scholarly journals Standardization of the Ayurvedic Formulation Haridra Khanda Using High-Performance Thin-Layer Chromatography-Densitometry

2008 ◽  
Vol 91 (5) ◽  
pp. 1162-1168 ◽  
Author(s):  
Kedar Kumar Rout ◽  
Sagarika Parida ◽  
Sagar Kumar Mishra

Abstract The present study aimed to standardize the Ayurvedic preparation Haridra Khanda containing Curcuma longa as a major ingredient. Various physicochemical parameters such as alcohol-soluble extractive, water-soluble extractive, total ash, and acid-insoluble ash were determined according to the Ayurvedic Pharmacopoeia of India. Microscopic evaluation of the formulation revealed the presence of various diagnostic cell structures of C. longa. Trace metal analysis indicated the absence of toxic metals such as As, Cd, Hg, and Pb. High-performance thin-layer chromatographic (HPTLC) fingerprint patterns at multiple wavelengths (254, 366, and 430 nm) identified the number of components present at each wavelength. The bioactive markers curcumin (C1), demethoxycurcumin (C2), and bisdemethoxycurcumin (C3) were quantified by using a simple, rapid, and efficient HPTLC method using plates precoated with silica gel 60F254 stationary phase. The instrumental precision [coefficient of variation (CV)] was 0.51, 0.64, and 0.79% and the repeatability of the method (CV) was 0.89, 1.11, and 0.95%, respectively, for C1 to C3. Limits of detection and quantitation for compounds C1 to C3 were 20, 20, and 15 ng and 50, 40, and 50 ng, respectively. Response was a linear function in the ranges of 50350, 40240, and 50300 ng with correlation coefficient (r) 0.9998, 0.9995, and 0.9992, respectively, for C1 to C3. The mean recovery values of 99.63 (C1), 98.65 (C2), and 98.97% (C3) indicated the excellent accuracy of the method. It is shown that HPTLC can be applied successfully for the marker evaluation of the formulation containing C. longa.

Author(s):  
Nondita Prasad ◽  
Balbir Singh ◽  
Diksha Puri

  Objective: Justicia gendarussa Burm. (family Acanthaceae) commonly known as nilinirgundi, is found in Southern India possesses multifarious biological activities due to large range of phytoconstituents. The present study is designed to evaluate the various pharmacognostic parameters of the leaves of J. gendarussa, found in Dehradun district of Uttarakhand for its authentication.Methods: Fresh leaves were taken for the morphological and microscopical (histology and powder) evaluation. Physicochemical parameters (ash values, extractives values, florescence analysis, microbial contamination, and loss on drying) were also performed. Phytochemical screening and thin-layer chromatographic fingerprinting of extracts were also performed to check the presence of various phytoconstituents.Results: The microscopy of the leaves evinced the presence of anisocytic stomata, cuboidal calcium oxalate crystals, cystoliths, multicellular covering trichomes, starch grains and oil globules. The quantitative estimation of total ash, acid insoluble, and water soluble ash values were 13.8%, 1.2%, and 4.5% w/w, respectively. The alcohol soluble and water soluble extractives were estimated as 11.45% and 15.67% w/w, respectively. Foreign organic matter and loss on drying values obtained were 0.23% and 11.2% w/w. Phytochemical screening of petroleum ether, chloroform, methanol and aqueous extracts ascertained the presence of alkaloids, phenolic compounds, saponins, tannins, carbohydrates, flavonoids, glycosides, steroids and triterpenoids. The thin-layer chromatography (TLC) profiling of different extracts revealed the presence of potential compounds which can be further isolated with the help of high-performance liquid chromatography or high-performance TLC.Conclusion: The results of this study provide suitable standards for the authentication of this plant. In the present study, there are certain variations observed from the evaluations done on the same species by other research groups. The probable reason suggested for such disparity is due to the environmental and geographical differences in the locations of the plant collected.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Beata Polak ◽  
Emilia Pajurek

AbstractThe separation of some water- and fat-soluble vitamins via micellar systems of reversed-phase high-performance thin-layer chromatography (HPTLC) and pressurized planar electrochromatography (PPEC) was subjected to research. Hence, the influence of the mobile phase composition (surfactant and acetonitrile concentration, eluent buffer pH) on the migration distances and zone separation of some vitamins (thiamine, riboflavin, niacin, pyridoxine, cyanocobalamin, folic acid, ergocalciferol and α-tocopherol) was investigated. Our results indicated that the applied technique has an impact on the solute order. Comparing the system capacity of HPLC and PPEC (measured as height of the theoretical plate) for the mobile phase systems with and without surfactant shows differences, especially for fat-soluble vitamin. The variances and reproducibilities (% RDS) values of the vitamin are less in PPEC than in TLC. Moreover, the migration distances of water-soluble vitamins are longer than fat-soluble ones. Overall, eluent consisting of 50% acetonitrile, 18.75 mM SDS, the buffer of pH 6.99 via the PPEC technique was most appropriate for determining the investigated vitamins in the artificial mixture and the two commercially available vitamin combinations.


