Validated high performance thin layer chromatographic analysis of leaves and flower extracts of

is a significant therapeutic plant has a place with family apocynaceae contains in excess of 70 distinct sorts of chemotherapeutic agents and alkaloids which help in treating different illnesses. For the most part, it is known as Vincarosea, Ammocallisrosea and Lochnerarosea. There are numerous or more than 400 alkaloids present in plant, which are used as flavor, agrochemicals, pharmaceuticals, fragrance, ingredients, food addictives, and pesticides. To develop a validated high performance thin layer chromatographic method for the analysis of leaves and flower extracts of Sample solutions were applied onto the plates with automatic TLC sampler Linomat V (Camag, Muttenz, Switzerland) and were controlled by WinCATS software. Plates were developed in 10 x 10cm twin trough glass chamber (Camag, Muttenz, Switzerland). A CAMAG TLC scanner was used for scanning the TLC plates. Pre-coated silica gel aluminium plates 60F254. For vincristine, simultaneous estimation of vincristine was performed by HPTLC on a silica gel plate using toluene-methanol-diethylamine (8.75: 0.75: 0.5, v/v/v) as the mobile phase. The method was validated as per the ICH guidelines. The Rf value was found to be 0.76 for flower and 0.80 for leaves at 250 nm which shows the presence of vincristin in . In this research paper, a validated HPTLC Method has been developed for the analysis of leaves and flower extracts

A Validated HPTLC Densitometric Method for The Quantitative Determination of Ubidecarenone in Bulk and in Capsule Formulation

A new, simple, precise, accurate and rapid high performance thin layer chromatographic method has been developed and validated for the estimation of ubidecarenone in bulk and in capsule formulation. The chromatographic separation was performed on aluminium TLC plates precoated with silica gel 60F254 as a stationary phase and methanol:water (7:3) as a mobile phase. Detection was performed densitometrically in the absorbance mode at 280nm for the evaluation of chromatograms. The system has given well sharp peak of ubidecarenone (Rf=0.51±0.02). The linearity of the method was established in the range of 1-6 ng/µL with correlation coefficient (r2) of 0.9995. The method was validated for precision, accuracy, robustness, ruggedness, LOD, and LOQ as per ICH guidelines. The limit of detection was found to be 0.0392 ng/µL, whereas the limit of quantitation was found to be 0.1189 ng/µL. The percentage label claim for ubidecarenone in the capsule formulation was found to be 99.96±0.4703. The accuracy of the method was confirmed by recovery studies. The percentage recovery was found to be in the range of 100.10-101.45% for ubidecarenone. The % RSD value was found to be less than 2. The low %RSD value indicates that there is no interference due to excipients used in the formulation. Hence, the developed method was found to be simple, precise, accurate, and rapid for the analysis of ubidecarenone in bulk and pharmaceutical formulation and it can be effectively applied for the quality control analysis of ubidecarenone in bulk and pharmaceutical formulation.

Studies on Natural Products Using Monoclonal Antibodies: A Review

An immunoblotting system (“eastern blotting”) was developed for small-molecule herbal medicines like glycosides, with no conjugation function to the membrane. Briefly, the crude extracts of herb medicines were developed by thin-layer chromatography (TLC). The small-molecule herbal medicines on TLC plates were transferred to polyvinylidene fluoride (PVDF) or polyethersulfone (PES) membranes by heating. Antigen components were divided into two categories based on their function, i.e., their membrane recognizing (aglycone part) and fixing (sugar moiety) abilities. This procedure allows for the staining of only target glycosides. Double eastern blotting was developed as a further staining system for two herb medicines using a set of MAbs and substrates.

Identification of phenolic compounds in the buds of Algerian Pistacia atlantica desf. Subsp. atlantica by antioxidant activity guided fractionation

Objectives A bio-guided selection and identification of the most active compounds obtained from Algerian Pistacia atlantica desf. Subsp. atlantica. Methods An antioxidant activity guided fractionation was performed on buds’ extract using extensive chromatographic and spectroscopic techniques. The antioxidant potentials of isolated compounds and other unpurified fractions were evaluated against DPPH radicals using TLC plates and test tubes. Results The results showed that all isolated compounds and fractions exhibited eminent DPPH scavenging potential. Two coumarins (7-ethoxycoumarin and 7-hydroxy-5-methoxycoumarin) and two flavonoids (3′,5,7-Trihydroxy-4′-methoxyflavanone and 5,6,7,4′-tetrahydroxyflavonol-3-O-rutinoside) were isolated for the first times from the titled subspecies. Conclusions These results confirm that the species of P. atlantica is far from being exhausted of active compounds, especially polyphenols.

STABILITY-INDICATING HPTLC METHOD FOR DETERMINATION OF DACLATASVIR IN PHARMACEUTICAL DOSAGE FORM

A simple, specific, precise, and accurate stability-indicating assay method using high performance thinlayer chromatography (HPTLC) is described for estimation of daclatasvir dihydrochloride (DCV) in bulk drug and in the tablet dosage form. The separations were achieved on prepared TLC plates precoated with silica gel 60 F254. The mobile phase developed and optimized for bringing out the separation involves toluene: methanol in the ratio of (6:4 V/V). The densitometric scanning wavelength selected was 312.0 nm. The compact bands of DCV were obtained at RF value of 0.702±0.032. The method developed was able to separate peaks of all the degradation products formed in ICH prescribed stress conditions with sufficient difference in their RF values. The developed method was validated for linearity and range, specificity, precision, accuracy, and robustness, and the results were found to be within acceptance criteria. The reliability of the method was evaluated when it was applied for the estimation of DCV in pharmaceutical tablet formulation and assay results gave good recovery.

