THE METABOLITE FINGERPRINTS, ANTIMALARIAL ACTIVITIES AND TOXICITIES OF ARTOCARPUS CHAMPEDEN STEMBARK FROM VARIOUS REGIONS IN INDONESIA

In Indonesia, cempedak (Artocarpus champeden Spreng) stembark from family of moraceae had been traditionally used for malarial treatment. Difference in the location of growth could cause the difference of metabolite fingerprints. As a result, there might be different toxicity and antimalarial activity in the same plants. The goal of this study was to obtain the fingerprints of the metabolites found in A. champeden stembark from different parts of Indonesia in order to authenticate and control the extract's quality. Fingerprints were performed using the HPTLC-Densitometry technique, in vitro toxicity and antimalarial activity were also determined using MTT assay and HRP2 assay. The correlation between metabolite fingerprints, toxicity and antimalarial activity was analysed using chemometrics tools: Principle Component Analysis (PCA), Partial Least Square (PLS) and Hierarchical Clustering Analysis (HCA). As a result, there is significant difference between fingerprints and toxicity profiles of A. champeden (p<0.05), whereas for antimalarial profiles, there is no significant difference between of them (p>0.05). Meanwhile, the nutrients (copper, zinc and manganese) are suspected to be responsible for the metabolite content. Besides morachalcone-A, compounds with Rf values ​​of 0.66 and 0.63 can be proposed as additional markers because they have responsibility for antimalarial activity and toxicity.

Short communication extraction and phytochemical analysis of Hyptis spicigera leaves

Plants are recognized in the pharmaceutical industry for their broad structural diversity as well as their wide range of pharmacological activities, which is due to their biologically active compounds known as phytochemicals. The present study reports the extraction, thin layer chromatography and screening of phytochemical constituent of Hyptis spicigera leaves. The thin layer chromatography of the leave extract shows 11 bands with Rf values of 0.03, 0.06, 0.09, 0.12, 0.17, 0.19, 0.20, 0.23 and 0.31 respectively. Qualitative phytochemical screening showed the presence of alkaloids, flavonoids, steroids, emodins, and cardiac glycoside while phenols, tannins, terpenoids, tri terpenoids and anthraquinones were absent. The presences of these phytochemicals showed that Hyptis spicigera leaves may be useful for medicinal purpose.

Extraction and Purification of Secondary Metabolites of Paris polyphylla (Smith) Using Column Chromatography.

Paris polyphylla Smith is an erect and herbaceous plant. It has rich chemical constituents such steroidal saponins, phytosterols, flavonoids, and alkaloids that possess antimicrobial and anticancer activities. In the current investigation, we examined the effect of different solvents in the extraction yield and further purification via column chromatography. The ground powdered rhizomes of P. polyphylla was extracted with 100% hexane, 100% ethyl acetate, 70% ethanol, and 70% methanol. The extracts were filtered, evaporated using a rotary evaporator, concentrated, and measured. Subsequently, the solvent with high extraction yield was further purified using column chromatography. Ethanol produced the highest extraction yield as compared to the other solvents. About 30 fractions were eluted which was pooled into four fractions based on the Rf values and bands observed through thin layer chromatography (TLC) analysis.

Preliminary Phytochemical Screening, Isolation, Characterization, Structural Elucidation and Antibacterial Activities of Leaves Extracts Rhus Vulgaris (Kimmo)

Background: Rhus vulgaris commonly known as sumac, a plant that is known to possess different therapeutic values including antioxidant and antibacterial activities. Medicines from plants contributed largely to human health. The aim of this study was to screen the phytochemical constituents, isolate, elucidate the structure and antibacterial activity of methanol extract from the leaves of Rhus vulgaris.Methods: The methanolic extract of Rhus vulgaris was subjected to column chromatography and eluted with solvent mixture of methanol: chloroform (1:8) ratio. The eluted fractions were run in the TLC mobile phase with the different solvent ratio. Based on the TLC profile the fractions with similar Rf values were pooled together. The structure of the isolated compound was characterized based on the spectral data (IR, 1H NMR, 13C NMR, and DEPT) and extracts from Rhus vulgaris has been shown to have antibacterial activity were tested against four strains bacteria Streptococcus aureus(gram-positive) and Escherichia coli, Salmonella typhimurium, and K. pneumoniae (gram-negative) using Agar well diffusion method.Result: The results showed that the methanol extracts were active against all the tested bacteria. The structure of this compound 1-p-tolyl pentadeca-7,9-dien-1-ol was characterized by means of 1H NMR, 13C NMR, and IR spectral data.Conclusion: Therefore, it is concluded that the use of herbal plants and their recipes are the major source of drugs in a traditional medicinal system to cure different diseases.

