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Current Perspectives and Methods for the Characterization of Natural Medicines
Published in Rohit Dutt, Anil K. Sharma, Raj K. Keservani, Vandana Garg, Promising Drug Molecules of Natural Origin, 2020
Muthusamy Ramesh, Arunachalam Muthuraman, Nallapilai Paramakrishnan, Balasubramanyam I. Vishwanathan
High-performance thin-layer chromatography (HPTLC) is an advanced version of thin-layer chromatography. It is a sophisticated, and automated analytical technique to conduct qualitative and quantitative analysis. It is also known as flatbed chromatography. It enables multiple sample handling and complicated separation. The chromatogram of HPTLC is easily visible and the samples are detected using a UV/visible or fluorescence detector. It employs a pre-coated plate in the analysis of samples. The layer thickness is ~100–200 µM. The sample volume is low and the efficiency is high in comparison to TLC. The basic principle of HPTLC based separation of components is adsorption. HPTLC used in the pharmaceutical industry, food analysis, clinical laboratory, natural product characterization, etc. In some cases, HPTLC found to superior over the HPLC technique. Tien Do et al. compared the HPLC and HPTLC for the efficient isolation of marine secondary-metabolites from the extracts of Mentha spicata L. and Fallopia multiflora. The parameters like chromatographic separation, purity, and quantity of isolated compounds, the solvent consumption, the duration and cost of operations were considered for the comparison. The results have shown HPTLC as a superior method over the HPLC method (Do et al., 2014). The HPTLC-based characterized phytoconstituents and marine compounds are listed in Table 2.2.
Herbal Drug Development: Challenges and Opportunities
Published in Megh R. Goyal, Durgesh Nandini Chauhan, Plant- and Marine-Based Phytochemicals for Human Health, 2018
Bhagyashree Kamble, Neelam Athawale, Anand Gugale, Ashika Advankar, Ashwini Ghagare, Shankar Katekhaye, Abhishek Kulkarni, Priyanka Kanukuntla
HPTLC enables us to handle the most complicated separation. HPTLC method has been reported for quantitative estimation of many markers in various plants such as swertiamarin in different marketed polyherbal formulations.13 Simultaneous estimation of diosgenin and levodopa has been done by HPTLC. The proposed HPTLC method has provided faster and effective quantitative control for analysis of diosgenin and levodopa.69 Advantages of HPTLC include: extremely flexible design, multiple detections can takes place at a time, cost effective and time saving, process monitoring is very easy, results can be easily compared.
Fingerprinting Techniques for Herbal Drugs Standardization
Published in Ravindra Kumar Pandey, Shiv Shankar Shukla, Amber Vyas, Vishal Jain, Parag Jain, Shailendra Saraf, Fingerprinting Analysis and Quality Control Methods of Herbal Medicines, 2018
Ravindra Kumar Pandey, Shiv Shankar Shukla, Amber Vyas, Vishal Jain, Parag Jain, Shailendra Saraf
High Performance Thin Layer Chromatography (HPTLC) is the common fingerprinting method mainly used to analyze compounds with low or moderate polarities. The HPTLC technique is widely used in the pharmaceutical industry for quality control of herbs and health products, identification and detection of adulterants, substituents in the herbal products, and also helps in the identification of pesticide contents and Mycotoxins. The HPTLC method has several advantages, which are as follows: Several samples can be run simultaneously by the use of a smaller quantity of the mobile phase as compared to HPLC.Mobile phases of pH 8 and above can be used for HPTLC.Repeated detection (scanning) of the chromatogram with the same or different conditions is possible.HPTLC has been investigated for simultaneous assay of several components in a multi-component formulation. With this technique, authentication of various species of plant as well as the evaluation of stability and consistency of their preparations is possible.
Analytical method development, validation and forced degradation studies for rutin, quercetin, curcumin, and piperine by RP-UFLC method
Published in Drug Development and Industrial Pharmacy, 2021
Shanmugam Ramaswamy, Kuppusamy Gowthamarajan, Lalitha Priyanka Dwarampudi, Mahendran Bhaskaran, Madhuri Kadiyala
Standardization of herbal medicines is one of the main problems in the development of herbal medicines [17]. In addition to the standardization of herbal medicines by HPLC, the UFLC was the most important due to its high pressure limit, selectivity, and sensitivity [14]. The major disadvantage associated with herbal method development is that multiconstituents and unavailability of reference standard. Most of the herbal substances are been standardized with the high-performance thin-layer chromatography (HPTLC). The main limitation associated with HPTLC is that the reference standard and sample requirement is high when compared to that of RP-UFLC and multi sample analysis is not possible. RP-UFLC methods provide the cost effective, stable validation results when compared to that of HPTLC. The greater advantage is that the methods developed by RP-UFLC can be successful used in pharmacokinetic in human and animal studies with accurate results. The UFLC can withstand pump pressures up to 6800 psi [44] and can be used for the qualitative and quantitative estimation of herbal ingredients.
Quality Control Standardization and Evaluation of Antimicrobial Potential of Daruhaldi (Berberis aristata DC) Stem Bark
Published in Journal of Dietary Supplements, 2020
Mohammad Irfan Khan, Md. Azizur Rahman, Mohammad Khalid, Mohammad Khushtar, Md. Mujahid
The solvent system for the HPTLC analysis was chosen on the basis of improved separation between the spots. The selected solvent system showed better separation between the spots. All the sample constituents in the HPTLC chromatograms were clearly separated without any tailing and diffuseness. The chromatograms revealed numerous spots (Figure 2). Methanolic extract of Berberis aristata stem bark in the solvent system of toluene:ethyl acetate:formic acid (15:10:2) revealed 5 and 6 spots at UV 254 and 366 nm, respectively, with max Rf values in the range of 0.32 to 0.91 indicating the occurrence of 6 different constituents in it. Chloroform extract of Berberis aristata stem bark in toluene:ethyl acetate:glacial acetic acid (10:5:1) revealed 12 and 8 spots at UV 254 and 366 nm, respectively, with max Rf values in the range of 0.16 to 0.84 indicating the occurrence of 12 different constituents. In the HPTLC study, the B. aristata extract showed spots corresponding to selected standard berberine. Hence, quantitative estimation of this marker compound was performed in the selected solvent system. The calibration plot was found to be linear, and the Rf value of berberine was found to be 0.31. The berberine content in B. aristata stem bark was calculated from the peak area of HPTLC chromatogram of berberine in methanolic extract of B. aristata stem bark and standard plot (Figure 3). It was found to be 0.032 mg/g (w/w).
Consumption of Decorticated Pulses Ensures the Optimum Intake of Isoflavones by the Urban Indian Population
Published in Nutrition and Cancer, 2019
Anuradha Deorukhkar, Laxmi Ananthanarayan
The high-performance thin-layer chromatography (HPTLC) analysis was performed using winCATS Planar Chromatography Manager software. 4 µL concentrated extract samples were applied with 100 µL Hamilton syringe on 20 × 10 cm F254 TLC silica gel plates on LINOMAT 5 applicator. The plate was developed in CAMAG twin trough chambers with mobile phase toluene: ethyl acetate: acetone: formic acid = (20:4:2:1 v/v) as described by Yuan et al. (20). Developed TLC plates were scanned at a wavelength of 254 nm in CAMAG TLC Scanner 3. The concentration of isoflavones in the injected sample was determined from calibration curves obtained with genistein and daidzein standards.