HPTLC

Sitagliptin chemically is (3R) -3-amino-1-[3- (trifluoromethyl)-6,8-dihydro-5h- [1,2,4] triazolo [3,4-c] pyrazin-7-yl]-4-(2,4,5-trifluorophenyl) butan-1-one (Fig. 1), an oral anti-diabetic agent that blocks Dipeptidyl peptidase-4 (DPP-4) activity. Sitagliptin increased incretin levels (GLP-1 and GIP) which inhibit glucagon release, in turn decreases blood glucose, but more significantly increases insulin secretion. The present work describes development and validation of a new simple, accurate, precise and stability indicating HPTLC method for the determination of sitagliptin in tablet dosage form. The chromatographic separation was achieved by using Toluene: Ethyl acetate: Methanol (3: 6: 1 v/v/v) as mobile phase and UV detection at 238 nm. The developed method was validated with respect to linearity, accuracy, precision, limit of detection, limit of quantitation and robustness as per ICH guidelines. The drug was subjected to stress condition of acid hydrolysis, alkali hydrolysis, photolysis, thermal degradation. Results found to be linear in concentration range of 100-500 ng/band. The developed method can be used for the quantification of bulk drug as well as in formulation. Keywords: Sitagliptin, HPTLC, Degradation Studies

Alogliptin Benzoate is a novel hypoglycemic drug that belongs to dipeptidylpeptidase-4 inhibitor class which stimulates glucose dependent insulin release. The Present work describes development and validation of a new simple, accurate, precise and stability indicaing HPTLC method for the determination of alogliptin benzoate in tablet dosage form. The chromatographic separation was achieved by using Chloroform: Methanol 3:7 v/v as mobile phase and UV detection at 275 nm. The developed method was validated with respect to linearity, accuracy, precision, limit of detection, limit of quantitation and robustness as per ICH guidelines. The drug was subjected to stress condition of acid hydrolysis, alkali hydrolysis, photolysis, thermal degradation. Results found to be linear in concentration range of 500-2500 ng/band. The developed method can be used for the quantification of bulk drug as well as tablet dosage form

Triphala is an important Ayurvedic formulation containing, Haritaki (Terminalia chebula Retz),Vibhitaki (Terminalia bellarica Roxb),Amalaki (Emblica officinalis L.) as a ingredients. All the constituents are available and prepared according to the reference present in Yoga Ratnakara Madhyama Khanda. Triphala (Haritaki, Vibhitaki and Amalaki) classified as an important medicine of the Rasayana and Cakshusya group which is mainly indicated in Prameha, Sthoulya and Kustha and also it is believed to promote health, immunity and longevity. Though they are individually very potent drug and have their own specific mode of action on different disease conditions. Triphala the sense of its Rasa, Vipaka and to some extent its Prabhava is similar to its three contents but its Virya, Doshagnata and Guna are not exactly similar to the composing three drugs. This is due to Samyoga Samskara by which the clinical efficacy of particular drug changes several methods are adopted to prepare Triphala some uses equal proportions (1:1:1) some in different proportions (1:2:4). Till date there is only one scientific work has been carried out and less publication of 1:2:4 combination forms of Triphala. In the present study the Yavakuta formulation of Triphala was subjected to Pharmacognostical (microscopic), HPTLC, and pharmaceutical (evaluation of various physiochemical parameters) evaluation in order to prepare a preliminary profile of the formulation. The sample was subjected for various phytochemical parameters like water soluble extractive (55.8%w/w), alcohol soluble extractive (42.2% w/w), ash value (0.89% w/w), loss on drying (11.39% w/w), and the pH (5.0), HPTLC. The HPTLC, solvent system was Toluene:ethyl acetate (9:1), showed the presence of 5 spots at 254 nm and 2 spots at 366 nm. Thus the physiochemical and microscopic characters achieved may provide guidelines for standardization of formulation, Triphala Yavakuta combine formulation (1:2:4).

