Investigation of ethanol, acetone and aqueous azadirachta indica A. Juss (NEEM) extracts against plasmodium falciparum

Malaria is a public health concern, causing significant morbidity and mortality annually due to the emergence of drug-resistance parasites. Therefore, the need for novel antimalarial agents with novel mechanisms of action and broad therapeutic potential is critical. In this study, Azadirachta ind...

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Main Author:	Ghazali, Siti Zulaiha
Format:	Thesis
Language:	English
Published:	2022
Subjects:	R Medicine
Online Access:	http://eprints.usm.my/54955/1/SITI%20ZULAIHA%20BINTI%20GHAZALI-FINAL%20THESIS%20P-NFM000319%28R%29%20PWD_-24%20pages.pdf
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spelling	my-usm-ep.549552022-09-28T01:16:07Z Investigation of ethanol, acetone and aqueous azadirachta indica A. Juss (NEEM) extracts against plasmodium falciparum 2022-02 Ghazali, Siti Zulaiha R Medicine Malaria is a public health concern, causing significant morbidity and mortality annually due to the emergence of drug-resistance parasites. Therefore, the need for novel antimalarial agents with novel mechanisms of action and broad therapeutic potential is critical. In this study, Azadirachta indica A. Juss (Neem), a long-used herbal remedy has been chosen due to the scarcity of data on Malaysian Neem on its antimalarial property. Thus, this study aimed to determine in vitro antimalarial activity of Malaysian’s Neem leaves crude extracts against laboratory-adapted, sensitive (3D7) and chloroquine-resistance (W2) P. falciparum strains. At first, all the collected leaves were taxonomized by The Forest Research Institute Malaysia (FRIM) and further authenticated molecularly before extraction with ethanol, acetone and aqueous followed by a phytochemical screening of the extracts. DPPH radical scavenging is used to determine antioxidant activity before GC-MS profiling. The brine shrimp lethality test (BSLT) and a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay against the primary human umbilical vein endothelial cells (HUVEC) were used for toxicity assay. The antimalarial activity was determined using the malarial SYBR Green I fluorescence-based assay against both strains. The study was extended with a green synthesis of Neem-silver nanoparticles (AgNPs) and further evaluated on their antimalarial property. Our data showed that crude extract produced by acetone (15.55 ± 0.04%) and ethanol (9.20 ± 0.05%) using the soxhlet method produces more yield compared to aqueous extract (5.86 ± 0.06%). However, aqueous extract contained the highest total phenolic and flavonoid content. Meanwhile, the brine shrimp lethality test and MTT assay showed all extracts are not toxic with LC50 > 1000 ppm and CC50 values > 30 g/mL, respectively. Whereas, all extracts showed a moderate antimalarial effect against 3D7. However, acetone and aqueous extract showed inactive antimalarial activity against W2 (IC50 > 50 ug/mL). On the other hands, when Neem combined with nano silver (Neem-AgNO3), it showed a 4-folds increment of antimalaria activity against 3D7 (IC50 = 8.815 ± 0.230) and W2 (IC50 = 23.110 ± 0.088). Thus, this study showed promising antimalarial activity of Malaysian Neem when combining with nanoparticle and should be further evaluated of its property to overcome the chloroquine resistance in malaria treatment. 2022-02 Thesis http://eprints.usm.my/54955/ http://eprints.usm.my/54955/1/SITI%20ZULAIHA%20BINTI%20GHAZALI-FINAL%20THESIS%20P-NFM000319%28R%29%20PWD_-24%20pages.pdf application/pdf en public masters Universiti Sains Malaysia Pusat Pengajian Sains Perubatan
institution	Universiti Sains Malaysia
collection	USM Institutional Repository
language	English
topic	R Medicine
spellingShingle	R Medicine Ghazali, Siti Zulaiha Investigation of ethanol, acetone and aqueous azadirachta indica A. Juss (NEEM) extracts against plasmodium falciparum
description	Malaria is a public health concern, causing significant morbidity and mortality annually due to the emergence of drug-resistance parasites. Therefore, the need for novel antimalarial agents with novel mechanisms of action and broad therapeutic potential is critical. In this study, Azadirachta indica A. Juss (Neem), a long-used herbal remedy has been chosen due to the scarcity of data on Malaysian Neem on its antimalarial property. Thus, this study aimed to determine in vitro antimalarial activity of Malaysian’s Neem leaves crude extracts against laboratory-adapted, sensitive (3D7) and chloroquine-resistance (W2) P. falciparum strains. At first, all the collected leaves were taxonomized by The Forest Research Institute Malaysia (FRIM) and further authenticated molecularly before extraction with ethanol, acetone and aqueous followed by a phytochemical screening of the extracts. DPPH radical scavenging is used to determine antioxidant activity before GC-MS profiling. The brine shrimp lethality test (BSLT) and a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay against the primary human umbilical vein endothelial cells (HUVEC) were used for toxicity assay. The antimalarial activity was determined using the malarial SYBR Green I fluorescence-based assay against both strains. The study was extended with a green synthesis of Neem-silver nanoparticles (AgNPs) and further evaluated on their antimalarial property. Our data showed that crude extract produced by acetone (15.55 ± 0.04%) and ethanol (9.20 ± 0.05%) using the soxhlet method produces more yield compared to aqueous extract (5.86 ± 0.06%). However, aqueous extract contained the highest total phenolic and flavonoid content. Meanwhile, the brine shrimp lethality test and MTT assay showed all extracts are not toxic with LC50 > 1000 ppm and CC50 values > 30 g/mL, respectively. Whereas, all extracts showed a moderate antimalarial effect against 3D7. However, acetone and aqueous extract showed inactive antimalarial activity against W2 (IC50 > 50 ug/mL). On the other hands, when Neem combined with nano silver (Neem-AgNO3), it showed a 4-folds increment of antimalaria activity against 3D7 (IC50 = 8.815 ± 0.230) and W2 (IC50 = 23.110 ± 0.088). Thus, this study showed promising antimalarial activity of Malaysian Neem when combining with nanoparticle and should be further evaluated of its property to overcome the chloroquine resistance in malaria treatment.
format	Thesis
qualification_level	Master's degree
author	Ghazali, Siti Zulaiha
author_facet	Ghazali, Siti Zulaiha
author_sort	Ghazali, Siti Zulaiha
title	Investigation of ethanol, acetone and aqueous azadirachta indica A. Juss (NEEM) extracts against plasmodium falciparum
title_short	Investigation of ethanol, acetone and aqueous azadirachta indica A. Juss (NEEM) extracts against plasmodium falciparum
title_full	Investigation of ethanol, acetone and aqueous azadirachta indica A. Juss (NEEM) extracts against plasmodium falciparum
title_fullStr	Investigation of ethanol, acetone and aqueous azadirachta indica A. Juss (NEEM) extracts against plasmodium falciparum
title_full_unstemmed	Investigation of ethanol, acetone and aqueous azadirachta indica A. Juss (NEEM) extracts against plasmodium falciparum
title_sort	investigation of ethanol, acetone and aqueous azadirachta indica a. juss (neem) extracts against plasmodium falciparum
granting_institution	Universiti Sains Malaysia
granting_department	Pusat Pengajian Sains Perubatan
publishDate	2022
url	http://eprints.usm.my/54955/1/SITI%20ZULAIHA%20BINTI%20GHAZALI-FINAL%20THESIS%20P-NFM000319%28R%29%20PWD_-24%20pages.pdf
_version_	1747822301002334208

Investigation of ethanol, acetone and aqueous azadirachta indica A. Juss (NEEM) extracts against plasmodium falciparum

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