Extraction of Squalene From Tea Leaves (Camellia sinensis) and Its Variations With Leaf Maturity and Tea Cultivar

Sheng, Yue Yue; Xiang, Jing; Wang, Kai Rong; Li, Ze Yu; Li, Kai; Lu, Jinmei; Ye, Jian Hui; Liang, Yue; Zheng, Xin

doi:10.3389/fnut.2022.755514

Cited by 8 publications

(7 citation statements)

References 42 publications

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“…The quantification of squalene content was conducted following the methodologies of Shen et al [45] and Rothblat et al [46], with minor adjustments. In brief, the hairy roots were subjected to overnight drying in an oven set at 60 • C. For the subsequent steps, 200 mg of the dried sample was employed.…”

Section: Determination Of Squalene Contentmentioning

confidence: 99%

Enhancing Centelloside Production in Centella asiatica Hairy Root Lines through Metabolic Engineering of Triterpene Biosynthetic Pathway Early Genes

Alcalde,

Palazon,

Bonfill

et al. 2023

Plants

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Centella asiatica is a medicinal plant with a rich tradition of use for its therapeutic properties. Among its bioactive compounds are centellosides, a group of triterpenoid secondary metabolites whose potent pharmacological activities have attracted significant attention. Metabolic engineering has emerged as a powerful biotechnological tool to enhance the production of target compounds. In this study, we explored the effects of overexpressing the squalene synthase (SQS) gene and transcription factor TSAR2 on various aspects of C. asiatica hairy root lines: the expression level of centelloside biosynthetic genes, morphological traits, as well as squalene, phytosterol, and centelloside content. Three distinct categories of transformed lines were obtained: LS, harboring At-SQS; LT, overexpressing TSAR2; and LST, simultaneously carrying both transgenes. These lines displayed noticeable alterations in morphological traits, including changes in branching rate and biomass production. Furthermore, we observed that the expression of T-DNA genes, particularly aux2 and rolC genes, significantly modulated the expression of pivotal genes involved in centelloside biosynthesis. Notably, the LS lines boasted an elevated centelloside content but concurrently displayed reduced phytosterol content, a finding that underscores the intriguing antagonistic relationship between phytosterol and triterpene pathways. Additionally, the inverse correlation between the centelloside content and morphological growth values observed in LS lines was countered by the action of TSAR2 in the LST and LT lines. This difference could be attributed to the simultaneous increase in the phytosterol content in the TSAR2-expressing lines, as these compounds are closely linked to root development. Overall, these discoveries offer valuable information for the biotechnological application of C. asiatica hairy roots and their potential to increase centelloside production.

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Section: Determination Of Squalene Contentmentioning

confidence: 99%

Enhancing Centelloside Production in Centella asiatica Hairy Root Lines through Metabolic Engineering of Triterpene Biosynthetic Pathway Early Genes

Alcalde,

Palazon,

Bonfill

et al. 2023

Plants

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“…However, the domesticated varieties that are sold as houseplants are mostly smaller than their natural ones. Large, leathery leaves on Croton nigritanus range in different sizes (Yue et al, 2022).. They can be linear or spherical in some cases, and can be of different sizes.…”

Section: Introductionmentioning

confidence: 99%

“…When grown inside, it hardly ever blooms, according to Wen et al, (2018), Squalene is a triterpene with molecular formula C30H50, in which there are six isoprene units. Squalene was first identified in shark liver extract, and later found in the other tissues of shark and vegetable oils such as olive oil (Yue et al, 2022). Due to the high market demand for squalene, investigation to find other sources of squalene is very essential (Ardhyni et al, 2022).…”

Section: Introductionmentioning

confidence: 99%

ISOLATION OF SQUALENE FROM DICHLOROMssETHANE EXTRACT OF CROTON NIGRITANUS LEAF

Abdullahi,

Nkemakonam,

Garba

et al. 2024

FJS

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A precursor of steroids with a variety of biological activities is squalene. The purpose of the study was to identify and describe the industrial/medical chemical found in Croton nigritanus. Using the solvents such as; dichloromethane, ethyl acetate, and n-hexane, the procedure entails a liquid-liquid extraction stage using the partition method and a solid-liquid extraction step using the maceration method. For separation and purification, gravity column chromatography (GCC) and thin-layer chromatography were employed. The isolated chemical's identity was determined using FTIR, 1H NMR and GC-MS spectroscopy. The inquiry successfully extracted and analyzed the major chemical compound which was identified as Squalene.

