Isolierung, Strukturaufklärung, Quantifizierung und Formulierung der Saponine und Flavonoide der Samen von Glinus Lotoides

Suitable extraction methods for the seeds of Glinus lotoides have been developed and the major saponins and flavonoids have been isolated. In this, four new hopane-type saponins, glinusides F, G, H and K, and the known succulentoside B as well as the two known flavones, 5,7,4'f-trihydroxyflavone-6,8-di-C-glucoside (vicenin-2) and 5,7,4'f-trihydroxy-flavone-8-C-sophoroside (vitexin-2''-O-glucoside), have been isolated from the seeds of the plant. Based on the spectral analyses including 2D NMR and HRESI mass spectroscopy, the new structures have been characterized as 3alpha-O-alpha-Dxylopyranosyl-6beta-O-alpha-D-xylopyranosyl-16alpha-O-alpha-D-xylopyranosyl-22-hydroxy-hopane (glinuside F), 3alpha-O-beta-L-rhamnopyranosyl-(1->2)-alpha-D-xylopyranosyl-6beta,16alpha-dihydroxy-22-O-beta-L-rhamnopyranosyl-hopane (glinuside G), 3alpha-O-beta-l(TM)hamnopyranosyl-( 1->2)-alpha-D-xylopyranosyl-6beta-O-alpha-D-xylopyranosyl-16alpha-hydroxy-22-O-beta-L-rhamnopyranosyl-hopane (glinuside H) and 3alpha-O-beta-L-rhamnopyranosyl-(1->2)-alpha-D-xylopyranosyl-6beta-O-alpha-D-xylopyranosyl-16alpha-O-alpha-D-xylopyranosyl-22-hopane (glinuside I). The total flavonoids and saponins of the seeds of G. lotoides in the crude extracts and tablet formulations thereof have been quantified by reversed-phase high performance liquid chromatographic (RP-HPLC) methods with UV detection. The saponins were analyzed after acid hydrolysis in 3M HCl at 100 °C for 1 hour. Vicenin-2 and mollugogenol B have been isolated and used as reference substances for the quantification of total flavonoids and saponins, respectively. The identities of vicenin-2 and mollugogenol B have been confirmed using UV, MS and NMR spectral analyses and comparison with respective published data. The purity of the isolated mollugogenol B has been determined by TLC, HPLC and UV spectrophotometry as 98%, calculated from HPLC peak area. The molar absorptivity of the methanol solution of mollugogenol B at a concentration of 0.948 mg/100ml was found to be 3.98, 4.16 and 4.12 at ƒÉmax of 260sh, 251 and 243 nm, respectively, which is in good agreement with literature values, indicating the purity of mollugogenol B. Similarly, the purity of vicenin-2 has been determined as 97% using HPLC, which is confirmed by TLC and UV methods. Satisfactory separation of the components of the flavonoids and saponins compounds has been obtained in less than 30 and 25 min, for the flavonoids and saponins, respectively. The HPLC methods were validated for linearity, repeatability, limits of detection (LOD) and quantification (LOQ). Repeatability (inter- and intra-day, n = 6 and 9, respectively) showed less than 2% relative standard deviation (RSD). The LOD and LOQ were found to be 0.075 and 0.225 mg/mL, respectively, for vicenin-2 and 0.027 and 0.082 mg/100mL, respectively, for mollugogenol B. The total amounts of saponins of the crude extract and tablet formulation have been determined as glinuside G equivalent (mol. Wt. 900) which approximates the average molecular weight of the saponins of G. lotoides. The content of flavonoids and saponins of six single tablets has been