Validasi Metode Analisis Dalam Penetapan Kadar Benzo(a)piren Pada Ikan Bakar

VALIDASI METODE ANALISIS DALAM PENETAPAN KADAR BENZO(A)PIREN PADA IKAN BAKAR Windi Astuti1), Sri Sudewi1), Henki Rotinsulu1) 1)Program Studi Farmasi FMIPA UNSRAT Manado, 95115 ABSTRACT Benzo (a) pyrene is carcinogenic compounds formed by incomplete combustion of organic compounds during processing. This research was conducted by validating analytical methods used in the determination of the levels of benzo (a) pyrene and determines levels of benzo (a) pyrene in grilled fish. Determination of Benzo (a) was carrid out by High Performance Liquid Chromatography (HPLC). Optimization of benzo (a) pyrene was carrid out with a combination of mobile phase, acetonitrile : water (90:10) and (85:15) with a flow rate of 1,0 and 1,5 mL/min. Validation of analysis methods was carrid out by the linearity test, accuracy test, precision test and limits of detection test and quantity limit (LOD & LOQ) using a standard solution of benzo (a) pyrene. The results showed that the choosen condition of mobile phase, acetonitrile : water (90:10) and flow rate 1,0 mL/min. The linearity value resulted r2 = 0,9017. Accuracy test gives an average value of 100,00 recovery; 101,73 and 97,67%. an intraday precision test result of 0,01; 0,54 and 0,00% and interday; 0,01; 0,02 and 0,01%. Value of limit of detection and limit of quantity (LOD & LOQ) was 13,53 and 45,11μg/L. The levels of benzo (a) pyrene in grilled fish in restaurant X in Manado City 15,397 µg/L. Validation of analytical methods has already defined in accordance with the terms. The level of benzo (a) pyrene in grilled fish can be determined by using High Performance Liquid Chromatography (HPLC). The result level of Benzo (a) pyrene of no-treatment group and treatment group without wrapping with variable time 60 minutes are exceeds 10 µg which is maximum limit set Food and Drug Regulatiory Agency Republic of Indonesia (FDRA RI). Keywords: Validation of analysis methods, grilled fish, benzo (a) pirene, HPLC. ABSTRAK Benzo(a)piren merupakan senyawa karsinogenik yang terbentuk akibat pembakaran tidak sempurna dari senyawa organik selama proses pengolahan. Penelitian ini dilakukan dengan memvalidasi metode analisis yang digunakan dalam penetapan kadar benzo(a)piren dan mengetahui kadar benzo(a)piren dalam ikan bakar. Metode penentuan Benzo(a)piren dilakukan dengan Kromatografi Cair Kinerja Tinggi (KCKT). Optimasi penetapan benzo(a)piren dilakukan dengan kombinasi Fase gerak Asetonitril : Aquades (90:10) dan (85:15) dengan laju alir 1,0 dan 1,5 mL/menit . Validasi metode analisis dilakukan dengan uji linearitas, uji akurasi, uji presisi dan uji batas deteksi dan batas kuantitas dengan menggunakan larutan standar benzo(a)piren. Hasil penelitian menunjukkan bahwa kondisi optimum terpilih pada fase gerak Asetonitril : Aquades (90:10) dan laju alir 1,0 mL/menit. Nilai linearitas yang dihasilkan r2 = 0,9017. Uji akurasi memberikan nilai rata-rata perolehan kembali 100,00; 101,73 dan 97,67%. Uji presisi memberikan hasil pada intraday 0,01; 0,54 dan 0,00% dan interday 0,01; 0,02 dan 0,01%. Uji batas deteksi dan batas kuantitas memberikan hasil yaitu 13,53 dan 45,11µg/L. Kadar benzo(a)piren yang dihasilkan pada ikan bakar di rumah makan kota manado yaitu 15,397 µg/L. Validasi metode analisis sesuai dengan syarat yang sudah ditetapkan. Kadar benzo(a)piren pada ikan bakar bisa ditetapkan dengan menggunakan menggunakan Kromatografi Cair Kinerja Tinggi (KCKT). Kadar benzo(a)piren yang dihasilkan oleh rumah makan di kota manado melebihi dari batas maksimum yang ditetapkan Badan Pengawasan Obat dan Makanan Republik Indonesia (BPOM RI) yaitu sebesar 10 µg/L. Kata Kunci : Validasi metode analisis, Benzo(a)piren, ikan bakar, KCK

