Here, we describe the use of a fluorescence based lateral flow competition assay for the screening of four classes of drugs, viz, Δ9-tetrahydrocannabinol (THC), cocaine (through the detection of benzoylecgonine, BZE), opiates (through the detection of morphine, MOR) and amphetamine (AMP) present in the sweat of a fingerprint. The Drug Screening Cartridge was specifically developed for fingerprint sample collection and analysis. For this study, the cut-offs were set at: 190, 90, 68 and 80 pg/fingerprint for THC, BZE, MOR and AMP, respectively. Working with three UK coroners, the Drug Screening Cartridge, together with its fluorescence reader, was applied to the detection of drugs in the sweat of a fingerprint from deceased individuals. The study shows that there was sufficient sweat present on the fingertips to enable analysis and that the Drug Screening Cartridge could detect the presence, or absence, of each drug. The presence of the drugs was confirmed using LC–MS-MS analysis of a second fingerprint sample collected simultaneously. Excellent correlation was achieved between the results obtained from the Drug Screening Cartridge and the LC–MS-MS analysis of the fingerprint samples obtained from 75 individuals. The accuracy of the results was: 99% for THC; 95% for BZE; 96% for MOR and 93% for AMP. The results obtained using the Drug Screening Cartridge were also compared to toxicological analysis of blood and urine samples with good correlation. The accuracy of the results between the Drug Screening Cartridge and blood was: 96%, 92%, 88% and 97% for THC, BZE, MOR and AMP, respectively. The comparison with urine showed an accuracy ranging between 86% and 92%. This fingerprint sample method has a collection time of just 5 s and a total analysis time of <10 mins. These results show that the lateral flow Drug Screening Cartridge is an excellent screening test to provide information on drug use from the sweat in a single fingerprint sample.
An accurate, sensitive, selective and precise stability indicating Isocratic RP-LC method and UVvisible Spectroscopic method were developed for the quantitative determination of enzalutamide in bulk and synthetic mixture. The HPLC was carried out by reversed-phase technique on Sun fire C18, 5 µm column with mobile phase containing methanol: water (70:30). The flow rate was 1.0 ml/min and effluents were monitored at 234 nm with help of photodiode array (PDA) detector. UV spectroscopic determination was carried out at an absorption maximum of 234 nm using methanol as a solvent. The linearities were in the range of 4-14 µg/ml for UV-visible spectroscopic method and 0.03-20 µg/ml for RP-LC method, respectively. Validation of proposed method has been carried out with respect to linearity, accuracy, precision, specificity and robustness. Stock solution of EZA was subjected to acid and alkali hydrolysis, chemical oxidation, dry heat degradation and photolytic degradation and quantification has been carried out by proposed RP-LC method. Enzalutamide is susceptible to acidic and basic hydrolysis while it is stable towards chemical oxidation, dry heat and photolytic stress condition. Statistical comparison of both the method has been carried out by Student's F-test showed no significant difference between the results obtained by the two methods. Due to sensitivity, rapidity and accuracy of methods, we believe that the both proposed methods will be useful for the routine quality control analysis and quantification of drug in bulk and from synthetic mixture.