found to be between 95 and 103% for flavonoids and 94 to 98% for saponins. The validated HPLC methods could be employed to standardize a laboratory produced purified extract, which could be used as a secondary standard for the routine quality control. Accordingly, the secondary standard (extract F) has been found to contain 10% vicenin-2 and 21.3% total flavonoids calculated as vicenin-2 equivalent. The amount of saponins in the secondary standard was 6.3% mollugogenol B and 12.4% total aglycons calculated as mollugogenol B, 14.2% glinuside G and 25.4% total saponins calculated as glinuside G equivalent. Admixing of the crude liquid extract of the seeds of G. lotoides with Aeroperl(TM) 300 Pharma, as an inert carrier material, and subsequent drying the mixture has provided a non-adherent and free-flowing powder. The required amount of Aeroperl(TM) 300 Pharma has been optimized as 30%, based on the physicochemical properties such as particle size distribution, surface morphology, moisture content and sorption kinetics of the dry extract preparations containing 10, 20, 30, 40, 50 and 60% w/w Aeroperl(TM) 300 Pharma. A tablet formulation containing granules of the dry extract preparation (947 mg), Avicel(TM) PH101 (363 mg) and Ac-Di-sol (90 mg) provided the best tablet properties such as disintegration time of 2.4 min at a tablet hardness of 73 N. Granulation by roller compaction of the dry extract preparation has improved the disintegration time of the tablets. Oblong tablets which could administer the traditional dose as a single tablet have been developed and enteric coated. Enteric coated tablets of G. lotoides with pharmacopoeial requirements could be prepared using methacrylic acid/ ethylacrylate co-polymer (Eudragit(TM) L 100/55 or Kollicoat(TM) MAE 100P). Propylene glycol and acetyl tributyl citrate could be used as plasticizers at concentrations of about 11% in the dry film.Glinus lotoides wird in Athiopien traditionell als Anthelminthikum eingesetzt. Das Ziel der vorliegenden Arbeit ist es, Auszuge von G. lotoides herzustellen, diese zu charakterisieren und zu standardisieren. In einem weiteren Abschnitt sollen aus den Auszügen Arzneiformulierungen hergestellt werden. Um die Ziele der Arbeit zu erreichen, muss ein standardisiertes Extraktionsverfahren entwickelt werden. Des Weiteren ist ein umfangreiches Screening der Inhaltsstoffe durchzuführen, was die Isolierung und Identifizierung der pharmakologisch aktiven Substanzen einschliesst. Die Isolierung und die Strukturaufklarung der Saponine und der Flavonoide als wirksame Inhaltsstoffgruppen von G. lotoides liefern vier neue Hopan-Saponine, die Glinuside F, G, H und K, das bekannte Succulentosid B sowie zwei bekannte Flavonoidglykoside; 5,7,4'-trihydroxyflavon-6,8-di-C-glucosid (Vicenin-2) und 5,7,4'-trihydroxy-flavon-8-C-sophorosid (Vitexin-2"-O-glucosid). Die Strukturen der Saponine werden mittels NMR, HRESI Massenspektroskopie und Zucker-Analytik als 3beta-O-beta-D-xylopyranosyl-6alpha-O-beta-D-xylopyranosyl-16beta-O-beta-D-xylopyranosyl-22-hydroxy-hopan (Glinusid F), 3beta-O-alpha-L-rhamnopyranosyl-(1->2)-beta-D-xylopyranosyl-6alpha,16beta-dihydroxy-22-O-alpha-L-rhamnopyranosyl-hopan (Glinusid G), 3beta-O-alpha-l(TM)hamnopyranosyl-( 