Penetapan Kadar β-karoten dalam Buah Tomat (Lycopersicum esculentum Mill.) Berdasarkan Ketinggian Tempat Tumbuhnya dengan Metode Spektrofotometri UV-Vis

The quality of secondary metabolites in plants is determined by the altitude where they grow, in tomato plants secondary metabolites that have the potential as antioxidant activity are caused by β-carotene. β-carotene is a red-orange pigment that is very abundant in plants and fruits. β-carotene is an organic compound and is classified as a terpenoid, β-carotene is also one of the antioxidants that can prevent disease. The purpose of this study was to determine the level of β-carotene in tomatoes based on the altitude where they grew. The sample used in this study was Tomato Fruit (Lycopersicum esculentum Mill.) which was taken at an altitude of ±1206, ±845, ±548 and ±76 masl. Qualitative testing using Fourier Transform Infra Red (FTIR) and Thin Layer Chromatography (TLC), the mobile phases used are chloroform and ethyl acetate (7:3), the Rf values of the samples and comparisons are not much different. Quantitative testing using UV-Vis Spectrophotometry method at a wavelength of 461 nm. The results showed that the four positive samples contained β-carotene. The levels of β-carotene in the samples studied were sample A (±1206 masl) as much as 5.642 mg/100 gr, sample B (±845 masl) as much as 7.986 mg/100 gr, sample C (±548 masl) as much as 11.128 mg/100 gr and sample D (±76 masl) as much as 3.792 mg/100 gr. From this study, it was found that the highest β-carotene content was found in sample C (±548 masl) and the lowest β-carotene level was found in sample D (±76 masl). Environmental factors such as light, temperature, pH, altitude, and temperature greatly affect the content of β-carotene.Keywords: Determination of rates; β-carotene; tomatoes; UV-Vis spectrophotometry AbstrakKualitas metabolit sekunder dalam tumbuhan salah satunya ditentukan oleh ketinggian tempat tumbuhnya, dalam tanaman tomat metabolit sekunder yang berpotensi sebagai aktivitas antioksidan salah satunya disebabkan oleh β-karoten. β-karoten adalah pigmen berwarna merah-orange yang sangat berlimpah pada tanaman dan buah-buahan. β-karoten merupakan senyawa organik dan diklasifikasikan sebagai suatu terpenoid, β-karoten juga merupakan salah satu antioksidan yang dapat mencegah penyakit. Tujuan dari penelitian ini adalah untuk mengetahui kadar β-karoten dalam buah tomat berdasarkan ketinggian tempat tumbuhnya. Sampel yang digunakan dalam penelitian ini adalah Buah Tomat (Lycopersicum esculentum Mill.) yang diambil pada ketinggian ±1206, ±845, ±548 dan ±76 mdpl. Pengujian secara kualitatif menggunakan metode Fourier Transform Infra Red (FTIR) dan Kromatografi Lapis Tipis (KLT), fase gerak yang digunakan yaitu berupa kloroform dan etil asetat (7:3) diperoleh nilai Rf sampel dan pembanding yang tidak jauh berbeda. Pengujian secara kuantitatif menggunakan metode Spektrofotometri UV-Vis pada panjang gelombang 461 nm. Hasil penelitian menunjukkan bahwa dari keempat sampel positif mengandung β-karoten. Kadar β-karoten dalam sampel yang diteliti yaitu sampel A (±1206 mdpl) sebanyak 5,642 mg/100 gr, sampel B (±845 mdpl) sebanyak 7,986 mg/100 gr, sampel C (±548 mdpl) sebanyak 11,128 mg/100 gr dan sampel D (±76 mdpl) sebanyak 3,792 mg/100 gr. Dari penelitian ini diketahui bahwa kadar β-karoten tertinggi terdapat pada sampel C (±548 mdpl) dan kadar β-karoten terendah terdapat pada sampel D (±76 mdpl). Faktor lingkungan seperti cahaya, suhu, pH, ketinggian tempat, dan temperature sangat berpengaruh terhadap kandungan β-karoten.Kata kunci: Penetapan kadar; β-karoten; buah tomat; spektrofotometri UV-Vis

Isolation of Arbutin from Leaves and Fruits of Buni (Antidesma Bunius L. Spreng) As Tyrosinase Enzym Inhibitor

