INTRODUCTION:

Pumpkin (Cucurbita moschata), a member of the Cucurbitaceae family and the order Violales, is a widely recognized plant known for its distinctive creeping and climbing growth habits^1,2. It possesses a resilient wedge-shaped root that exhibits resistance to low temperatures. With its large heart-shaped leaves that can extend up to 25cm in length, the plant produces unisex flowers in vibrant shades of yellow to orange^3,4.

Despite being commonly referred to as a vegetable, pumpkins are scientifically classified as fruits and hold significant nutritional value. They are rich in essential components such as water, protein, sugars, fiber, calcium, potassium, copper, vitamins, and saturated fatty acids^5,6. The seeds of the pumpkin plant have attracted considerable attention due to their diverse range of bioactive substances and compounds, making them valuable in the development of medicinal and functional food products^6-8. Extensive research has been conducted to extract and isolate these bioactive compounds, driven by their numerous health benefits^8,9. Various industries, including food, cosmetics, and pharmaceuticals, have recognized the potential applications of these compounds.

Pumpkin consumption offers a multitude of benefits, encompassing immune system enhancement (pumpkin exhibits antioxidant properties primarily attributed to its carotene compounds, which can help reduce the risk of degenerative and cancerous diseases), eye health maintenance (consuming pumpkin supports healthy vision and provides protection against related ailments), antioxidant activity (the presence of antioxidants in pumpkin contributes to its ability to counteract oxidative stress), vitamin A source (pumpkin serves as a rich source of vitamin A, including alpha-carotene and beta-carotene, antiglycemic effects (Pumpkin demonstrates properties that aid in regulating blood sugar levels), anti-cholesterol and antihypertensive properties (Pumpkin consumption has been associated with the reduction of cholesterol and blood pressure levels)^10-15. Additionally, pumpkin exhibits antibacterial, anti-inflammatory, antifungal, and antitumor activities, making it a promising candidate for various therapeutic applications. Pumpkin seeds are also utilized in the treatment of prostate and bladder problems. Moreover, in the food industry, pumpkin can be employed as a natural coloring agent^14-16. Carotenoids, particularly beta-carotene, play a crucial role in mediating the biological effects of pumpkin. Within the human body, beta-carotene undergoes breakdown facilitated by beta-carotene deoxygenase systems present in the gastric mucosa. This process converts beta-carotene into retinal, which is further transformed into retinol (vitamin A)^16-24. Vitamin A plays a vital role in vision, cell differentiation, glycoprotein synthesis, and mucus secretion from epithelial cells. Furthermore, it offers protective effects against cataracts, macular degeneration, arthritis, and osteoporosis while promoting skin health^24-35.

This study seeks to provide valuable insights into the composition of pumpkin seeds, particularly their content of biologically active substances. Pumpkin seeds have gained significant attention as a potential source of compounds with medicinal and functional food properties, owing to their remarkable richness in biologically active compounds. In addition to their potential health benefits, extensive research has been dedicated to developing effective extraction methods to obtain a substantial yield of these biologically active compounds. This research holds promising implications for various industries, including the industrial, food, cosmetic, and pharmaceutical sectors, where these compounds can be utilized.

MATERIALS AND METHODS:

Materials:

HCl solution, acetone, dichloromethane, ethanol, distilled water, sodium carbonate solution, gallic acid standard solution, and Folin-Ciocalteu reagent were used as reagents and solvents. All reagents were purchased from ShamLap (Syria).

Sample Collection:

Pumpkin (Cucurbita moschata) fruits were obtained from a local farm, ensuring ripeness and uniformity.

Carotenoid Extraction:

To begin the extraction process, 100mg of pumpkin pulp was precisely measured and transferred to a blender. 4 mL of an extraction solvent composed of 70% acetone and 30% dichloromethane was added to the blender. The contents were thoroughly mixed and blended at 3500 rpm for 10minutes. To ensure proper solubilization. Subsequently, subject the mixture to blending at a speed of 3500rpm for 10 minutes. Following this step, the resulting mixture was filtrated to remove any precipitate. Next, the filtrate obtained was transferred to a calibration flask with a capacity of 10mL. To achieve the desired volume, the extraction solvent was carefully added to the flask until it reached the calibration line.

