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  • Extraction, Fatty Acid Profile and Elemental Analysis of Gongronema latifolium (Utazi) Leaf

  • Photo S. I. Okonkwo* Photo V. E. Mmuo Photo E. C. Okafor Photo V.O. Offiah Photo C. K. Okonkwo Photo P. O. Okwuego Photo V. S. Okonkwo Photo A. T. Kene-Okonkwo

  • 1Departments of Pure and Industrial Chemistry, Chukwuemeka Odumegwu Ojukwu University, Uli, Anambra State, Nigeria.
    2Department of Chemistry, School of Science Laboratory Technology, Federal Polytechnic, Oko, Anambra State, Nigeria. 
    3Departments of Pure and Industrial Chemistry, Chukwuemeka Odumegwu Ojukwu University, Uli, Anambra State, Nigeria 
    4Departments of Pure and Industrial Chemistry, Chukwuemeka Odumegwu Ojukwu University, Uli, Anambra State, Nigeria
    5Department of Diagnostic Medical Sonography and Ultrasound Technology, Ace Institute of Technology, Elmhurst, New York, USA.
    6Departments of Pure and Industrial Chemistry, Chukwuemeka Odumegwu Ojukwu University, Uli, Anambra State, Nigeria
    7Department of Medical Biochemistry, Chukwuemeka Odumegwu Ojukwu University, Uli, Anambra State, Nigeria, ???
    8Tansian University Oba, Anambra State, Nigeria.

Abstract

Medicinal plants are well known support for traditional medicine system that are used to mitigate human diseases. The African continent and culture have provided evidence of the efficacy of indigenously grown plants as remedies against diseases. The effectiveness of these plants are as a result of the presence of secondary metabolites. In this study, the fatty acid profile, the active components and elemental (mineral) composition of Gongronema latifolium (Utazi) leaf were evaluated. Extraction was conducted using Ultrasound and Soxhlet extractor.The isolation and elemental compositions were determined using TLC and AAS respectively. The results of the fatty acid profile analysis indicated that oleic acid; (C18:1) was contained highest with a value of 21.61135 µg/mL ± 0.664, followed by palmitic acid (C16:0) which had 16.054 µg/mL ± 0.677, followed by mystic acid (C14:0) (14.5081 µg/mL ± 1.355) and the least contained fatty acid was docosahexaneoic acid (24:0) (7.2603 µg/mL ± 0.604). The total saturated fatty acid content of the leaf extract of Gongronema latifolium was 72.771 µg/mL, which was higher than 46.6336 µg/mL of the total amount of unsaturated fatty acid. The Retention factor (Rf) value of the active components in aqueous extract of the TLC analysis using acetone: water: ammonium in a ratio of 90:3:7 produced four components with four colours which were 1.32 (yellow), 1.24 (gray), 1.36 (greenr), and 1.40 (brown), respectively. The results of the elemental analysis showed that K was the highest contained element with 8.975 ppm, followed by S; (8.245 %), Zn; (2.4662 ppm), and Ca; (1.6035 ppm), respectively. The least contained elements were As; (0.014 ppm) and Pb; (0.00165 ppm), respectively. The results of the analysis suggest that Gongronema latifolium leaf contains a diverse range of fatty acids, reasonable concentration of essential elements, but with less quantities of some heavy metals which are potential health hazards upon accumulation in the body. Ultrasound Assisted Extraction gave a yield of Crude Extract which produced 12-15%(~19-24g) and Bioactive Compounds Flavonoids (Kaempferol, quercetin) Alkaloids, Saponins, Polyphenols, terpenoids.

Keywords

Fatty Acids, TLC, UAE, Elements, Gongronema latifolium, GC-FID, AAS, Soxhlet extractor, Extraction.

