Effect of Extraction Techniques on Phytochemicals and Antioxidants Activity of Garcinia quaesita Leaves

Garcinia quaesita is an endemic plant in Sri Lanka with a wide array of domestic and medicinal values, yet very limited studies have been reported. The goal of this study was to investigate the phytochemicals and antioxidants capacity of leaves of G. quaesita under different extraction techniques in order to monitor the effect of extraction techniques on the above parameters. Water was used as the solvent and four extraction methods, namely sonication (EN01, one-hour,RT, 40 kHz), Soxhlet (EN02, six-hours, 105 ºC), maceration with agitation (EN03, six-hours, RT, 1000 rpm), and maceration with agitation upon heating (EN04, six-hours, 60 ºC, 1000 rpm), were applied. Phytochemical screenings and quantification of polyphenolics (TPC), flavonoids (TFC), tannins (TTC), terpenoids (TC), saponins (SC), and alkaloids (AC) were performed using standard methods, whereas antioxidants capacity was assessed using FRAP and DPPH assays. The results revealed that G. quaesita contains a wide range of phytochemicals and the TPC, TTC and SC profiles appeared to be higher in the extraction process of EN04 (60.73 ± 0.06 mg GAE/g, 60.12 ± 0.06 mg TAE/g, and 257.64 ± 0.72 mg SE/g, respectively) whereas those were lower in the technique EN01. In addition, TFC and TC levels were high in EN02 (3.02 ± 0.00 mg QE/g and 17.75 ± 0.04 mM LE/g respectively) and besides, TFC was low in method EN03, whereas TC was low in EN01. Interestingly, AC was found to be higher in EN03 (1.16 ± 0.03 mg AE/g) and lesser in EN02. Total antioxidant capacity ( FRAP value) was calculated to be higher in extraction method EN04 (72.08 ± 0.00 mg Trolox Eq/g) whereas it was lower in extraction method EN01. The method, EN03 gave the lower IC50 value in DPPH radical scavenging assay (10.49 ± 0.12 mg/ml), indicating high scavenging activity. In conclusion, the extraction technique EN04 is well suitable to extract polyphenolics and antioxidants from G. quaesita leaves. Though G. quaesita leaves contain a diverse range of bioactive compounds and antioxidants properties, it is important to choose the correct extraction technique based on the objectives of the desired research work, as the number of phytochemicals extracted, and antioxidant capacity varies depending on the extraction technique used. Full paper submission of ICIET 202

Phytochemistry and medicinal properties of Psidium guajava L. leaves: A review

Psidium guajava L. (Myrtaceae), also known as guava, is a medicinal tree native to tropical America that has been introduced and is widely available in many countries. Almost all plant parts of P. guajava have a long history of being used to treat a variety of ailments, in addition to applications as foods. Guava leaves are used as both medicine and food purposes, and there are numerous scientific reports on their medicinal uses, chemical composition and pharmacological properties. Cancer, blood pressure, diarrhea, bowel irregularities, diabetes, cough, cold, constipation, dysentery, scurvy, weight loss, improves skins tonicity are some of the diseases treated with guava leaves. Polyphenols, flavonoids, saponins, tannins, terpenoids, glycosides, flavones, cardiac glycosides, cardenolides, phlobatanins, steroids and other classes of bioactive compounds have been identified from the leaves. The primary chemical constituents of guava leaves are phenolic compounds, iso-flavonoids, gallic acid, catechin, quercetin, epicathechin, rutin, naringenin, kaempferol, caryophyllene oxide, p-selinene etc. Several studies have demonstrated its pharmacological activities including antioxidant, antimicrobial, antidiabetic, antitumor, anticancer, antidiarrheal, healing, cytotoxic, hepatoprotective, anti-inflammatory, antimalarial/ anti-plasmodial, dental plaque, antiglycative and many more. This review is aimed on compiling all the literature reported on pharmacological activities and phytochemical compositions of guava leaves as a support to the scientific community for further studies and to provide scientific data to validate its traditional uses