PENETAPAN KADAR BENZO(A)PIREN PADA DAGING BABI BAKAR DENGAN MENGGUNAKAN HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC)

PENETAPAN KADAR BENZO(A)PIREN PADA DAGING BABI BAKAR DENGAN MENGGUNAKAN HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC) Alfin Syahrin M. Ruslan1), Sri Sudewi1), Henki Rotinsulu1) 1)Program Studi Farmasi FMIPA UNSRAT Manado, 95115 ABSTRACT Benzo(a)pyrene (BAP) is a prototype compound of polycyclic aromatic hydrocarbons (PAH). BAP and oter PAH produced primarily by the incomplete combustion or pyrolysis of organic materials in the enironvement. The purpose of this study was to determine the benzo(a)pyrene compound during the treatment and time of processing as well as the devreasing levels of benzo(a)pyrene in roasted pork wrapped with banana leaves and aluminium foil which done by High Liquid Performance Chromatography (HPLC) analytical methods. HPLC analysis used in assay of the benzo(a)pyrene assay was done with a mixture of acetonitrile mobile phase : destilled water (90:10 and 85:15) with a flow rate of 1.0 and 1.5 mL/min. Result of the research on the levels of benzo(a)pyrene in roasted pork without wrapping with 30, 45, 60 minutes were 6.588; 8.432; 26488 g/L, respectively. The content of compound of benzo(a)pyrene in roasted pork wrapped in banana leaves with 30, 45 and 60 minutes were 6.016; 7.068 and 7.159 mg/L respectively. The levels of benzo(a)pyrene compound in grilled pork wrapped with aluminum foil grilled in 30, 45 and 60 minutes were 0.470; 0.817 and 7.015 mg/L, respectively. Decreasing levels of benzo(a)pyrene in the group of wrapping with banana leaves with parameters of 30, 45, and 60 minutes have decreased the levels of 8, 16, and 72 %, then the group of wrapping aluminum foil have decreased the benzo(a)pyrene with parameter 30, 45, and 60 minutes were 92, 90 and 73 %. Benzo(a)pyrene compound for roasted pork wrapped in aluminum foil could reduce the levels of benzo(a)pyrene more than the roasted pork wrapped with banana leaves. Keywords : Benzo(a)pyrene, grilled pork chops, HPLC analysis ABSTRAK Benzo(a)piren (BaP) merupakan senyawa prototipe polisiklik aromatik hidrokarbon (PAH). BaP dan PAH lainnya diproduksi terutama oleh pembakaran tidak sempurna atau pirolisis bahan organik di lingkungan. Penelitian ini bertujuan untuk mengetahui senyawa benzo(a)piren pada perlakuan dan waktu pengolahan serta mengetahui penurunan kadar benzo(a)piren pada daging babi bakar yang dibungkus daun pisang dan almunium foil dilakukan dengan metode analisis High Liquid Performace Cromatography (HPLC) yang digunakan dalam penetapan kadar Benzo(a)piren pada daging babi bakar dengan variable waktu pengolahan. Optimasi penetapan kadar benzo(a)piren dilakukan dengan kombinasi Fase gerak Asetonitril : Aquades (90:10) dan (85:15) dengan laju alir 1,0 dan 1,5 mL/menit. Hasil penelitian yang dilakukan kadar benzo(a)piren dalam daging babi bakar bakar tanpa pembungkusan dengan waktu 30, 45 dan 60 menit yaitu 6,588;  8,432 dan 26,488  µg/L. Kadar senyawa benzo(a)piren dalam daging babi  bakar yang dibungkus menggunakan daun pisang dengan waktu 30, 45 dan 60 menit yaitu 6.016; 7,068 dan 7,159  µg/L. Kemudian untuk kadar senyawa benzo(a)piren pada daging babi bakar yang dibungkus menggunakan allumunium foil dengan waktu 30, 45 dan 60 menit yaitu 0,470;  0,817  dan 7,015 µg/L. Penurunan kadar benzo(a)piren pada kelompok pembungkusan daun pisang dengan parameter waktu 30, 45, dan 60 menit memiliki penurunan kadar sebesar 8, 16, dan 72%, kemudian kelompok pembungkusan allumunium foil penurunan kadar benzo(a)piren dengan parameter waktu 30, 45, dan 60 menit sebesar 92, 90, dan 73%. Kandungan senyawa Benzo(a)piren dengan perlakuan yang dibungkus dengan alumunium foil dapat menurunkan kadar Benzo(a)piren lebih banyak dibandikan dengan perlakuan yang di bungkus daun pisang.   Kata Kunci : Benzo(a)piren, daging babi bakar, HPL

