hptlc method
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2022 ◽  
Vol 8 (4) ◽  
pp. 270-279
Anup K Chakraborty ◽  
Harshita Tripathi ◽  
Sarita Karole ◽  
Kavita R Loksh

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

Abstract A systematic DoE and Analytical Quality by Design (AQbD) approach was utilized for the development and validation of a novel stability indicating high-performance thin–layer chromatographic (HPTLC) method for Rivaroxaban (RBN) estimation in bulk and marketed formulation. A D-optimal design was used to screen the effect of solvents, volume of solvents, time from spotting to development and time for development to scanning. ANOVA results and Pareto chart revealed that toluene, methanol, water and saturation time had an impact on retention time. The critical method and material attributes were further screened by Box-Behnken design (BBD) to achieve optimal chromatographic condition. A stress degradation study was carried out and structure of major alkaline degradant was elaborated. According to the design space, a control strategy was used with toluene: methanol: water (6:2:2) and the saturation time was 15 min. A retention factor (RF) of 0.59 ± 0.05 was achieved for RBN using chromatographic plate precoated with silica gel at detection wavelength 282 nm with optimized conditions. The linear calibration curve was achieved in the concentration range of 200–1,200 ng/band with r 2 > 0.998 suggesting good coordination between analyte concentration and peak areas. The quadratic model was demonstrated as the best fit model and no interaction was noted between CMAs. The optimized HPTLC method was validated critically as stated in International Conference on Harmonization (ICH) Q2 (R1) guideline and implemented successfully for stress degradation study of RBN. The developed HPTLC method obtained through AQbD application was potentially able to resolve all degradants of RBN achieved through forced degradation study. The obtained results demonstrate that a scientific AQbD approach implementation in HPTLC method development and stress degradation study drastically minimizes the number of trials in experiments, ultimately time and cost of analysis could be minimized.

N Sunitha ◽  
S Manohar Babu ◽  
B Appa Rao

Lip balm was prepared and its antioxidant activity was determined by PFRAP, CUPRAC, Flourimetry methods and results of emission spectra was examined and fournd to possess almost same concentration of 0.3 ug/ml. And the ascorbic acid was determined by HPTLC method and the Rvalue of the lip balm was found to be 0.98 and is near to the abscorbic acid sample. And hence proved that beetroot lip balm was found to possess antioxidant activityThe anti oxidant activity was determined and was found to be simple, novel and economical.

N Sunitha ◽  
S Manohar Babu ◽  
B Appa Rao

Sweet corn fibres of about 15 gm were extracted with methanol for 5 hours in heating mantle at 40C and filtered and allowed to dry. The dried gel was further analyzed for estimation of Retinyl palmitate by spectrophotometrically by laboratory method and found to be 140 mg/kg. The dried extract gel was further estimated for ascorbic acid both by UV and HPTLC and found to be linear in the range of 1-5 ug/ml and 5-10ug/ml, correlation coefficient was found to be 0.997 and 0.998 and the amount of ascorbic acid was found to be 338 ng/ml and 9.9 ng/ml by UV and HPTLC respectively. The method was found to be linear.

Kavitha J ◽  
Bonda Vismitha ◽  
Kokilambigai KS ◽  
Seetharaman R ◽  
Lakshmi KS

Oxetacaine is a potent local anesthetics used to relieve pain associated with peptic ulcer. The current method details about a rapid, accurate and precise HPTLC technique for the assessment of Oxetacaine in Pharmaceutical formulation. Chromatographic resolution was carried out on precoated HPTLC plates (Silica gel 60 F254 on Aluminum plate) employing methanol: water: glacial acetic acid (8: 1.8: 0.2 v/v/v) as mobile phase. Densitometric assessment was carried out at 220nm [Camag TLC Scanner III with winCATs software (version – 1.3.4)]. The drug was identified with a Rf value of 0.61. The reliability of the projected method was ascertained by evaluating various validation parameters as per ICH guidelines. The proposed technique can evaluate ten or more formulation units concurrently in a single run and affords a more rapid and cost-effective QC tool for regular analysis of Oxetacaine in pharmaceutical formulations.

