Evaluation of Skin Anti-aging Potential of Citrus reticulata Blanco Peel.

BACKGROUND: The peel of Citrus reticulata Blanco is traditionally used as tonic, stomachic, astringent, and carminative. It is also useful in skin care. OBJECTIVE: To study the anti-aging potential of alcoholic extracts of C. reticulata Blanco peel using in vitro antioxidant and anti-enzyme assays. MATERIALS AND METHODS: Plant extracts were obtained by Soxhlation (CR HAE- Hot Alcoholic Extract of Citrus reticulata) and maceration method (CR CAE- Cold Alcoholic Extract of Citrus reticulata). Qualitative and quantitative phytochemical analysis was performed. Further, in vitro antioxidant, anti-enzyme, and gas chromatography-mass spectrometry (GC-MS) analyses were performed. RESULTS: Total phenolic and flavonoid contents of CR HAE were found to be higher than CR CAE. EC50 value of CR HAE and CR CAE for 1,1-Diphenyl-2-picrylhydrazyl, Superoxide anion, and 2, 2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) assays were 250.33 ± 40.16 µg/ml and 254.73 ± 15.78 µg/ml, 221.27 ± 11.25 µg/ml and 354.20 ± 23.79 µg/ml, and 59.16 ± 2.17 µg/ml and 59.12 ± 6.21 µg/ml, respectively. Oxygen radical absorbance capacity values for CR HAE and CR CAE were found to be 1243 and 1063 µmoles 6-hydroxy-2,5,7,8-tetra methylchromane-2-carboxylic acid equivalent/g of substance, respectively. Anti-collagenase and anti-elastase activities were evaluated for both CR HAE and CR CAE. EC50 values of CR HAE and CR CAE for anti-collagenase and anti-elastase were 329.33 ± 6.38 µg/ml, 466.93 ± 8.04 µg/ml and 3.22 ± 0.24 mg/ml, 5.09 ± 0.30 mg/ml, respectively. CR HAE exhibited stronger anti-collagenase and anti-elastase activity than CR CAE. GC-MS analysis of CR HAE was carried out because CR HAE exhibited higher antioxidant and anti-enzyme potential than CR CAE. CONCLUSION: C. reticulata peel can be utilized in anti-wrinkle skin care formulations. SUMMARY: Skin anti-aging potential of Citrus reticulata Blanco peel was evaluated throughIn vitro antioxidant and anti-enzyme assaysTwo types of extraction were performed and extracts were subjected to qualitative and quantitative phytochemical analysis. Extract obtained by Soxhlation (CR HAE) showed higher total phenolic and flavonoid contents than extract obtained by maceration (CR CAE)CR HAE demonstrated strong DPPH and Superoxide free radical scavenging activity whereas, ABTS scavenging activity of both the extracts were found to be similar. Oxygen Radical Absorbance Capacity (ORAC) of CR HAE was found to be more; indicating its strong antioxidant potentialIn vitro collagenase and elastase enzyme inhibition activities were evaluated for both the extracts and CR HAE showed strong anti-collagenase and antielastase potential indicating its anti-aging abilityGC-MS analysis of CR HAE revealed the presence of various compounds mainly including Polymethoxyflavones. CR HAE exhibited promising antioxidant and anti-enzymatic activity and can be used as a potent antiwrinkle agent in anti-aging skin care formulations. Abbreviation Used: ECM: Extracellular matrix, UV: Ultra violet, ROS: Reactive Oxygen Species, MMP: Matrix metalloproteinase, Chc: Clostridium histolyticum collagenase, DPPH: 2, 2-diphenyl-1-picrylhydrazyl, GC-MS: Gas Chromatography-Mass Spectroscopy, RT: Room Temperature, µg GAE/ mg: Microgram Gallic acid equivalent / milligram, W/V: Weight by Volume, µg QE/ mg: Microgram Quercetin equivalent / milligram, CR HAE: Hot Alcoholic Extract of Citrus reticulata Blanco, CR CAE: Cold Alcoholic Extract of Citrus reticulata Blanco, EC50: Half Maximal Effective Concentration, PMS NADH: Phenazine methosulfate nicotinamide adenine dinucleotide, NBT: Nitroblue tetrazolium, DMSO: Dimethyl sulfoxide, APS: Ammonium Persulphate, AAPH: 2,2 -azobis(2-amidino-propane) dihydrochloride, TROLOX: (±) 6-hydroxy-2,5,7,8-tetramethyl chromane-2-carboxylic acid, ORAC: Oxygen Radical Absorbance Capacity, FALGPA: N-[3-(2-Furyl) acryloyl)]-Leu-Gly-Pro-Ala, SANA: Succinyl-Ala-Ala-Ala-p-nitroanilide, Rf: Retardation Factor, MSD: Mass Selective Detector.