Introduction

The search for herbal cures using plants is a common practice in Africa, particularly in Nigeria, (Akpanabiatu et al., 2005).  Plants contain some organic compounds which provide definite physiological action in the human body, (Edoga et al., 2005). These compounds are synthesized by secondary metabolism of living organisms. Secondary metabolites are widely used in the human therapy, scientific research and countless other areas, (Vasu et al., 2009; Gabriel et al., 2007).  The availability of plants as direct therapeutic agents make them more attractive when compared to modern medicine, (Agbo and Ngogang, 2005; Agbo et al., 2005).  Gongronema latifolium (G. latifolium) is one of the plants believed to have enormous metabolites. It is a non-woody climbing herbaceous plant from the family of Asclepiadaceae, (Osuagwu et al., 2013). It is a perennial shrub, widespread in the tropical and subtropical regions, especially in Africa, (Morebise et al., 2002). In South-Eastern and South-Western Nigeria, G. latifolium is called ‘Utazi’ and ‘Arokeke’ respectively, (Agbo et al., 2005). The leaf blade is broadly ovate to almost circular with a deep cordate base and an acuminate apex (Balogun et al., 2016). G. latifolium is grown in family farms and use to prepare meals due to its nutritional importance, (Owu et al., 2012; Ugochukwu et al., 2003). It is taken as a purgative against intestinal worms, stomach pain (Onike, 2010), and fever, diarrhea and ulcers (Akuodor et al., 2010). G. Latifoluim leaves are used traditional by the Ikales of Ondo state of Nigeria to treat malaria, nausea and anorexia (Oliver, 1996; Morebise and Fafunso 1998; Morebise et al., 2002). It is widely used for the treatment of asthma (Essien et al., 2007). According to Schneider et al., (1993); Cora and Bruce, (2000); Nwanjo et al., (2006); Tripoli et al., (2007); Offor  et al; (2015); Mgbeje et al., (2019), the phytochemical studies of G. latifolium leaves showed the presence of glycosides, alkaloids, saponin, tannin, flavonoids, essential oils, and pregnanes. Egbung et al., (2011), further reported the presence of mineral elements (Cr, Cu, Se, Zn, Na, Ca, P, Co and Fe) and vitamins (A, C, riboflavin, niacin, amino acids and thiamine). It is also a rich source of iron, (Eleyinmi, 2007). In a separate studies conducted by Sylvester et al., (2015); Ugochukwu and Babady (2003); Edet et al., (2004); Edet et al., (2011), rats were subjected to Streptozotocin-induced diabetes mellitus, and treated with G. Latifolium leaf extracts. The results showed a lowered blood glucose of the diabetic rats. Edim et al., (2012); Adeleye et al., (2011); Enyi-Idor et al., (2012), reported that the aqueous and ethanolic extracts together with the essential oil from G. latifolium leaves showed moderate inhibitory activity against Staphylococcus sp., Escherichia coli, Salmonella sp., etc. Eze and Nwanguma (2013), proposed the potential of G. latifolium extract for food preservation. Fatty acids are important compounds of our diet, (Ahmad, 2017; De Vogli et al., 2011). Health benefits of fats and oils includes; cardiovascular, (Saravanan, et al., 2010), cognitive, (Yehuda et al., 1999; McNamara, 2006). Fats are made up of saturated and unsaturated fatty acids.  These saturated fats and trans fats, with sedentary life, are associated with an increase in obesity, which are risk factors for cardiovascular, hypertension, stroke, and coronary artery diseases, (Chauhan, et al., 2016). The consequences of unhealthy dietary guidelines are becoming a global issue (Bauer et al., 2012). WHO recommends that the replacement of saturated fatty acids such as palmitic acid; C16:0, with polyunsaturated fatty acids can decreases LDL (low-density lipoprotein) cholesterol levels and increase HDL (high-density lipoprotein), (WHO, 1996). A mineral is a chemical element required as an essential nutrient by organisms, other than C, H2, N2, O2. These elements are classified as minerals in the four groups of essential nutrients; the others are vitamins, essential fatty acids, and essential amino acids, (Msuya and Mamiro, 2018; Murray et al., 2010). Important trace elements, necessary for mammalian life, includes Fe, Co, Cu, Zn, Mn, Mo, I, and Se (Bowman and Russell, 2011; Onuegbu, et al., 2011). bioactive compound recovery. Ultrasound-assisted extraction (UAE) is an emerging technique known for enhancing the yield  of bioactive compounds through acoustic cavitation, which improves solvent penetration into plant matrices. UAE has been successfully applied in phytochemical extraction, improving the recovery of lipids, flavonoids, and polyphenols while maintaining their bioactivity. Given the therapeutic potential of Gongronema latifolium, incorporating UAE alongside According to Nitin Mehta.,et al(2022) compared different extraction methods, including UAE, highlighting UAE as a faster and solvent-efficient alternative with comparable results to traditional methods. Ultrasound-Assisted Extraction and the Encapsulation of Bioactive Compounds In an article by Jinchao shen and Xueguang Shiao(2005),it discusses the fundamentals and mechanisms of UAE, emphasizing its efficiency in extracting bioactive components from plant materials. According to S.I Okonkwo et al.,(2010)Saponin was extracted from methanol extract of Ocimum gratissimum leaves,identifying it using High performance liquid chromatography and applying it in the synthesis of nano emulsion

According to Chinaza Amala Ezeilo,Sylvia Ifeyinwa Okonkwo et al(2020) Determination of the concentration of heavy metals(Cu,Pb ,Cd,Hg,As and Zn)were assessed using Atomic absorption spectroscopy to evaluate potential health hazard associated with this metals

  1. Experimental

2.1. Sample Collection.

Fresh leaves of G. Latifolium were collected at the botanical garden of the department of Science Laboratory Technology, Federal Polytechnic, Oko. Orumba North L.G.A. Anambra State, Nigeria. The leaves were rinsed thoroughly using clean water, air dried at room temperature and ground into powder using electric blender. The ground sample was kept in an air light container for analysis.