In the era of increasing demand for indigenous medicines, maintaining quality standards is the need of the hour. Standardisation of compound formulations is lagging behind because of absence of reference standards. Manjishthadi Ghanavati is an important Ayurvedic formulation containing Manjishtha (Rubia cordifolia Linn.), Haritaki (Terminalia chebula Retz.), Amalaki (Emblica officinalis Gaertn.), Bibhitaki (Terminalia bellerica Roxb.), Katuki (Picrohriza kurroa Royle. ex Benth), Vacha (Acorus calamus Linn.), Daruharidra (Barberis aristata DC) Guduchi (Tinospora cordifolia Willd.) and Nimba (Azadirechta indica A Juss.). All the constituents are available and prepared according to the reference present in Sharangadhara Samhita Madhyama Khanda Chapter 2/136. Hence the present study was undertaken to standardize the compound Ayurvedic formulation through Pharmacognostical and pharmaceutical evaluation. The sample was subjected for various phytochemical parameters like water soluble extractive (49.72%w/w), alcohol soluble extractive (41.28 % w/w), ash value (10 % w/w), loss on drying (10.85 % w/w), the pH (6.0), Hardness(4 kg/cm2) and Disintegration time (32 min.). The HPTLC, solvent system was Toluene: ethyl acetate (9:1), showed the presence of 7 spots at 254nm and 2 spots at 366nm. Thus, the physiochemical and microscopic characters achieved may provide guidelines for standardization of formulation Manjishthadi Ghanavati.

Gutika Anjana is an important Ayurvedic formulation containing Gairika (Ochre), Saindhava (Sodii chloridum), Pippali (Piper longum Linn.) and Shunthi (Zingiber officinale  Roxb.) as main ingredient. All the constituents are available and prepared according to the reference present in Sushruta Samhita Uttaratantra Vataabhishyanda Pratishedha. Till date no work has been carried out to standardize the formulation. Hence the present study was undertaken to standardize the compound Ayurvedic formulation through Pharmacognostical and pharmaceutical evaluation. The sample was subjected for various Phytochemical parameters like water soluble extractive (32.8%w/w), alcohol soluble extractive (32.3% w/w), ash value (8.4% w/w), loss on drying (14.25% w/w), the pH (6), HPTLC. The HPTLC, solvent system was Toluene:ethyl acetate (9:1), showed the presence of 11 spots at 254nm and 13 spots at 366nm. Thus the physiochemical and microscopic characters achieved may provide guidelines for standardization of formulation, Gutika Anjana.  

Cefixime, cefpodoxime and cefepime are cephalosphrin antibiotics used widely in the different infectious diseases. Newer HPTLC methods were developed for their estimation from individual dosage forms due to versatile applications and advantages. The HPTLC chromatogram of cefixime was developed by using a mobile phase ethylene acetate: methanol: water (4.5:5:0.5%v/v) and scanning wavelength was 292nm. The Rf value was 0.58±0.02. The mobile phase optimized for cepodoxime was methanol: ethyl acetate: toluene (1.5:3:5.5% v/v), with the Rf value 0.53±0.02. The cefixime was retained using methanol :water : chloroform (6:3:1%v/v), on a  silica gel G60 F254 aluminium sheet and  scanning wavelength kept at 285nm. The Rf value was 0.44. The methods were optimized validated as per guidelines and successfully applied for individual dosage form containing  of each of cephalosporin.

The present study was design to standardize and develop scientific data for identification and quality control of Oleo Resin of Pine. The proximate analysis and nature of Oleo Resin of Pine (ORP) was confirmed by organoleptic, physiochemical characterization and fluorescence analysis. The preliminary phytochemical analysis was done by qualitative chemical tests. The total chemical components were confirmed by TLC and HPTLC analysis in suitable solvent system. Identification of chemical compounds, their molecular weight and structure were detected by GC-MS and the functional groups were detected by FTIR. Thermal analysis was carried out by DSC. The organoleptic and physiochemical data showed characteristic features of ORP. The preliminary phytochemical analysis of hydroalcholic extract of ORP showed presence of carbohydrates, glycosides, alkaloid, phytosterols, terpenes, saponins and phenols. The DSC thermogram of powder of ORP showed enthalpy of transition more than zero for all the 4 endothermic peaks. The GC-MS chromatogram of ORP revealed the presence of two compounds, Longicyclene at RT 21.11 and Longifolene at RT 22.03. TLC and HPTLC fingerprinting of ORP showed separation of components in Toluene: Ethyl acetate (93:7) mobile phase at 254, 366, 550nm.The FTIR spectrogram of ORP showed the seven characteristic bands assigned to different functional groups. The observations and results of the study have provided evidence based scientific data for standardization of ORP which will serve as reference standards to establish its identity, purity and also help to minimize adulteration and substitution. Keywords: Standardization; Oleo Resin of Pine; HPTLC; GCMS; FTIR; DSC

