Extraction of industrial hemp (Cannabis sativa L.)

U okviru ovog rada izvršeno je ispitivanje različitih procesa ekstrakcije herbe industrijske konoplje. Za ekstrakciju određenih bioaktivnih jedinjenja primenjene su klasične tehnike ekstrakcije, maceracija i Soxhlet ekstrakcija, dok su od savremenih metoda primenjene ultrazvučna ekstrakcija, mikrotalasna ekstrakcija, ekstrakcija vodom u subkritičnom stanju i ekstrakcija ugljendioksidom u superkritičnom stanju. Praćenjem uticaja ulaznih promenjivih (procesnih parametara) na odabrane izlazne promenjive (sadržaj bioaktivnih jedinjenja ili antioksidativnu aktivnost) za svaku tehniku ekstrakcije pojedinačno utvrđeni su optimalni uslovi ekstrakcije. Nakon izvršenog poređenja odabrana je najadekvatnija tehnika ekstrakcije za pripremu tečnog ekstrakta koji će biti transformisan u suvi ekstrakt primenom metode sušenja raspršivanjem. Prilikom dobijanja suvog ekstrakta ispitan je uticaj različite koncentracije nosača na kvalitet ekstrakata i efikasnost sušenja. Dobijeni ekstrakti su okarakterisani u pogledu fizičkih i hemijskih osobina. Sadržaj kanabinoida u tečnim ekstraktima suve herbe industrijske konoplje ispitanih u ovoj doktorskoj disertaciji je bio u opsegu od 0,4314 mg/mL CBD-a i 0,0364 mg/mL THC-a (u optimalnom ekstraktu dobijenom maceracijom) do 1,0420 mg/mL CBD-a i 0,0430 mg/mL THC-a (u optimalnom ekstraktu dobijenom mikrotalasnom ekstrakcijom). U ekstraktima koji su nepolarnog karaktera i koji su dobijeni primenom Soxhlet ekstrakcije sadržaj CBD-a i THC-a je bio 64,40 i 2,90 mg/g, dok je primenom superkritične ekstrakcije dobijen sadržaj CBD-a i THC-a u opsegu od 76,1193 do 163,1111 mg/g i od 4,1746 do 6,5803 mg/g. Tečni ekstrakti dobijeni primenom vode i etanola kao ekstragensa bili su bogati flavonoidima i fenolima, a utvrđeno je i da imaju značajnu antioksidativnu aktivnost, dok su ekstrakti dobijeni Soxhlet ekstrakcijom i ekstrakcijom ugljendioksidom u superkritičnom stanju bili bogati lipofilnim jedinjenjima i terpenima. Ovako dobijeni ekstrakti predstavljaju visokovredne proizvode koji imaju veliki potencijal za upotrebu u farmaceutskoj industriji.Various extraction processes of the aerial parts of industrial hemp were examined. From classic extractions, maceration and Soxhlet extraction were applied, while from novel methods, ultrasound-assisted extraction, microwave-assisted extraction, subcritical water extraction and supercritical carbon dioxide extraction were applied. By monitoring the influence of the input variables on the selected output variables for each extraction, the optimal extraction conditions were determined separately. After a comparison of all the extraction processes tested, microwave-assisted extraction was selected to prepare the liquid extract for spray drying. The effect of different carrier concentrations was examined during the preparation of the dry extract. The extracts obtained were characterized in terms of physical and chemical properties. The content of cannabinoids in liquid extracts of dry Cannabis sativa herba examined ranged from 0.4314 mg/mL for CBD and 0.0364 mg/mL for THC in the optimum extract obtained by maceration to 1.0420 mg/mL for CBD and 0.0430 mg/mL for THC in the optimum extract obtained by microwave-assisted extraction. The CBD and THC content in extract obtained by Soxhlet extraction was 64.4000 mg/mL and 2.9000 mg/mL, respectively, while the highest content of CBD and THC in the supercritical carbon dioxide extract was 163.11 and 6.5803 mg/g. In addition to cannabinoids, the liquid extracts obtained were rich in phenols and flavonoids, and have considerable antioxidant activity, while the extracts obtained by Soxhlet extraction and supercritical carbon dioxide were rich in lipophilic compounds and terpenes. The extracts mentioned above are high value products which have great potential for implementation in the pharmaceutical industry

Effect of extraction solvent on total polyphenols content and antioxidant activity of Cannabis sativa L.