1->2)-beta-D-xylopyranosyl-6alpha-O-beta-D-xylopyranosyl-16beta-hydroxy-22-O-alpha-L-rhamnopyranosyl-hopan (Glinusid H) and 3beta-O-alpha-L-rhamnopyranosyl-(1->2)-beta-D-xylopyranosyl-6alpha-O-beta-D-xylopyranosyl-16beta-O-beta-D-xylopyranosyl-22-hopan (Glinusid I) aufgeklart. Die Gesamtsaponin- und Favonidgehalte der Samen von G. lotoides werden mittels RPHPLC in den Extrakten und in den Tabletten bestimmt. Die quantitative Bestimmung der Saponine wird nach saurer Hydrolyse (3M HCl, 100 °C fur 1 Stunde) durchgeführt, wobei Mollugogenol B entstehen. Für die quantitative Bestimmung der Flavonoide wird Vicenin-2 als externer Standard eingesetzt. Mittels DC und HPLC unter verschiedenen Bedingungen wird eine mindestens 97 %ige Reinheit des isolierten Vicenin-2 und Mollugogenol B gefunden. Die Berechnung der Konzentrationen erfolgt über die Peakflachen von Vicenin-2 und von Mollugogenol B nach der Methode des externen Standards. Der Gesamtflavonoidgehalt und der Gesamtsaponingehalt von G. lotoides werden über die Gesamtpeakfläche des jeweiligen Chromatogramms ermittelt. Im Rahmen der Validierung der Gehaltsbestimmungsmethode erfolgt die Bestimmung von Selektivität, Wiederfindung, System- und Methodepräzision, Bestimmungsgrenze und Nachweisgrenze. Als Sekundärstandard wird der aufgereinigte Extrakt F charakterisiert. Der Flavonoid- und Saponingehalt der Tabletten beträgt zwischen 95 und 103 % für die Flavonoide und zwischen 94 und 98 % für die Saponine. Eine zur Formulierung von Tabletten geeignete Extraktzubereitung wird durch Mischung des Extraktes mit Aeroperl(TM) 300 Pharma als Träger-Material hergestellt. Die benötigte Menge von Aeroperl(TM) 300 Pharma wird durch Untersuchungen zur Wasseraufnahme und REM-Untersuchungen optimiert. Hierbei ergibt eine Extraktzubereitung mit 30% (m/m) Aeroperl(TM) 300 Pharma die besten Eigenschaften im Hinblick auf die weitere Verarbeitung. Zusätzlich, durch Walzen-Kompaktierung der Extrakt-Aeroperl-Mischung wird die Fliessfähigkeit der Tablettiermischung und der Zerfall der Tabletten verbessert. Eine geeignete Tablettenformulierung, die 947 mg Extraktzubereitung, 363 mg Avicel(TM) PH101, und 90 mg Ac-Di-sol enthält, wird durch mehrere Optimierungsschritte entwickelt. Diese Tabletten zeigen einen Zerfall der Tabletten innerhalb von 2,4 Minuten bei einer Harte von 73 N. Zusätzlich erhalten die Oblong-Tabletten einen Magensaftresistenten Überzug, der den Arzneibuchanforderungen entspricht. Dieser Überzug wird mittels der hier für geeigneten Filmbildner Eudragit(TM) L 100/55 oder Kollicoat(TM) MAE 100P hergestellt. Der Filmüberzug enthält Propylenglykol und Acetyltributylcitrat als Weichmacher. Durch die Prüfung der Zerfallzeit wird sichergestellt, dass die überzogenen Tabletten in künstlichen Magensaft Ph. Eur. (0,1 M Salzsäure, pH 1,0) innerhalb 2 Stunden nicht zufallen jedoch anschliessend in künstlichen Darmsaft (Phosphat-Pufferlosung, pH 6,8) innerhalb von 15 Minuten zufallen. Zusammenfassend kann festgestellt werden, dass es gelungen ist, einen Auszug von G. lotoides herzustellen, dieser zu einer zur Formulierung von Tabletten geeigneten Extraktzubereitung zu verarbeiten, eine Tablettenformulierung zu entwickeln und die Tabletten mit einem magensaftresistenten Überzug zu versehen