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“…seed. An extensive study encompassing 104 genotypes of 30 Amaranthus species revealed squalene concentrations ranging from 10.4 to 73.0 g/kg in amaranth oil [13].…”

Section: Introductionmentioning

confidence: 99%

Extraction and Purification of Squalene from Virgin Olive Oil via Catalytic Transesterification and Molecular Distillation

Manikyam,

Mani

2024

Euro. J. Med. Plants.

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Squalene, a highly valuable compound abundant in various natural sources, shows great potential in pharmaceutical, cosmetic, and nutraceutical applications. This research article outlines a comprehensive methodology for extracting and purifying squalene from virgin olive oil, a rich source of the compound. The extraction process begins with degumming, which involves heating the olive oil to 60-70°C to reduce viscosity, followed by the addition of 2-3% warm water to hydrate phospholipids. Food-grade phosphoric acid is then added to react with the phospholipids, forming precipitates. The mixture is stirred for 20-30 minutes and allowed to rest for an additional 20-30 minutes, enabling impurities to settle. The upper layer of degummed oil is separated via decantation or centrifugation and washed with warm water for pH adjustment. Next, transesterification is performed by mixing 100 ml of virgin olive oil with 25% methanol (w/w) and a catalyst (0.5% sodium methoxide or PTSA), and heating the mixture to 80-90°C under reflux for 1-2 hours. Following transesterification, the solvent and acetone are distilled out, and acetone precipitation is repeated 2-3 times to remove unsaponified matter, which is then filtered and evaporated. The concentrated oil undergoes molecular distillation at 180°C and 0.0033 bar pressure for 1 hour, yielding the distillate and residue for further analysis. Qualitative analysis using Thin Layer Chromatography (TLC) involves Merck TLC plates with silica gel 60 F254 and a hexane: chloroform (9:1) mobile phase. Spots are developed with a 10% HCl solution, confirming the presence of squalene with an RF value of 0.93. Quantitative analysis via High-Performance Thin-Layer Chromatography (HPTLC) employs Merck TLC plates with cyclohexane as the mobile phase and CAMAG at 254 nm and 366 nm wavelengths, revealing a squalene purity of 67% and a recovery rate of 69.8%. The initial purification through transesterification facilitated the conversion of ester groups, yielding squalene-rich fractions, while acetone precipitation effectively removed saponified matter. Molecular distillation further enhanced squalene purity. TLC analysis confirmed the qualitative presence of squalene, and HPTLC provided precise quantitative measurements. The obtained squalene purity of 67% significantly enriches the initial content in virgin olive oil, though further optimization could enhance purity and yield. Complementary techniques like GC-MS or HPLC could validate the purification process. This study presents an efficient, replicable procedure for extracting and purifying squalene from virgin olive oil, with significant implications for pharmaceutical, cosmetic, and nutraceutical industries. The findings support a sustainable and ethical shift towards vegetable-derived squalene, meeting market demands while ensuring high-quality production.

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Extraction of Squalene From Tea Leaves (Camellia sinensis) and Its Variations With Leaf Maturity and Tea Cultivar

Cited by 8 publications

References 42 publications

Enhancing Centelloside Production in Centella asiatica Hairy Root Lines through Metabolic Engineering of Triterpene Biosynthetic Pathway Early Genes

Enhancing Centelloside Production in Centella asiatica Hairy Root Lines through Metabolic Engineering of Triterpene Biosynthetic Pathway Early Genes

ISOLATION OF SQUALENE FROM DICHLOROMssETHANE EXTRACT OF CROTON NIGRITANUS LEAF

Extraction and Purification of Squalene from Virgin Olive Oil via Catalytic Transesterification and Molecular Distillation

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