ÖZAmaç: Yüksek spesifiklikte spektral ayırıcılığı, kolay bulunan çözücülerin kullanımı, ekonomik ve çevre dostu oluşu ve ekstraksiyona gerek duymaması sebebiyle birinci türev senkron spektroflorimetri yönteminin üstün olduğu bulundu. Gereç ve Yöntemler: Bu çalışmada farmasötik preparatlardan klonazepam (CLO) ve paroksetin hidroklorürün (PH) eş zamanlı tayini için basit, hassas ve zamandan tasarruf sağlayan birinci türev senkronize spektroflorimetri yöntemi geliştirilmiştir. Bulgular: CLO, 512.79 nm'deki (PH'nin sıfır olduğu dalga boyu noktasında) emisyon dalga boyunda tayin edilmiştir. Benzer şekilde PH ise 336.00 nm'deki (CLO'nun sıfır olduğu dalga boyu noktasında) emisyon dalga boyunda ölçüm yapılarak tayin edilmiştir. Birinci türev için pik yüksekliğine karşı çizilen konsantrasyon grafiği CLO için 1-5 µg/mL, PH için 5-25 µg/mL aralığında doğrusal olarak bulunmuştur. Yöntemin istatistiksel validasyonu ICH kılavuzlarına uygun olarak istatistiksel gerçekleştirilmiştir. Teşhis sınırı CLO ve PH için sırasıyla 0.055 ve 0.033 µg/mL, tayin sınırı ise 0.169 ve 0.102 µg/mL'dir. Önerilen yöntem kullanılarak piyasa preparatlarındaki yüzde geri kazanım sonuçları CLO ve PH için sırasıyla %100.45 ile %99.38 arasında ve yüzde bağıl standart sapma değerleri de kesinlik ve doğruluk çalışmalarında 2'den daha düşük bulunmuştur. Sonuç: Bu spektroflorimetri yönteminin, basit spektrumlar, yüksek seçicilik ve düşük girişim gibi birçok avantajı olduğu bulunmuştur. Yüksek duyarlılığından dolayı, bu yöntem CLO ve PH'nin birlikte formüle edilmiş dozaj formlarından analizi için uygundur. Anahtar kelimeler: Birinci türev senkronize spektroflorimetri, klonazepam, paroksetin hidroklorür Objectives: First derivative synchronous spectrofluorimetry has been found to be superior because of its highly specific spectral discrimination and readily available solvent, it is economical, eco-friendly, and lacks an extraction procedure. Materials and Methods:In the present study, a new simple, sensitive, and time-saving first derivative synchronous spectrofluorimetry method has been developed for simultaneous estimation of clonazepam (CLO) and paroxetine hydrochloride (PH) in pharmaceutical dose forms. Results: CLO was determined at the emission wavelength of 512.79 nm (zero-crossing wavelength point of PH). Similarly, PH was measured at 336.00 nm (zero-crossing wavelength point of CLO). The first derivative amplitude-concentration plots were rectilinear over the range of 1-5 µg/ mL for CLO and 5-25 µg/mL for PH. The method was validated statistically as per the ICH guidelines. The limits of detection were 0.055 and 0.033 µg/mL and quantification limits were 0.169 and 0.102 µg/mL for CLO and PH, respectively. The percentage recovery in commercial formulation was found to be in the range 100.45% and 99.38% for CLO and PH, respectively, by the proposed method, and percent relative standard deviation values for precision and accuracy studies were found to be less than 2. Conclusion: This spectrofluorimetry method has been found to have several ad...
Objective A impressionable, discriminatory and precise stability indicating high performance thin layer chromatographic method has been developed and validated for the estimation of of Enzalutamide in bulk and synthetic mixture. Method The method engaged HPTLC aluminium plates pre-coated with silica gel 60F-254 as the stationary phase while the solvent system was ethyl acetate: toluene (4.5:5.5, v/v). The Rf value of enzalutamide was detected to be 0. 39 ± 0. 005 and the densitometric analysis were carried out in absorbance mode at 246 nm. The linear regression analysis data for the calibration plots presented a virtuous linear relationship for enzalutamide over a concentration range of 20 - 1000ng/band. Results The limit of detection and limit of quantification for enzalutamide was found to be 9.05 and 27.43ng/band. Enzalutamide was imperilled to acid and alkali hydrolysis, chemical oxidation, dry heat degradation and photolytic degradation. The degraded product peaks were well resolved from the pure drug peak with substantial difference in their Rf values. Conclusion Stressed samples were assayed using developed TLC technique. Suggested method was validated with respect to linearity, accuracy, precision and robustness. The method was successfully applied to the estimation of enzalutamide in synthetic mixture.
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