Open Access Maced J Med Sci electronic publication ahead of print,
published on December 20, 2019 as https://doi.org/10.3889/oamjms.2019.838
_______________________________________________________________________________________________________________________________
Open Access Maced J Med Sci. 1
ID Design Press, Skopje, Republic of Macedonia
Open Access Macedonian Journal of Medical Sciences.
https://doi.org/10.3889/oamjms.2019.838
eISSN: 1857-9655
Basic and Clinical Medical Researches in Vietnam
Transformation Chlorophyll a of Spirulina platensis to Chlorin e6
Derivatives and Several Applications
Hoa Thi Hai Bui
1
, Tam Thi Pham
1
, Hien Thi Thu Nguyen
1
, Trung Minh Do
2
, Vu Thi Nga
3
, Nguyen Duy Bac
2
, Vu Thi Bich
Huyen
4
, Hai Minh Le
5
, Quang Canh Tran
6*
1
Hanoi Open University, Hanoi, Vietnam;
2
Vietnam Military Medical University, Hanoi, Vietnam;
3
Institute for Research and
Development, Duy Tan University, 03 Quang Trung, Danang, Vietnam;
4
Department of Genetics and Biochemistry, Faculty
of Biology, Hanoi National University of Education;
5
Vinh University, Nghe An, Vietnam;
6
Center for Hygiene and Food
Safety, Haiduong Medical Technical University, Hai Duong, Vietnam
Citation: Bui THH, Pham TT, Nguyen TTH, Trung DM,
Nga VT, Bac ND, Huyen VTB, Le MH, Tran QC.
Transformation Chlorophyll a of Spirulina platensis to
Chlorin e6 Derivatives and Several Applications. Open
Access Maced J Med Sci.
https://doi.org/10.3889/oamjms.2019.838
Keywords: Spirulina platensis; Chlorin e6 trimethylester;
Photodynamic therapy; Hela cell; Antibacterial
*Correspondence: Quang Canh Tran. Center for
Hygiene and Food Safety, Haiduong Medical Technical
University, Hai Duong, Vietnam. E-mail:
tranquangcanh68@gmail.com
Received: 23-Sep-2019; Revised: 20-Nov-2019;
Accepted: 21-Nov-2019; Online first: 20-Dec-2019
Copyright: © 2019 Hoa Thi Hai Bui, Tam Thi Pham,
Hien Thi Thu Nguyen, Trung Minh Do, Vu Thi Nga,
Nguyen Duy Bac, Vu Thi Bich Huyen, Hai Minh Le, Quang
Canh Tran. This is an open-access article distributed
under the terms of the Creative Commons Attribution-
NonCommercial 4.0 International License (CC BY-NC 4.0)
Funding: This study was funded by a project (No. KYTH-
86) from the Ministry of Education and Training of
Vietnam
Competing Interests: The authors have declared that no
competing interests exist
Abstract
BACKGROUND: Spirulina platensis contains a large amount of chlorophylls, chlorophyll a, that are starting
materials to synthesize functionalized chlorins. Chlorin e6 (Ce6) as well as its derivatives are second generation
sensitizers using in photodynamic therapy (PDT) of various cancers. In this study, we transfer chlorophyll a of S.
platensis to Ce6 derivatives and determine their several applications.
AIM: We aimed to evaluate the effects of Ce6 derivatives to treat cancer cells.
METHODS: Ce6 trimethylester was created from methyl pheophorbide a2 in S. platensis provided by the Hidumi
Company, Nghe An province, Viet Nam. Hela cells were incubated with Ce6 trimethylester and the irradiated with
the diode laser dose of 1.2 J/cm
2
/min through the system of filters £ 650 nm. MTT assay and clonogenic assay
were used to determine survival rate and cloning efficiency of cells. Antimicrobial effect of Ce6 trimethylester with
halogen light were studied with Propionibacterium acnes VTCC 0218 and Staphylococcus aureus VTCC 0173.
RESULTS: From dry biomass (700 g) of S. platensis, after extracting chlorophyll a and methanolysis, 4.2 g of
methyl pheophorbide a was obtained. The reaction to give Ce6 trimethylester with 82% yield was performed with
potassium hydroxide (KOH) in MeOH/THF/CHCl3. After irradiation with a 650 nm laser at 1.2 J, the cell viability in
all samples decreased with Ce6 trimethylester treatment, the survival declining trend of Hela cells treated with
Ce6 trimethylester were proportional when concentration of Ce6 trimethylester increased. The rate of colony
formation was declined as the concentration of Ce6 trimethylester treated was increased. The growth of both S.
aureus and P. acnes can be inactivated by Ce6 trimethylester PDT. The MIC99 value against P. acnes VTCC
0218 and S. aureus VTCC 0173 of Ce6 trimethylester with halogen light was 1.25 μg/ml.
CONCLUSION: The Ce6 trimethylester from S. platensis cultivated in Viet Nam could be used as a potential
photosentizer for photodynamic therapy for treatment of cancer and acne.
Introduction
Spirulina platensis has been used for
production nutritional supplements, functional foods,
cosmetics as well as other biomaterials [1], [2].
Spirulina contains the very large amount of
chlorophylls, specially chlorophyll a [3]. Chlorophylls
are starting materials to synthesize functionalized
chlorins that have the ability to improve the
photodynamic activity [4]. Chlorophyll a, as a natural
chlorin, is a potential renewable resource to product
chlorin photosensitizers [5] which can use for
photodynamic therapy to treat cancer. Chlorins, that
are a class of tetrapyrroles derived from plant, are
potential photosensitizers because they can absorb
remarkably strongly in the light with high phototoxic
red spectrum [6].
Photodynamic therapy (PDT), is a treatment
therapy for cancer, uses some photosensitizers [6].
The photosensitizer molecules have singlet state with
two electrons with opposite spins. They are activated
by absorption a photon of light with appropriate
wavelength to transfer from the excited singlet state to
more stable excited triplet state. The photosensitizers
in triplet state can participate in photochemical
reactions with oxygen to form reactive oxygen species
(ROS) which can destroy pathogenic bacteria, tumors,