PROFIL METABOLIT BERBAGAI EKSTRAK DAUN Chrysophyllum cainito L. MENGGUNAKAN UPLC-QTOF-MS/MS

ABSTRACT Chrysophyllum cainito L. is a plant which empirically used as traditional medicine. The pharmacological effect of C. cainito is caused by secondary metabolite activity contain in the leaves. The aim of this research was to know the metabolites profile in n-hexane extract, ethyl acetate extract, and methanol extract of C. cainito leaves using UPLC-QToF-MS/MS. Dried powder of C. cainito leaves was extracted with n-hexane, ethyl acetate, and methanol with gradual extraction using Ultrasonic Assisted Extraction (UAE). Each extract was prepared with methanol and DCM solvent then injected 5 µl into UPLC-QToF-MS/MS and analyzed with Masslynx 4.1 softwares and Chemspider. The result showed that there were 28 compounds from n-hexane extract with diethyltoluamide as major compound, 47 compounds from ethyl acetate extract with loliolide as major compound, and 34 compounds from methanol extract with eplerenone as major compound. Based on literature study, there were also several compounds that likely having activity as phytoestrogens. Keyword: Metabolite profiling, chrysophyllum cainito L., UPLC QToF-MS/MS     ABSTRAK Chrysophyllum cainito L. merupakan tumbuhan yang secara empiris digunakan sebagai obat tradisional. Efek farmakologi tersebut disebabkan adanya aktivitas dari berbagai senyawa metabolit sekunder yang terkandung dalam daun C. cainito. Tujuan dari penelitian ini untuk mengetahui profil metabolit ekstrak n-heksana, ekstrak etil asetat, dan ekstrak metanol daun C. cainito menggunakan UPLC-QToF-MS/MS. Serbuk kering daun C. cainito diekstraksi secara bertingkat menggunakan n-heksana, etil asetat, dan metanol dengan metode UAE. Masing-masing ekstrak dipreparasi dengan metanol dan DCM lalu diinjeksikan sebanyak 5 µl ke dalam UPLC-QToF-MS/MS, kemudian dianalisis dengan software Masslynx 4.1 dan Chemspider. Hasil menunjukkan profil metabolit dari masing-masing ekstrak daun C. cainito, yaitu ekstrak n-heksana dengan 28 senyawa dan diethyltoluamide sebagai senyawa mayor, 47 senyawa terkandung dalam ekstrak etil asetat dengan senyawa mayor loliolide, dan 34 senyawa terkandung dalam ekstrak metanol dengan senyawa mayor yaitu eplerenone. Dari studi literatur diketahui terdapat beberapa senyawa yang memiliki aktivitas sebagai fitoestrogen. Kata Kunci: Metabolite profiling, Chrysophyllum cainito L., UPLC QToF-MS/M

Metabolite profiling of compounds from Sargassum polycystum using UPLC-QToF-MS/MS

Background: There are many types of seaweed that have high economic value. Brown seaweed (Sargassum polycystum) can be used as a raw material in the industry and as a medicinal plant. Maintaining the quality of a compound requires an analytical method that can identify the diversity of metabolome profiles. Objective: This investigation seeks to discover the metabolite profile of S. polycystum from Sumenep, Madura Island, Indonesia, utilizing the UPLC-QToF MS/MS equipment. Materials and Methods: The extract was further fractioned using n-hexane, ethyl acetate, and water. The metabolite profiling of extract and fractions used the UPLC-QToF-MS/MS instrument. It was produced with SPE and then introduced into the MS Xevo G2-S QToF detector of the ACQUITY UPLC® H-Class System. The findings of the UPLC-QToF-MS/MS analysis were processed with the MassLynx 4.1 software to obtain chromatogram data and m/z spectra of each observed peak, which were then validated using the ChemSpider and MassBank databases. Results: Based on the results of metabolite profiling using UPLC-QToF-MS/MS, the 96 % ethanol extract of S. polycystum indicated a total of 61 compounds, the n-hexane fraction indicated a total of 55 compounds, the ethyl acetate fraction indicated a total of 67 compounds, and the water fraction indicated a total of 49 compounds. Conclusion: There are 232 compounds in the extract and a fraction of S. polycystum consisting of 168 known compounds and 64 unknown compounds

Multi-residue analysis of pharmaceuticals in Belgian surface water : a novel screening-to-quantification approach using large-volume injection liquid chromatography coupled to high-resolution mass spectrometry

