Isolation and Identification of DPPH Radical (2,2-diphenyl-1-pikrylhidrazyl) Scavenging Active Compound in Ethyl acetat fraction of Piper acre Blume

Piper acre Blume, known as Black Betel (local name), is a plant that is widely used by the people of East Kalimantan, especially in Samarinda, for the treatment of illness. Leaves (3-4 months old) are collected from Samarinda, extracted, fractionated, and monitored by DPPH antiradical activity. The isolation of the Piper acre Blume is performed on the active fraction, and the structure identification is based on spectroscopic data of the compound.  The leaves were dried, pulverized, and macerated with MeOH. Dried MeOH extract was obtained upon evaporation of the solvent. The extract was then fractionated by vacuum liquid chromatography (vlc), eluted gradually by solvents having different polarities (n-hexane, ethyl acetate and methanol). The fractions obtained were monitored using TLC [n-hexane: ethyl acetate (3: 1 v/v)] that was visualized by UV254 nm, UV366 nm and DPPH. The isolation was performed by preparative TLC [SiO2, n-hexane: ethyl acetate (3: 1)] on ethyl acetate fraction that showed the highest DPPH antiradical value. A single compound was obtained, and it appeared as a round spot and pure according to TLC performances at 3 different solvent systems. The isolated Piper acre Blume compound displayed the IC50 value on the anti-radical DPPH (measured at λ 520 nm) as 10.41µg/mL. The IR spectrum (KBr) showed –OH band (3450 cm-1), aliphatic bands [alkene, 3010 cm-1; alkana 2900 cm-1), an aromatic overtone bands (1900-200 cm-1) and a strong C=O band (1725 cm-1). The NMR (1H- and 13C-) (mono and 2D) indicated the present of a p-di-substituted aromatic signals (δ, 7.54 and 7.52, d, J =6 Hz, 1 H each), 2 methyl (δ, 0.96, d, J = 7.0 Hz, 6 Hs), a triplet signal (δ, 4.22 ppm). Other signals of CH- and CH2 were shown as m signals at δ, 1.64 and 1.34 ppm.  Based on those data, the compound was identified as isoamyl p-OH benzoate that is grouped as parabens used as a preservative in the pharmaceutical preparations. In conclusion, the anti-radical (DPPH) active compound present in the leaves of Piper acre Blume is identified as isoamyl p-OH benzoate, having IC50 value anti-radical DPPH 10,41µg/mL

Modeling and Optimization of Mitragyna speciosa Extraction using Box Behnken Design

The extraction of kratom (M. speciosa) leaf powder was optimized with preliminary extraction to be further optimized with the Box-Behnken experimental design. The individual and interactive effects of process variables (sample-to-solvent ratio, extraction time, solvent concentration) were assessed. The preliminary extraction results showed that ultrasound-assisted extraction (UAE) and methanol were chosen for further optimization. The experimental data were analyzed by Pareto analysis of variance (ANOVA) and second-order polynomial models were developed using multiple regression analysis. The model developed showed a good fit with the experimental data with a high coefficient of correlation (R2) and predictive ability (predicted R2). An optimization study was performed and the optimal extraction conditions were sample-to-solvent ratio value 1.5:10; extraction time of 10 minutes, and methanol concentration of 100%

Phytochemical and Pharmacological Activities of Curcuma purpurascens Blume, A Review

Curcuma sp. is generally used for medicine, starch sources, preservatives, dyes and cosmetics. The use of Curcuma spp. for medical has increased because there have been many studies related to its active ingredients, such as flavonoids, essential oils, tannins, quinones, and terpenoids, as well as pharmacological activities, including wound healing, antioxidants, antifungal, anticancer, gastroprotective, and hepatoprotective. Curcuma purpurascens Blume is a species of Curcuma from family Zingiberaceae and used for traditional medicine. This article focuses on reviewing the literatures on C. purpurascens and discussing its morphology, phytochemical content, and pharmacological aspects. The method used to review this article was by exploring several databases such as Scopus, Pub Med, and Google Scholar to identify and download original articles and research journals related to the morphology, phytochemical content, and biological activity of Curcuma purpurascens Blume. The result of this review will later provide information about the uses and presence of Curcuma purpurascens Blume which is still rarely studied so further study related to its pharmacological activity tests and active compound as natural medicines can be explored

