HPTLC

A simple and sensitive high performance thin layer chromatography (HPTLC) method has been developed for the quantitative estimation of Donepezil HCL in its bulk and tablet dose tablet formulation (5 mg). Donepezil HCL was chromatographed on silica gel 60 F254 TLC plate using  methanol : toluene (2:8, v/v) as mobile phase. Donepezil HCL showed Rf value 0.54 + 0.008 and scanned at 245 nm using a camag TLC scanner 3. The method was validated in terms of linearity (100 – 800 ng/spot), precision (system precision = 0.0123 and method precision = 0.0084), accuracy (100.3 ± 0.76) and specificity. The limit of detection and limit of quantification for Donepezil HCL were found to be 1.72 ng/spot and 2.07 ng/spot, respectively. The developed method was successfully used for the assay of Donepezil HCL tablet formulation. This method also contain forced degradation studies for standard and tablet. The method was found to be simple, sensitive, specific, accurate and precise and can be used for the routine quality control testing of Donepezil HCL in tablet dosage form.

Ayurveda, being as ancient system of medicine, which comprises hundreds of Kalpanas of pharmaceutical preparations but only few of these Kalpanas are in practice. In this modern era many good preparations have been extinguished in the passage of time among these, Varti Kalpana is main one. Now-a-days, when human life is very fast, we should be ready to make necessary changes in our science according to the need of hour. In present study Nimbadi Yonivarti is selected for the management of Swetapradara which has been mentioned as Anubhuta Yoga by Vd. Devisharan Garga Ayurveda Upadhyaya in “Nari Roganak. The present study was aimed at setting up a standard profile of Nimbadi Yonivarti which was prepared using pharmacognostically authenticated raw drugs followed by subjecting it to detailed pharmacognostical, physicochemical and phytochemical (including Thin Layer Chromatography) analysis as per standard protocol. The observations were systematically recorded. Pharmacognostical findings (crystals, epicarp cells, trichom, etc.) confirm the ingredients present in the finished product. Identified phytochemical components (Carbohydrates, tannins, fanolic compound, glycosides, steroids, flavonoids.etc.) support the intended action of the formulation in vaginal discharge.

A new simple, accurate, precise and selective stability-indicating high performance thin layer chromatographic (HPTLC) method has been developed and validated for the determination of Dapoxetine hydrochloride in bulk and in formulation. Chromatographic separation was performed on aluminum plate precoated with Silica Gel 60 F254 using Ethyl Acetate: Methanol (9:1 v/v) as the mobile phase with saturation time 20 min, followed by densitometric scanning at 239 nm. This system was found to give compact spot for Dapoxetine hydrochloride (Rf value 0.78 ± 0.005) and specificity in accordance with international conference on harmonization (ICH) prescribed stress conditions. The calibration curve was found to be linear between 100-700 ng/band. The proposed method was found to be accurate, precise, reproducible, specific and sensitive and applicable for the determination of Dapoxetine in bulk and in formulation. The drug was subjected to stress condition of hydrolysis (acid, base and neutral), oxidation, photolysis and thermal degradation.

To elucidate the terpenoid profile of Cucumis melo (L). fruit extract using high performance thin layer chromatography (HPTLC). The ethanolic extract prepared from the fruit of Cucumis melo (L). using Soxhlet apparatus. n-hexane: ethyl acetate (7.2: 2.9) was employed as mobile phase for terpenoids. 3µl of test solution and 2µl of standard solution was loaded as 6mm band length in the 3 x 10 Silica gel 60F254 TLC plate using Hamilton syringe and CAMAG LINOMAT 5 instrument.  HPTLC analysis was done using CAMAG HPTLC system equipped with automatic TLC sampler IV, TLC scanner 3, REPROSTAR 3 with 12 bit CCD camera for photo documentation, winCATS Planer Chromatography software. The Rf value of the different compounds present in the extract was found to 0.06, 0.21 and 0.93 of peak 1, 2 and 3 respectively. Among them, peak 1 was found to be terpenoid compounds. C. melo fruit extract showed the presence of terpenoids and it was confirmed from the chromatogram after derivatization.  HPTLC profile of terpenoids has been chosen here to reveal the diversity existing at biochemical level in C. melo. Such finger printing is useful in differentiating the species from the adulterant and act as a biochemical marker for this medicinally important plant in the pharmaceutical industry and plant systematic studies.

Two simple, sensitive and precise HPTLC and RP-HPLC methods were developed for the estimation of berberine from Coscinium fenestratum, and its formulation. For the determination of berberine by HPTLC method, precoated silicagel 60F254 on aluminium sheets and a mobile phase system comprising of n-butanol: glacial acetic acid : water (8:1:1 % v/v/v ) was selected. After development the plate was scanned and quantified at 350 nm. Linearity was found in the concentration range of 10 to 50 ng/spot (r=0.9992). Limit of detection was found to be 5 ng/spot and limit of quantification was found to be 10 ng/spot. In RP-HPLC method, separation was achieved on a Phenomenex, Luna, C18 column (150 x 4.6mm internal diameter, 5µ particle size) using a mobile phase consisting of potassium dihydrogen phosphate (pH - 2.5) (A)  : acetonitrile (B), where B was run in gradient programme (20% for 0.01-20min, 50% for 20-25min, 50% for 25-26min, 20% for 26-30min), at a flow rate of 1ml/min and the elute was monitored at 220nm. The calibration curve was obtained in the range of 100 - 500 µg/mL. The slope, intercept and correlation coefficient values were found to be 57588, 6041and 0.9959, respectively. The method was validated in compliance with ICH guidelines. Low relative standard deviation and good % recovery values of both the methods showed that the developed methods were highly precise, accurate and can be employed for the routine analysis of formulations containing berberine.