Phenol Extraction:

For the extraction and analysis of phenols, the seeds of both white and red pumpkin plants were subjected to a drying process in shaded conditions until a firm weight was achieved. The dry seeds were then peeled and ground into a powder form. A total of 25g of the powdered pulp was accurately weighed and transferred to a conical flask. Next, 100ml of a 70% ethanol and 30% water mixture was added to the flask containing the pulp powder. The contents were thoroughly stirred, and the pH was adjusted to 3 using hydrochloric acid. Subsequently, the flask was placed in an ultrasonic bath operating at a temperature of 37°C and a frequency of 40kHz. The ultrasonic treatment lasted for two hours, allowing efficient extraction of the phenols present in the sample. The ultrasonic waves were generated at a power of 132 watts.

After the extraction process, the resulting mixture was filtered at a speed of 2500 cycles for 15 minutes. The filtrate was then subjected to evaporation using a rotary evaporator under vacuum conditions. The volume of the evaporated solution was adjusted to 2ml. Consequently, an extract labeled as (1) was obtained, containing the concentrated phenolic compounds for further analysis.

High-Performance Liquid Chromatography (HPLC) Analysis:

The analysis was conducted using the American Agilent HPLC system. The experimental HPLC conditions for the analysis: C18 column (250×4.6mm, particle size of 5 μm), wavelength of UV/Vis detector (454nm for carotenoid determination and 254 for phenol determination), column temperature (30°C), flow rate (0.9mL/min), injection volume (20μL). To identify most phenols, a PDA detector was employed, which measured wavelengths of 254nm and 320nm.

Carotenoid Determination:

To evaluate the linearity of HPLC method, a standard solution of beta-carotene with a concentration of 100 mcg/mL was prepared. The standard solution was prepared by weighing 1mg of beta-carotene standard and dissolving it in 3mL of dichloromethane. The volume was then completed to 10 mL using acetone in a titration flask. From this solution, a series of beta-carotene titrations was prepared, yielding concentrations of 0.3, 1.5, 3, 4.5, 6, and 9mcg/mL. Subsequently, the prepared solutions were injected into the device, and the Pearson coefficient was calculated. The resulting equation of the line was determined as follows: Y= 0.1763× + 0.0243. The coefficient of determination (R2) was determined, yielding a value of 0.997. Through the analysis we conclude that the carotene content in red pumpkin is much higher than in yellow pumpkin.

Extraction Yield Calculation:

The extraction process involved three successive extractions of a pumpkin sample. Subsequently, the sample was subjected to re-extraction after the addition of standard beta-carotene spiking at three different concentrations. Each concentration underwent three re-extractions. The recovery was determined for each concentration based on the previous table. It is worth noting that the extraction yield obtained was 103.51%. This indicates that the extraction method for beta-carotene from pumpkin demonstrates a highly favorable yield.

Table 1. recovery of beta-carotene spiked at three different concentrations

Original amount contained in 0.1g of sample (mcg)	Added amount of beta-carotene to 0.1 g of sample (mcg)	Total theoretical quantity in 0.1g Sample (mcg)	Total quantity measured in 0.1 g of sample (mcg)	Recovery
12.51	1.4	13.91	14.08	101.11
12.51	2.8	15.31	15.89	103.8
12.51	5.6	18.11	19.13	105.6

Quantification of Carotene in Yellow and Red Pumpkin Pulp:

The carotene content in yellow and red pumpkin pulp was measured. The quantity of carotene in 1 g of sample was determined for each pulp type.

