Validirane RP-RP-HPLC i TLC metode za simultano određivanje tamsulozin hidroklorida i finasterida u istom dozirnom pripravku

Reversed phase high-performance liquid chromatography (RP-HPLC) and thin-layer chromatography (TLC) methods have been developed and validated for simultaneous estimation of tamsulosin hydrochloride and finasteride in bulk drug and in combined dosage forms. RP-HPLC separation was achieved on a Phenomenex C18 column using methanol/0.02 mol L-1 ammonium acetate buffer/triethylamine (79.9 + 20 + 0.1, V/V/V) (pH 9.2) as mobile phase. The TLC separation was achieved on an aluminium-backed layer of silica gel 60F254 using toluene/methanol/triethylamine (9 + 1.5 + 1, V/V/V) as eluent. Quantification was achieved with photodiode array (PDA) detection at 235 nm over the concentration range 0.5–16 and 150 µg mL1 with mean recovery of 99.8 ± 0.9 and 100.0 ± 0.8 % for tamsulosin hydrochloride and finasteride, respectively, by the RP-HPLC method. Quantification was achieved with UV detection at 270 nm over the concentration range 100–2000 ng per spot and 250–5000 ng per spot with mean recovery of 98.9 ± 0.9 and 99.6 ± 0.7 % for tamsulosin hydrochloride and finasteride, respectively, by the TLC method. Both methods are simple, precise, accurate and sensitive and are applicable to the simultaneous determination of tamsulosin hydrochloride and finasteride in bulk drug and in combined dosage forms.U radu su opisani razvoj i validacija inverzno fazne kromatografije visoke učinkovitosti (RP-HPLC) i tankoslojne kromatografije (TLC) za simultano određivanje tamsulozin hidroklorida i finasterida kao čistih supstancija i u kombiniranim tabletama. Za RP-HPLC odjeljivanje korištena je Phenomenex C18 kolona (250 mm, 4,6 mm, 5 µm) i metanol/0,02 mol L–1 pufer s amonijevim acetatom/trietilamin (79,9+20+0,1, V/V/V) (pH 9,2) kao pokretna faza, pri protoku 1 mL min-1. TLC odjeljivanje rađeno je na silikagelu 60F254 na aluminijskoj podlozi, koristeći toluen/metanol/trietilamin (9+1,5+1, V/V/V) kao eluens. Za detekciju u RP-HPLC metodi korištena je fotodioda (PDA) pri 235 nm te je provedena kvantitacija u koncentracijskom području 0,5–16 µg mL–1 i 1–50 µg mL–1, uz srednji analitički povrat od 99,8 ± 0,9 % za tamsulozin hidroklorid i 100,0 ± 0,8 % za finasterid. Za kvantitaciju u TLC metodi korištena je UV detekcija pri 270 nm u koncentracijskom području 100–2000 ng po točki za tamsulozin hidroklorid i 250–5000 ng po točki za finasterid, uz srednji analitički povrat od 98,9 ± 0,9, odnosno 99,6 ± 0,7 %. Obje metode su jednostavne, precizne, točne i osjetljive i mogu se primijeniti za simultano određivanje tamsulozin hidroklorida i finasterida kao čistih supstancija i u kombiniranim dozirnim oblicima

Measurement of tamsulosin in human serum by liquid chromatography-tandem mass spectrometry

AbstractA simple, sensitive and robust method to extract tamsulosin from human serum, and quantify by liquid chromatography–tandem mass spectrometry (LC–MS/MS) was developed and validated and is applicable as a measure of compliance in clinical research. Tamsulosin was extracted from human serum (100μL) via liquid–liquid extraction with methyl tert-butyl ether (2mL) following dilution with 0.1M ammonium hydroxide (100μL), achieving 99.9% analyte recovery. Internal standard, d9-finasteride, was synthesised in-house. Analyte and internal standard were separated on an Ascentis® Express C18 (100mm×3mm, 2.7μm) column using a gradient elution with mobile phases methanol and 2mM aqueous ammonium acetate (5:95, v/v). Total run-time was 6min. Tamsulosin was quantified using a triple quadrupole mass spectrometer operated in multi-reaction-monitoring (MRM) mode using positive electrospray ionisation. Mass transitions monitored for quantitation were: tamsulosin m/z 409→228 and d9-finasteride m/z 382→318, with the structural formulae of ions confirmed by Fourier transform ion cyclotron resonance mass spectrometry (within 10ppm). The limit of quantitation was 0.2ng/mL, and the method was validated in the linear range 0.2–50ng/mL with acceptable inter- and intra-assay precision and accuracy and stability suitable for routine laboratory practice. The method was successfully applied to samples taken from research volunteers in a clinical study of benign prostatic hyperplasia

The Isolation of Photoactive Chloroplasts from Leaves of the Mangrove, Kondelio condel Druce

A method for isolating photoactive chloroplasts from mangrove leaves was investigated. Chloroplasts with the Hill reaction activity(photoreduction of ferricyanide) were isolated from the leaves of a mangrove plant, Kandelia candel Druce, by employing PEG in the grinding medium. Some characteristics of these chloroplasts are presented in this paper. Grinding medium for isolating photoactive chloroplasts is consisted of 1M sucrose,10%(W/V) PEG 4000, 0.025 M sodium iso-ascorbate, 0.05 M magnesium chloride and 0.05 M histidine buffered at pH 6.5 with HCI. Chloroplasts isolated without PEG reduces ferricyanide considerably in the dark. However, ferricyanide reduction in the dark was decreased by addition of PEG in the grinding medium. A possibility of tannins as the reductant is examined

Validated spectrofluorimetric method for the determination of tamsulosin in spiked human urine, pure and pharmaceutical preparations

A novel, sensitive and selective spectrofluorimetric method was developed for the determination of tamsulosin in spiked human urine and pharmaceutical preparations. The proposed method is based on the reaction of tamsulosin with 1-dimethylaminonaphthalene-5-sulfonyl chloride in carbonate buffer pH10.5 to yield a highly fluorescent derivative. The described method was validated and the analytical parameters of linearity, limit of detection (LOD), limit of quantification (LOQ), accuracy, precision, recovery and robustness were evaluated. The proposed method showed a linear dependence of the fluorescence intensity on drug concentration over the range 1.22 x 10(-7) to 7.35 x 10(-6)M. LOD and LOQ were calculated as 1.07 x 10(-7) and 3.23 x 10(-7)M, respectively. The proposed method was successfully applied for the determination of tamsulosin in pharmaceutical preparations and the obtained results were in good agreement with those obtained using the reference method. Copyright (c) 2013 John Wiley & Sons, Ltd