The ever growing number of emerging micropollutants such as pharmaceuticals requests rapid and sensitive full-spectrum analytical techniques. Time-of-flight highresolution mass spectrometry (TOF-HRMS) is a promising alternative for the state-ofthe- art MS/MS instruments because of its ability to simultaneously screen towards a virtually unlimited list of suspect compounds and to perform target quantification. The challenge for such suspect screening is to develop a strategy which minimizes the false negative rate without restraining numerous false positives. At the same time, omitting laborious sample enrichment through large-volume injection ultraperformance liquid chromatography (LVI-UPLC) is advantageous avoiding selective preconcentration. A novel suspect screening strategy was developed using LVI-UPLC-TOF-MS aiming the detection of 69 multi-class pharmaceuticals in surface water without the a priori availability of analytical standards. As a novel approach, the screening takes into account the signal intensity-dependent accurate mass error, hereby assuring the detection of 95% of pharmaceuticals present in surface water. Subsequently, the validation and applicability of the full-spectrum method for target quantification of the 69 pharmaceuticals in surface water is discussed. Analysis of five Belgian river water samples revealed the occurrence of 17 pharmaceuticals in a concentration range of 17 ng L-1 up to 3.1 μg L-1

Antidiabetic Effect of Actinodaphne angustifolia and Profiling of Bioactive Metabolites using UPLC-QToF/ESI-MS Method

Diabetes mellitus adalah gangguan kronik yang menyebabkan paras glukosa darah meningkat akibat kekurangan insulin, sama ada sepenuhnya atau sebahagian. Kami menyiasat sifat antidiabetik Actinodaphne angustifolia dalam model tikus dan mengenal pasti fitokimia bioaktif dengan menggunakan kaedah UPLC-QTOF/ ESI-MS. Struktur pankreas tikus, profil lipid dan glukosa darah dinilai selepas intervensi selama satu minggu. Analisis UPLC-QTOF/ESI-MS telah dijalankan untuk mengenal pasti flavonoid dan terpenoid dalam ekstrak daun. Ekstrak Actinodaphne angustifolia dengan ketara meningkatkan lipoprotein berketumpatan tinggi (HDL) (p<0.05) sambil mengurangkan jumlah kolesterol (TC), lipoprotein berketumpatan rendah (LDL) dan glukosa darah. Tambahan pula, struktur seni tisu pulau pankreas juga pulih dengan baik berbanding kumpulan kawalan. Sebanyak 45 flavonoid dan 109 sebatian terpenoid telah dikenal pasti menggunakan analisis berasaskan UPLC-QTOF/ESI-MS dan kajian tambahan perlu dijalankan untuk mengenal pasti agen antidiabetik yang berpotensi. Diabetes mellitus is a chronic disorder that causes elevated blood glucose levels due to a lack of insulin, either completely or partially. We investigated the antidiabetic property of Actinodaphne angustifolia in a rat model and identified the bioactive phytochemicals by using the UPLC-QTOF/ESI-MS method. The rats’ pancreatic structures, lipid profile and blood glucose were assessed after a one-week intervention. UPLC-QTOF/ESI-MS analysis was conducted to identify flavonoids and terpenoids in the leaf extract. Actinodaphne angustifolia extract markedly increased the high-density lipoprotein (HDL) (p<0.05) while reducing total cholesterol (TC), low-density lipoprotein (LDL) and blood glucose. Furthermore, the tissue architecture of pancreatic islets was also well recovered as compared to the control group. A total of 45 flavonoids and 109 terpenoid compounds were identified using UPLC-QTOF/ESI-MS-based analysis and additional studies should be undertaken to identify the potential antidiabetic agents

Tissue biochemical diversity of 20 gooseberry cultivars and the effect of ethylene supplementation on postharvest life

The European gooseberry (Ribes uva-crispa) is still an understudied crop with limited data available on its biochemical profile and postharvest life. A variety of polyphenols were detected in the skin and flesh of 20 gooseberry cvs, representing mainly flavonol glycosides, anthocyanins and flavan-3-ols. In contrast, gooseberry seeds were for the first time characterised by the presence of considerable amounts of hydroxycinnamic acid glycosides tentatively identified by UPLC-QToF/MS. All cvs examined represented a good source of vitamin C while being low in sugar. Furthermore, the postharvest stability of bioactives was explored by supplementation of exogenous ethylene in air at 5 °C. Results suggest a low sensitivity of gooseberries to ethylene. The overall quality of gooseberries remained stable over two weeks, showing potential for extended bioactive life