    1. Extraction of Oil from Utazi Leaves

Soxhlet Extraction Method: Round boiling flask (500mL) was dried in oven at 105-1100C for about 15 minutes and transferred into a desiccator and allowed to cool. The 500mL flask was filled with n-hexane solvent. 160g of powdered G. latifolium leaf was weighed and inserted into the thimble of the soxhlet apparatus with cotton wool underneath to serve as filter. The apparatus was assembled on the boiling flask. It was allowed to stand on electric hot plate at temperature of 60-750C, and refluxed for about 4 times for five repeated extraction. Extract from the flask was collected and emptied into a rotator evaporator at temperatures of 40-600C to separate the n-hexane solvent from the extracted oil. The extracted oil was collected and stored in a container for analysis.

Ultrasound Assisted Extraction

160 g of Gongronema latifolium leaf powder was accurately weighed and placed into a 2 L extraction flask.1.6 L of 80% ethanol was added to the flask.The ultrasonic probe was immersed into the solution.The ultrasound system was set to 70% amplitude with a pulse mode (5s ON, 5s OFF) for 30 minutes while maintaining a temperature of 50°C using a water bath.The extract was filtered using a vacuum filtration system.The filtrate was concentrated using a rotary evaporator at 40°C under reduced pressure.The concentrated extract was freeze-dried to obtain a powdered form.

    1. Preparation of Sample for Fatty Acid Profile: The sample oil (5mL) was pipette into beaker containing 20mL of n-hexane, shaked very well and poured into a separating funnel and allowed to stand for 30 minutes. The n-hexane layer and the sample were separately collected and stored for fatty acid profile analysis with GC-FID.
    2. Aqueous Extraction of Sample (Maceration Method): Five hundred milliliter (500mL) of distilled water was poured into a flat bottom flask, 20 g of the ground G. latifolium leaf was inserted into the flask, then thoroughly stirred. The mixture was placed on magnetic stirrer, five glass beads were inserted and the mixture allowed to stir for 5 minutes. It was covered, kept and stirred using magnetic stirrer for 5 minutes daily for 2days. The mixture was filtered using filter paper. The filtrate was poured into a conical flask and placed on a water bath at below 50 0C. The flask base was immersed in the water, and the mixture allowed to concentrate. After that, the filtrate was allowed to cool down and kept in the beaker for TLC analysis.
    3. TLC Isolation: The TLC plates were measured 5cm and the concentrated filtrate of G. latifolium was dotted on the plate using a stirring rod, and allowed to stay for some minutes to enable it. The solvent phase (acetone: water: ammonium in a ratio of 90:3:7), was measured into the beaker and the spotted plates were placed to stand in the beaker containing the solvent mixture for 1 hour, after which the plates were removed and allowed to dry. The colours were noted and the retention factor measurements were taken. Rf-value = Distance from origin to component spot Distance from origin to solvent front
    4. Digestion of Sample for Elemental Analysis: The samples (2g) were weighed into a digestion flask and 20mL of aqua regia acid mixture (65mL conc. HNO3; 8mL perchloric acid; 2mL conc. H2SO4) was added into it and sealed with a cork. The flask was placed under fume cupboard and heat applied with the aid of electric hot plate at temperature of 600C and allowed to heat until clear digest was obtained. The clear digest was diluted to 100mL with distilled water and filtered into reagent bottle for analysis of heavy metal with Atomic adsorption spectrophotometer (AAS), according to (AOAC, 2003).
    5. Determination of Sulphur Content: The cake sample (1g) was mixed with 5.0g of NaCO3 in a crucible. The mixture was preheated at 4000C for 30mins in an electric furnace, and then fused at 9500C. After the fusion, the crucible was allowed to cool and was placed on its side in a 250mL beaker. Enough deionized water barely to cover the contents of the crucible was added and the beaker was heated at a temperature just below boiling on a hot plate, until the melt was thoroughly decimated. The crucible was then removed and washed with deionized water. At this point 20mL of 6M HCl was added to neutralize the NaCO3 and to make the solution slightly acidic. This was filtered into a 100mL volumetric flask and the volume made up to the mark with deionized water. The solution was brought to boiling and 10mL of 10% BaCl2.2H2O was slowly added to precipitate. The solution was allowed to cool and was filtered. The residue was washed with deionized water. The ashless filtered paper was ignited at low temperature (400C), and the precipitate weighed. The percentage sulphur in the precipitate was calculated from the expression below. Calculation: % Sulphur = Weight loss ×13.17Weight of Sample
    6. Determination of Nitrogen: The sample (1g) was weighed into a 300mL kjehdal flask (gently to prevent the sample from touching the walls of the side of each) and then the flask was stoppered and shaken. Then 0.5g of the kjedahl catalyst mixture was added. The mixture was heated cautiously in a digestion rack on electric hot plate until a clear solution appeared. The clear solution was then allowed to stand for 30 minutes and allowed to cool. After cooling about 100mL of distilled water was added to avoid caking and then 5mL of the filtrate and 5mL of 40% NaOH was transferred to the kjedahl distillation apparatus. A 250mL receiver beaker containing 10mL of 10% boric acid and indicator mixture containing 5 drops of Bromocresol blue and 1 drop of methylene blue was placed under a condenser of the distillation apparatus so that the tap was about 20cm inside the solution. Then 5mL of 40% sodium hydroxide was added to the digested sample in the apparatus and distillation commenced immediately until 50 drops get into the receiver beaker, after which it was titrated to pink colour using 0.01N hydrochloric acid. Calculations: % Nitrogen = Titre value ×
        0.01×
       14×
       4.
  2. RESULTS