Several studies have shown that plant extractive substances have the potential as active compounds inhibiting the enzyme tyrosinase. Arbutin is an enzyme inhibitor of tyrosinase, known as a popular whitening agent used in cosmetics because of its effectiveness in overcoming skin hyperpigmentation. The purpose of this study was to conduct a qualitative and quantitative analysis of arbutin on Buni Leaves and Fruits (Antidesma bunius L. Spreng). The raw simplicia used are mature and young buni leaves, green, red and purple buni fruits. The extraction method is maceration using methanol as solvent. The initial screening for arbutin content was carried out using thin layer chromatography (TLC) and dichlormethan:methanol 50:50 used as mobile phase. Isolation of arbutin content was carried out using Preparative TLC with the same eluent. Qualitative and quantitative analysis were performed using High Pressured Liquid Chromatography (HPLC) with mobile phase of acetonitrile: water 60:40. The tyrosinase enzyme inhibition activity test was then carried out in vitro using 96-well microplate, l-tyrosine and l-dopa were used as substrate at a wavelength of 492 nm. The Rf values obtained ??for mature buni leaves and green buni fruits, respectively 0.61 and 0,62. The retention time of HPLC chromatogram respectively 2,784 minutes and 2,758 minutes. Arbutin levels in leaves and fruits are 7.9 mg / g and 2 mg / g. The activity of the enzyme tyrosinase of mature buni leaves on L-dopa and L-tyrosine substrate were respectively stated as IC50 values ??of 88.7191ppm and 101.33347 ppm. The activity of the enzyme tyrosinase of the green buni fruit on L-dopa and L-tyrosine substrate respectively stated IC50 values ??of 198,0293 ppm and 246,1296 ppm.

Application of Central Composite Design for screening and Optimization of HPTLC method for simultaneous quantitation of Aprepitant, Dexamethasone and Ondansetron in their synthetic mixtures

New HPTLC method was developed and optimized for estimation of Ondansetron (OND), Dexamethasone (DEX) and Aprepitant (APT) in laboratory prepared ternary mixtures by using Central composite design (CCD). The independent variables used for the optimization were the acetone content in mobile phase (%mL), distance of developing solvent (cm) and saturation time (min). HPTLC Separation was performed on Precoated silica gel F254 aluminum plate (10X10 cm, 100μm thickness) with a mobile phase consisting of chloroform: methanol: acetone: ethyl acetate: ammonia (9:4:2:5:0.2 % v/v/v/v). Quantification of OND, APT and DEX were achieved based on a Densitometric analysis over the concentration range of 200-1200 ng/band, 500-1000 ng/band and 1000-2000 ng/band, respectively, at 254nm. The method was yielded dense and well-resolved bands at Rf values of 0.54± 0.02, 0.79±0.02 and 0.23±0.01 for OND, APT and DEX, respectively. The linear regression analysis for the calibration plots produced r2= 0.9997, r2= 0.9998 and r2=0.9997 for OND, APT and DEX, respectively. The method was validated according to the ICH guidelines. The robustness test was determined that the selected factors have an insignificant effect on the responses. The results indicated that the method is suitable for the routine quality control testing of OND, APT and DEX in their bulk form.

Development of thin-layer chromatographic method for determination of caffeine in black, green, and white tea

Caffeine is naturally present in tea and coffee giving the pleasant and stimulant effect. Several different types of teas, black, green, and white teas bought in market were analysis for caffeine content. The boiled sample tea was filtered through filter paper. Lead(II) acetate was used to separate tannins from caffeine followed by filtration through filter paper with a black ribbon. The liquid-liquid extraction was carried out using dichloromethane (3×5 mL) and sodium sulfate as a drying agent. The TLC method was performed on Merck precoated silica gel plates 5×10 cm (60F254, 200 μm) using either methanol or dichloromethane as solvents and the mobile phases were glacial acetic acid and ethyl acetate (95:5, v/v), while the second one was consisted of ethyl acetate and ethanol (80:20, v/v), respectfully. The Rf values were 0.36 and 0.86 for the first and the second mobile phase, respectively, in comparison to the standard caffeine. The values for pH of boiled sample teas were in the range from 4.85 to 5.80. The most abundant tea sample for caffeine was determined in green tea bought in the grocery store for health nutrition (2.04 %). The yield for tea samples from green market, white tea and two tea black samples were 0.06, 0.71, 0.07, and 0.05%, respectively. The developed TLC method can be used for determination of caffeine content in tea samples.