Mitragyna speciosa: Opioid Addiction Treatment and Risk of Use

Kratom (Mitragyna speciosa (Korth.) Havil.) is a plant that originated from the rainforest in Southeast Asia, mainly grows in Thailand, Malaysia, and Indonesia. Kratom has been used traditionally as an herbal remedy for the treatment of various illnesses. Kratom gained notoriety due to its potential as an analgesic, opiate withdrawal treatment, anxiolytic, antidepressant, and antidiabetic with an unclear risk of addiction and toxicity fueled by a false sense of security due to its identity as a member of the coffee family. This article is a narrative review on kratom to highlight its pharmacological and toxicological properties, and the analytical method of Kratom, especially its potential as an opioid withdrawal therapy and its risk of abuse

Profil Proliferasi Sel Limfosit Benalu Batu (Begonia medicinalis) Asal Kabupaten Morowali UTARA Provinsi Sulawesi Tengah

The plant of Benalu Batu (Begonia medicinalis) from the North Morowali regency has been used empirically and is known to have anticancer activity. The study aims to determine the profile of the activity of lymphocytes cell proliferation of extracts and fractions of Benalu Batu in vitro as well to measure the correlation of total flavonoid content against the stimulating index of lymphocytes cell proliferation. The extraction of simplicia by using the maceration method with methanol solvent. The liquid-liquid partition method is used in the fractionation of methanol extracts with hexane, ethyl acetate, and water solvents, sequentially. Test of lymphocytes cell proliferation using MTT reduction (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) method with concentration series 10, 20, 50, and 100 μg/mL. The total of flavonoids content of extracts and fractions were determined by using colorimetric method. The stimulation index (IS) is used in measuring the activity of lymphocytes cell proliferation. The test results showed that the methanol extracts, hexane fraction, ethyl acetate, and water may increase the proliferation of lymphocytes in the entire test concentration. The concentration of test influences on increasing the stimulation of the proliferation of lymphocytes cells. A test concentration of 100 μg/mL has the highest stimulation index on each test sample and its stimulant index value of hexane fraction, ethyl acetate fraction, methanol fraction and water fraction were 10.12, 6.56, 4.82, and 4.17, respectively. Total flavonoids content and stimulation indices have very low correlation (r = 0,082) and concentrations of 10 μg/mL have the highest correlation coefficient (r2 = 0.18). The results showed that the hexane and ethyl acetate fractions can be developed as immunostimulant materials for the anticancer supportive therapeutic agents despite having a very low correlation to the flavonoid levels.Tumbuhan Benalu Batu (Begonia medicinalis) asal kabupaten Morowali Utara telah digunakan secara empiris dan diketahui memiliki aktivitas antikanker. Penelitian ini bertujuan untuk mengetahui profil aktivitas proliferasi sel limfosit dari ekstrak dan fraksi Benalu Batu secara in vitro serta mengukur korelasi kadar flavonoid total terhadap indeks stimulasi proliferasi sel limfosit. Ekstraksi simplisia Benalu Batu menggunakan metode maserasi dengan pelarut metanol. Metode partisi cair-cair digunakan dalam fraksinasi ekstrak metanol dengan pelarut n-heksana, etil asetat dan air secara berurutan. Uji proliferasi sel limfosit menggunakan metode MTT reduction (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) dengan seri konsentrasi 10, 20, 50 dan 100 µg/mL. Kadar flavonoid total ekstrak dan fraksi-fraksi ditetapkan berdasarkan metode kolorimetri. Indeks stimulasi (IS) digunakan dalam pengukuran aktivitas proliferasi sel limfosit. Hasil pengujian menunjukkan bahwa ekstrak metanol, fraksi n-heksana, etil asetat dan air dapat meningkatkan proliferasi sel limfosit pada seluruh konsentrasi uji. Konsentrasi uji memiliki pengaruh terhadap peningkatan indeks stimulasi proliferasi sel limfosit. Konsentrasi uji 100 µg/mL memiliki indeks stimulasi tertinggi pada setiap sampel uji dan nilai indeks stimulasi dari fraksi n-heksana, fraksi etil asetat, ekstrak metanol dan fraksi air adalah 10,12; 6,56; 4,82 dan 4,17 secara berurutan. Kadar flavonoid total dan indeks stimulasi memiliki korelasi yang sangat rendah (r=0,082) dan konsentrasi 10 µg/mL memiliki koefisien korelasi tertinggi (r2=0,18). Hasil penelitian menunjukkan bahwa fraksi n-heksana dan fraksi etil asetat dapat dikembangkan sebagai bahan imunostimulansia guna agen terapi suportif antikanker meskipun memiliki korelasi yang sangat rendah terhadap kadar flavonoid