Table 1: Shows the Results of the Fatty Acid Content of Gongronema latifolium leaf.

 

Fatty Acid (µg/mL)

 

Concentration

Lauric Acid

C12:0

9.44165±0.681

Mystric Acid

C14:0

14.5081±1.355

Pentadecaoic Acid

C15:0

8.7489±0.0016

Palmitic Acid

C16:0

16.0541± 0.677

Palmitoloic Acid

C16:1

12.2206±0.380

Heptadecanoic Acid

C17:0

8.7542±4.073

Stearic Acid

C18:0

7.30985±0.682

Oleic Acid

C18:1

21.61135±0.664

Linoleic Acid

C18:2

12.8016±1.660

Docosahexaenoic Acid

C24:0

7.2603±0.604

Oil Content (%)

 

2.9835 ± 0.9417

Table 2: Results of the TLC Analysis of the Active Components in Aqueous Extract of Gongronema latifolium (Utazi) Leaf

 

Rf-Value (cm)

Mean % Conc. ± S. D

Colours

1.24

1.32

1.36

1.40

3.15 ± 0.685

2.05 ± 0.212

3.70 ± 0.565

3.60 ± 0.848

Gray

Yellow

Green

Brown

Table 3: Results of the yield and bioactive components using Ultrasound Assisted Extraction

 

Parameter

Values

Crude Extract Yield

12-15%(~19-24g)

Bioactive Compounds

Flavonoids(Kaempferol,quercetin),Alkaloids,Saponins,Polyphenols,terpenoids

Figure 1: The Results of the Elemental Content of G. latifolium Leaf.