Estimation of the total phenolic contents in the fruit of the red and white pumpkin plant:

Total phenolic contents were determined using a colorimetric method based on the Folin-Ciocalteu assay. The assay relies on the formation of a blue complex between phosphomolybdate and phosphorus tungstate in an alkaline medium in the presence of phenolic compounds. The intensity of the resulting color is directly proportional to the concentration of phenolic compounds in the sample. To perform the colorimetric titration, the absorbance was measured at a wavelength of 755nm using a spectrophotometer. Gallic acid was used as the calibration standard to quantify the polyphenols present in the pumpkin fruit. The extraction of phenols and flavonoids from the pumpkin fruit involved separately weighing 50g of white pumpkin and red pumpkin samples and placing them in conical flasks. A 70% ethanol and 30% water mixture (100mL) was added to each flask, and the pH was adjusted to 3 using hydrochloric acid. The flasks were subjected to ultrasonic wave bath treatment at 37°C and a frequency of 40 kHz for two hours. The extracts were then filtered, evaporated, and adjusted to a final volume of 5mL. To determine the phenol and flavonoid content in each extract using the Folin-Ciocalteu method, a series of gallic acid standards dissolved in ethanol (75%) was prepared to construct the titration curve. The absorbance of the extract and the series of gallic acid standards mixed with Folin-Ciocalteu reagent and sodium carbonate was measured at 760nm after incubation at 40°C for 30 minutes. As shown in Figure 1, the equation of the calibration curve was Y=0.0096X+0.0023 and R² has a value of 0.9987, demonstrating very good linearity.

Figure 1. Plot of absorbance vs concentration (mg/100 mL) for gallic acid using Folin-Ciocalteu method.

Determination of total flavonoid content:

The total flavonoid content in the methanolic extract of white and red pumpkin fruit pulp, as well as in the seeds, was determined. The aluminium chloride chromatography method was used, and a series of quercetin standards dissolved in ethanol (75%) was prepared to construct the titration curve. The extracts and quercetin dilutions were mixed with 95% ethanol, aluminium chloride reagent, sodium acetate, and distilled water. After incubating for 30 minutes at room temperature, the absorbance of the reaction was measured at a wavelength of 415nm using a double-beam spectrophotometer. The total flavonoid content in the extracts was reported in terms of quercetin equivalents (mg/g extract).

Determination of volatile organic compounds in red pumpkin seeds and white pumpkin:

Soxhlet extraction was performed on crushed pumpkin seeds of white and red varieties. Each type of seed, weighing 20grams, was placed in a Soxhlet apparatus, and extraction was carried out using 300ml of absolute ethanol for approximately three hours. The resulting extraction solution was then evaporated using a rotary evaporator under vacuum until dry. The dried extract was dissolved in 2ml of methanol and stored in an opaque container for further analysis using GC-MS technology.

The oil extract obtained from the pumpkin seeds was subjected to analysis on the Agilent 6970 GC-MS system. The analytical column used was DB-35 MS with a length of 30 meters. Helium was used as the carrier gas at a flow rate of 0.9ml/min. The thermal program for analysis started at an initial temperature of 75°C for 2 minutes, followed by an increase of 2 degrees per minute until reaching 150°C. The temperature was then increased at a rate of 4 degrees per minute until reaching 250°C, where it was held for 5 minutes. The fragmentation energy used for the analysis was 70 electron volts.

Statistical Analysis:

Data obtained from the experiments were subjected to appropriate statistical analysis using software.

RESULTS:

Carotenoid content in pumpkin pulp:

Figure 2 demonstrates the content of β-carotene in yellow and red pumpkin pulp.

Figure 2. Content of β-carotene in yellow and red pumpkin pulp.

Content of phenols in pumpkin seeds:

Phenolic compounds show high antibacterial, antibacterial, anti-tumor and antifungal activity. The Content of phenolic compounds in orange and white pumpkin seeds was determined using the HPLC method and the results are presented in Table 2.

Table 2. Content of phenols in orange and white pumpkin seeds.

Phenols	Retention time	Concentration μg/mL
Phenols	Retention time	Orange pumpkin seed	White pumpkin seed
Caffeic acid	7.3	12.27	6.17
Vanillic acid	12.4	8.11	4.87
Syringic acid	16.57	6.98	5.34
Ferulic acid	24.58	7.23	4.19
p-Coumaric acid	26.49	9.24	2.88
Quercetin	28.19	5.76	1.87
Rutin	31.14	8.51	6.15

Total phenolic contents in the fruit of the red and white pumpkin plant:

Total phenolic contents in the fruit of the red and white pumpkin plant were determined using a colorimetric method based on the Folin-Ciocalteu assay and the results are shown in Figure 3.

