Progress in the analysis of phytocannabinoids by HPLC and UPLC (or UHPLC) during 2020-2023.

INTRODUCTION: Organic molecules that bind to cannabinoid receptors are known as cannabinoids. These molecules possess pharmacological properties similar to those produced by Cannabis sativa L. High-performance liquid chromatography (HPLC) and ultra-performance liquid chromatography (UPLC, also known as ultra-high-performance liquid chromatography, UHPLC) have become the most widely used analytical tools for detection and quantification of phytocannabinoids in various matrices. HPLC and UPLC (or UHPLC) are usually coupled to an ultraviolet (UV), photodiode array (PDA), or mass spectrometric (MS) detector. OBJECTIVE: To critically appraise the literature on the application of HPLC and UPLC (or UHPLC) methods for the analysis of phytocannabinoids published from January 2020 to December 2023. METHODOLOGY: An extensive literature search was conducted using Web of Science, PubMed, and Google Scholar and published materials including relevant books. In various combinations, using cannabinoid in all combinations, cannabis, hemp, hashish, C. sativa, marijuana, analysis, HPLC, UHPLC, UPLC, and quantitative, qualitative, and quality control were used as the keywords for the literature search. RESULTS: Several HPLC- and UPLC (or UHPLC)-based methods for the analysis of phytocannabinoids were reported. While simple HPLC-UV or HPLC-PDA-based methods were common, the use of HPLC-MS, HPLC-MS/MS, UPLC (or UHPLC)-PDA, UPLC (or UHPLC)-MS, and UPLC (or UHPLC)-MS/MS was also reported. Applications of mathematical and computational models for optimization of protocols were noted. Pre-analyses included various environmentally friendly extraction protocols. CONCLUSION: During the last 4 years, HPLC and UPLC (or UHPLC) remained the main analytical tools for phytocannabinoid analysis in different matrices.

Chemical Constituents, Antioxidant, Anti-Tyrosinase, Cytotoxicity, and Anti-Melanogenesis Activities of Etlingera elatior (Jack) Leaf Essential Oils.

Essential oils of plants have been used widely in cosmetic preparations. Being both perfuming and active ingredients, the functions of essential oils mean they are high-value ingredients. In this study, the leaf of Etlingera elatior (Jack) or Torch ginger was used. The essential oils (EO) were prepared by conventional hydrodistillation (HD) and microwave-assisted hydrodistillation (MAHD). The volatile compounds of EOs were analyzed by gas chromatography spectroscopy (GC-MS). The antioxidant activities by means of DPPH radical scavenging and ferric-reducing antioxidant power (FRAP) were determined. The inhibition of tyrosinase activity was investigated. The cytotoxicity was performed against human fibroblast cell lines (NIH/3T3) and melanoma cell lines (A375 and B16F10). The decreasing melanin content was measured in melanoma cell lines. The resulting essential oils were detected for 41 compounds from HD extraction dominants by terpenes, namely sesquiterpenes (48.499%) and monoterpenes (19.419%), while 26 compounds were detected from MAHD with the fatty alcohols as the major group. The higher antioxidant activities were found in HD EO (IC50 of 16.25 ± 0.09 mg/mL from DPPH assay and 0.91 ± 0.01 mg TEAC/g extract from FRAP assay). The survival of normal fibroblast cell lines remained at 90% at 500 µg/mL HD EO, where the EO possessed the half-maximal toxicity dose (TD50) of 214.85 ± 4.647 and 241.128 ± 2.134 µg/mL on B16F10 and A375 cell lines, respectively. This could suggest that the EO is highly selective against the melanoma cell lines. The melanin content was decreased at the half-maximum efficacy (IC50) at 252.12 ± 3.02 and 253.56 ± 3.65 in the A375 and B1610 cell lines, respectively, which were approximately 2.8-fold lower than kojic acid, the standard compound. The results of this study evidence the use of Etlingera elatior (Jack) leaf as a source of essential oil as an active agent in cosmetics.

Polysaccharides from Volvariella volvacea Mushroom: Extraction, Biological Activities and Cosmetic Efficacy.