The Effect of Ursolic Acid from Plantago Lanceolata Leaves on Leukocytes Migration and Chemokines Level

Initially considered as a normal body response to injury, inflammation is currently known as a major event contributing to the development of many human disorders. Many drugs and bioactive molecules have been discovered from medicinal plants and the number is still growing by time. Among those medicinal plants used in folk medicines, Plantago lanceolata is used to cure inflammatory-related diseases. In our previous study, we showed that the n-hexane insoluble fraction of P. lanceolata leaves (HIF) demonstrated a potent anti-inflammatory activity by inhibiting leukocytes migration in mice. This study aimed to identify the anti-inflammatory compound from the HIF and to investigate the effect on the chemokines level. P. lanceolata leaves were initially macerated with dichloromethane. The dried extract was partitioned using n-hexane to obtain n-hexane soluble fraction (HSF) and n-hexane insoluble fraction (HIF). Both fractions were evaluated for their anti-inflammatory activities in thioglycollate-induced leukocyte migration. The active fraction (HIF) was subjected to preparative thin-layer chromatography (TLC) to isolate the major compound. The structure of the compound was identified based on NMR, IR, and Mass spectra. Moreover, we investigated the effect of the compound on the level of chemokines responsible for leukocytes migration. The active compound was identified as ursolic acid, based on its spectral data. Ursolic acid at the dose of 30, 60, and 120mg/kg BW inhibited leukocyte migration and reduced chemokines level (IL-8 and MCP-1)

Antiinflamatory Activity of Methanolic Extract of Mangifera Casturi in Thioglycollate-induced Leukocyte Migration on Mice

Kasturi (Mangifera casturi) is a plant originally from Kalimantan, mainly in South Kalimantan. Although there is no report regarding the USAge of the fruit in traditional healings, current research demonstrate that methanolic extract of Mangifera casturi fruit demonstrated high antioxidant activity. As oxidant plays crucial role in inflammatory related diseases, we investigate the antiinflammatory effect of Mangifera casturi fruit methanolic extract. To assess the antiinflammatory activity, we used thioglycollate-induced leukocyte migration in mice. Male balb/c mice were pretreated (i.p) with Mangifera casturi fruit methanolic extract followed by thioglikolat (i.p) to induce leukocyte migration. After 4,5 hours, the mice were sacrificed and the number of leukocytes were counted using hemocytometer under light microscope. Our experiment indicates that Mangifera casturi fruit methanolic extract significantly exhibited antiinflammatory activity by inhibiting the migration of leukocytes induced by thioglycollate. However, the activity of the extracts was lower than the positive control, indomethacin. Further purification was required to obtain the active compound with the activity comparable to indomethacin. Thin layer chromatography (TLC) analysis demonstrates that the methanolic extract of Mangifera casturi fruit contains terpenoid and phenolic compounds

Effects of Antioxidant, Anti-Collagenase, Anti-Elastase, Anti-Tyrosinase of The Extract and Fraction From Turbinaria decurrens Bory.

Brown macroalgae (BM) which contain fucoxanthin exhibited high antioxidant activity. This study was performed to examine antioxidant, anti-collagenase, anti-elastase, anti-tyrosinase activities, and effect on cell viability of Human Dermal Fibroblast adult (HDFa) of BM, Turbinaria decurrens Bory. T. decurrens dried powder were macerated by ethanol to obtain extract (ETD) and was fractination by column chromatography to obtain fraction (FTD). Fucoxanthin content was determined using HPLC. The antioxidant activities, anti-collagenase, anti-elastase, and tyrosinase inhibitory assay were performed. The effect of ETD and fucoxanthin standard on cell viability were conducted on HDFa cell-induced by hydrogen peroxide (H2O2). The HPLC analysis showed that ETD and FTD contain fucoxanthin of 284.9±3.3µg/g and of 653.4±30.6µg/g dry-weight, respectively. The antioxidant assay showed that ETD and FTD produced high antioxidant activity by ferric reducing antioxidant power (FRAP) and b-carotene bleaching (BCB) methods that were comparable to fucoxanthin. ETD exhibited significantly higher tyrosinase inhibitory than kojic acid (p<0.01), while FTD had a comparable effect to kojic acid. The result also revealed that ETD and FTD produced anti-elastase and anti-collagenase (matrix metalloproteinase-1 (MMP-1). Fucoxanthin and ETD were able to maintain cell viability on HDFa cell-induced H2O2. This study suggests that T. decurrens may be effective to prevent skin aging and wrinkle formation, possibly through the antioxidant activity and maintain cell viability of fibroblast.