In this study, water and various concentrations of ethanol in water (30 %, 50 %, 70 %, and 90 %) were used as a solvent in the extraction of two different samples of hemp (Cannabis sativa L.). The extraction yield, total phenols content, total flavonoids content, antioxidant activity, and reductive capacity were determined in the obtained extracts. The extraction yield was from 8.16 to 19.56 %, the content of total phenols was in the range from 5.85 to 17.05 mg GAE/g dw, and content of total flavonoids was in the range from 5.85 to 9.25 mg CE/g dw. Antioxidant activity was tested by DPPH assay and EC50 values were from 0.1331 to 0.7563 mg/mL, while EC50 values obtained by reducing power test ranged from 0.4450 to 1.1980 mg/mL. Ethanol/water mixture (50 %) was determined to be the best solvent for the extraction of phenolic compounds from both hemp samples. Total phenols content in 50 % ethanolic extracts were 17.05 mg/g dw and 9.25 mg/g dw for young and mature hemp, respectively

Subcritical water extraction of antioxidants from aerial parts of hemp (Cannabis sativa L.)

Hemp (Cannabis sativa L.) is an herbaceous annual dioecious plant recognizable for their characteristic spiky leaves from the Cannabaceae family. Cannabis sativa L. is a plant with more than 480 compounds which can be divided into diverse phytochemical classes such as cannabinoids, terpenoids, flavonoids, noncannabinoid phenols, hydrocarbons, nitrogen-containing compounds, carbohydrates. The most studied class is the cannabinoids, but the presence of other bioactive components, especially polyphenols is signifiifcant. The extraction of polyphenolic compounds of hemp was carried out by subcritical water. The effects of different extraction temperatures (120⁰C – 220⁰C) on the extraction yield, total phenol content, total flavonoid content, and antioxidant activity, determinate by DPPH method, and reductive capacity, were investigated. Extraction time was 10 minutes. In obtained extracts content of total phenols was in the range from 1.00 to 2.26 mg GAE/mL extract, the content of total flavonoids was in the range from 0.48 to 0.81 mg CE/mL extract. Antioxidant activity was from 0.0006 to 0.0017 ml extract/ml reaction mixture, while the reductive capacity was from 0.0014 to 0.0058 ml extract/ml reaction mixture

Zapremina kokičavosti i prinos zrna u dialelnom setu linija kokičara

Popping volume and yield are traits caused by several heredity factors. It is difficult to obtain superior genotypes for both traits but it is possible to develop genotypes with good popping volume and satisfactory yield. The hybrid ZPPL2 x ZPPL5 was superior in yield, heterosis and SCA for both yield and popping volume. As inbred ZPPL4 in all combinations has a good value for popping volume could be concluded that this inbred may be used as parent in further crosses. Analysis of variance of the combining ability indicating significant SCA effect for grain yield, and significant both GCA and SCA effects for popping volume. Therefore it can be stated that yield is influenced by non-additive and popping volume by both additive and non- additive gene effects. Analysis of variance of genetic components for popping volume indicates that the additive as well as dominant components significantly affected the inheritance of this trait in popcorn. The objective of this study was to evaluate heteorsis and combing ability for grain yield and popping volume in dialalel set of six maize inbred lines.Zapremina kokičavosti i prinos su svojstva pod uticajem nekoliko naslednih faktora. Teško je dobiti superiorne genotipove za oba svojstva ali je moguće dobiti genotip sa dobrom zapreminom kokičavosti i zadovoljavajućim prinosom. Hibrid ZPPL2 x ZPPL5 je superioran za prinos, heterozis i PKS za prinos i zapreminu kokičavosti. Kako linija ZPPL4 u svim kombinacijama ima dobre vrednosti za zapreminu kokičavosti može se zaključiti da ova linija se može koristiti kao roditelj u budućim ukrštanjima. Analiza varijanse za kombinacionu sposobnost ukazuje na značajan PKS efekat za prinos zrna, značajan PKS i OKS efekat za zapreminu kokičavosti. Prinos je pod uticajem neaditivnih a zapremina kokičavosti i aditivnih i neaditivnih gena. Analiza varijanse genetičkih komponenti za zapreminu kokičavosti ukazuje da aditivne kao i dominatne komponente značajno utiču na nasleđivanje ovog svojstva kod kokičara. Cilj ovih istraživanja bio je ispitivanje heteorzisa i kombinacionih sposobnosti za prinos zrna i zapreminu kokičavosti u dialelnom setu šest linija kukuruza