DISCUSSION

Fatty acids are essential compounds in man’s diet, and their esterification with glycerol molecules give rise to the main constituents of fats and oil, (Ahmad, 2017). The WHO recommends a daily fat intake equivalent to 20%-35% of total daily energy, (Eilander et al., 2015). The results of the determination of fatty acid profile of G. latifolium leaf shown in Table 1, indicated that oleic acid (C18:1) was the fatty acid with the highest value of 21.611 ± 0.664 µg/mL. Oleic acid is a monounsaturated fatty acid which has beneficial effects on cancer, autoimmune and anti-inflammatory diseases beside its ability to facilitate wound healing, (Sales-Campos, 2013). Also, in Table 1, palmitic acid (16:0) content of the G. latifolium leaf was 16.05 ± 0.677 µg/mL which was next in concentration to oleic acid. Although, palmitic acid is a saturated fatty acid but it has health benefits which includes supporting skin health, anti-inflammatory effects and potentially metabolic health benefits. On the other hand, too much of palmitic acid in proportion to other healthy fatty acids may increase the risk for cardiovascular disease, (Levy, 2020). The concentration of palmitc acid in this study was lesser than 36% of the total fatty acid obtained by Eleyinmi, (2007). Mystric acid was 14.508 ± 1.355 µg/mL as seen in Table 1. It showed that mystric acid (C14:0) was the third highest fatty acid in terms of concentration. It is a saturated fatty acid. Other fatty acids that were reasonably present in G. latifolium leaf were linoleic acid: 12.802 ± 1.660 µg/mL (C18:2), palmitoleic acid: 12.221 ± 0.380 µg/mL (C16:1) and the least contained fatty acid was docosahexaenoic acid: 7.260 ± 0.604 µg/mL (C24:0). WHO, (1996), stipulates that the replacement of saturated fatty such as palmitic acid by polyunsaturated fatty acids decreases low-density lipoprotein cholesterol levels and total cholesterol/high density lipoprotein acids rich diet decreases the risk of coronary heart disease. The total oil content of the G. latifolium leaf was 2.9835 ± 0.9417%. The amount of oil obtained in this study was higher than 0.3-1.25% obtained from the fresh leaves of allspice, (Krishnamoorthy, 2004). Generally, in Table 1, the total amount of saturated fatty acids content of G. latifolium leaf (72.0771 µg/mL) was higher than (46.6336 µg/mL) of unsaturated fatty acid in the plant leaf. However, these values in percentage were higher than what Eleyinmi, (2007), obtained in his work on chemical and antibacterial of G. latifolium leaf. He obtained 50.2% and 39.4% of saturated and unsaturated fatty acids respectively in the oil. Fatty acids are widely used as inactive ingredient in drug preparations and are used in lipid formulation as the carriers for active substance. In Table 2, the components isolated from the aqueous extract of G. latifolium leaf were four active components. The first component identified showed gray colour, with a retention factor (Rf) of 1.24 cm and a mean concentration of 3.15 ± 0.685 cm. The gray colour happens to be the least migrated component, followed by the component with yellow colour with the Rf value 1.32 cm and a mean concentration of 2.05 ± 0.212 cm. Green component had Rf value of 1.36 cm and a mean concentration of 3.70 ± 0.565 cm. Finally, brown colour was the most migrated with Rf value of 1.40 cm, and a mean concentration of 3.60 ± 0.848 cm. According to Ahmad and Wudil, (2013), the active components obtained from the TLC separation of Gongronema latifolium (Utazi) leaf was higher in aqueous extract than ethanol extracts. This was supported by Ezekwesili-Ofili and Okaka, (2019), who reported that the polarity of solvents can raise the solubility of active components. Hassan et al., (2015); Ismail et al., (2015), obtained Rf value of 0.86 cm with green colour of the TLC analysis of the leaves and flowers extract of senna siamea lam using ethyl acetate. Moreso, Sangeeta and Vrunda, (2016), disclosed the presence of flavones, flavonols, biflavonyl, kaempferol, etc., in the chromatographic separation of flavonoids in extracts of moringa and ocimum (leaf and flower). However, the disparity in the TLC separation could be due to the polarity differences of the solvent used for extraction and the solvent mixtures used for the TLC separation. But from the results of this study, the TLC analysis has given an idea of the components present in G. latifolium leaf, which may not have been exhaustively separated. The results of the elemental analysis presented in Figure 1, showed that K was highest contained element with a value of 8.975 ppm, followed by S; (8.245 %), Zn; (2.4663 ppm), Ca; (1.6035 ppm), N; (1.344 %), Na; (1.3375 ppm), and Fe; (0.859 ppm), respectively. The least contained elements according to Figure 1, were Mg; (0.5645 ppm), Mn; (0.1105 ppm), Cu; (0.1015 ppm), Cd; (0.023 ppm), As; (0.014 ppm), and Pb; (0.00165 ppm), respectively. Nwamkezie and Obiakor-Okeke, (2014), obtained 7.83mg/100g of Fe, 54.60 mg/100g of Mg, 78.5 mg/100g of K, and 68.30mg/100g of Ca respectively. In a study by Adeyeye et al., (2020), G. latifolium leaf contained 13.3 mg/100g of Fe, 4.71 mg/100g of Zn, 548 mg/100g of Mg, 659 mg/100g of K, 304 mg/100g of Ca, 1.91 mg/100g of Cu, 44.7 mg/100g of Na, 0.00016 mg/100g of Pb and 0.0008 mg/100g of Cd respectively. More so, Usoro et al., (2018), opined that G. latifolium leaf contained 471±12.08 mg/kg of K, 0.397±0.07 mg/kg of Zn, 143.8±8.13 mg/kg of Na, 130±7.45 mg/kg of Ca, 133±5.02 mg/kg of Mg, 1142±14.21 mg/kg of Fe, 0.16±0.009 mg/kg of Pb, and 0.23±0.003 mg/kg of Cd, respectively. In a similar vein, according to Roszyk, (2005), the mean sulphur content of chrysanthemum leaf ranged between 0.25 and 0.45 %, and this was further buttressed by Macz et al., (2001a), that the leaf sulphur concentration of 0.3-0.5 % is the acceptable limits. However, it is evident that the various result findings showed a similar trend in the concentration of the elements. This can be attested by the values obtained for K, Fe, Ca, Na, Pb, Cd and Zn. Furthermore, as shown in Figure 1, it is important to note that G. latifolium leaf is rich in macro-elements and therefore, suitable for the management of high blood pressures. It contains high amount of potassium which reduces blood pressure level and minimizes its effect in the heart (Carter, 2018). Also, both K and Na are needed in the body to regulate and control glucose absorption and assist protein retention during growth, (NRC, 1989). Children and adults can take G. latifolium leaf because it contains good amount of zinc and calcium which helps in building strong immunity, bones, teeth and also blood clothing, (Medline, 2018). Although, Zn concentration in this study was lower than the recommended dietary allowance of between 15-20 mg per day, (Fleck, 1976). Zinc has anti-inflammatory properties and support healthy thyroids. G. latifolium leaf contains small amount of iron; (0.0859 ppm). Iron helps in facilitating the oxidation of carbohydrates, protein and fats, (Adeyeye et al., 2014). It plays important roles in transporting oxygen and carbon dioxide in the blood. The human requirement of iron for children is between 10-15 mg, for women is 18 mg and 12 mg for men, (Fleck, 1976). It also contain magnesium, and manganese which are micro-nutrients the human bodies does not need in large quantities. WHO, (1996), stipulates 0.01-0.1 mg as acceptable standards heavy metals, and the lead concentrations were within permissible limits. Chronic exposure to lead is associated with kidney damage in adults, (Navas-Acien et al., 2009). Lead, arsenic, and cadmium are not needed in the body and their presence is an indication of pollution. Eleyinmi, (2007), disclosed that G, latifolium leaf contains 44.3% of free nitrogen extractives, and this is far higher than 1.344% obtained in this study. Nitrogen is a critical component of amino acids which are building blocks of protein, and a key element in the synthesis of nucleotides, that are essential for DNA and RNA production. UAE significantly reduced extraction time compared to Soxhlet and maceration while yielding a comparable or higher amount of bioactive compounds.The controlled temperature (50°C) prevented the degradation of thermolabile compounds.The pulse mode (5s ON, 5s OFF) ensured effective cavitation, improving solvent penetration and extraction efficiency.UAE used a lower volume of solvent compared to Soxhlet extraction, making it a more environmentally friendly approach.