Figure 3. Content of total phenols in total phenolic contents in the fruit of the red and white pumpkin plant.

Total flavonoid content in methanolic extract in white and red pumpkin fruit pulp and seeds:

The total flavonoid content in the methanolic extract of white and red pumpkin fruit pulp, as well as in the seeds, was determined. The total flavonoid content in the extracts was reported in terms of quercetin equivalents (mg/g extract). Figure 4 presents the results of the total flavonoid content.

Table 3. GC-MS analysis of orange pumpkin seed

No	RT	Name of the compound	Molecular Formula	Peak Area (%)
1	6.41	Propane, 1,1,3-triethoxy-	C₉H₂₀O₃	7.58
2	7.52	n-Hexadecanoic acid	C₁₆H₃₂O₂	5.14
3	9.24	Hexadecanoic Acid, Ethyl Ester	C₁₈H₃₆O₂	2.17
4	12.87	11, 14-Eicosadienoic acid, methyl ester	C₂₁H₃₈O₂	3.07
5	14.21	cis-Vaccenic acid	C₁₈H₃₄O₂	2.05
6	15.73	Linoleic acid ethyl ester	C₂₀H₃₆O₂	1.37
7	17.49	Ethyl Oleate	C₂₀H₃₈O₂	2.58
8	18.74	Hexadecanoic acid, 1-(hydroxymethyl)-1,2 ethanediyl ester	C₃₅H₆₈O₅	10.38
9	20.44	9-hexadecyn-1-ol	C₁₆H₃₀O	2.11
10	22.39	(R)-(-)-(Z)-14-Methyl-8-hexadecen-1-ol	C₁₇H₃₄O	13.1
11	24.71	9,12-Octadecadienoyl chloride, (Z,Z)- propanetriyl ester, (E,E,E)-	-----	17.59
12	25.81	9-Octadecenoic acid, 1,2,3-propanetriyl ester	C₅₇H₁₀₄O₆	19.19
13	26.11	Squalene	C₃₀H₅₀	21.33
14	29.31	1H-Purin-6-Amine,[(2Fluorophenyl)methyl	C₁₂H₁₀N₅	1.45
15	31.87	Gamma.-Tocopherol	C₂₈H₄₈O₂	1.42
16	33.64	1-Heptatriacotanol	C₃₇H₇₆O	1.21
17	34.77	Stigmasterol	C₂₉H₄₈O	3.69

Table 4. GC-MS analysis of white pumpkin seed

No	RT	Name of the compound	Molecular Formula	Peak Area (%)
1	6.32	Propane, 1,1,3-triethoxy	C₉H₂₀O₃	6.58
2	7.12	Hexadecanoic acid, 1-(hydroxymethyl)-1,2)- ethanediyl ester	C₃₅H₆₈O₅	4.76
3	8.14	Octadecenoic acid, 1,2,3-propanetriyl ester	C₁₈H₃₁ClO	9.14
4	8.97	Bicyclo[10.1.0]tridec-1-ene	C₁₃H₂₂	7.48
5	10.14	9-Octadecenoic acid,1,2,3-propanetriyl ester(E,E,E)	C₅₇H₁₀₄O₆	11.19
6	11.21	CIS-9-Hexadecenal	C₁₆H₃₀O	2.77
7	12.04	Triarachine	C₆₃H₁₂₂O₆	1.43
8	12.88	Tetratriacontane	C₄₄H₉₀	0.87
9	13.21	Eicosane	C₂₀H₄₂	5.84
10	14.97	Linoleic acid ethyl ester	C₂₀H₃₆O₂	0.17
11	16.14	Ethyl Oleate	C₂₀H₃₈O₂	1.42
12	17.54	Hexadecanoic acid, 1-(hydroxymethyl)-1,2 ethanediyl ester	C₃₅H₆₈O₅	8.64
13	219.83	9-hexadecyn-1-ol	C₁₆H₃₀O	4.82
14	22.47	(R)-(-)-(Z)-14-Methyl-8-hexadecen-1-ol	C₁₇H₃₄O	0.87
15	24.34	9,12-Octadecadienoic Acid (Z,Z)-, 2,3-Dihydroxypropyl Ester	C₂₁H₃₈O₄	4.11
16	25.19	9 Octadecenoic acid, 1,2,3-propanetriyl ester -	C₅₇H₁₀₄O₆	12.43
17	26.47	Squalene	C₃₀H₅₀	9.87
18	27.11	1H-Purin-6-Amine,[(2Fluorophenyl)methyl	C₁₂H₁₀N₅	0.98
19	29.62	Celidoniol, deoxy-	C₂₉H₆₀	3.87
20	31.11	Gamma.-Tocopherol	C₂₈H₄₈O₂	1.47
21	33.57	1-Heptatriacotanol	C₃₇H₇₆O	0.92
22	34.68	Stigmasterol	C₂₉H₄₈O	4.22
23	35.19	Chondrillasterol	C₂₉H₄₈O	6.71
24	36.45	Beta.-Sitosterol	C₂₉H₅₀O	5.92