Food insecurity in Farta District, Northwest Ethiopia: A community based cross–sectional study

Background Access to sufficient food is essential for household welfare as well as for accomplishing other development activities. Households with insufficient access to food often face other challenges related to food insecurity including poor health and a decline in productivity. These challenges can often create a vicious circle whereby households are unable to produce enough food even during a good crop season. Thus, this study aimed to determine the magnitude of food insecurity and its determinants in rural households of Farta District, Northwest Ethiopia. Methods A community based cross-sectional study was conducted from September to October 2012. Household heads were recruited using a multistage random sampling technique. Data were collected by face-to-face interviews using the Household Food Insecurity Access Scale (HFIAS) tool after verbal informed consent. Data were entered to Epi info 2002 and exported to SPSS version 16 for analysis. Multiple logistic regressions were fitted and odds ratios with 95% confidence intervals were calculated to identify associated factors and control confounding effect. Results A total of 836 households were included in this study. Nearly three quarters of the households (70.7%) had food insecurity. Households headed by females (AOR = 3.18, 95% CI:1.08, 15.21), lack of education (AOR = 2.59, 95% CI: 1.46, 4.60), family size of 4-7 (AOR = 2.39, 95% CI: 1.21,4.70), family size of >7 (AOR = 13.23,95% CI:6.18, 28.32), few or absence of livestock (AOR = 5.60, 95% CI:1.28, 24.43), absence of income from off-farm activities (AOR = 3.12, 95% CI:1.53, 6.36), lack of irrigation (AOR = 3.54, 95% CI:2.14, 5.18) and lack of perennial income (AOR = 3.15, 95% CI:1.88, 5.27) were factors associated with food insecurity. Conclusions This study revealed that most households of the district were food insecure. Hence, the promotion of contraceptive use, off-farm employment activities and the development of small scale irrigation are important recommendations to reduce food insecurity

Frequency and percentage of ANC visits among urban and rural residents in Ethiopia in 2019.

Frequency and percentage of ANC visits among urban and rural residents in Ethiopia in 2019.</p

DHS datasets authorization letter.

BackgroundAlthough there have been consistent improvements in maternal mortality, it remains high in developing countries due to unequal access to healthcare services during pregnancy and childbirth. Thus, this study aimed to further analyze the variations in the number of antenatal care utilizations and associated factors among pregnant women in urban and rural Ethiopia.MethodsA total of 3962 pregnant women were included in the analysis of 2019 Ethiopian Demographic and Health Survey data. A negative binomial Poisson regression statistical model was used to analyze the data using STATA version 14.0. An incident rate ratio with a 95% confidence interval was used to show the significantly associated variables.ResultsOf the 3962 (weighted 3916.67) pregnant women, about 155 (15.21%) lived in urban and 848 (29.29%) rural residences and did not use antenatal care services in 2019. Women age group 20–24 (IRR = 1.30, 95%CI:1.05–1.61), 25–29 (IRR = 1.56, 95%CI:1.27–1.92), 30–34 (IRR = 1.65, 95%CI:1.33–2.05), and 35–39 years old (IRR = 1.55, 95%CI:1.18–2.03), attending primary, secondary, and higher education (IRR = 1.18, 95%CI:1.07–1.30), (IRR = 1.26, 95%CI:1.13–1.42) and (IRR = 1.25, 95%CI:1.11–1.41) respectively, reside in middle household wealth (IRR = 1.31, 95%CI:1.13–1.52), richer (IRR = 1.45, 95%CI:1.26–1.66) and richest (IRR = 1.68, 95%CI:1.46–1.93) increases the number of antenatal care utilization among urban residences.While attending primary (IRR = 1.34, 95%CI:1.24–1.45), secondary (IRR = 1.54, 95%CI:1.34–1.76) and higher education (IRR = 1.58, 95%CI:1.28–1.95), following Protestant (IRR = 0.76, 95%CI:0.69–0.83), Muslim (IRR = 0.79, 95%CI:0.73–0.85) and Others (IRR = 0.56, 95%CI:0.43–0.71) religions, reside in poorer, middle, richer, and richest household wealth (IRR = 1.51, 95%CI:1.37–1.67), (IRR = 1.66, 95%CI:1.50–1.83), (IRR = 1.71, 95%CI:1.55–1.91) and (IRR = 1.89, 95%CI:1.72–2.09) respectively, being married and widowed/separated (IRR = 1.85, 95%CI:1.19–2.86), and (IRR = 1.95, 95%CI:1.24–3.07) respectively were significantly associated with the number of antenatal care utilization among rural residences.ConclusionThe utilization of antenatal care is low among rural residents than among urban residents. To increase the frequency of antenatal care utilization, health extension workers and supporting actors should give special attention to pregnant women with low socioeconomic and educational levels through a safety-net lens.</div

Description of individual and household-level variables.

