Monitoring of cyanobacteria using derivative spectrophotometry and improvement of the method detection limit by changing pathlength

Effective monitoring tools and methods are needed for the early detection and management of cyanobacteria in water bodies to minimize their harmful impacts on the environment and public health. This research investigated changing the cuvette pathlength (10-, 50-, and 100-mm) to improve the detection of cyanobacteria using UV-Vis spectrophotometry with subsequent application of derivative spectrophotometry and Savitzky-Golay (S-G) transformation. A non-toxigenic strain of blue-green cyanobacteria, Microcystis aeruginosa (CPCC 632), and a green algae strain for comparison, Chlorella vulgaris (CPCC 90), were studied in a wide range of concentrations (955,000–1,855 cells/mL). In each concentration range, method detection limits were established with absorbance measurements and S-G first derivative of absorbance using 10-, 50-, and 100-mm cuvette pathlengths. Increasing the cuvette pathlength from 10 to 100 mm resulted in a 15-fold improvement in sensitivity with absorbance and a 13-fold improvement with S-G first derivative of absorbance for M. aeruginosa. Overall, adoption of 100 mm pathlength and application of S-G derivative spectra improved the method detection limit for M. aeruginosa from 337,398 cells/mL to 4,916 cells/mL, which is below the WHO guideline for low probability of adverse health effects (<20,000 cells/mL). Similarly, the detection limit for C. vulgaris was improved from 650,414 cells/mL to 11,661 cells/mL. The results also showed that spectrophotometry could differentiate M. aeruginosa from C. vulgaris based on the variations in their pigment absorbance peaks.

SIMULTANEOUS DETERMINATION OF PARACETAMOL AND TAPENTADOL IN TABLETS BY RATIO SPECTRA DERIVATIVE SPECTROPHOTOMETRY

The application of the ratio spectra derivative spectrophotometry to the simultaneous determination of Paracetamol (PCM) and Tapentadol (TAP) in combined pharmaceutical tablets is presented. The spectrophotometric procedure is based on the use of the first derivative of the ratio spectra obtained by dividing the absorption spectrum of the binary mixtures by a standard spectrum of one of the compounds. The first derivative amplitudes were measured at 220 and 232 nm for the assay of TAP and PCM, respectively. Calibration graphs were established for 1-5 μg mL-1 for TAP and 6.5-32.5 μg mL-1 for PCM in binary mixture. The detection limits for TAP and PCM were found 0.098 and 0.595 μg mL-1, respectively, while the quantification limits were 0.298 μg mL-1 for TAP and 1.805 μg/ml for PCM. The relative standard deviations were found to be less than 2%, indicating reasonable repeatability of both methods. The proposed methods were hence validated as per ICH guidelines and successfully applied to the determination of these drugs in commercial tablets.

Spectrophotometric Determination of Carbimazole and Its Major Impurity, Degradation Product and Metabolite: Methimazole -=SUP=-*-=/SUP=-

The present research work was carried out in order to develop simple, accurate and precise UV sprctrophotometric methods having comparable sensitivity as that of sophisticated chromatographic techniques. Two methods were developed namely first derivative spectrophotometry and ratio spectra derivative spectrophotometry for accurate determination of specified impurity methimazole (imp A) in presence of drug carbimazole. First derivative spectrophotometric method involves recording of zero order spectra of both the drugs carbimazole and methimazole and its mixture in the range of 200-400 nm and subsequent conversion of these spectra into first derivative spectra. The drugs carbimazole and methimazole were determined by using zero crossing wavelengths of 227 and 260 nm respectively. In the second approach, ratio spectra were recorded for carbimazole and methimazole by selecting appropriate divisor concentration and converted into first derivative spectra. The determination of carbimazole and methimazole were carried out at wavelength 226.2 and 257 nm, respectively. Both the methods were validated as per ICH guideline. The drugs carbimazole and methimazole showed linear response with good correlation coefficient and exhibited specificity, accuracy and precision within acceptable range. The second method of ratio spectra derivative spectrophotometry was found more sensitive as compare to first derivative spectrophotometry in detecting level of impurity methimazole up to 0.5% as per official specification. Hence, these developed methods can be used as alternative to sophisticated chromatographic technique for determination of assay and related impurity in bulk drug and formulation. Keywords: carbimazole, methimazole, first derivative spectrophotometry, ratio spectra derivative spectrophotometry.