Classical extraction of polyphenolic compounds from industrial hemp (Cannabis sativa L.)

Hemp (Cannabis sativa L.) is an herbaceous annual dioecious plant recognizable for their characteristic spiky leaves from the Cannabaceae family. C. sativa L. is a complex plant with more than 480 compounds which can be divided into diverse phytochemical classes such as cannabinoids, terpenoids, flavonoids, noncannabinoid phenols, hydrocarbons, nitrogen containing compounds, carbohydrates. The most studied class is the cannabinoids, but C. sativa is a significant source of polyphenols, also. The effect of different water/ethanol mixtures (30, 50, 70, and 90%) and pure water on the polyphenol content and antioxidant activity of C. sativa L. extracts obtained by classical extraction at room temperature for 24 h was investigated. Two different samples were used, aerial parts of young hemp and aerial parts of mature hemp. Young hemp present stage of plant growth before the plant has a reproductive organ, and a mature plant is one which is an incomplete stage of growth with flowers as reproductive organs. The extraction yield, qualitative and quantitative phenolic profiles, total phenols content, total flavonoids content, antioxidant activity, and reductive capacity were determinate in the obtained extracts. TP was from 5.76 to 17.05 mg GAE/g dw for aerial parts of young hemp, and from 5.00 to 10.48 mg GAE/g dw for aerial parts of mature hemp. TF content was from 2.37 to 4.82 mg CE/g dw and from 1.67 to 3.58 mg CE/g dw for aerial parts of young hemp and aerial parts of mature hemp, respectively. Sinapic acid, protocatechin acid, vanillic acid, syringic acid, epicatechin acid, ferulic acid, isovitexin, rutin, cinnamic acid, naringenin, and apigenin were detected in hemp extracts. Antioxidant activity in obtained extracts was determined by the DPPH assay. IC50 values were from 0.1600 to 0.6700 mg/mL and from 0.2500 to 1.7900 mg/mL for aerial parts of young and mature hemp, respectively. Reductive capacity was expressed by EC50 values. EC50 were in the range from 0.5800 to 0.9700 mg/mL for young hemp extracts, while for mature hemp extracts were from 0.7200 to 1.5800 mg/mL