Determination of volatile organic compounds in red pumpkin seeds and white pumpkin:

After the soxhlet extraction of crushed pumpkin seeds of white and red varieties, volatile organic compounds were estimated using GC-MS (Tables 3 and 4).

DISCUSSION:

This work aimed to provide a thorough examination of the phytochemical composition and the various health benefits associated with pumpkin (Cucurbita moschata). The discussion section of this article summarizes and discusses the key findings and implications of the research conducted. The research highlighted the rich nutritional value of pumpkins, which are classified as fruits and contain essential nutrients such as water, protein, fiber, vitamins, minerals, and bioactive compounds. The bioactive substances found in pumpkin seeds have gained significant attention due to their potential in medicinal and functional food products. One of the key benefits of pumpkin consumption is its ability to enhance the immune system. This can be attributed to the antioxidant properties of carotene compounds present in pumpkins. These compounds play a crucial role in reducing the risk of degenerative and cancerous diseases. Furthermore, pumpkin consumption supports eye health maintenance and provides protection against related ailments. The presence of antioxidants in pumpkins contributes to their ability to counteract oxidative stress, thus exhibiting antioxidant activity. Pumpkin is also recognized as a rich source of vitamin A, including alpha-carotene and beta-carotene. Vitamin A plays a vital role in vision, cell differentiation, glycoprotein synthesis, mucus secretion, and overall skin health. The article emphasizes the role of carotenoids, particularly beta-carotene, in mediating the biological effects of pumpkin. The study also highlights the antiglycemic effects of pumpkin, which aids in regulating blood sugar levels. Additionally, pumpkin consumption has been associated with the reduction of cholesterol and blood pressure levels, demonstrating anti-cholesterol and antihypertensive properties. Pumpkin exhibits various other activities, including antibacterial, anti-inflammatory, antifungal, and antitumor activities, making it a promising candidate for various therapeutic applications. The seeds of the pumpkin plant have been utilized in the treatment of prostate and bladder problems. Moreover, pumpkin can serve as a natural coloring agent in the food industry. The article further discusses the quantification of carotenoids using high-performance liquid chromatography (HPLC) technology and the concentration of carotene in yellow and red pumpkin pulp. The HPLC analysis provides insights into the presence and concentration of bioactive compounds in pumpkin samples. The methods section of the article describes the sample collection process, phytochemical extraction procedures, and HPLC analysis techniques used to evaluate the phytochemical composition of pumpkin. The results obtained from these experiments were subjected to appropriate statistical analysis. Overall, this comprehensive review provides valuable insights for researchers, industries, and health-conscious individuals interested in harnessing the potential of pumpkin for various applications in nutrition and healthcare. The findings of this study highlight the importance of incorporating pumpkin into the diet to promote overall health and well-being. Further research and exploration of pumpkin's bioactive compounds and their mechanisms of action are recommended for future investigations.

