63 ng g⁻¹ (anchovy) −3.30 μg g⁻¹ (spinach) and 2.67 ng g⁻¹ (salami) −0.51 μg g⁻¹ (lettuce), respectively.     

Time and incidence of ovulation and conception rates after incorporating estradiol cypionate into a timed artificial insemination protocol

Two experiments were conducted to determine the effect of estradiol cypionate (ECP), when incorporated into a conventional GnRH-PGF_2α-GnRH timed artificial insemination protocol (Ovsynch), on systemic estradiol (E₂), time and incidence of ovulation, luteal development, and conception rate in Holstein cows. Our objective was to determine if administration of 0.25 mg of ECP at the time of the second GnRH injection would effectively synchronize ovulation and increase conception rate. In Experiment 1, lactating Holstein cows (n = 23; 58.7 ± 1.2 d in milk) were synchronized with PGF_2α (at d −10). Ten days later, Ovsynch was initiated with the administration of 100 μg of GnRH (d 0) followed by PGF_2α on d 7. On d 9, cows were assigned randomly to be treated with either GnRH + 0.25 mg of ECP (OVS-ECP; n = 11) or GnRH and 1 mL of cottonseed oil (OVS-C; n = 12). Ovarian activity was monitored by ultrasonography on d 0, 7, and 9. To determine the time of ovulation, ultrasound examinations were conducted at 12 and 20 h posttreatment and then at least every 3 h until either 36 h posttreatment or ovulation was observed. Blood samples were collected on d 0, 7, 9, and 16 for progesterone analysis. Blood samples also were collected at the time of treatment (d 9, 0 h) and at 6, 12, 20, and 28 h for E₂ analysis. Incidence of ovulation did not differ between treatments. Mean ovulation time relative to the second GnRH administration was similar between treatments. Serum progesterone concentration did not differ between treatments at any time. Serum E₂ concentration was not different at the time of treatment (0 h); however, mean E₂ concentration was greater for the OVS-ECP group at 6 and 12 h after treatment compared with OVS-C. In Experiment 2, lactating dairy cows (n = 333) in 3 commercial herds were randomly assigned to OVS-ECP (n = 169) or OVS-C (n = 164). Cows were inseminated 22 to 24 h posttreatment. Conception rates did not differ between treatments. Estradiol cypionate treatment was successful in increasing serum E₂ when administered at the time of the second dose of GnRH in the Ovsynch protocol. Conception rates, however, were not affected by treatment.     

Sensitive and rapid determination of catechol in tea samples using mesoporous Al-doped silica modified electrode

A mesoporous Al-doped silica (Al/SiO₂) was synthesised according to the published method, and then used to modify the carbon paste electrode (CPE). The electrochemical behaviour of catechol was investigated. Compared with the unmodified CPE, the resulting mesoporous Al/SiO₂ modified CPE remarkably increases the peak currents of catechol, and greatly lowers the peak potential separation. Therefore, the mesoporous Al/SiO₂ exhibits catalytic activity to catechol and significantly improves the determining sensitivity. Based on this, a sensitive, rapid and convenient electrochemical method was proposed for the determination of catechol. The linear range is between 5.0 × 10⁻⁷ and 5.0 × 10⁻⁵ mol L⁻¹ with a correlation coefficient of 0.998. The limit of detection is as low as 1.0 × 10⁻⁷ mol L⁻¹. Finally, this novel method was employed to determine catechol in tea samples, which testified by high-performance liquid chromatography.     

Effect of PEF on microbial inactivation and physical-chemical properties of green tea extracts

The effects of pulsed electric fields (PEF) on (1) the inactivation of Escherichia coli and Staphylococus aureus in green tea beverage, and (2) the color, green tea polyphenols (GTP) content, and total free amino acids in green tea extracts were investigated. Green tea extract samples inoculated with E. coli and S. aureus were treated using a bench-scale PEF system at electric field strengths of 18.1, 27.4, and 38.4 kV/cm and total treatment times of 40, 80, 120, 160 and 200 μs. The inactivation of E. coli and S. aureus by PEF treatment at 38.4 kV/cm for 160 and 200 μs reached 5.6 and 4.9 log reductions, respectively. PEF processing caused no considerable changes in color, GTP and total free amino acids. The storage tests at 4 °C showed that synergistic effect of low temperature storage and the antimicrobial functionality of GTP resulted in a considerable reduction in the microoganisms of the PEF-treated tea beverage, extending its shelf-life to over 6 months at 4 °C.     

Short communication: Comparison of 3 solid digesta passage markers in dairy cows

This study investigated the usefulness of acid-detergent fiber-bound ¹⁵N [acid detergent insoluble (ADI)-¹⁵N] as a solid digesta passage marker in dairy cows compared with chromium (Cr) and ytterbium (Yb) (as labeled fiber or forage, respectively). Intrinsically (ADI-¹⁵N) or extrinsically (Cr, Yb) labeled alfalfa hay was pulse-dosed intraruminally to 7 lactating dairy cows. Following marker administration, spot fecal samples were collected for up to 72 h for marker analyses. Urine and milk samples were also collected and analyzed for Yb and Cr. Fecal marker excretion data were processed using 2-compartment mathematical age-dependent/age-independent (G_n→G₁) models. The rate of passage of the marker in the first, age-dependent compartment tended to be slower for Yb compared with Cr and ADI-¹⁵N, which resulted in a trend for longer mean retention time (MRT) in this compartment when Yb was used as a marker (19.0 h) compared with Cr and ADI-¹⁵N (14.5 and 13.9 h, respectively). The rate constant of marker disappearance for the second or age-independent compartment tended to be greater for Yb compared with Cr and ADI-¹⁵N, which led to a shorter MRT of Yb in this compartment (15.6) versus ADI-¹⁵N (32.1) and Cr (24.8 h). The cumulative MRT was greater for ADI-¹⁵N versus Cr and Yb (46.0, 39.3, and 34.4 h, respectively). Total MRT of marker tended to be greater for ADI-¹⁵N than for Yb (46.6 vs. 36.6 h, respectively). Urine and milk analyses data suggested no measurable losses of Yb along the digestive tract, but about 0.79% of Cr dosed intraruminally was secreted or excreted in milk and urine in the 48-h period following marker administration. Collectively, this study confirmed previous observations that ADI-¹⁵N can be used reliably as a solid digesta passage marker for ruminants, producing pre-duodenal and total-tract retention times similar to that of Cr-labeled fiber. Retention time in the age-independent compartment was underestimated when Yb was used as a marker, emphasizing the need to process forages to isolate fiber before labeling with Yb.