Ultrasonic-assisted extraction and UHPLC determination of ascorbic acid, polyphenols, and half-maximum effective concentration in Citrus medica and Ziziphus spina-christi fruits using multivariate experimental design.

This study aimed to determine the concentrations of ascorbic acid and polyphenols in fruits and peels of Citrus medica and Ziziphus spina-christi grown in Ethiopia. Conditions of ultrasound-assisted extraction (UAE) and ultra-high-performance liquid chromatography coupled with a diode array detector (HPLC-DAD) were optimized, using a multivariate experimental design. The optimum conditions of UAE were 15 min extraction time at 35 ℃, with 75 % aqueous methanol as solvent, and a fruit powder-to-solvent ratio (m/v) of 1:15. Among the different drying conditions investigated, freeze-drying was found to be appropriate for analyzing ascorbic acid, polyphenols, and antioxidant potential. The overall ranges, across the fruits and peels, of ascorbic acid, total polyphenols, and antioxidant potentials (EC50) obtained were 8.7 ± 1.4-91.2 ± 2.6 mg/100 g, 253.0 ± 6.3-764.1 ± 25.8 mg GAE/100 g and 2.4 ± 0.1-26.1 ± 2.9 mg/mL, respectively. This indicates that the fruits and peels of the studied plants are advantageous as sources of ascorbic acid and polyphenols.

Discrimination of the geographical origin of gluten-free teff grains from northwestern parts of Ethiopia by fatty acid analysis.

Teff (Eragrostis tef) is a gluten-free cereal, and the consumer also prefers teff due to its nutritional composition. Determining the geographical origin of teff is important to select the right product for consumers. The quality and consumer preference of teff varies based on their production origin; consequently, their prices differ significantly. This work studied the profile of fatty acids in seventy-two teff samples by using gas chromatography coupled with mass spectrometry (GC-MS) and identifying the markers to discriminate the geographical origin of teff depending on their production region. Principal component analysis (PCA) and linear discriminat Analysis (LDA) were used to visualize data trends, and construct classification models for teff samples according to their geographical origins. Thirty different fatty acids were detected in all of the collected teff samples. The total mean concentration of fatty acids ranged from 739.85 to 938.06 mg/100g across the six districts in the three zones (East Gojjam,Awi, and West Gojjam). Stearic acid,trans-vaccenic acid, linoleic acid, azelaic acid, and capric acid were the most discriminating fatty acids of teff grains between East Gojjam and West Gojjam zones, while palmitic, palmitoleic, and oleic acid discriminated Awi zone teff samples from the other zones. The recognition and prediction abilities of the LDA model for the classification of the production zones were 98.6 % and 94.4 %, respectively. Hence, the fatty acid profiles combined with multivariate data analysis too can be used in the determination of the geographical origin of teff grains.

Variations in elemental composition of rice (Oryza sativa L.) with different cultivation areas of Ethiopia.

Variations in the elemental composition of rice (Oryza sativa L.) grains, and the link with the growing soil, were investigated across the major production areas of Ethiopia (Fogera, Metema and Pawe). The elements (Ca, Mg, Fe, Zn, Mn, Cu, Ni, Cr, Cd and Pb) were determined by using flame atomic absorption spectroscopy (FAAS), after digesting samples through an optimized procedure with respect to volumes of reagents (HNO3, HClO4 and H2O2), temperature and time. The accuracy of the FAAS method was in the range of 87‒113%. The most abundant element in rice was Mg (414‒561 mg kg-1) followed by Fe (49.4‒168 mg kg-1), while in soil was Fe (11674‒12917 mg kg-1) followed by Mg (619‒709 mg kg-1). Chromium, Cd and Pb were all below the limit of quantitation of the method. The concentrations of the elements, except Zn in rice and Fe in soil, varied significantly (p < 0.05) with the growing region. Notably, rice from Fogera contained more than double Fe, while from Pawe less than half Cu than from the other region. Soils from the rice fields of Pawe, generally, had lower levels of the elements than from the other regions. The order of the abundances of the elements in soil was reflected in the rice grains, except for the reversal between Fe and Mg. However, elemental concentrations were higher in soil than in rice, indicating the absence of bioaccumulation by the rice grains. Furthermore, only copper exhibited a strong positive correlation (r = 0.991) between the rice grain and soil.