Analisis Natrium Diklofenak Dalam Sampel Jamu Pegal Linu Yang Dijual Di Kabupaten Semarang Secara Klt-Spektrofotometri Uv-Vis

Herbal medicine for rheumatic pain is one of the traditional medicinal products that are massively demanded by the public because it has many benefits. Medicinal Chemicals (MC) are often added to herbal medicine for rheumatic pain to strengthen their properties, one of which is Diclofenac Sodium. Based on the Decree of the Minister of Health of the Republic of Indonesia No. 246 of 2010, traditional medicines are prohibited from containing Medicinal Chemicals (MC). This study aims to analyze the content of Diclofenac Sodium Medicinal Chemicals (MC) in the herbal medicine for rheumatic pain which is sold in Semarang Regency. This type of research was conducted using a laboratory experimental method which descriptively describes the results of the study based on the data obtained. The research method consisted of organoleptic test, qualitative analysis and quantitative analysis of the samples of herbal medicine for aches and pains. Organoleptic test was carried out by tasting the taste, smelling the smell, seeing the color and feeling the dosage form of the Jamu Pegal Linu sample. Qualitative analysis was performed by Thin Layer Chromatography (TLC) and quantitative analysis was performed by UV-Vis Spectrophotometry with 3 samples considered positive. Samples B, D, and E which is sold in Semarang Regency were positive for Diclofenac Sodium based on the Rf values ​​obtained from the samples, namely 0.28, 0.3, and 0.3, which were almost the same as the standard Rf for Sodium Diclofenac, which was 0.26. The stationary phase used a Silica Gel 254 and the mobile phase used Ethyl Acetate and N-Hexane in a ratio of 25: 25. In quantitative analysis, a wavelength of 275 nm was obtained with a linear equation y = 0.0245x + 0.0989 and a value of r = 0.9994 with a concentration of obtained in samples B, D, E were 39.27%, 2.67% and 4.9%, respectively. ABSTRAK Jamu Pegal Linu merupakan salah satu produk obat tradisional yang banyak diminati oleh masyarakat karena memiliki banyak manfaat. Bahan Kimia Obat (BKO) sering ditambahkan pada Jamu Pegal Linu untuk menambah khasiatnya, salah satunya adalah Natrium Diklofenak. Berdasarkan Permenkes RI No. 246 tahun 2010, obat tradisional dilarang mengandung BKO. Penelitian ini bertujuan untuk menganalisis kandungan Natrium Diklofenak pada sediaan Jamu Pegal Linu yang dijual di Kabupaten Semarang. Jenis penelitian dilakukan menggunakan metode eksperimental laboratorium. Metode penelitian terdiri dari uji organoleptis, analisis kualitatif dan kuantitatif. Uji organoleptis dilakukan dengan dengan cara mengamati warna, bau, rasa dan bentuk sampel. Analisis kualitatif dilakukan dengan Kromatografi Lapis Tipis (KLT) dan analisis kuantitatif dilakukan dengan Spektrofotometri UV-Vis. Sampel B, D, dan E yang dijual di Kabupaten Semarang positif mengandung Natrium Diklofenak berdasarkan nilai Rf yang didapatkan dari sampel berturut-turut yaitu 0.28, 0.3, dan 0.3 mendekati nilai Rf baku Natrium Diklfenak yaitu 0.26. Fase diam menggunakan Silica Gel 254 dan fase gerak menggunakan Etil Asetat dan N-Heksan dengan perbandingan 25 : 25. Pada analisis kuantitatif diperoleh panjang gelombang 275 nm dengan persamaan garis linier y = 0,0245x + 0,0989 dan nilai r = 0.9994 dengan kadar pada sampel B, D, E berturut-turut adalah 39.27%, 2.67% dan 4.9%.

TLC and HPTLC Finger Printing Analysis of Cyperus rotundus (Linn.)

Cyperus rotundus (Linn.) is a versatile plant belonging to the family Cyperaceae, used in herbal medicines worldwide to cure various human ailments. The present study attempts to analyze the profiles of flavonoids in different extracts of C. rotundus with the help of thin-layer chromatography (TLC) and high-performance thin-layer chromatographic (HPTLC) fingerprint. Flower and stem extracts of C. rotundus were screens out with the help of TLC, and the Rf values were determined. HPTLC was used to quantify the flavonoid from flower extract of a plant at a 1.0 mg/mL concentration. It revealed the occurrence of flavonoids, especially quercetin in the ethanolic extract of C. rotundus flower, by using mobile phase toluene-ethyl acetate-formic acid (3:4:2.5 v/v), on a pre-coated plate of silica gel and quantified the amount of quercetin by densitometry absorbance mode at 257 nm. The limits of detection and quantification were 30.08 & 91.16 ng/mL, and the relative standard deviation ranged between 1.03 to 1.48 for intra-day and inter-day for HPTLC. The calibration range was 200-700 ng per band (r2 = 0.99321). Quercetin quantity in the ethanolic extract of the flower was found to be 0.011 mg/mL of the extract with an average recovery of 99.01–100.00%. Such fingerprinting is valuable in quality control and checking adulterants of natural drugs. Therefore, it can be helpful for the assessment of different marketable pharmaceuticals preparations.