AKTIVITAS ANTIINFLAMASI EKSTRAK METANOLIK BUAH MANGGA KASTURI (Mangifera casturi) MELALUI PENGHAMBATAN MIGRASI LEUKOSIT PADA MENCIT YANG DIINDUKSI THIOGLIKOLAT

Kasturi (Mangifera casturi) is a plant originally from Kalimantan, mainly in South Kalimantan. Although there is no report regarding the usage of the fruit in traditional healings, current research demonstrate that methanolic extract of Mangifera casturi fruit demonstrated high antioxidant activity. As oxidant plays crucial role in inflammatory related diseases, we investigate the antiinflammatory effect of Mangifera casturi fruit methanolic extract. To assess the antiinflammatory activity, we used thioglycollate-induced leukocyte migration in mice.  Male balb/c mice were pretreated (i.p) with Mangifera casturi fruit methanolic extract followed by thioglikolat (i.p) to induce leukocyte migration. After 4,5 hours, the mice were sacrificed and the number of leukocytes were counted using hemocytometer under light microscope.   Our experiment indicates that Mangifera casturi fruit methanolic extract significantly exhibited antiinflammatory activity by inhibiting the migration of leukocytes induced by thioglycollate. However, the activity of the extracts was lower than the positive control, indomethacin. Further purification was required to obtain the active compound with the activity comparable to indomethacin. Thin layer chromatography (TLC) analysis demonstrates that the methanolic extract of Mangifera casturi fruit contains terpenoid and phenolic compounds.Mangga kasturi (Mangifera casturi) adalah mangga  khas Kalimantan Selatan. Buah mangga   kasturi dilaporkan memiliki  aktivitas antioksidan dan potensial untuk pengobatan berbagai penyakit termasuk penyakit yang berhubungan dengan inflamasi. Penelitian ini bertujuan untuk menguji aktivitas antiinflamasi dari ekstrak metanol buah mangga kasturi melalui uji migrasi leukosit pada mencit yang diinduksi thioglikolat. Secara singkat, mencit diinduksi dengan thioglikolat untuk menaikkan jumlah leukosit, dan dihitung penghambatan migrasi leukosit oleh ekstrak metanolik buah mangga kasturi. Thioglikolat diberikan selama 4,5 jam (i.p.) sedangkan ekstrak metanolik buah mangga kasturi (i.p.) diberikan 30 menit sebelum pemberian thioglikolat. Keduanya diberikan  secara injeksi intra peritoneal (i.p.). Penghitungan jumlah leukosit dilakukan menggunakan haemositometer dengan bantuan mikroskop. Jumlah leukosit pada kelompok normal, indometasin, ekstrak dosis 625; 125; dan 2,5 g/Kg BB berturut-  turut adalah 38,24%; 11,28%; 65,24%; 19,72%; dan 7,18%. Analisis statistik dengan menggunakan uji post hoc multiple comparison Games Howell dengan tingkat kepercayaan 95% menunjukkan bahwa ekstrak metanolik buah mangga kasturi dosis 1250 dan 2500 mg/Kg BB mempunyai potensi antiinflamasi  melalui penghambatan migrasi leukosit pada mencit yang diinduksi thioglikolat. Aktivitas ekstrak metanolik buah mangga kasturi tersebut lebih lemah dibandingkan indometasin yang menberikan respon antiinflamasi yang sama pada dosis yang lebih kecil. Uji Kromatrografi Lapis Tipis (KLT) menunjukkan ekstrak metanolik buah mangga kasturi mengandung senyawa triterpenoid dan fenolik