CONCLUSSION:

This comprehensive review has provided valuable insights into the phytochemical composition and health benefits of pumpkin (Cucurbita moschata). Pumpkin, classified as a fruit, offers a rich source of essential nutrients, bioactive compounds, and minerals. The bioactive substances found in pumpkin seeds have garnered significant attention for their medicinal and functional food applications. Pumpkin consumption has been associated with numerous advantages, including immune system enhancement, eye health maintenance, antioxidant activity, vitamin A source, antiglycemic effects, and properties that help lower cholesterol and blood pressure levels. Moreover, pumpkin exhibits antibacterial, anti-inflammatory, antifungal, and antitumor activities, making it a promising candidate for various therapeutic applications.

The role of carotenoids, particularly beta-carotene, in mediating the biological effects of pumpkin has been emphasized. The article also discussed the quantification of carotenoids using high-performance liquid chromatography (HPLC) technology and highlighted the concentration of carotene in yellow and red pumpkin pulp. Furthermore, the extraction and analysis of phenols from pumpkin seeds were outlined.

Overall, this article serves as a valuable resource for researchers, industries, and health-conscious individuals interested in harnessing the potential of pumpkin for various applications in nutrition and healthcare. The findings emphasize the importance of incorporating pumpkin into a balanced diet to reap its numerous health benefits. Further research and development in utilizing pumpkin's bioactive compounds hold promising prospects for the food, cosmetics, and pharmaceutical industries.

REFERENCES:

1. Abuelgassim AO, Arabia S. The Effect of Pumpkin (Cucurbita pepo L) seed and L-arginine suplementation on serum lipid concentrations in atherogenic rats. Safar J. Tradit. Complement Altern Med. 2012; 9(1): 131–137. doi: 10.4314/ajtcam.v9i1.18.

2. Adams GG, Imran S, Wang S, Mohammad A, Kok S, Gray DA, Channell GA, Morris GA, Harding SE.The hypoglycaemic effect of pumpkins as anti-diabetic and functional medicines. Food Res. Int. 2011; 44(4): 862-867. https://doi.org/10.1016/j.foodres.2011.03.016.

3. Bandoniene D, Zettl D, Meisel T, Maneiko M. Suitability of elemental fingerprinting for assessing the geographic origin of pumpkin (Cucurbita pepo var. styriaca) seed oil. Food Chem. 2013; 136(3-4): 1533-1542. doi:10.1016/j.foodchem.2012.06.040.

4. Bardaa S, Ben Halima N, Aloui F, et al. Oil from pumpkin (Cucurbita pepo L.) seeds: evaluation of its functional properties on wound healing in rats. Lipids Health Dis. 2016; 15: 73. doi:10.1186/s12944-016-0237-0.

5. Bharti SK, Kumar A, Sharma NK, et al. Tocopherol from seeds of Cucurbita pepo against diabetes: validation by in vivo experiments supported by computational docking. J Formos Med Assoc. 2013; 112(11): 676-690. doi:10.1016/j.jfma.2013.08.003.

6. Butinar B, Bucar-Miklavcic M, Valencic V, Raspor P. Stereospecific analysis of triacylglycerols as a useful means to evaluate genuineness of pumpkin seed oils: lesson from virgin olive oil analyses. J Agric Food Chem. 2010; 58(9): 5227-5234. doi:10.1021/jf904542z.

7. Cândido FG, de Oliveira FCE, Lima MFC, Pinto CA, da Silva LL, Martino HSD, Dos Santos MH, Alfenas RCG. Addition of pooled pumpkin seed to mixed meals reduced postprandial glycemia: a randomized placebo-controlled clinical trial. Nutr Res. 2018; 56: 90-97. doi:10.1016/j.nutres.2018.04.015 .

8. Mazaheri Y, Torbati M, Azadmard-Damirchi S, Savage GP. Effect of roasting and microwave pre-treatments of Nigella sativa L. seeds on lipase activity and the quality of the oil. Food Chem. 2019; 274: 480-486. doi:10.1016/j.foodchem.2018.09.001.