Stability-Indicating Determination of Tedizolid Phosphate in the Presence of its Active Form and Possible Degradants

Tedizolid phosphate is an antibiotic prodrug that is metabolized into tedizolid which is used against various resistant bacterial strains. In this study, tedizolid phosphate was subjected to stress degradation conditions, namely, hydrolysis (neutral, acidic and alkaline), thermal, oxidative and photolytic ones. The prodrug was stable toward thermal and photolytic stress conditions, while it showed significant degradation upon applying oxidative and hydrolytic conditions. Two suggested chromatographic methods are described for separation and determination of tedizolid phosphate from the resulted degradation products. The first method is HPLC using Waters Xselect HSS C18 (250 × 4.6 mm, 5 μm) analytical column and mobile phase composed of phosphate buffer (50 mM, pH 6.5):acetonitrile (70:30, %v/v) pumped at flow rate of 1.0 mL/min with UV-detection at 300 nm. The second method is a TLC coupled with densitometric quantitation, precoated silica TLC-plates as a stationary phase and a mobile phase of methanol:butanol:ethyl acetate:ammonia (33%, w/v) (60:20:20:10,%v/v) were used. The chromatographed plates were scanned at 300 nm. The linearity was confirmed over concentration range of 1–100 μg/mL and 1–12 μg/band for HPLC and TLC-densitometric methods, respectively. Both methods were found to be suitable for determination of tedizolid phosphate in pure form and in its pharmaceutical formulations.

Cerenkov Luminescence Imaging in the Development and Production of Radiopharmaceuticals

Over the past several years there has been an explosion of interest in exploiting Cerenkov radiation to enable in vivo and intraoperative optical imaging of subjects injected with trace amounts of radiopharmaceuticals. At the same time, Cerenkov luminescence imaging (CLI) also has been serving as a critical tool in radiochemistry, especially for the development of novel microfluidic devices for producing radiopharmaceuticals. By enabling microfluidic processes to be monitored non-destructively in situ, CLI has made it possible to literally watch the activity distribution as the synthesis occurs, and to quantitatively measure activity propagation and losses at each step of synthesis, paving the way for significant strides forward in performance and robustness of those devices. In some cases, CLI has enabled detection and resolution of unexpected problems not observable via standard optical methods. CLI is also being used in analytical radiochemistry to increase the reliability of radio-thin layer chromatography (radio-TLC) assays. Rapid and high-resolution Cerenkov imaging of radio-TLC plates enables detection of issues in the spotting or separation process, improves chromatographic resolution (and/or allows reduced separation distance and time), and enables increased throughput by allowing multiple samples to be spotted side-by-side on a single TLC plate for parallel separation and readout. In combination with new multi-reaction microfluidic chips, this is creating a new possibility for high-throughput optimization in radiochemistry. In this mini review, we provide an overview of the role that CLI has played to date in the radiochemistry side of radiopharmaceuticals.

In Silico Plasma Protein Binding Studies of Selected Group of Drugs Using TLC and HPLC Retention Data

Plasma protein binding is an important determinant of the pharmacokinetic properties of chemical compounds in living organisms. The aim of the present study was to determine the index of protein binding affinity based on chromatographic experiments. The question is which chromatographic environment will best mimic the drug–protein binding conditions. Retention data from normal phase thin-layer liquid chromatography (NP TLC), reversed phase (RP) TLC and HPLC chromatography experiments with 129 active pharmaceutical ingredients (APIs) were collected. The stationary phase of the TLC plates was modified with protein and the HPLC column was filled with immobilized human serum albumin. In both chromatographic methods, the mobile phase was based on a buffer with a pH of 7.4 to mimic physiological conditions. Chemometric analyses were performed to compare multiple linear regression models (MLRs) with retention data, using protein binding values as the dependent variable. In the course of the analysis, APIs were divided into acidic, basic and neutral groups, and separate models were created for each group. The MLR models had a coefficient of determination between 0.73 and 0.91, with the highest values from NP TLC data.

Stability-indicating UHPLC and TLC-densitometric Methods for the Determination of Tiamulin Fumarate

Two novel stability-indicating methods were developed for the determination of tiamulin fumarate. in the presence of its degradation products. The first was UHPLC-UV method. Efficient separation was achieved by isocratic elution with a mobile phase of 0.1% aqueous ortho-phosphoric acid (pH 3.5 ± 0.5) and methanol (20:80, v/v) with UV detection at 210 nm. Linearity was obtained in the range of 0.5-10.0 µg mL-1 with mean accuracy of 100.40 ± 0.71. The second method was a TLC-densitometric evaluation of a thin-layer chromatogram of the intact drug using a mobile phase of methanol: pentanol: ethyl acetate: 16.5% ammonia (5:4:2:4, by volume). The TLC-plates were scanned densitometrically at 220 nm where Rf values were 0.58, 0.48 and 0.74 for tiamulin F, its acidic and oxidative degradants, respectively. Moreover, the plates were sprayed with 16% sulfuric acid, heated at 105°C for 10 min. where a yellow-coloured band appeared corresponding to the intact drug was scanned densitometrically at 450 nm. While the bands of the two degradants were no longer observed anymore. Tiamulin F was determined in the range of 1.0-10.0 μg/band with mean accuracy of 100.27% ± 1.47 at 220 nm and 99.93% ± 1.38 at 450 nm. The proposed methods were successfully applied for the determination of drug in marketed oral solution. The obtained results were statistically analyzed and found to be in accordance with those obtained by a reported method.