Short analytical validation of method for homocysteine analysis in plasma using tandem mass spectrometry

Homocistein je aminokiselina koja u ljudskom organizmu nastaje u metioninskom ciklusu. Hiperhomocisteinemija, povišena plazmatska koncentracija homocisteina, uključena je u patogenezu brojnih poremećaja od kojih su najznačajnije kardiovaskularne, neurodegenerativne, neuropsihijatrijske bolesti, preeklampsija i neke kongenitalne malformacije. Klasična homocistinurija je nasljedni metabolički poremećaj kod kojeg je uslijed mutacija enzima uključenih u metioninski ciklus također prisutna hiperhomocisteinemija. Tandemska spektrometrija masa je analitička metoda koja tijekom posljednjih godina sve više ulazi u rutinsku upotrebu u medicinsko-biokemijskom laboratoriju. To je robusna metoda visoke točnosti, preciznosti, specifičnosti i osjetljivosti koja zahtijeva mali volumen uzorka, kratko je vrijeme analize i niska je cijena analize po uzorku. Homocistein se do sada određivao imunokemijskom metodom nefelometrije, no literaturni i empirijski podaci prikupljeni tijekom rada u laboratoriju upućuju na činjenicu da su rezultati dobiveni tandemskom spektrometrijom masa točniji i pouzdaniji. Tijekom provođenja kratke analitičke validacije metode tandemske spektrometrije masa za određivanje plazmatske koncentracije homocisteina utvrđeno je da su zadovoljeni kriteriji prihvatljivosti za točnost, preciznost i linearnost. Usporedbom s postojećom metodom nefelometrije Passing-Bablokovom regresijskom analizom pokazano je da postoje proporcionalno i konstantno odstupanje između metoda, tj. da one nisu usporedive. Usprkos činjenici da metode nisu usporedive, zbog mnogih prednosti koje uključuju analitičke karakteristike i nižu cijenu analize po uzorku nova je metoda uvedena u rutinski rad.Homocysteine is an amino acid which is formed during the methionine cycle in the human body. Hyperhomocysteinemia, elevated plasma homocysteine concentration, is involved in the pathogenesis of many disorders, including cardiovascular, neurodegenerative, neuropsychiatric diseases, preeclampsia and some congenital malformations. Classical homocysteurinuria is a hereditary metabolic disorder which also causes hyperhomocysteinemia due to the mutation of enzymes involved in the methionine cycle. Tandem mass spectrometry is an analytical method that increasingly entered routine use over the past few years. It is a robust method of high accuracy, precision, specificity and sensitivity that requires a small sample volume, short time of analysis and low costs of analysis per sample. Homocysteine has so far been determined by the immunochemical method nephelometry but the literature and empirical data collected during laboratory work indicate that the results obtained by tandem mass spectrometry are more accurate and reliable. During the short analytical validation of the tandem spectrometry method for determining total plasma homocysteine concentration, it was found that the acceptance criteria for accuracy, precision and linearity were fulfilled. Comparison with the existing nephelometry method with the Passing-Bablok regression analysis showed that there is a proportional and constant deviation between methods, what means that they are not comparable. Despite the fact that the methods are not comparable, due to many advantages that include analytical characteristics and lower costs of analysis per sample, a new method is introduced in routine use