Microwave-assisted extraction of cannabidiol and

Cannabis contains 66 cannabinoids which are divided into 10 subclasses: cannabigerol (CBG) type, cannabichromene (CBC) type, cannabidiol (CBD) type, Δ9 -tetrahydrocannabinol (THC) type, Δ8 -THC type, cannabicyclol (CBL) type, cannabielsoin (CBE) type, cannabinol (CBN) and cannabinodiol (CBND) types, cannabitriol (CBT) type, and miscellaneous types subclass. In industrial cannabis, the allowed content of THC, which is the carrier of psychoactive activity, is different in different countries. According to the European Monitoring Centre for Drugs and Drug Addiction in the European Union countries, THC is allowed to be 0.2% expressed on dry matter, while in Serbia it is allowed to contain 0.3%. In most countries of the European Union, cannabis is legalized for medical purposes. Some contries allow its use only in the form of ready-made preparations containing isolated or chemically synthesized certain cannabinoids, while others allow the use of the extract. The extraction of CBD and THC of industrial cannabis was carried out by a microwaveassisted technique, an emerging environmentally-friendly technology. The effects of different extraction parameters (ethanol concentration (30-70%), extraction time (10-30 min), and liquid/solid ration (5-15 mL/mg)) on the extraction of CBD and THC were investigated using a response surface methodology. In obtained extracts content of CBD was in the range from 0.034 to 0.064 mg/mL, and content of THC was in the range from 0.224 to 1.842 mg/mL. Experimental results were described by the second order polynomial model. Model was estimated using analysis of variance (ANOVA). The optimization process carried out in order to obtain the most optimal content of desired types of cannabinoids

Microwave-assisted extraction of antioxidants from the areal parts of cannabis and process modelling

Nowadays, cannabis (Cannabis sativa L.) is one of the most controversial medicinal plants. The usage of cannabis and its derived products is based on the biological activity of the present pharmacologically active compounds; firstly cannabinoids and then many others: phenols, terpenoids, fatty acids, tocopherols. Today, cannabinoids are in the focus of scientific and medical research. That is why this research was focused on phenols and other bioactives in cannabis. Microwave-assisted extraction was applied for the extraction of phenolic compounds from areal parts of cannabis, sort Helena. The effects of different extraction parameters (ethanol concentration (30-70 %), extraction time (10-30 min), and liquid/solid ratio (5-15 mL/mg)) on the extraction yield, total phenols, total flavonoid content, antioxidant activity, and reductive capacity were investigated as part of the analysis of the extraction process. Box-Behnken's design was chosen as the experimental design. The content of total phenols in extracts was from 10.34 to 15.79 mg EGK/g of the drug and the content of total flavonoids was from 4.97 to 9.68 mg CE/g of the drug. The experimental results were described by the second order polynomial model. The model was estimated using the analysis of variance (ANOVA). The optimization process was carried out for all parameters simultaneously, to obtain the maximum yields of polyphenol compounds, antioxidant activity, and reductive capacity

Dissolving and water absorption ability of spray-dried willow gentian extract

Roots of the willow gentian (Gentiana asclepiadea L., Gentianaceae) are used in folk medicine to treat gastrointestinal diseases.Spray drying is the most commonly used technique for microencapsulation of plant extracts in the pharmaceutical, cosmetic and food industries. The choosing of suitable carriers is one of the critical points in the development of microencapsulated powder with the desired physicochemical properties.The extract of the willow gentian roots was spray-dried using different carriers (maltodextrin (MD) and whey protein (WP)at concentrations of 20, 40, and 60%; pectin (P), starch (S), and gelatin (G) at concentrations of 2.5, 5, and 7.5%).The aim of this study was to examine the ability of the obtained powdersto dissolveand absorb water.For this purpose, the values of water solubility (WSI) and water absorption index (WAI) were determined.WSI is a measure of the powder degradation and dissolving in water, while WAI represents an indicator of water absorption ability. It is desirable that instant tea powders show high WSI and low WAI values.WSI values ranged from 71.15% (7.5% P) to 91.94% (7.5% G) indicating good powder disintegration properties. Concentration-dependent decrease in WSI values was observed for WP, P and S, while the opposite trend was observed in the case of MD.The carrier-free powdershowed relatively low WAI value (13.32%). Powders with 20% MD and 5% S showed slightly lower WAI values (12.77 and 12.72%, respectively). On the other hand, powders with 5 and 7.5% P showed remarkably high WAI values of 91.59 and 91.49%, respectively. A concentration-dependent increase in WAI was observed in powders encapsulated withMD and WP.All encapsulated powders showed acceptable disintegration properties (high WSI), while in terms of water absorption (low WAI) powders with 20% MD and 5% S were the most suitable