9. Pirintsos S, Panagiotopoulos A, Bariotakis M, et al. From Traditional Ethnopharmacology to Modern Natural Drug Discovery: A Methodology Discussion and Specific Examples. Molecules. 2022; 27(13): 4060. doi:10.3390/molecules27134060.

10. Wane A, Thallaj NK, Mandon D. Biomimetic interaction between Fe(II) and O2: effect of the second coordination sphere on O2 binding to Fe(II) complexes: evidence of coordination at the metal centre by a dissociative mechanism in the formation of mu-oxo diferric complexes. Chemistry. 2009; 15(40): 10593-10602. doi:10.1002/chem.200901350.

11. Thallaj NK, Machkour A, Mandon D, Welter R. Square pyramidal geometry around the metal and tridentate coordination mode of the tripod in the [6-(3′-cyanophenyl)-2-pyridylmethyl] bis (2-pyridylmethyl) amine FeCl2 complex: a solid state effect. New J. Chem. 2005; 29(12): 1555-1558. https://doi.org/10.1039/B512108F.

12. Thallaj NK, Mandon D, White KA. The design of metal chelates with a biologically related redox-active part: Conjugation of riboflavin to bis (2-pyridylmethyl) amine ligand and preparation of a ferric complex. Eur. J. Inor. Chem. 2007; (1): 44-47. https://doi.org/10.1002/ejic.200600789.

13. Thallaj NK, Orain PY, Thibon A, Sandroni M, Welter R, Mandon D. Steric congestion at, and proximity to, a ferrous center leads to hydration of α-nitrile substituents forming coordinated carboxamides. Inorg Chem. 2014; 53(15): 7824-7836. doi:10.1021/ic500096h.

14. Thallaj NK, Przybilla J, Welter R, Mandon D. A ferrous center as reaction site for hydration of a nitrile group into a carboxamide in mild conditions. J Am Chem Soc. 2008; 130(8): 2414-2415. doi:10.1021/ja710560g.

15. Malek ZS, Sage D, Pévet P, Raison S. Daily rhythm of tryptophan hydroxylase-2 messenger ribonucleic acid within raphe neurons is induced by corticoid daily surge and modulated by enhanced locomotor activity. Endocrinology. 2007; 148(11): 5165-5172. doi:10.1210/en.2007-0526.

16. Malek ZS, Pévet P, Raison S. Circadian change in tryptophan hydroxylase protein levels within the rat intergeniculate leaflets and raphe nuclei. Neuroscience. 2004; 125(3): 749-758. doi:10.1016/j.neuroscience.2004.01.031.

17. Malek ZS, Dardente H, Pevet P, Raison S. Tissue-specific expression of tryptophan hydroxylase mRNAs in the rat midbrain: anatomical evidence and daily profiles. Eur J Neurosci. 2005; 22(4): 895-901. doi:10.1111/j.1460-9568.2005.04264.x.

18. Malek ZS, Labban LM. Photoperiod regulates the daily profiles of tryptophan hydroxylase-2 gene expression the raphe nuclei of rats. Int J Neurosci. 2021; 131(12): 1155-1161. doi:10.1080/00207454.2020.1782903.

19. Machkour A, Thallaj NK, Benhamou L, Lachkar M, Mandon D. The coordination chemistry of FeCl3 and FeCl2 to bis[2-(2,3-dihydroxyphenyl)-6-pyridylmethyl](2-pyridylmethyl)amine: access to a diiron(III) compound with an unusual pentagonal-bipyramidal/square-pyramidal environment. Chemistry. 2006;12(25):6660-6668. doi:10.1002/chem.200600276.

20. Isbera M, Abbood A, Ibrahim W. Weight and Content Uniformity of Warfarin Sodium Half Tablets. Research J. Pharm. and Tech. 2016; 9(3): 215-218. doi: 10.5958/0974-360X.2016.00039.1

21. Abbood A, Layka R. Weight and content uniformity Study of captopril half-tablets. Research J. Pharm. and Tech. 2017; 10(6): 1621-1626. doi: 10.5958/0974-360X.2017.00285.2.