Estudo fitoquímico, atividades biológicas e propriedades antioxidantes da espécie vegetal Chusquea pinifolia Ness (Ness)(Poaceae)

Orientador: Dr. Obdulio Gomes MiguelCoorientador: Dr. Vinícius Bednarczuk de OliveiraDissertação (mestrado) - Universidade Federal do Paraná, Setor de Ciências da Saúde, Programa de Pós-Graduação em Ciências Farmacêuticas. Defesa : Curitiba, 22/03/2018Inclui referências: p.76-82Resumo: A espécie Chusquea pinifolia pertence à família Poacea, é uma espécie endêmica encontrada nos estados de Minas Gerais, São Paulo, Rio de Janeiro e Paraná, localizada em campos de altitude na região de alta montanha. O ambiente onde é encontrada e a falta de estudo sobre a espécie foi o que motivou o desenvolvimento deste trabalho que tem como principal objetivo a identificação de compostos e a avaliação das atividades biológicas presentes nos constituintes químicos. Para o estudo da espécie foram obtidos extratos brutos para folha e caule utilizando etanol como solvente. Posteriormente os extratos brutos foram fracionados com solventes de polaridade crescente para obtenção das frações hexano, clorofórmio, acetato de etila e hidroalcoólica remanescente. Foram feitos doseamentos de polifenóis e flavonoides totais. As atividades biológicas realizadas foram a de toxicidade frente à Artemia salina, atividade hemolítica e atividade antioxidante (DPPH, complexo fosfomolibdênio). Isolou-se um triterpeno identificado como Friedelina da fração hexano do caule, estudos realizados apontam significativa atividade analgésica desta substância, e da fração clorofórmio das folhas foi isolado um flavonoide identificado como Tricin, avaliado como composto majoritário do extrato bruto das folhas. Nenhuma das frações dos extratos brutos causou toxicidade frente ao microcrustáceo Artemia salina. Na atividade hemolítica a fração hexano do caule, nas concentrações de 500, 800 e 1000 ?g/mL, causou hemólise de média 60% em relação ao controle. Na atividade antioxidante pelo método DPPH, a fração acetato de etila do caule foi a que apresentou melhor atividade, com IC50 de 21?g/mL, no complexo fosfomolibdênio a fração clorofórmio tanto para folha como para o caule expressaram atividades respectivamente de 18,23% e 41,08%, comparada ao controle vitamina C. Conclui-se que a espécie Chusquea pinifolia necessita de mais estudos com finalidade de verificar suas atividades farmacológicas já que os resultados demonstram baixa toxicidade e boa atividade antioxidante nos modelos analisados. Palavras-chave: Friedelina. Tricin. Artemia salina. Hemólise. Complexo fosfomolibdênio.Abstract: The species Chusquea pinifolia belongs to the family Poacea, is a species found in the states of Minas Gerais, São Paulo, Rio de Janeiro and Paraná, located in high altitude fields in the high mountain region. Due to the environment where it is found and the lack of study on a species in which the development of the work has as main objective the identification of an evaluation of the biological activity present in the chemical constituents. For the study of the species with obtained crude extracts for leaf and stem, use as solvent. Subsequently the crude extracts were fractionated with solvents of increasing polarity to obtain the remaining hexane, chloroform, ethyl acetate and hydroalcoholic fractions. Doses of polyphenols and total flavonoids were made. As biological activities performed the toxicity to Artemia salina, hemolytic activity and antioxidant activity (DPPH, phosphomolybdenum complex).It was isolated a triterpene identified as Friedelina of the hexane fraction of the stem, studies carried out indicate significant analgesic activity of this substance, and of the chloroform fraction of leaves was isolated a flavonoid identified as Tricin, evaluated as the major compound of the raw extract of the leaves. None of the fractions from the crude extracts caused toxicity to the microcrack Artemia salina. In the hemolytic activity, the hexane fraction of the stem, at concentrations of 500, 800 and 1000 ?g /mL, caused a mean hemolysis of 60% in relation to the control. In the antioxidant activity by the DPPH method, a ethyl acetate fraction of the stem was the one that presented the best activity, with IC50 of 21 ?g/mL, phosphomolybdenum complex the chloroform fraction for both leaf and stem respectively expressed 18.23% and 41.08% compared to vitamin C control.It is concluded that the species Chusquea pinifolia requires further studies to verify its pharmacological activities since the results demonstrate low toxicity and good antioxidant activity in the analyzed models. Keywords: Friedelina. Tricin. Artemia salina. Hemolysis. Fossomolybdenum complex