Polysaccharides from Volvariella volvacea (VVP) were investigated for their cosmetic-related activities and in vivo efficacy for use as a multifunctional active cosmetic ingredient. Three different polysaccharide extraction methods, including hot water shaking (HS), microwave-assisted (MA) and ultrasonic-assisted (UA), were used. Extractable yield, polysaccharide contents and biological activities, including antioxidant, anti-tyrosinase and anti-elastase activities, were compared. The polysaccharides from HS provided the highest extraction yield (15.58 ± 0.96% w/w) and the highest beta-glucan content (18.80 ± 0.81% w/w). The HS polysaccharides also possessed the highest inhibitory effects toward lipid peroxidation (IC50 of 0.0378 mg/mL), tyrosinase (51.46 mg KAE/g), and elastase (604.21 ± 73.66 mg EGCG/g). The cytotoxicity of the VVP was determined for safe use. A cosmetic gel cream containing VVP was developed and 0.2% VVP formulation was observed to be the most stable in color. UV protection factors, skin irritation by single patch test, and in vivo efficacy, including skin moisturization, anti-wrinkle and whitening, were measured. The VVP showed no cytotoxicity against human dermal skin fibroblast. The gel cream containing VVP provided less sun protection factor; however, it significantly exhibited the skin benefits of increasing moisture, gross elasticity, net elasticity, and skin firmness. Improvements to skin roughness, scaliness, wrinkles and in melanin content were also depicted gradually along 8 weeks. V. volvacea, therefore, could be a good source for polysaccharides being used as a moisturizing, anti-wrinkle, and whitening agent in cosmetic preparations.

The Feasibility of Utilizing Cultured Cordyceps militaris Residues in Cosmetics: Biological Activity Assessment of Their Crude Extracts.

Solid-based residues (SBRs) left from harvesting the fruiting bodies of cultured Cordyceps mushrooms are a challenge to sustainability. Therefore, in this study, the SBRs from the cultivation of Cordyceps militaris (C. militaris) via solid-state fermentation (SSF) were employed to prepare crude extracts, with the aim of considering their possible use in cosmetics. The SBRs obtained from cultivation with solid media containing defatted rice bran mixed with barley, white rice, Riceberry rice, and wheat were named SBR-B, SBR-R, SBR-Rb, and SRB-W, respectively. They were extracted with solvents of differing polarity and then evaluated for their total phenolic content (TPC), total flavonoid content (TFC), and total carbohydrate content (TCC). In addition, antioxidant and tyrosinase inhibitory activities, photoprotection, and cytotoxicity were also assessed. The results revealed that the total bioactive contents and biological capacities of crude SBR extracts were significantly influenced by the types of SBR and extraction solvent (p < 0.05). The SBR-B extracted with hot water exhibited the highest antioxidant activity (66.62 ± 2.10, 212.00 ± 3.43, and 101.62 ± 4.42 mg TEAC/g extract) when assayed by DPPH, ABTS, and FRAP methods, respectively, whereas tyrosinase inhibitory activity (51.13 ± 1.11 mg KAE/g extract) with 90.43 ± 1.96% inhibition at 1 mg/mL was excellently achieved by SBR-Rb extracted by 50% (v/v) ethanol. Correlations between bioactive contents in the crude extracts and their biological activities were mostly proven at a strong level (p < 0.01). The capability of the crude extracts to absorb UV over the range of 290-330 nm disclosed their potential roles as natural UV absorbers and boosters. Cytotoxicity analysis using fibroblast cell lines tested with hot water and 50% (v/v) ethanolic SBR extracts demonstrated safe use within a concentration range of 0.001-10 mg/mL. Interestingly, their fibroblast proliferative capacity, indicating anti-aging properties, was highly promoted. The chemical composition analyzed via LC-MS/MS techniques showed that seven phenolic acids and four flavonoids were identified in the crude SBR extracts. Furthermore, the other compounds present included nucleosides, nucleobases, amino acids, sugars, phospholipids, alkaloids, organic acids, vitamins, and peptides. Therefore, it is emphasized that SBRs from C. militaris can be a prospective source for preparing crude extracts employed in cosmetics. Lastly, they could be further utilized as multifunctional ingredients in cosmetics and cosmeceuticals.

Simultaneous quantification of sulforaphene and sulforaphane by reverse phase HPLC and their content in Raphanus sativus L. var. caudatus Alef extracts.

A simple, rapid and precise HPLC assay was developed for the well-known anti-cancer isothiocyanates-sulforaphene (SE) and sulforaphane (SF). The analytical system comprised RP-C18 column with isocratic 5% THF-95% water. High resolution was obtained (and eluted) of two distinct HPLC peaks of similar structures SE and SF analytes (at 23.01±0.02 and 25.65±0.03 min, respectively). The respective LOD vs. LOQ for SE and SF was 0.34 and 0.36 µg/ml vs. 1.02 and 1.08 µg/ml. This assay had the best linearity and accuracy. The recoveries were in the range of 96.83-101.17%. SF and SE were quantified in the pod of Raphanus sativus L. var. caudatus Alef extracts (2253.05±246.18 and 111.94±16.49 µg/g in the crude extract, respectively), while only SE was detected in the stem (1105.14±243.10 µg/g crude extract), as SF was lower than the detection limit. The validated method thus minimized and expedited simultaneous SE and SF analysis.