Recent Applications of High Performance Thin Layer Chromatography and Derivative Spectrophotometry in Pharmaceutical Analysis

At present, no one can imagine drug development, marketing and post-marketing without rigorous quality control at each stage. Only modern, selective, accurate and precise analytical methods for determination of active compounds, their degradation products and stability studies are able to assure the appropriate amount and purity of drugs administered every day to millions of patients all over the world. For routine control of drugs simple, economic, rapid and reliable methods are desirable. The major focus of current scrutiny is placed on high-performance thin layer chromatography and derivative spectrophotometry methods, which fulfill routine drug estimation’s expectations [1-4]. The present paper reveals state-of-the-art and possible applications of those methods in pharmaceutical analysis between 2010 and 2018. The review shows advantages of high-performance thin layer chromatography and derivative spectrophotometry, including accuracy and precision comparable to more expensive and time-consuming methods as well as additional fields of possible applications, which contribute to resolving many analytical problems in everyday laboratory practice.

Comparison of UV- and Derivative-Spectrophotometric and HPTLC UVDensitometric Methods for the Determination of Amrinone and Milrinone in Bulk Drugs

Background: Spectrophotometry and thin layer chromatography have been commonly applied in pharmaceutical analysis for many years due to low cost, simplicity and short time of execution. Moreover, the latest modifications including automation of those methods have made them very effective and easy to perform, therefore, the new UV- and derivative spectrophotometry as well as high performance thin layer chromatography UV-densitometric (HPTLC) methods for the routine estimation of amrinone and milrinone in pharmaceutical formulation have been developed and compared in this work since European Pharmacopoeia 9.0 has yet incorporated in an analytical monograph a method for quantification of those compounds. Methods: For the first method the best conditions for quantification were achieved by measuring the lengths between two extrema (peak-to-peak amplitudes) 252 and 277 nm in UV spectra of standard solutions of amrinone and a signal at 288 nm of the first derivative spectra of standard solutions of milrinone. The linearity between D252-277 signal and concentration of amironone and 1D288 signal of milrinone in the same range of 5.0-25.0 μg ml/ml in DMSO:methanol (1:3 v/v) solutions presents the square correlation coefficient (r2) of 0,9997 and 0.9991, respectively. The second method was founded on HPTLC on silica plates, 1,4-dioxane:hexane (100:1.5) as a mobile phase and densitometric scanning at 252 nm for amrinone and at 271 nm for milrinone. Results: The assays were linear over the concentration range of 0,25-5.0 μg per spot (r2=0,9959) and 0,25-10.0 μg per spot (r2=0,9970) for amrinone and milrinone, respectively. The mean recoveries percentage were 99.81 and 100,34 for amrinone as well as 99,58 and 99.46 for milrinone, obtained with spectrophotometry and HPTLC, respectively. Conclusion: The comparison between two elaborated methods leads to the conclusion that UV and derivative spectrophotometry is more precise and gives better recovery, and that is why it should be applied for routine estimation of amrinone and milrinone in bulk drug, pharmaceutical forms and for therapeutic monitoring of the drug.

Quantitative analysis of mixed pigments for Chinese paintings using the improved method of ratio spectra derivative spectrophotometry based on mode

Background Mixed pigment analysis is an important and complex subject in preserving and restoring Chinese paintings since the colors observed by our naked eyes or instruments such as hyperspectral cameras are usually a mixture of several kinds of pigments. The purpose of this study was to explore a more effective method to confirm the type of every pure pigment and their proportion in pigment mixtures on the surface of paintings. Methods Two endmember extraction algorithms were adopted to identify the types of pigments and an improved method of ratio spectra derivative spectrophotometry was used to determine their proportion. Main works (1) Extracting the pure pigment components from mixed spectrum by adopting two blind source separation algorithms: Independent Component Analysis and Non-negative Matrix Factorization; (2) matching the separated pure spectrum with the pigment spectral library built in our laboratory to determine the pigment type; and (3) calculating the proportions of mixed pigments using the Ratio Spectra Derivative Spectrophotometry based on Mode, which is improved based on the original algorithm. Finally, a comparison was made with two abundance inversion algorithms: Least Squares Algorithm and Minimum Volume Simplex Analysis. The correlation coefficient and root mean square error were used to provide evidence for accuracy evaluation. Conclusions (1) Non-negative matrix factorization is more suitable for endmember extraction; and (2) Ratio spectra derivative spectrophotometry based on mode is more suitable for abundance inversion.