A simple, precise and accurate high performance thin layer chromatography (HPTLC) method have been developed for the quantification of strychnine and brucine.  The strychnine and brucine are isolated from biological samples and herbal formulations using a liquid–liquid extraction procedure.  The clean-up procedure is performed using an acid solution.  The method has a linear range of 45 – 1000 ng/spot for strychnine and  95 – 1000 ng/spot for brucine.  The method precision was found to be 0.86 - 2.61 (% RSD) and 0.77 - 2.46 (% RSD) for strychnine and brucine respectively.  Accuracy of the method was checked by recovery studies conducted at three different concentration levels and the average percentage recovery was found to be 101.34 % for strychnine and 101.23 % for brucine.

A simple validated high performance thin layer chromatographic method was developed for the determination of Aceclofenac in presence of its degradant. Separation of Aceclofenac from the degradant could be achieved using aluminium backed silica gel 60 F254 plate with toluene: ethyl acetate: glacial acetic acid, (6:4:0.02v/v) as mobile phase. Densitometry analysis was carried out at 282 nm. The method showed high sensitivity with good linearity over the concentration range of 0.5 – 4 µg/spot. The method was successfully applied to the analysis of pharmaceutical formulation containing Aceclofenac with excellent recovery. The LOD and LOQ were found to be 0.1 and 0.5 µg/spot. Aceclofenac was subjected to hydrolytic, oxidative, thermal and photolytic degradation. It was found that the drug was highly susceptible to acid hydrolysis. Kinetic investigation of the drug followed pseudo-first order reaction. From the Arrhenius plot the activation energy was found to be 13.19 kcal/mole. Statistical analysis revealed that the developed method is accurate and reliable. Hence it can be used for routine quality control analysis of Aceclofenac in tablet formulation.

A simple, accurate and precise high performance thin layer chromatographic (HPTLC) method has been developed for the estimation of Acenocoumerol in bulk drug and dosage form. The method employed silica gel 60 F254 precoated plates as stationary phase and mixture of toluene: isopropyl alcohol:formic acid (7.5:2:0.5) as mobile phase. Densitometric scanning was performed at 290 nm using Camag TLC scanner 3 with WINCAT software of version 1.4.3 Camag. Beer’s law was obeyed in the concentration range of 30ng/spot-90ng/spot. The Retention factor for Acenocoumerol was found to be 0.68. The limit of detection and limit of quantitation were found to be 5 ng/spot & 15 ng/spot respectively. The % RSD of intra-day variation and inter day variation were found to be 1.10 and 1.12 respectively. As per ICH guidelines the results of the analysis were validated in terms of linearity, precision, accuracy, limit of detection and limit of quantification, and were found to be satisfactory. The proposed method can also be used for routine quality control to accurately determine Acenocoumerol in bulk and dosage form.

A simple, sensitive, accurate and precise high performance thin layer chromatographic method was developed and validated for simultaneous determination of efavirenz (EFA), emtricitabine (EMT) and tenofovir (TEN) in combined tablet formulation and forced degradation studies were performed as per ICH guidelines. Precoated silica gel 60F 254 was used as stationary phase and the mobile phase used was chloroform: methanol (90:10), gives high resolution for each drug. The densitometric evaluation of each drug was carried out at 262 nm. The developed method was simple, accurate and is suitable for analysis of the drugs and degradation products in stability studies of samples.

The objective of this work was to develop and validate simple, rapid and accurate chromatographic methods (A and B) for determination of Desvenlafaxine succinate in solid dosage form. In method A - RP-HPLC method was based on Reversed Phase High Performance Liquid Chromatography, on ODS C18 RP column (150 mm × 4.6 mm i.d., 5 µ), using Methanol : 50 mM Phosphate buffer (pH 8.0): Acetonitrile (50:40:10 % v/v) as the mobile phase, at a flow rate of 1 mL/min at ambient temperature. Quantification was achieved by UV detection at 225 nm over a concentration range of 5-25 µg/mL for Desvenlafaxine succinate. The mean retention time for Desvenlafaxine succinate was found to be 4.80 min. The amount of Desvenlafaxine succinate estimated as percentage label claim was found to be 99.83 ± 1.1093. In method B - HPTLC method was based on TLC separation of the drug using silica gel 60 F 254 aluminium sheets and Chloroform : Methanol : Water (60:30:10 v/v/v) as mobile phase. Detection was carried out at 226 nm over the concentration of 1 - 3.5 µg/mL Desvenlafaxine Succinate. The mean Rf value of Desvenlafaxine succinate was found to be 0.63. The amount of Desvenlafaxine succinate was estimated as percentage label claim found to be 101.57 ± 0.92668. Both of these methods were found to be simple, precise, accurate, selective and could be successfully applied for determination of pure laboratory prepared mixture and tablet. Key words: RP-HPLC, HPTLC, Desvenlafaxine succinate, marketed formulation.