CONCLUSION

Fatty acid profile and elemental analysis of Gongronema latifolium (Utazi) leaf have provided valuable insight into the composition of this important traditional plant. The study identified a range of important fatty acids, including oleic and linoleic acids, as well as palmitic and mystric acids. Essential elements were also identified, but more importantly, is the presence of heavy metals which are likely to pose danger to health on accumulation. Ultrasound-assisted extraction (UAE) is a rapid and efficient method for extracting bioactive compounds from Gongronema latifolium. UAE provided a higher yield in a shorter time while reducing solvent consumption, making it a sustainable alternative to conventional methods.

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        19. De Vogli, R., Kouvonen, A., and Gimeno, D (2011). "Globesization": Ecological Evidence on the                      Relationship between Fast Food Outlets and Obesity among Advanced Economies. Critical     Public Health, 21(4):395-402
        20. Edet, E.E., Akpanabatu, M.I., Uboh, F.E., Edet, .T.E., Eno, .A.E., and Itam, .E.H. (2011) Gongronema    latifolium    Crude    Leaf   Extract    Reverses    Alterations    Haematological    Induces   and   Weight-loss   in   Diabetic   Rats.   Journal   of Pharmacology and Toxicology, 6(2): 135-143
        21. Egbung, .G, Atangwho, .I.J, Iwara, .I.A, and Eyong, .U.E. (2011). Micronutrient and Phytochemical Composition of Root, Bark and Twig Extracts of Gongronema latifolium. Journal of Medicine Sciences, 2(11): 1185-1188.
        22. Eilander, A., Harika, R.K., and Zock, P.L. (2015). Intake and Sources of Dietary Fatty Acids in                         Europe: Are Current Population Intakes of Fats aligned with Dietary Recommendations. European Journal of Lipid Science and Technology, 117(9): 1370-1377.    
        23. Eleyinmi, A.F. (2007). Chemical Composition and Antibacterial Activity of Gongronema latifolium. Journal of Zhejiang UniversityScience B 8(5):352-358.
        24. Edim, .E.H, Egomi, .U.G, Ekpo, .U.F, and Archibong, .E.U. (2012). A Review on Gongronema                           latifolium (Utazi): A Novel Antibiotic against Staphylococcus aureus related Infections. International Journal of Biochemistry and Biotechnology, 1(8):204-208.
        25. Edoga, H.O., Okwu, D.E., Mbaebie, B.O.  (2005). Phytochemicals Constituents of Some Nigerian                        Medicinal Plants. Afri. J. Biotecnol., 4(7): 685-688
        26. Enyi-Idoh, .K, Utsalo, .S. Arikpo, .G. and Eja, .M. (2012). Tme-dependent Evaluation of the Anti-           bacterial and
        27. Phytochemcal Properties of Vernonia amygdalina and Gongronema     latifolium. The Internet Journal of Herbal and Plant Medicine. 1(2):111-119.
        28. Essien, .J.P, Ebong, .G.A, and Akpan, .E.J. (2007) Antioxidant and Antitussive Properties of                           Gongronema latifolium leaves used locally the Treatment of Fowl Cough in Nigeria.      Journal of Applied Sciences and Environmental Management, 11(4): 47-50.
        29. Ezekwesili-Ofili, J. O. and Okaka, A. N. C. (2019). Herbal Medicines in African Traditional                              Medicine; Philip F.B.; Ed. IntechOpen, DOI: 10.5772/80348.
        30. Eze, .S.O, and Nwanguma, .B.C. (2013). Effects of Tannin Extract from Gongronema latifolium          Leaves on Lipoxygenase Cucumeropsis Manii Seeds. Journal of Chemistry 1(3):1-7
        31. Fleck, H. (1976). Introduction to Nutrition. 3rded. New York, USA: Macmillian, pp: 207-219.
        32. Gabriel, A., Agbor, D.K. and Julius, E.O. (2007). Medicinal Plants can be Good Sources of                               Antioxidants: Case Study in Cameroon. Pakistan Journal of Biological Science 10 (4):       537-544.
        33. Hassan, R.I.L., Seyedmohamm, B., and Khodarahmi, R.I. (2015). Therapeutic Potentials of the                      Most Studied Flavonoids: Highlighting Antibacterial and Anti-diabetic Functionalities 2(6):43-56
        34. Ismail, A.H., Idris, A.N., Amina, M.M., Ibrahim, A.S. and Audu, S.A. (2015). Phytochemical                        Studies and Thin Layer Chromatography of Leaves and Flower Extracts of Senna Siamea      lam for Possible Biomedical Applications. Journal of Pharmacognosy and Phytotherapy 7(3):18-26.
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        38. Medline, P. (2018). Calcium in Diet: Medline Plus Medical Encyclopedia. Retrieved September              23, 2018, from https:// Medline Plus. gov/ency/ article/002412.htm Bowman, H. U. and Russell, C. I. (2011), Assessment of Constraints to Mango Consumption in Selected Communities of River State, Nigeria, International Journal of Research in Applied Natural and Social Sciences,  2(3):31-32.
        39. McNamara, R.K. and Carlson, S.E. (2006) Role of Omega-3 Fatty Acids in Brain Development                          and Function: Potential Implications for the Pathogenesis and Prevention of Psychopathology. Prostaglandins, Leukotrienes and Essential Fatty Acids, 75 (1): 329-349.
