ISOLATION AND DETERMINATION CARPAINE ALKALOID IN PAPAYA (CARICA PAPAYA L.) LEAF EXTRACT BY THIN-LAYER CHROMATOGRAPHY SCANNER

Objective: This research was conducted to isolate the alkaloid carpaine by chromatography method and to determine it quantitatively by Thin Layer Chromatography Scanner. Methods: Dried leaves were macerated with ethanol 70% and fractionated with dichloromethane. Isolation of carpaine alkaloid from the dichloromethane fraction was carried out by column chromatography and preparative thin-layer chromatography according to the Rf value in Thin Layer Chromatography (TLC) after exposure by Dragendorff reagent. Results: The content of carpaine alkaloid was 7.5 mg with Rf 0.58 and dichloromethane: methanol (9.2:0.8) as eluent. Validation showed the linearity (R2) 0.9988, the limit of detection(LOD) was 0.05 ppm, the Limit of Quantification (LOQ) was 0.19 ppm, the recovery from 98.93-102.43%, and the % coefficient of variation was 0.16%. Conclusion: Carpaine alkaloid in papaya leaf extract was 10.52%.

Analisis Sidik Jari Kromatografi Lapis Tipis Tanaman Anting-Anting (Acalypha indica L.)

Anting-anting plant (Acalypha indica L.) is a plant that the community has used as raw material for herbal medicine. Anting-anting plants contain chemical compounds, so it is difficult to ensure these plants' safety and quality control. The research objective was to validate the method by knowing the stability, specificity, precision, and rigidity of the chromatographic profile of alkaloid compounds in the TLC analysis of the anting-anting extract. Extraction was carried out using ethyl acetate solvent for 20 minutes with a frequency of 42 kHz. The eluent used is cyclohexane: toluene: diethylamine (75:15:10) with Dragendorff reagent. The method of validation was acceptable according to the criteria except for the stability test of the analyte during chromatography. The instability of the analyte is indicated that not all spots form a straight diagonal line. The specificity of the method was carried out by comparing the fingerprint patterns of the leaves of the anting-anting with ekor kucing (Acalypha hibsida Farm.) and teh-tehan plants (Acalypha siamensis). The fingerprint profile showed a yellow stain (Rf = 0.39) on the leaves of the anting-anting, which did not appear in the fingerprint profiles of the two comparison samples. Keywords: anting-anting, fingerprint analysis, thin layer chromatography, method validation Anting-anting (Acalypha indica L.) merupakan tanaman yang telah dimanfaatkan masyarakat sebagai bahan baku obat herbal. Anting-anting mengandung banyak senyawa kimia sehingga sulit menjamin keamanan dan pengendalian mutu dari tanaman tersebut. Penelitian ini dilakukan untuk validasi metode dengan mengetahui kestabilan, spesifitas, presisi, dan ketegaran profil kromatografi senyawa alkaloid pada analisis KLT ekstrak anting-anting. Ekstraksi dilakukan menggunakan pelarut etil asetat selama 20 menit dengan frekuensi 42 kHz. Eluen yang digunakan adalah sikloheksana : toluena : dietilamin (75:15:10) dengan reagen Dragendorff. Validasi metode dapat diterima sesuai kriteria kecuali uji stabilitas analit selama kromatografi. Ketidakstabilan analit ditunjukkan dengan seluruh noda yang dihasilkan tidak membentuk garis diagonal yang lurus. Spesifitas pada metode dilakukan dengan membandingkan pola sidik jari daun anting-anting dengan daun ekor kucing (Acalypha hibsida Farm) dan daun teh-tehan (Acalypha siamensis). Profil sidik jari menunjukkan bahwa terdapat noda berwarna kuning kehijauan (Rf = 0,39) pada daun anting-anting yang tidak muncul pada profil sidik jari kedua sampel pembanding tersebut. Kata kunci: anting-anting, analisis sidik jari, kromatografi lapis tipis, validasi metode

Deteksi Golongan Senyawa Ekstrak Kasar Metabolit Ekstrasel Mikroalga Laut Spirulina sp. Sebagai Agen Antioksidan

Microalgae excrete antioxidant compounds as a defense system to protect themselves from the danger of ultraviolet rays. These compounds also can be used as the organic materials of cosmetics or medicines. This study aimed to determine the antioxidant activity of marine microalgal extracellular metabolite extract of Spirulina sp.. Extracellular metabolites were extracted from the residual media filtrate from the harvesting. The qualitative antioxidant test’s results using the thin-layer chromatography technique and 2,2-diphenylpycrilhydrazil (DPPH) reagent showed antioxidant activity. Moreover, the component identification using ninhydrin and Dragendorff reagent in thin layer chromatography test showed alkaloid and peptide compounds. To support the results, the identification using infrared spectrum analysis showed the peaks at 1117 cm-1 (C-N and C-C stretching), 1458 cm-1 (C-H bending of methyl group), 1635 cm-1 (C=O stretching of amide group), and 3454 cm-1 (N-H stretching of amine and amide groups).