DETERMINACIÓN DE LOS NIVELES DE 3,5,4´–TRANS TRIHIDROXIESTILBENO (TRANS-RESVERATROL) POR CROMATOGRAFÍA LÍQUIDA DE ALTA RESOLUCIÓN EN VINOS TINTOS BORGOÑA PERUANOS - 2012

RESEÑA HISTÓRICA DEL VINO EN EL PERÚ Y UVA BORGOÑA PRODUCCIÓN GENERAL DEL VINO PROPIEDADES TERAPÉUTICAS DEL VINO PRINCIPALES COMPONENTES DEL VINO TRANS-RESVERATROL CARACTERÍSTICAS QUÍMICAS DEL TRANS-RESVERATROL BIOSÍNTESIS DE TRANS-RESVERATROL METABOLISMO Y BIODISPONIBILIDAD DEL TRANS-RESVERATROL EFECTO QUIMIO-PROTECTOR METODOLOGÍAS PARA LA DETERMINACIÓN DE TRANS-RESVERATROL CROMATOGRAFÍA LIQUIDA DE ALTA RESOLUCIÓN FUNDAMENTO DE SEPARACIÓN INSTRUMENTACIÓN VALIDACIÓN ANALÍTICA LINEALIDAD EXACTITUD PRECISIÓN LIMITE DE DETECCIÓN LIMITE DE CUANTIFICACIÓ

Determinación de polifenoles: Quercetina, ácido cafeico, ácido cumarico en vinos de la provincia de Castilla

Se determinó cuantitativamente de los polifenoles en los vinos de la región Arequipa específicamente de Ácido Cafeico, Acido p-Cumárico y Quercetina los cuales afectan en gran magnitud en las características organolépticas de los vinos. Las cantidades de Ácido Cafeico, Acido p-Cumárico y Quercetina se analizaron en 7 vinos. Para poder determinar la cantidad de estos compuestos fenólicos en el matriz vino se tuvo que realizar una investigación experimental cuantitativa mediante HPLC el cual se tuvo que validar el método a usar para poder afirmar que los resultados obtenidos sean veraces y confiables; entre los parámetros, a usar se tomó en cuanta la linealidad, precisión, exactitud y sensibilidad. Los vinos analizados se elaboraron en casas vitivinícolas sur de la Región Arequipa; principalmente a partir de frutos ricos en flavonoles. Los vinos tintos se elaboraron principalmente a partir de uvas en la provincia de Castilla distrito de Aplao. La cantidad de Ácido Cafeico osciló entre 1.5 - 4.65 mg L-1, el Ácido p-Cumárico entre 1.32 – 19.34 mg L-1. Y la cantidad de Quercetina fue de 5.21 – 8.43 mg L-1. Se encontraron niveles inferiores a los presentados en vinos de otras nacionalidade