        40. Mgbeje, B.I., Umoh, E.U. and Emmanuel-Ikpeme, C., (2019). Comparative Analysis of Phytochemical Composition of Four Selected Tropical Medicinal Plants Namely: Ocimumgratissimum, Piper guineense, Gongronema latifolium and Vernonia amygdalina.            Journal of Complementary and Alternative Medicinal Research, 7(3): 1-11.
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  25. Edoga, H.O., Okwu, D.E., Mbaebie, B.O.  (2005). Phytochemicals Constituents of Some Nigerian                        Medicinal Plants. Afri. J. Biotecnol., 4(7): 685-688
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  28. Essien, .J.P, Ebong, .G.A, and Akpan, .E.J. (2007) Antioxidant and Antitussive Properties of                           Gongronema latifolium leaves used locally the Treatment of Fowl Cough in Nigeria.      Journal of Applied Sciences and Environmental Management, 11(4): 47-50.
  29. Ezekwesili-Ofili, J. O. and Okaka, A. N. C. (2019). Herbal Medicines in African Traditional                              Medicine; Philip F.B.; Ed. IntechOpen, DOI: 10.5772/80348.
  30. Eze, .S.O, and Nwanguma, .B.C. (2013). Effects of Tannin Extract from Gongronema latifolium          Leaves on Lipoxygenase Cucumeropsis Manii Seeds. Journal of Chemistry 1(3):1-7
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  33. Hassan, R.I.L., Seyedmohamm, B., and Khodarahmi, R.I. (2015). Therapeutic Potentials of the                      Most Studied Flavonoids: Highlighting Antibacterial and Anti-diabetic Functionalities 2(6):43-56
  34. Ismail, A.H., Idris, A.N., Amina, M.M., Ibrahim, A.S. and Audu, S.A. (2015). Phytochemical                        Studies and Thin Layer Chromatography of Leaves and Flower Extracts of Senna Siamea      lam for Possible Biomedical Applications. Journal of Pharmacognosy and Phytotherapy 7(3):18-26.
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  36. Levy, J. (2020) Palmitic Acid: Benefits vs. Risks: What you Need to Know. Dr. Axe-Nutrition-                        Fats and Oils.
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  38. Medline, P. (2018). Calcium in Diet: Medline Plus Medical Encyclopedia. Retrieved September              23, 2018, from https:// Medline Plus. gov/ency/ article/002412.htm Bowman, H. U. and Russell, C. I. (2011), Assessment of Constraints to Mango Consumption in Selected Communities of River State, Nigeria, International Journal of Research in Applied Natural and Social Sciences,  2(3):31-32.
  39. McNamara, R.K. and Carlson, S.E. (2006) Role of Omega-3 Fatty Acids in Brain Development                          and Function: Potential Implications for the Pathogenesis and Prevention of Psychopathology. Prostaglandins, Leukotrienes and Essential Fatty Acids, 75 (1): 329-349.
  40. Mgbeje, B.I., Umoh, E.U. and Emmanuel-Ikpeme, C., (2019). Comparative Analysis of Phytochemical Composition of Four Selected Tropical Medicinal Plants Namely: Ocimumgratissimum, Piper guineense, Gongronema latifolium and Vernonia amygdalina.            Journal of Complementary and Alternative Medicinal Research, 7(3): 1-11.
  41. Morebise .O, and Fafunso, .M.A. (1998). Antimicrobial and Phytotoxic Activities of Saponin                               Extracts from Two Nigerian Edible Medicinal Plants. Biokemistri. 8(2): 69-77.
  42. Morebise, O, Fafunso, .M.A, Makinde, .J.M, Olafide, .O.A, and Awe, .E.O. (2002). Anti-inflammatory Properties of Leaves of Gongronema latifolium. Phytotherapy Research, 16(1): 575-577.
  43. Msuya, P. and Mamiro, L. (2018). Prevention and Therapy of Cancer by Dietary Monoterpenes.                        The Journal of Nutrition 129 (3): 775S–778S.
  44. Murray, J., Licciardello, C., Russo, M.P., Chiusano, M. L., Carletti, G., Recupero, G. R., Marocco,                      A. (2010). Use of a Custom Array to Study Differentially Expressed Genes during Blood     Orange (Citrus sinensis L. Osbeck) Ripening, Journal of Plant Physiology, 167, (1): 301–  310.
  45. Navas-Acien, E.M., Rodríguez, E.D., Uretra C.D., and Romero, S.N (2009). Concentrations of                   Cadmium and Lead in different Types of Milk,” Zeitschrift Lebensmitteluntersuchung      und-Forschung. 8 (3):162-168.
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  49. Nwanjo, .H.U., Okafor, M.C., Oze, G.O., (2006). Anti-lipid Peroxidative Activity of Gongronema                       latifolium in Streptozotocin Induced Diabetic Rats. Niger J Physiol Sci.21 (1): 61-65.
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  54. Osuagwu, .A.N, Ekpo, .I.A, Okpako, .E.C, and, Ottoho, .E. (2013). The Biology, Utilization and                        Phytochemical Composition of the Fruits and Leaves of Gongronema latifolium Benth. Agrotechnology, 2(115), pp.1-4.
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S. I. Okonkwo
Corresponding author