Design of a nanoplatform for treating pancreatic cancer

Doctor of PhilosophyDepartment of ChemistryStefan H. BossmannPancreatic cancer is the fourth leading cause of cancer-related deaths in the USA. Asymptomatic early cancer stages and late diagnosis leads to very low survival rates of pancreatic cancers, compared to other cancers. Treatment options for advanced pancreatic cancer are limited to chemotherapy and/or radiation therapy, as surgical removal of the cancerous tissue becomes impossible at later stages. Therefore, there's a critical need for innovative and improved chemotherapeutic treatment of (late) pancreatic cancers. It is mandatory for successful treatment strategies to overcome the drug resistance associated with pancreatic cancers. Nanotechnology based drug formulations have been providing promising alternatives in cancer treatment due to their selective targeting and accumulation in tumor vasculature, which can be used for efficient delivery of chemotherapeutic agents to tumors and metastases. The research of my thesis is following the principle approach to high therapeutic efficacy that has been first described by Dr. Helmut Ringsdorf in 1975. However, I have extended the use of the Ringsdorf model from polymeric to nanoparticle-based drug carriers by exploring an iron / iron oxide nanoparticle based drug delivery system. A series of drug delivery systems have been synthesized by varying the total numbers and the ratio of the tumor homing peptide sequence CGKRK and the chemotherapeutic drug doxorubicin at the surfaces of Fe/Fe₃O₄-nanoparticles. The cytotoxicity of these nanoformulations was tested against murine pancreatic cancer cell lines (Pan02) to assess their therapeutic capabilities for effective treatments of pancreatic cancers. Healthy mouse fibroblast cells (STO) were also tested for comparison, because an effective chemotherapeutic drug has to be selective towards cancer cells. Optimal Experimental Design methodology was applied to identify the nanoformulation with the highest therapeutic activity. A statistical analysis method known as response surface methodology was carried out to evaluate the in-vitro cytotoxicity data, and to determine whether the chosen experimental parameters truly express the optimized conditions of the nanoparticle based drug delivery system. The overall goal was to optimize the therapeutic efficacy in nanoparticle-based pancreatic cancer treatment. Based on the statistical data, the most effective iron/iron oxide nanoparticle-based drug delivery system has been identified. Its Fe/Fe₃O₄ core has a diameter of 20 nm. The surface of this nanoparticle is loaded with the homing sequence CGKRK (139-142 peptide molecules per nanoparticle surface) and the chemotherapeutic agent doxorubicin (156-159 molecules per surface), This nanoplatform is a promising candidate for the nanoparticle-based chemotherapy of pancreatic cancer

Nanoplatforms for highly sensitive fluorescence detection of cancer-related proteases

Numerous proteases are known to be necessary for cancer development and progression including matrix metalloproteinases (MMPs), tissue serine proteases, and cathepsins. The goal of this research is to develop an Fe/Fe3O4 nanoparticle-based system for clinical diagnostics, which has the potential to measure the activity of cancer-associated proteases in biospecimens. Nanoparticle-based "light switches" for measuring protease activity consist of fluorescent cyanine dyes and porphyrins that are attached to Fe/Fe3O4 nanoparticles via consensus sequences. These consensus sequences can be cleaved in the presence of the correct protease, thus releasing a fluorescent dye from the Fe/Fe3O4 nanoparticle, resulting in highly sensitive (down to 1 x 10(-16) mol l(-1) for 12 proteases), selective, and fast nanoplatforms (required time: 60 min)

Peptide nanosponges designed for rapid uptake by leukocytes and neural stem cells

The structure of novel binary nanosponges consisting of (cholesterol-(K/D)nDEVDGC)3-trimaleimide units possessing a trigonal maleimide linker, to which either lysine (K)20 or aspartic acid (D)20 are tethered, has been elucidated by means of TEM. A high degree of agreement between these findings and structure predictions through explicit solvent and then coarse-grained molecular dynamics (MD) simulations has been found. Based on the nanosponges' structure and dynamics, caspase-6 mediated release of the model drug 5(6)-carboxyfluorescein has been demonstrated. Furthermore, the binary (DK20) nanosponges have been found to be virtually non-toxic in cultures of neural progenitor cells. It is of a special importance for the future development of cell-based therapies that DK20 nanosponges were taken up efficiently by leucocytes (WBC) in peripheral blood within 3 h of exposure. The percentage of live cells among the WBC was not significantly decreased by the DK20 nanosponges. In contrast to stem cell or leucocyte cell cultures, which have to be matched to the patient, autologous cells are optimal for cell-mediated therapy. Therefore, the nanosponges hold great promise for effective cell-based tumor targeting