A simple, rapid, accurate and precise high performance thin-layer chromatography (HPTLC) method was developed and validated for simultaneous estimation of alogliptin and metformin active pharmaceutical ingredients and fixed dose combination. Alogliptin and metformin densitograms were developed on silica gel 60 F254 HPTLC plates with chloroform: methanol: 0.5 % ammonium sulphate (4:4:2 %v/v) as mobile phase. Densitometric quantification was performed at 254 nm. For Alogliptin and Metformin Rf values were found as 0.66 and 0.44, respectively. The linearity curves of Alogliptin and Metformin were obtained in the concentration range of 100-500 ng/spot and 4000-20000 ng/spot by area with correlation co-efficient of 0.998 and 0.995 for Alogliptin and Metformin, respectively. Limit of detection was found to be 2 ng and 40 ng/spot for Alogliptin and Metformin, respectively; lowest possible quantity to be quantified by the proposed method was found to be 6 ng and 130 ng per spot for Alogliptin and Metformin, respectively. The method was validated for precision, accuracy, specificity and robustness. The developed method was validated and found to be selective, specific and suitable for application in pharmaceutical analysis of these drugs in bulk and fixed dose combination.

To develop a new, economical, precise and accurate stability indicating HPTLC method was developed and validated for the determination of Adapalene in bulk drug. The present study deals with development and validation of stability indicating HPTLC method for estimation of Adapalene. Chromatographic separation was performed on aluminium plate pre-coated with Silica Gel 60 F254 using Tetrahydrofuran: 2-Propanol: Water (3:3:3 v/v/v) as a mobile phase. The wavelength selected for densitometric scanning was 230 nm. Regression plots revealed linear relationship in the concentration range of 20-120 ng spot-1. The Rf value of Adapalene was found to be 0.76 (±0.02).The LOD and LOQ were found to be 3.15 and 9.57 ng spot-1respectively. The method was validated as per International Conference on Harmonization (ICH) guidelines, demonstrating to be accurate and precise within the corresponding linearity range of titled analytes. Inherent stability of the drug was studied by exposing drug to acid, alkali, oxidative, photolytic and thermal conditions. Relevant degradation was found to take place under these conditions. The proposed method has been validated as per ICH Q2 (R1) guidelines. This method can be used for routine quality control analysis of Adapalene in bulk drug.

Arbutin and quercetin are anti-cancer agents, extracted from leaves and flowers of Origanum majorana L which can be simultaneously estimated using a rapid, specific and sensitive high – performance thin layer chromatographic technique using silica gel G60F254 as the stationary phase and butyl acetate: methanol: formic acid: toluene (8: 1.5: 5: 0.5) as mobile phase for the separation. The method was validated for linearity, precision and recovery. Linearity was established in the range 300-900 ng/spot for arbutin and 150-450 ng/spot for quercetin, respectively. The % RSD values of repeatability of application, repeatability of measurement, intra-day and inter day precision studies of arbutin and quercetin were found to be below 1 for 500 and 250 ng/spot of arbutin and quercetin, respectively. The recovery of the drugs were found to be 99.8 and 100.2%, proves that the developed method was highly accurate. Hence the proposed validated method could be applied for the simultaneous analysis of arbutin and quercetin in methanolic extract of Origanum majorana L as well as for its marketed formulation.