Karakterisasi Senyawa Metabolit Sekunder Arak Tradisional Bali dan Koktail Menggunakan Skrining Fitokimia, Spektrofotometer UV-Vis dan Kromatografi Cair Kinerja Tinggi-Spektrometri Massa

Arak tradisional bali merupakan minuman beralkohol yang diperoleh dari hasil fermentasi nira yang berasal dari tanaman siwalan. Arak koktail kecarum adalah minuman beralkohol yang terbuat dari arak dengan penambahan&nbsp; berbagai macam rempah-rempah atau herbal. Pengembangan metode analisis kandungan senyawa kimia pada minuman beralkohol pada umumnya adalah analisis untuk menentukan kandungan alkohol, tetapi penelitian mengenai analisis metabolit sekunder pada minuman beralkohol belum banyak dilakukan secara signifikan. Berdasarkan hal tersebut, penelitian ini bertujuan mengidentifikasi senyawa metabolit sekunder yang terdapat dalam arak tradisonal bali dan arak koktail kecarum menggunakan skrining fitokimia, spektrofotometer UV-Vis dan kromatografi cair kinerja tinggi-spektrometri massa (UPLC-QTOF-MS/MS). Sampel arak yang digunakan adalah arak tradisional bali yang didapatkan dari kabupaten Karang Asem, sedangkan arak koktail kecarum diperoleh dari daerah Sanur, Bali. Analisa metabolit sekunder pada arak tradisional dan arak koktail kecarum dilakukan dengan skrining fitokimia, identifikasi, dan konfirmasi senyawa metabolit sekunder menggunakan spektrofotometer UV-Vis dan kromatografi cair-spektrometri massa (UPLC-QTOF-MS/MS). Berdasarkan hasil skrining fitokimia, arak tradisional bali tidak mengandung senyawa metabolit sekunder, sedangkan arak koktail kecarum mengandung senyawa metabolit sekunder golongan senyawa fenolik yang terdiri dari flavonoid, tanin dan kumarin, golongan senyawa terpen yang terdiri dari minyak atsiri, saponin, sterol, dan triterpenoid. Hasil uji kuantitatif menggunakan metode spektrofotometer UV-Vis diperoleh kadar senyawa fenolik total sebanyak 335,77 mg GAE/mL; kadar senyawa flavonoid total sebanyak 620,17 mg QE/mL; dan kadar senyawa tanin total sebanyak 416,63 mg TAE/mL. Berdasarkan hasil identifikasi senyawa metabolit sekunder menggunakan kromatografi cair kinerja&nbsp; tinggi-spektrometri massa (UPLC-QTOF-MS/MS) diperoleh bahwa arak koktail kecarum mengandung 24 senyawa metabolit sekunder&nbsp; golongan senyawa fenolik yang terdiri dari senyawa flavonoid dan fenol

Antidiabetic effect of actinodaphne angustifolia and profiling of bioactive metabolites using UPLC-QToF/ESI-MS method

Diabetes mellitus is a chronic disorder that causes elevated blood glucose levels due to a lack of insulin, either completely or partially. We investigated the antidiabetic property of Actinodaphne angustifolia in a rat model and identified the bioactive phytochemicals by using the UPLC-QTOF/ESI-MS method. The rats’ pancreatic structures, lipid profile and blood glucose were assessed after a one-week intervention. UPLC-QTOF/ESI-MS analysis was conducted to identify flavonoids and terpenoids in the leaf extract. Actinodaphne angustifolia extract markedly increased the high-density lipoprotein (HDL) (p<0.05) while reducing total cholesterol (TC), low-density lipoprotein (LDL) and blood glucose. Furthermore, the tissue architecture of pancreatic islets was also well recovered as compared to the control group. A total of 45 flavonoids and 109 terpenoid compounds were identified using UPLC-QTOF/ESI-MS-based analysis and additional studies should be undertaken to identify the potential antidiabetic agents

Metabolite Profiling of 96% Ethanol Extract from Marsilea crenata Presl. Leaves Using UPLC-QToF-MS/MS and Anti-Neuroinflammatory Predicition Activity with Molecular Docking