Description of individual and household-level variables.</p

A histogram showing the number of antenatal care visits among urban residents in Ethiopia, 2019.

A histogram showing the number of antenatal care visits among urban residents in Ethiopia, 2019.</p

Factors associated with the number of ANC service utilizations between urban and rural communities in Ethiopia, 2019.

Factors associated with the number of ANC service utilizations between urban and rural communities in Ethiopia, 2019.</p

STROBE-checklist.

BackgroundAlthough there have been consistent improvements in maternal mortality, it remains high in developing countries due to unequal access to healthcare services during pregnancy and childbirth. Thus, this study aimed to further analyze the variations in the number of antenatal care utilizations and associated factors among pregnant women in urban and rural Ethiopia.MethodsA total of 3962 pregnant women were included in the analysis of 2019 Ethiopian Demographic and Health Survey data. A negative binomial Poisson regression statistical model was used to analyze the data using STATA version 14.0. An incident rate ratio with a 95% confidence interval was used to show the significantly associated variables.ResultsOf the 3962 (weighted 3916.67) pregnant women, about 155 (15.21%) lived in urban and 848 (29.29%) rural residences and did not use antenatal care services in 2019. Women age group 20–24 (IRR = 1.30, 95%CI:1.05–1.61), 25–29 (IRR = 1.56, 95%CI:1.27–1.92), 30–34 (IRR = 1.65, 95%CI:1.33–2.05), and 35–39 years old (IRR = 1.55, 95%CI:1.18–2.03), attending primary, secondary, and higher education (IRR = 1.18, 95%CI:1.07–1.30), (IRR = 1.26, 95%CI:1.13–1.42) and (IRR = 1.25, 95%CI:1.11–1.41) respectively, reside in middle household wealth (IRR = 1.31, 95%CI:1.13–1.52), richer (IRR = 1.45, 95%CI:1.26–1.66) and richest (IRR = 1.68, 95%CI:1.46–1.93) increases the number of antenatal care utilization among urban residences.While attending primary (IRR = 1.34, 95%CI:1.24–1.45), secondary (IRR = 1.54, 95%CI:1.34–1.76) and higher education (IRR = 1.58, 95%CI:1.28–1.95), following Protestant (IRR = 0.76, 95%CI:0.69–0.83), Muslim (IRR = 0.79, 95%CI:0.73–0.85) and Others (IRR = 0.56, 95%CI:0.43–0.71) religions, reside in poorer, middle, richer, and richest household wealth (IRR = 1.51, 95%CI:1.37–1.67), (IRR = 1.66, 95%CI:1.50–1.83), (IRR = 1.71, 95%CI:1.55–1.91) and (IRR = 1.89, 95%CI:1.72–2.09) respectively, being married and widowed/separated (IRR = 1.85, 95%CI:1.19–2.86), and (IRR = 1.95, 95%CI:1.24–3.07) respectively were significantly associated with the number of antenatal care utilization among rural residences.ConclusionThe utilization of antenatal care is low among rural residents than among urban residents. To increase the frequency of antenatal care utilization, health extension workers and supporting actors should give special attention to pregnant women with low socioeconomic and educational levels through a safety-net lens.</div

Sociodemographic characteristics disparity of antenatal care service utilization between urban and rural communities in Ethiopia, 2019.

Sociodemographic characteristics disparity of antenatal care service utilization between urban and rural communities in Ethiopia, 2019.</p

Study population and sampling procedures to identify the final sample size in the 2019 EMDHS.

Study population and sampling procedures to identify the final sample size in the 2019 EMDHS.</p