Multielement analysis coupled with chemometrics modelling for geographical origin classification of teff [Eragrostis tef (Zuccagni) Trotter] grains from Amhara Region, Ethiopia.

Teff [Eragrostis tef (Zuccagni) Trotter] is an indigenous crop in Ethiopia, and Amhara region is the predominant teff producing region in the country. This study was aimed at developing an analytical methodology useful to determine the geographical origin of teff produced in the Amhara Region, based on multielement analysis combined with multivariate statistical techniques. For this, a total of 72 teff grain samples were collected from three zones (West Gojjam, East Gojjam, and Awi) and analysed for K, Na, Mg, Ca, Mn, Cu, Fe, Co, Ni, Zn, Cr, and Cd contents using inductively coupled plasma-optical emission spectroscopy (ICP-OES). The digestion and ICP-OES analysis method were accurate, with percentage recovery ranging 85.5 to 109% across the different metals analysed. Principal component analysis (PCA) and linear discriminant analysis (LDA) were applied to discriminate samples based on their production regions. Magnesium, Ca, Fe, Mn, and Zn were the most discriminating elements among the samples. The LDA model provided 96% correct classification of samples into production regions and varietal types, with an average prediction ability of 92%. Hence, the multielement analysis combined with statistical modeling can be used in the authentication of the geographical origin and varietal type of teff from Amhara region.

Trace Element Levels in Nails of Residents of Addis Ababa Are Shaped by Social Factors and Geography.

The Akaki catchment in Ethiopia is home to Addis Ababa and about five million people. Its watercourses receive a variety of wastes released by the residents and industries. River water is being used for irrigation, livestock watering, and other domestic purposes. This study tested the hypothesis that the river pollution would be reflected in higher levels of trace elements in the nails of residents living in Akaki-Kality Sub-City in the downstream, as compared to those living in Gullele Sub-City in the upstream of the Akaki catchment. Samples were taken and subsequently analysed for metals using inductively coupled plasma optical emission spectrometry (ICP-OES). The mean concentrations of Fe, Zn, Cu, Mn, Ni, Cr, Pb, and As in nails from Akaki-Kality were 488 ± 49, 106 ± 10, 5.2 ± 0.3, 13 ± 1.5, 11 ± 8, 2.2 ± 0.3, 0.09 ± 0.01, and 0.16 ± 0.01 µg/g, respectively. Likewise, the concentrations of Fe, Zn, Cu, Mn, Ni, Cr, Pb, and As in nails from Gullele were 1035 ± 135, 251 ± 10, 6.6 ± 0.4, 31 ± 3.7, 7.4 ± 1.7, 2.0 ± 0.3, 0.63 ± 0.01, and 0.25 ± 0.01 µg/g, respectively. Co and Cd were not detected. Contrary to the initial hypothesis, higher metal levels were found in nails of residents living in the upstream rather than the downstream area of the catchment. In particular, the concentrations of Fe (p = 0.000), Zn (p = 0.01), and Mn (p = 0.000) were significantly elevated in nails from Gullele and also high in comparison with internationally reported values. Besides, geography and social factors, especially education level, correlated to trace metals in nails. Most of the elements were significantly lower in the nails of individuals with a university degree compared to those who were illiterate or only completed primary school.

Urinary 8-OHdG level is not affected by geography and trace elements in nail of residents of Addis Ababa: It is shaped by interactions between different social factors.