22. Chbani D, Abbood A, Alkhayer M. Determination of Nitrite and Nitrate Ions levels in some types of processed meats marketed locally. Research J. Pharm. and Tech. 2018; 11(4): 1442-1447. doi: 10.5958/0974-360X.2018.00269.X.

23. Abbood A, Malek Z, Al-Homsh Y, Thallaj N. In vitro Study for Antibiotic resistance of bacteria causing Urinary Tract Infection from Syrian adults. Research J. Pharm. and Tech. 2022; 15(10): 4727-2. doi: 10.52711/0974-360X.2022.00794.

24. Abbood A, Malek Z, Thallaj N. Antibiotic resistance of urinary tract pathogens in Syrian children. Research J. Pharm. and Tech. 2022; 15(11): 4935-9. doi: 10.52711/0974-360X.2022.00829.

25. Labban L, Thallaj N, Malek Z. The implications of E-cigarettes or" vaping" on the nutritional status. Journal of Medical Research and Health Sciences, 2019; 2(11): 784-787. https://doi.org/10.15520/jmrhs.v2i11.128.

26. Khatib O, Alshimale T, Alsaadi A, Thallaj N. The Global Impact of HIV: A Comprehensive Review. IJAPSR. 2024; 4(3) 6–19. Apr., doi: 10.54105/ijapsr.C4040.04030424.

27. Deeb DN, Aldiab D, Sulaeman S. Effect of Storage Conditions on Bacteriological Growth in Infant Cereals. Research J. Pharm. and Tech. 2021; 14(2): 667-672. doi: 10.5958/0974-360X.2021.00119.0.

28. Alsubot S, Aldiab D. 5-hydroxymethylfurfural Levels in Coffee and Study of some effecting factors. Research J. Pharm. and Tech. 2019; 12(9): 4263-4268. doi: 10.5958/0974-360X.2019.00733.9.

29. Asaad RA. Hormone Receptor Status and its Relation to C-Reactive Protein and other Prognostic factors in Breast Cancer in Jableh- Syria. Research J. Pharm. and Tech. 2017; 10(9): 3003-3010. doi: 10.5958/0974-360X.2017.00532.7.

30. Asaad RA, Abdullah SS. Breast Cancer Subtypes (BCSs) Classification according to Hormone Receptor Status: Identification of Patients at High Risk in Jableh- Syria. Research J. Pharm. and Tech. 2018; 11(8): 3703-3710. doi: 10.5958/0974-360X.2018.00680.7.

31. Besherb S, Alallan L, Hassan Agha MA, AIshamas I, Thallaj N. Influence of soil salinity on the chemical composition of essential oil of Rosmarinus Officinalis in Syria, Research J. Pharm. and Tech. 2024; 17(4). DOI: 10.52711/0974-360X.2024.00358.

32. Thallaj N, Hasan Agha M, , Khatib C, Karaali A, Moustapha A, Labban L. Evaluation of Antimicrobial Activities and Bioactive Compounds of Different Extracts Related to Syrian Traditional Products of Damask Rose (Rosa damascena). 2020; 7(5); 1-21. DOI: 10.4236/oalib.1106302

33. Labban L, Kudsi M, Malek Z, Thallaj N. Pain Relieving Properties of Ginger (Z. officinale) and Echinacea (E. angustifolia) Extracts Supplementation among Female Patients with Osteoarthritis. A Randomized Study. Advances in Medical, Dental and Health Sciences. 2020; 3(3): 45-48. DOI: 10.5530/amdhs.2020.3.11.

34. Labban L , Thallaj N, Al Masri M. The Nutritional Value of Traditional Syrian Sweets and Their Calorie Density. 2019; 3(4): 40-47 . doi: 10.11648/j.wjfst.20190304.11

35. Labban L, Thallaj N, Labban A. Assessing the Level of Awareness and Knowledge of COVID 19 Pandemic among Syrians, Archives of Medicine. 2020: 12(2:8): 1-5. DOI: 10.36648/1989-5216.12.2.309.

Received on 18.03.2024 Modified on 04.06.2024

Research J. Pharm. and Tech 2024; 17(10):4915-4921.

DOI: 10.52711/0974-360X.2024.00756