Development of determination methods of quetiapine fumarate for forensic-pharmaceutical purposes

Quetiapine fumarate (an antipsychotic) is part of numerous generic drugs that are in fairly wide demand among the population, therefore, more and more data appear on the counterfeiting and smuggling of funds, as well as non-medical use, which are life-threatening for the population and explain the high prevalence of the active ingredient as object of forensic examination. The aim. To develop an algorithm for conducting a forensic pharmaceutical examination and propose a method for determining quetiapine fumarate for forensic pharmaceutical purposes. Materials and methods. All studies were performed using reagents that meet the EP, USP and USPU requirements, Class A glassware and qualified devices. Identification by IR spectroscopy was performed in the range from 500 to 4000 cm-1 on the device “Nicolet 380 FT-IR Spectrometer by Thermo Fisher Scientific” using a prefix “Smart Perfomer” with a ZnSe crystal. The UV absorption spectra of the solutions were recorded using a Specord 205 spectrophotometer from Analytik Jena AG (Germany). TLC was performed on Merck chromatographic plates (silica gel 60G F254, Germany). The following systems were used as mobile phases: hexane – acetone – 25 % ammonia solution (60: 40: 2); methanol – 25 % ammonia solution (100: 1.5), hexane – acetone – 25 % ammonia solution (50: 45: 5). Detection was performed under UV light (254 nm), followed by spraying with Dragendorff reagent. Analysis by gas chromatography with mass detection was performed using a GC gas chromatograph with a mass spectrometric detector GCMS-QP2020. Data were analyzed using the program: GCMSsolution, LabSolutions Insight (Shimadzu Corporation, Tokyo, Japan). Results. An algorithm for conducting a forensic pharmaceutical examination in accordance with the current legislation of Ukraine has been developed, methods for determining quetiapine for forensic pharmaceutical purposes have been proposed. Conclusions. The developed methods for determining quetiapine meet the requirements of the current legislation of Ukraine and the Ministry of Justice of Ukraine. The obtained data prove the high sensitivity and reproducibility of the methods and prove the possibility of their introduction into the practice of forensic examination

A spot test for determination of residual TBA levels in 18F-radiotracers for human use using Dragendorff reagent

A TLC spot test for determining residual TBA levels in positron emission tomography (PET) radiotracer formulations has been developed using Dragendorff reagent. Proof-of-concept is demonstrated by quantification of residual TBA in 6-[18F]fluoro-DOPA.

New biological findings of ethanol and chloroform extracts of fungi Suillellus rubrosanguineus and Tylopilus felleus

The aim of the research was to determine some basic biological activities of less biomedically studied but commonly known two fungi from the Boletaceae family Suillellus rubrosanguineus and Tylopilus felleus, which grow in the forests of Middle Europe. The cytotoxicity tests of the ethanol and chloroform extracts were carried out using NIH-3T3 and MCF-7 cell lines. The presence of alkaloids in the extracts was assessed by the reaction with Dragendorff reagent. In all of the extracts the positive reaction with the reagent was observed. In general, the extracts from Suillellus rubrosanguineus were more cytotoxic than the extracts from Tylopilus felleus and exhibited no selectivity of activities on healthy and cancer cell lines. However, the extracts from Tylopilus felleus proved to be selectively cytotoxic for cancer cell line. Tylopilus extracts or their isolated bioactive compounds could be considered for further study in pre-clinical experiments.

Antimicrobial Activity and Identification The Compounds of Methanol Extract from The Pleurotus Ostreatus Fruiting Body

<p><em>Pleurotus ostreatus</em> is an edible mushroom that also has potential as medicinal values. In this study, fruiting body of <em>P. ostreatus</em> was tested for its ability to inhibit the growth of fungi and bacteria. The fruiting body powder of <em>P. ostreatus </em>was extracted using methanol by maceration method. Analysis of this compound was done by using anisaldehid sulfuric acid, Dragendorff reagent, and FeCl₃. Using the agar well diffusion technique, the extracts were tested for their antimicrobial activity against <em>Staphylococcus aureus </em>(Gram positive), <em>Enterobacter aerogenes</em> (Gram negative) and <em>Candida albican</em> (yeast). The spot results on TLC using crude extract of <em>P. ostreatus</em> is terpenoids. Zone of inhibition for the various extracts varied between 10.9 - 23.2 mm. Ten miligrams extract exhibit maximum antimicrobial activity against most of the tested pathogens </p>

Quantification of cocaine and its adulterants (lidocaine and levamisole) using the Dragendorff reagent allied to paper spray ionization mass spectrometry

The combination of paper chromatography with PS(+)MS showed be promise to quantify cocaine, lidocaine and levamisole.

Phytochemical investigation of the tomatillo fruit (Physalis ixocarpa Brot., Solanaceae)

The occurence of alkaloids, witanolides, saponosides and flavonoids was investigated in the fruits of tomatillo (<i>Physalis ixocarpa</i> Brot.) cv. Bujna and Rendidora. The tomatillo is commonly cultivated in Mexico and has been introduced on a small scale in Poland. The chromatographic analysis of alkaloids showed only trace amounts of compounds reacting with the Dragendorff reagent. In the fraction of quaternary alkaloids soluble in water, only choline was found. Witanolides were not detected in tomatillo fruits. The saponosides were found only in trace amounts and their hemolytic indexes as well as saponification numbers were very low. Flavonoids were represented by the derivatives of quercetine differring with their sugar moieties attached to the hydroxyl group at C-3. It is concluded that the toxic compounds which would be harmful to human health were not found in tomatillo fruits. The presence of choline in them, as well as of flavonoids belonging to the vitamin P group is advantageous for health.