LIBRARY WORK – LIPIDS (GROUP 2)

AUTHOR(S) YEAR TITLE METHODS RESULTS

PUBLISHED
Yan Li, 2014 A comparative Since the concept of using algae to make The results obtained for Soxhlet extraction
Forough study: the impact of fuels was firstly discussed in the 1940s [1], of microalgal lipids showed a significant
Ghasemi different lipid a major focus for research, development and difference in extraction efficiency between
Naghdi, and extraction methods on commercialization has become the hexane and the mixture of hexane and
Peer M current microalgal cultivation of algae for the production of oil ethanol in both, total lipids and total
Schenk lipid research (lipid)-based products, in particular FAMEs, as well as each individual fatty acid
biodiesel through lipid transesterification. (P < 0.05, Figure 1A and B). As ethanol is a
Algal lipids can be divided into two major polar solvent, it can extract more polar lipids
types: polar lipids such as phospholipids and likely penetrate the cell wall, hence
and glycolipids, and neutral/non-polar lipids making triacylglycerides (TAGs; neutral
such as mono-, di- and tri-acylglycerides lipids) more available for the non-polar
and carotenoids based on their solvent hexane. The lipid extraction yield in
physiochemical characteristics [2, 3]. Some the mixture was nearly three times higher
of these substances have been intensively than when using hexane alone (Figure 1A).
studied, not only as biofuel feedstock, but Coincident with the reports of Ryckebosch
also as beneficial food additives and other et al. [11] and Lewis et al. [10], it seems that
high-value products (e.g., eicosapentaenoic extraction solvents containing a mixture of a
acid (C20:5 n-3, EPA), docosahexaenoic polar and a nonpolar solvent could extract
acid (C22:6 n-3, DHA) and other long-chain higher amounts of lipids and also some
polyunsaturated fatty acids. Therefore, there other compounds (e.g., pigments,
is mounting interest on investigation of carbohydrates and algaenans) [26].
microalgal potential for production of food Interestingly, this conclusion contradicts the
commodities and fatty acids bound as study of Shen et al. [27] stating that 1:1
triglycerides for nutraceutical efficacy in (v/v) of hexane and ethanol had less lipid
recent decades [6]. Significant advances yields than hexane on Scenedesmus
have been made in upstream processing to dimorphus and Chlorella protothecoides.
generate cellular biomass for lipid yields. Regardless of the biological difference of
However, as part of the downstream these algal species and its resulting different
process, lipid extraction continues to be a lipid class compositions, the contradiction is
significant challenge towards the possibly also related to the different
commercial production of microalgal oil proportions of hexane:ethanol in the mixture
production, even though a multitude of (3:1 vs. 1:1). A similar result was obtained
extraction methods have been described in using other mixtures as well, such as
the literatures. For microalgal oil extraction, chloroform-methanol [11] and hexane-
although an appropriate technique of cell hydroalcoholic solution [28] where different
disruption is a prerequisite [7, 8], the ratios of solvents also resulted in the
efficient extraction of lipids is highly different extraction efficiencies on
dependent on the polarity of the organic microalgal lipid extraction. Therefore, it is
solvent or solvent mixture used [9, 10]. In implied that only appropriate proportions of
general, solvent mixtures containing a polar polar and nonpolar solvents could achieve
and a non-polar solvent could extract a higher yields of lipid compared with single
greater amount of lipids [11]. For example, solvent extraction.
a combination of chloroform (non-polar),
methanol (polar) and water, known as the
Bligh & Dyer method, has been used for
lipid extraction from a wide range of
biological samples [11]. However, concerns
about biosafety issues using extraction
solvents has driven a demand for
biocompatible and less or non-toxic
solvents (e.g., dichloromethane) [12].