The objective of this study was to evaluate the association between exposure to heavy metals and oxidative DNA damage among residents living in the potentially more polluted downstream (Akaki-Kality) area of Addis Ababa, in comparison to the upstream area (Gullele). For this, 8-hydroxy-2'-deoxyguanosine (8-OHdG) was used as a biomarker for oxidative DNA damage and heavy metals (Fe, Zn, Mn, Cu, Ni, Cr, Pb, As) as indicators of exposure. The concentrations of heavy metals in nails were determined using inductively coupled plasma optical emission spectroscopy (ICP-OES), and 8-OHdG in urine using Enzyme-Linked with Immunosorbent Assay (ELISA), from 95 residents of the two areas, upstream and downstream. The urinary 8-OHdG concentration was not significantly different (p = 0.05) between the two Sub-Cities, with mean of 18.50 ± 4.37 ng/mg Creatinine in Akaki-Kality and 17.30 ± 5.83 ng/mg Creatinine in Gullele. Also, there were no statistically significant (p = 0.05) difference among the different demographic groups according to gender, age, educational status, body mass index or habit of alcohol consumption. However, the interactions of sex with age, sex with alcohol consumption and alcohol consumption with education were found to affect the urinary 8-OHdG levels of residents of the two Sub-Cities. The mean concentrations (µg/g) of the elements were 488 and 1035 for Fe, 106 and 251 for Zn, 13.0 and 31.2 for Mn, 5.23 and 6.63 for Cu, 11.2 and 7.39 for Ni, 2.23 and 2.02 for Cr, 0.09 and 0.63 for Pb; and 0.16 and 0.25 for As, in nail samples from Akaki-Kality and Gullele, respectively. The determined concentrations of the heavy metals in nails were not significantly associated (p = 0.05) with the corresponding urinary levels of 8-OHdG. Hence, the observed 8-OHdG might have been caused by environmental exposure to toxic substances other than the analyzed heavy metals.

Absence of significant association of trace elements in nails with urinary KIM-1 biomarker among residents of Addis Ababa in Upper Awash Basin, Ethiopia: a cross-sectional study.

The Akaki River in the Upper Awash Basin, which flows through Addis Ababa, the capital city of Ethiopia, has been highly polluted by sewage from factories and residential areas. A population-based cross-sectional study was used to assess the association between trace elements and kidney injury from residents living in polluted areas downstream (Akaki-Kality) versus upstream (Gullele) in Sub-Cities of Addis Ababa. A total of 95 individuals (53 from Akaki-Kality and 42 from Gullele) were included in the study. Kidney injury molecule 1 (KIM-1), lead, arsenic, cadmium, cobalt, lead, manganese, zinc, iron, copper, chromium and nickel were evaluated in residents' urine and nail samples. A large proportion (74%) of the sample population contained KIM-1, including 81% residents in Akaki-Kality and 64% residents in Gullele. KIM-1 was, however, not significantly different (p = 0.05) between the two Sub-Cities, with median of 0.224 ng/mL in Akaki-Kality and 0.152 ng/mL in Gullele. Most of the analyzed elements, except Pb, As, Cd and Co, were found in all of the nail samples, with median (µg/g) in the range of 442‒714 Fe, 97.0‒246 Zn, 11.6‒24.1 Mn, 4.49‒5.85 Cu, 1.46‒1.66 Cr and 1.22‒1.41 Ni. The high incidence of KIM-1 indicates a potential for long term renal tubular damage among residents of the Sub-Cities. The concentrations of the elements in nails were, however, not significantly associated (p = 0.05) with the corresponding levels of KIM-1 in urine. Hence, the observed KIM-1 might be related to exposure to toxic substances or factors other than those included in this study.

Elemental and Proximate Compositions of Sesame Seeds and the Underlying Soil from Tsegede, Ethiopia.