Departments of Pure and Industrial Chemistry, Chukwuemeka Odumegwu Ojukwu University, Uli, Anambra State, Nigeria.

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V. E. Mmuo
Co-author

Department of Chemistry, School of Science Laboratory Technology, Federal Polytechnic, Oko, Anambra State, Nigeria.

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E. C. Okafor
Co-author

Departments of Pure and Industrial Chemistry, Chukwuemeka Odumegwu Ojukwu University, Uli, Anambra State, Nigeria.

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V.O. Offiah
Co-author

Departments of Pure and Industrial Chemistry, Chukwuemeka Odumegwu Ojukwu University, Uli, Anambra State, Nigeria.

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C. K. Okonkwo
Co-author

Department of Diagnostic Medical Sonography and Ultrasound Technology, Ace Institute of Technology, Elmhurst, New York, USA.

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P. O. Okwuego
Co-author

Departments of Pure and Industrial Chemistry, Chukwuemeka Odumegwu Ojukwu University, Uli, Anambra State, Nigeria.

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V. S. Okonkwo
Co-author

Department of Medical Biochemistry, Chukwuemeka Odumegwu Ojukwu University, Uli, Anambra State, Nigeria.

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A. T. Kene-Okonkwo
Co-author

Tansian University Oba, Anambra State, Nigeria.

S. I. Okonkwo*, V. E. Mmuo, E. C. Okafor, V.O. Offiah, C. K. Okonkwo, P. O. Okwuego, V. S. Okonkwo, A. T. Kene-Okonkwo, Extraction, Fatty Acid Profile and Elemental Analysis of Gongronema latifolium (Utazi) Leaf, Int. J. of Pharm. Sci., 2025, Vol 3, Issue 3, 1244-1256. https://doi.org/10.5281/zenodo.15020463

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