Veena Devi Singh ◽  
Vijay Kumar Singh ◽  
Sanjay J Daharwal

New HPTLC method was developed and optimized for estimation of Ondansetron (OND), Dexamethasone (DEX) and Aprepitant (APT) in laboratory prepared ternary mixtures by using Central composite design (CCD). The independent variables used for the optimization were the acetone content in mobile phase (%mL), distance of developing solvent (cm) and saturation time (min). HPTLC Separation was performed on Precoated silica gel F254 aluminum plate (10X10 cm, 100μm thickness) with a mobile phase consisting of chloroform: methanol: acetone: ethyl acetate: ammonia (9:4:2:5:0.2 % v/v/v/v). Quantification of OND, APT and DEX were achieved based on a Densitometric analysis over the concentration range of 200-1200 ng/band, 500-1000 ng/band and 1000-2000 ng/band, respectively, at 254nm. The method was yielded dense and well-resolved bands at Rf values of 0.54± 0.02, 0.79±0.02 and 0.23±0.01 for OND, APT and DEX, respectively. The linear regression analysis for the calibration plots produced r2= 0.9997, r2= 0.9998 and r2=0.9997 for OND, APT and DEX, respectively. The method was validated according to the ICH guidelines. The robustness test was determined that the selected factors have an insignificant effect on the responses. The results indicated that the method is suitable for the routine quality control testing of OND, APT and DEX in their bulk form.

Ankita Gaikwad ◽  
Madhuri Shelar ◽  
Ganesh Andhale ◽  
Jyoti Kadam

Resveratrol is a plant compound that work as antioxidant and it also has anti-aging properties whereas Piperine is a alkaloid and it majorly used in spices.  A HPTLC (High performance thin layer chromatography) study is conducted for estimation of Resveratrol and Piperine. The Mobile phase used was Chloroform: Ethyl Acetate (50:50 v/v). Rf Value of Resveratrol 0.59 and Piperine 0.79 was found. Stationary phase of Silica gel 60 F254 was used. Densitometric analysis was performed at 325 nm. The method was found to be linear. Recovery (ranging from 97% to 102.24 %), limit of detection (40.26 ng/spot, 1.64 ng/spot respectively for resveratrol & piperine), limit of quantification (122.02 ng/spot, 4.98 ng/spot respectively for resveratrol & piperine) and precision (≤ 2.00%) were found to be satisfactory. Validation performed with linearity, accuracy, precision, specificity, robustness, limit of detection and limit of quantification. Every parameter found within the range. The developed method allows to confirm that an accurate and reliable potency measurement of a pharmaceutical preparation can be performed.

Planta Medica ◽  
2021 ◽  
Lisa-Anna-Maria Pihan ◽  
Samuel Peter ◽  
Günter Vollmer ◽  
Beat Meier ◽  
Evelyn Wolfram

AbstractThe genus Sideritis (Lamiaceae) comprises around 150 species, of which many are popular herbal remedies in Mediterranean folk medicine. Already mentioned by Dioscorides and Theophrastus, the “ironwort” or “Greek mountain tea” has been receiving increased attention in recent years. A European Union herbal monograph and assessment report (HMPC) has been issued, covering the species Sideritis scardica, S. clandestina, S. raeseri, and S. syriaca. This study presents results of a first pharmacognostic examination of the botanical and phytochemical differences among and between these emerging commercial species, and other, less studied species. An HPTLC method is proposed for normal phase separation of the species; this means applying two mobile phases on silica plates and subsequent derivatization with natural product reagent (NP/PEG) for visualization of phenolic compounds and anisaldehyde for a broader detection. With the help of selected reference compounds, a system suitability test was established for proper chromatographic separation. The method was applied to specimens from botanical gardens and commercial raw material in order to test its suitability for differentiation and authentication. The HPTLC analysis also includes, for the first time, S. hyssopifolia and other less used Sideritis species. The results might enable the development of a validated phytochemical fingerprint authentication procedure for quality assurance of Sideritis herba.

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