The proximate compositions and elemental contents of sesame (Sesamum indicum) seeds and the underlying soil from different cultivation areas of Tsegede, Ethiopia, were investigated. The ash, protein, fiber, fat, and carbohydrate contents of the sesame seeds were determined following standard methods. Essential major (Ca and Mg) and trace metals (Fe, Zn, Cu, Mn, and Ni) in the sesame seeds and the underlying soil were determined by using flame atomic absorption spectroscopy. The sesame seeds contained high levels of fat (52.9 ± 1.5%), followed by protein (23.5 ± 0.9%). The seeds contained 525 ± 1 and 453 ± 38 mg/kg of Mg and Ca, respectively. Iron was the most abundant (37.8 ± 1.4 mg/kg) of the trace metals, followed by Zn (14.6 ± 2.2 mg/kg) and Cu (7.26 ± 0.84). Manganese and Ni were found only in minute amounts. The concentrations of the trace metals varied significantly across the different cultivation areas. Similar to the sesame seeds, iron was found in higher amounts (212.6 ± 2.6 mg/kg) in the underlying soil followed by Zn and Cu, which were both 28.8 mg/kg. The amounts of the trace elements, Fe, Cu, and Zn found in the soils were about 2 to 6 times higher than that found in the sesame seeds. Whereas, the concentrations of Mg and Ca present in the seeds were comparable with that determined in the soils. Correlation analysis indicated that the Fe and Cu contents of the sesame seeds are negatively influenced by the amounts of Mg and Ca present in the soil. Ca in the seeds was also negatively associated with the Mg levels in soil.

Determination and health risk assessment of trace elements in the tap water of two Sub-Cities of Addis Ababa, Ethiopia.

Water is an essential component of all living things on earth and the contamination of water by heavy metals can cause detrimental health effects. This study aimed to determine the health risk posed by trace elements (Fe, Zn, Cu, Mn, Ni, Cr, Cd, Co, Pb, and As) present in the drinking water supplies of Gullele and Akaki-Kality Sub-Cities, upstream and downstream parts of Addis Ababa, respectively. The concentrations of the potentially toxic trace elements in the water samples were determined using inductively coupled plasma optical emission spectroscopy (ICP-OES). The highest concentration of the heavy metals was observed for Iron. Cadmium and cobalt were not detected in any of the tap water samples. Samples from Gullele contained higher levels of Fe and Mn, 220.3 ± 0.17 and 19.78 ± 0.08 µg/L, respectively compared to Akaki-Kality, 38.87 ± 0.14 and 2.08 ± 0.01 µg/L, respectively. Conversely, tap water from Akaki-Kality contained significantly higher levels of As than that from Gullele. Additionally, Cr and Ni were detected only in samples from Akaki-Kality, which might be due to the various industries in the area. The highest incremental lifetime cancer risk was found for arsenic, with values for children and adults in Akaki-Kality 2.50 × 10-4 and 4.50 × 10-4, respectively. Likewise, in Gullele Sub-City, it was 5.00 × 10-5 and 1.00 × 10-4 for adults and children, respectively. The results indicate that carcinogenic risk occurrence is probable from As in both studied areas.

Combining Multi-Element Analysis with Statistical Modeling for Tracing the Origin of Green Coffee Beans from Amhara Region, Ethiopia.

Characterization of coffee terroirs is important to determine authenticity and provide confidence for consumers to select the right product. In this regard, Amhara Region, which is located at the northwestern part of Ethiopia, produces various local coffee types with distinct cup qualities. The coffees are, however, not yet registered with certification marks or trademarks for indications of their geographical origins. This study was aimed at developing analytical methodology useful to determine the geographical origin of green coffee beans produced in Amhara Region based on multi-element analysis combined with multivariate statistical techniques. For this, a total of 120 samples of green coffee beans, collected from four major cultivating zones (West Gojjam, East Gojjam, Awi, and Bahir Dar Especial Zones) were analyzed for K, Mg, Ca, Mn, Fe, Cu, Fe, Co, Ni, Zn, Si, Cr, Cd, and Pb using inductively coupled plasma-optical emission spectroscopy. The elemental analysis data were subjected to principal component analysis (PCA) and linear discriminant analysis (LDA). PCA was used to explore the natural groupings of samples and the discriminatory ability of elements. Accordingly, the elements K, Mg, Ca, and Na were found to be the main discriminators among samples. LDA provided a model to classify the coffee samples based on their production zones with an accuracy of 94.2% and prediction ability of 93.4%. Thus, the elemental composition of green coffee beans can be used as a chemical descriptor in the authentication of coffee produced in Amhara Region.

GC-MS profiling of fatty acids in green coffee (Coffea arabica L.) beans and chemometric modeling for tracing geographical origins from Ethiopia.