Phytoestrogen is a group of compounds that can replace the estrogen function in the body. One of its roles was as anti-neuroinflammatory by inhibiting the microglia M1 polarity activation. Marsilea crenata Presl. is a plant that suspected to contain phytoestrogens. The aim of this research was to determine the metabolite profile of 96% ethanol extract of M. crenata using UPLC-QToF-MS/MS, and prediction the anti-neuroinflammatory activity of compounds with molecular docking. The 100 ppm of 96% ethanol extract in DCM and methanol were injected 5 µl each into the UPLC-QToF-MS/MS, and then analyzed by Masslynx 4.1 software to determine the compounds. The result of metabolite profiling shows a total 59 compounds in both DCM and methanol. Molecular docking was done with Autodock 4.2.6. After being analyzed, there are 3 compounds that are predicted to have activities similar to 17?-estradiol, they are prochlorperazine, 12-Aminododecanoic acid, and 1-methyl-2-[(4-methylpiperazin-1-yl)methyl]benzimidaol-5-amine hydrochloride. The results showed that the three compounds were predicted to be phytoestrogen compounds from M. crenata leaves, which have potential as anti-neuroinflammatory

Uptake and depuration of cyanotoxins in the common blue mussel Mytilus edulis.

Cyanobacteria produce a variety of secondary metabolites that possess - amongst other things - antifungal, antibacterial and antiviral properties. Being primary producers, they are also a vital component within the food web. However, certain strains also produce toxic metabolites such as the hepatotoxins microcystin (MC) and nodularin (NOD). Their toxicity - in combination with the increasing global occurrence - has resulted in a drinking water guideline limit of 1 μg L-1 being issued by the World Health Organisation (WHO). However, these toxins are not only present in water, but can be accumulated by fish and shellfish. Currently, no regulations regarding cyanotoxin-contaminated seafood have been established, despite similar toxicity to routinely-monitored marine toxins such as domoic acid (DA). To facilitate regular monitoring, a high-performance liquid chromatography photo diode array (HPLC-PDA) analysis method for the detection of DA was optimised to enable the simultaneous detection of DA and nine cyanotoxins. This method was then utilised to determine cyanotoxin concentration in laboratory cyanobacteria strains. To assess the accumulation and depuration of cyanotoxins in the common blue mussel Mytilus edulis, three feeding trials were performed. During these, mussels were exposed to two cyanobacteria strains - Nodularia spumigena KAC66 and Microcystis aeruginosa PCC 7813 - both individually and simultaneously. A rapid dose-dependent accumulation of cyanotoxins was observed with maximum concentration of 3.4 -17 μg g-1 ww accumulated by M. edulis, which was followed by a much slower depuration. During the final feeding trial with the two cyanobacteria strains, cyanotoxins were still detectable following twenty-seven days of depuration. Mortality in all studies was 7% or less, indicating that most mussels were unaffected by the maximum dose of 480 μg L-1 NOD (feeding study 1), 390 μg L-1 MC (feeding study 2), or 130 μg L-1 total cyanotoxins (feeding trial 3). Mortality in negative control tanks was lower throughout all three feeding trials ( < 1 - 2.6%). Consumption of a typical portion size (twentty mussels) would result in ingestion of cyanotoxins at levels significantly higher than the WHO recommended tolerable daily intake (TDI) of 2.4 μg NOD and/or MCs for a 60 kg adult. This value was exceeded not only during the exposure period (maximum levels 270 - 1370 μg cyanotoxins per twenty mussels), but also at the end of the depuration period 39-600 μg cyanotoxins per twenty mussels. These results illustrated that cyanotoxin monitoring of seafood should be considered not only during, but also following bloom events. In an attempt to investigate the cyanotoxin budget of the experimental system, not only mussels but also cyanobacteria cultures, the tank water and the mussel faeces were also analysed for their cyanotoxin content. Results showed that large quantities of MCs and NOD were unaccounted for during all exposure trials. The combined effect of cyanotoxin metabolism in M. edulis, biotic and/or abiotic degradation, protein binding, and losses during the extraction and analysis were thought to have contributed to the unaccounted cyanotoxin fraction. Mussel flesh was analysed for the presence of glutathione or cysteine conjugates; however, there was no evidence of their occurrence in the samples tested. Due to these discrepancies in the toxin budget of the system, the introduction of correction factors for the analysis of cyanotoxins in M. edulis was suggested in order to protect the general public