Preparation and characterization of standardized pomegranate extract-phospholipid complex as an effective drug delivery tool.

Punicalagins, a pair of anomeric ellagitannins, present in Punica granatum (Pomegranates) are known to possess excellent antioxidant activity in vitro, but poor oral bioavailability. The reasons cited for poor bioavailability are their large molecular size, poor lipophilicity, and degradation by colonic microflora into less active metabolites. The objective of the present research work was to complex the standardized pomegranate extract (SPE) with phospholipid to formulate standardized pomegranate extract-phospholipid complex (SPEPC), characterize it and check its permeability through an ex vivo everted gut sac experiment. SPEPC was prepared by mixing SPE (30% punicalagins) and soya phosphatidylcholine (PC) in 1:1 v/v mixture of methanol and dioxane and spray-drying the mixture. The complex was characterized by infrared spectroscopy, differential scanning calorimetry, X-ray diffraction, and scanning electron microscopy. It was evaluated for its octanol solubility, dissolution, and permeability by everted the gut sac technique. The characterization methods confirmed the formation of complex. Increased n-octanol solubility of the complex proved its increased lipophilicity. Dissolution studies revealed that the phospholipid covering may prevent the punicalagins to be released in gastro-intestinal tract, thus preventing their colonic microbial degradation. SPEPC showed better apparent permeability than SPE in an everted gut sac technique. Hence, it could be concluded that phospholipid complex of SPE may be of potential use in increasing the permeability and hence the bioavailability of punicalagins.

LC-MS/MS as a tool for identification of bioactive compounds in marine sponge Spongosorites halichondriodes (Dendy 1905).

The sensitivity, specificity and selectivity of liquid chromatography/mass spectrometry tandem mass spectrometry (LC-MS/MS) make it an essential tool for the characterization and identification of low molecular compounds such as fatty acids, sterols, cholastane derivatives, nucleosides etc. In the current work, the marine sponge Spongosorites halichondriodes (order Halichondrida, Family Halichondriidae); a particularly rich source of cytotoxic compounds is studied for the initial characterization of bioactive compounds. The composition of ethyl acetate and butanol extracts were subjected to LC-MS and LC-MS/MS. Many novel sterol derivatives compounds which were not reported in any marine sponge mainly belonged to the group of C25-C28 saturated and unsaturated esters like 3ß, 4ß, 7α, 12α-tetrahydroxy-5ß-cholan-24-oic acid methyl ester, 7 α, 12 ß-dihydroxy-5 ß-cholan24-oic acid methyl ester, novel isocoumarin citrinolactone A, a triterpenoid glycyrrhetinic acid as well as other unknown compounds in this species such as nucleoside inosine was identified. Other compound investigated was 3ß, 6ß, 7α-trihydroxy-5ß-cholan-24-oic acid methyl ester. All the sterol ester derivatives are reported here for the first time in marine sponge belonging to family Halichondriidae. However, the literature report supports the occurrence of 3ß-hydroxy sterols which is considered as a biomarker for this family.

Preliminary study on crystal dissolution activity of Rotula aquatica, Commiphora wightii and Boerhaavia diffusa extracts.

Several Ayurvedic plants are known to have activity against diverse urinary crystals. The traditional knowledge of Ayurveda, collective clinical experience in arthritis and the earlier experimental studies on urinary crystals led to the selection of three plants, viz. Rotula aquatica, Commiphora wightii Bhandari syn. C.mukul. and Boerhaavia diffusa for screening anticrystal activity against basic calcium phosphate (BCP), calcium pyrophosphate (CPPD) and monosodium urate monohydrate (MSUM). The effects of each plant were assayed on microcrystals in 24-well microplates in vitro. Our results show that the aqueous extracts of only R. aquatica and C. wightii have shown crystal dissolving activity against MSUM.