BACKGROUND: This study was aimed at the development of objective analytical method capable of verifying the production region of the coffee beans. One hundred samples of green coffee (Coffea arabica L.) beans from the major producing regions, comprising various sub-regional types, were studied for variations in their fatty acid compositions by using gas chromatography coupled with mass spectrometry. Principal component analysis (PCA) was used to visualize data trends. Linear discriminant analysis (LDA) was used to construct classification models. RESULTS: Twenty-one different fatty acids were detected in all of the samples. The total fatty acid content varied from 83 to 204 g kg-1 across the regions. Oleic, linoleic, palmitic, stearic and arachidic acids were identified as the most discriminating compounds among the production regions. The recognition and prediction abilities of the LDA model for classification at regional level were 95% and 92%, respectively, and 92% and 85%, respectively, at sub-regional level. CONCLUSION: Fatty acids contain adequate information for use as descriptors of the cultivation region of coffee beans. Chemometric methods based on fatty acid composition can be used to detect fraudulently labeled coffees, with regard to the production region. These can benefit the coffee production market by providing consumers with products of the expected quality. © 2019 Society of Chemical Industry.

Single-step isolation of embelin using high-performance countercurrent chromatography and determination of the fatty acid composition of seeds of Embelia schimperi.

Embelin (2,5-dihydroxy-3-undecyl-p-benzoquinone) is known for its potent anthelmintic activity, but also for wound-healing, antidiabetic, anticonvulsant, antitumour, anti-inflammatory, analgesic, hepatoprotective, antioxidant, antibacterial and antispermatogenic activities. A high-performance countercurrent chromatography method was developed for the purification of embelin from an extract of the seeds of Embelia schimperi fruit. The optimized solvent system (n-hexane-ethylacetate-ethanol-water, 7:3:7:3) resulted in the isolation of 13.9 mg of embelin, directly from 100 mg of crude extract, in a single step within a short time (40 min). Although the compound appeared to be completely pure when analysed by ultra-performance liquid chromatography (UPLC) with photo diode array detection, the purity was established as ~90% by UPLC-mass spectrometry. Furthermore, we report the fatty acid composition of the seeds of E. schimperi fruit. Nine fatty acids were quantified from the fruit seed extract by gas chromatography-mass spectrometry, with linoleic (46.4%), palmitic (21.5%) and oleic (19.6%) acids making up the largest proportions.

Levels of selected metals in leaves of Cannabis sativa L. cultivated in Ethiopia.

BACKGROUND: Cannabis sativa L. is one of the illicit drug bearing plants. Cannabis products are the most widely trafficked drugs worldwide. The highest levels of cannabis production in the world take place in the African continent. A small volume of cannabis is produced in rural areas of Ethiopia, of which a small portion is exported to neighboring countries and the majority is consumed at home. The literature survey revealed that there is no report on the metal contents in cannabis cultivated in Ethiopia. The main objective of this study is to determine the level of selected metals in leaves of Cannabis sativa L. cultivated in Ethiopia. METHODS: Cannabis sativa L. samples were collected from Metema (Amhara Region), Mekelle (Tigray Region), Sheshemene (Oromia Region) and Butajira (South Nations Nationality and Peoples (SNNP) Region) of Ethiopia. After proper sample pretreatment, the volumes of reagents used, digestion temperature and digestion time were optimized and using the optimized conditions the levels of metals were determined by flame atomic absorption spectrometry. RESULTS: The accuracy of the optimized procedure was evaluated by analyzing the digest of the spiked samples with standard solution and the percentage recoveries varied from 88 to 103%. The levels of metals determined (µg/g dry weight) were in the ranges Ca (657-1,511), Zn (321-380), Ni (124-172), Cu (122-176), Cd (3-10), Pb (8-10), and Cr (4-8). Zn was with the highest concentration among trace metals. CONCLUSION: A statistical analysis of variance (ANOVA) at 95% confidence level indicated that there is a significant difference in the levels of all the metals between the four sample means except Pb. The results indicate that the content of Pb and Cd exceeds the permissible amount for medicinal plants which form the raw materials for the finished products set by World Health Organization (WHO).