2008 ◽  
Vol 91 (5) ◽  
pp. 1179-1185 ◽  
Author(s):  
Harsha Pulpati ◽  
Yogesh S Biradar ◽  
Mandapati Rajani

Abstract Peganum harmala Linn. seed is a reputed drug of the Indian systems of medicine. We report a simple high-performance thin-layer chromatography (HPTLC) densitometric method for the quantification of 4 alkaloids, viz., harmine, harmaline, vasicine, and vasicinone from P. harmala seed. The 4 compounds were resolved in ethyl acetatemethanolammonia (7 1 0.3, v/v/v) mobile phase. The method was validated for precision, repeatability, and accuracy. Instrumental precision was 0.27, 1.53, 0.39, and 1.15% [relative standard deviation (RSD)] and repeatability of the method was 1.01, 0.79, 0.13, and 1.62% RSD for harmine, harmaline, vasicine, and vasicinone, respectively. Accuracy of the method was checked by a recovery study conducted at 3 different levels, and the average recovery was 97.88% for harmine, 97.69% for harmaline, 98.38% for vasicine, and 98.28% for vasicinone. The 4 compounds were quantified in P. harmala seed using the method, and it was found to contain 0.44, 0.096, 0.25, and 0.0007% (w/w) of each, respectively. The proposed HPTLC densitometric method for the quantification of the 4 compounds was found to be simple, precise, specific, sensitive, and accurate. It can be used for routine quality control of P. harmala seed and has the ability to quantify the 4 marker compounds in the formulations containing P. harmala. It also can be used to quantify any of these marker compounds in other herbal drugs.


2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Poonam Kushwaha ◽  
Babita Shukla ◽  
Jyotsana Dwivedi ◽  
Sumedha Saxena

Abstract Background In the present study, an HPTLC (high-performance thin-layer chromatography) method was developed for the quantitative determination and validation of the curcumin in the methanolic fraction of Curcuma longa L. For achieving good separation of curcumin, the mobile phase of chloroform:methanol (97:3) was used. The densitometric analysis of curcumin was performed at 420 nm in reflection/absorption mode. Results Linearity of the method was obtained in the range of 100‒600 ng per spot. During analysis, the methanolic fraction of the C. longa showed the presence of a quantifiable amount of curcumin. The content of curcumin was found to be 1.5% (per dry weight). Conclusions The method is specific, simple, precise, and accurate. The obtained data can have used for the routine analysis of the reported biomarkers in crude drugs and extracts. The quantification and the method validation of curcumin have not yet been reported in C. longa which can be utilized for the proper standardization of the plant.


Author(s):  
Hetal Janani ◽  
Jayanta Kumar Maji ◽  
Ashish Verma

Objective: The combination between the stem bark powder of Chirabilva and Veerataru is used ethno folklore for the management of anti-convulsion by tribal people. To standardise the evidently used polyherbal formulation in a systemic way for supporting the identity, quality, purity, safety and efficacy concern with modern technique.Methods: Microscopic anatomical examination and powder microscopy were performed fresh and dried plant materials respectively. The compound polyherbal formulation is processed by organoleptic characterization, macro-microscopic evaluation, physicochemical, phytochemical testing, DNA fingerprinting and high-performance thin-layer chromatography (HPTLC) profiling employing a standard methodology. Chromatographic fingerprinting after visualisation the data are exploited by multivariate chemometric technique.Results: Results of the experiment provided diagnostic characteristics to identify quality and purity and standardise the polyherbal formulation along with respective ingredients. The RAPD analysis of Dichrostachys cinerea and Holoptelea integrin folia showed some similar bands at the same base pair indicate the presence of genetical identity may take into consideration of the control the ingredients. HPTLC technique utilised to distinguish the ingredient and polyherbal formulation based on the presence or absence of certain target phytochemical (flavonoid, polyphenol, etc.) constituents, manifested as peaks or bands from the chemical fingerprint profiles. Visualized chromatographic profile of polyherbal formulation along with its constituents applying the multivariate chemometric technique is easily discriminated of respective retardation factor in principal component score space.Conclusion: The findings from this study will provide systemic evaluation for this anticonvulsant formulation and also serve as a master document to control the quality of polyherbal formulation.Keywords: Anticonvulsant, Chemometrics, DNA fingerprinting, HPTLC, Pharmacognosy


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