Alternative solvent methods for lipid
extraction thereby have been studied; for
instance, saponification has resulted in
significant lipid recoveries from several
types of microalgae [7, 13–16]. In recent
years, supercritical fluid technology has
been adopted for microalgal oil extraction,
especially for pharmaceutical and
neutraceutical bioproducts. In comparison
with liquid solvent extractions, the
supercritical fluid carbon dioxide (ScCO2)
technique offers several advantages, such as
no toxicity, no oxidation or thermal
degradation of extracts, high diffusivity and
easy separation of desired bioproducts [16–
18]. However, it has been reported that lipid
yield using ScCO2 extraction was much
lower than employing the Bligh & Dyer
method on heterotrophically cultured
microalgae of Crypthecodinium cohnii[19].
At present, comparative economics of
technical and physiochemical methods for
oil extraction have not been accomplished
on microalgae cells. Given the large
diversity of microalgae species, the ability
to successfully and effectively extract oil
from cellular biomass becomes paramount
in determining the yield and suitability
across oleaginous strains [18, 20]. However,
the current research attention towards oil
extraction from microalgae has been
predominantly focused on the potential
energy efficiency and cost. effectiveness of
the methods themselves. Despite the
differences in extraction efficiency obtained
depending on different extraction methods
[10–12, 21, 22], there is little attention on
the bias potentially derived from different
extraction methods, in particular when
screening optimal microalgal species for
lipid-based bioproducts. Due to the lack of a
standard extraction method for fatty acids
(FA) analysis, therefore, the motivation
behind this study was to investigate the
potential impact of different lipid extraction
methods on microalgal lipid research. The
present work includes a comparative study
of lipid extractions from lyophilised
biomass of the oleaginous green alga
Tetraselmis sp. Soxhlet extraction was
conducted for lipid recovery using either
single solvents or mixtures. In addition,
algal biomass was used for five different
extraction methods that were successfully
used for efficient algal lipid extraction in
previous studies. These parallel extraction
methods were: (1) the monophasic ternary
system of chloroform:methanol:water, one
of the most commonly used methods for
lipid extractions [23]; (2) a less hazardous
solvent mixture of
dichloromethane:methanol [12]; (3) another
alternative solvent mixture of propan-2-
ol:cyclohexane:water recommended by
Schlechtriem et al. [24]; (4) direct
saponification using KOH in ethanol [7]
and (5) supercritical CO2 extraction [25].
 We discuss and draw some parallels with
these extractions to highlight the differences
on extractable lipid production and
hydrolysed fatty acid methyl ester profiles
on microalgae cells.
Dania Akram 2023 Lipid extraction Autoclaved distilled water was used to Lipid yield in each strain was measured
Kiyani, Shafia and analysis of make dilutions of already sterilized stock using Bligh and Dyer method. Among all
Maryam, and microalgae strain chemicals. For solutions, 10% ammonium the algal species, Pectinodesmus (PHM3)
Hussnain pectinodesmus PHM3 solution, 2 N sodium hydroxide (NaOH) showed highest average lipid yield of 23%
Ahmed Janjua for biodiesel solution, 5% ferric chloride solution per gram. Due to this reason, all further
production (FeCl3), ethanol, H2SO4 solution, hexane experiments were conducted with
and methanol were purchased from Sigma Pectinodesmus (PHM3). Lipid yield from
Aldrich. Chloroform, 10% ferric chloride DHSYM, DHS, DHM1, DHM3 and PHM3
solution and 10% NaOH solution were strains. The highest lipid yield is of
purchased from Fischer Scientific. Pectinodesmus (PHM3). In this study
Deionized water, distilled water was combination of chemical and mechanistic
purchased locally. Folch solution used for techniques (Fig. 3) were used to enhance the
lipid extraction was prepared using lipid yield. Highest amount of lipid
chemical grade chloroform and methanol extracted was through continuous agitation
which were mixed in 2:1 (v/v) ratio in a using Folch solution as the highest surface
sterile bottle. The bottle was sealed and kept area contact between microalgae biomass
in the dark, at room temperature. and solution occurs through continuous
agitation. The second highest lipid levels
extracted were from continuous agitation in
ethanol solvent