High-performance Thin-layer Chromatography Plates Research Articles

Isolation and high-performance thin-layer chromatographic analysis of bioactive lupenone: A lupane triterpenoid from Diospyros melanoxylon

Lupenone was isolated for the first time from the stem bark of Diospyros melanoxylon and characterized with the help of physical and spectroscopic data. A high-performance thin-layer chromatography (HPTLC) method has been developed and validated for the quantification of lupenone in D. melanoxylon stem bark. HPTLC analysis was performed on HPTLC plates by using a binary mobile phase of n-hexane—ethyl acetate (8.2:1.8, v/v). It was quantified at 395 nm after derivatization with methanol—sulfuric acid reagent. The limits of detection and quantification were found to be 40 ng and 100 ng per spot, respectively. The linear regression analysis data for the calibration plot in the range of 100–500 ng spot−1 showed a good linear relationship between peak area and concentration (r 2 = 0.9997). The instrumental precision was 1.01% (coefficient of variation [CV]) and the repeatability of the method was 2.17% (CV). The method was validated for precision, recovery, and repeatability as per the International Conference on Harmonization guidelines. The developed HPTLC method is very accurate and precise and has been successfully used for the estimation of lupenone in D. melanoxylon stem bark.

Development and validation of a high-performance thin-layer chromatography method for the determination of artesunate and amodiaquine in tablet formulations

A simple, cost-effective, precise, accurate, and rapid planar chromatographic method was developed and validated for the separation and determination of amodiaquine and artesunate in tablet formulations. Both compounds were determined using high-performance thin-layer chromatography plates and a mobile phase composed of toluene, acetonitrile, methanol, ammonium acetate, and triethylamine in the ratio 10:5:3:1:0.5 (% v/v). Amodiaquine was evaluated densitometrically at a detection wavelength of λ = 345 nm, whereas artesunate was determined fluorimetrically at λ = 503 nm. The method was linear in the concentration ranges of 0.093–0.280 μg spot−1 for artesunate and 0.250–1.250 μg spot−1 for amodiaquine, respectively.

Evaluation of the effect of extraction solvent and organ selection on the chemical profile of Astragalus spinosus using HPTLC- multivariate image analysis.

The evaluation of extraction protocols for untargeted and targeted metabolomics was implemented for root and aerial organs of Astragalus spinosus in this work. The efficiency and complementarity of commonly used extraction solvents, namely petroleum ether, methylene chloride, ethyl acetate and n-butanol were considered for method evaluation using chemometric techniques in conjunction with new, simple, and fast high performance thin layer chromatography (HPTLC) method for fingerprint analysis by extracting information from a digitalized HPTLC plate using ImageJ software. A targeted approach was furtherly implemented by developing and validating an HPTLC method allowing the quantification of three saponin glycosides. The results of untargeted and targeted principle component analysis (PCA) and hierarchical cluster analysis (HCA) revealed that the apparent saponins profile seems to depend on a combined effect of matrix composition and the properties of the selected solvent for extraction, where both the biological matrix of the investigated plant organs, as well as the extraction solvent can influence the precision of metabolite abundances. Although, the aerial part is frequently discarded as waste, it is shown hereby that it has similar chemical profile compared to the medicinal part, roots, yet a different extraction solvents pattern is recognized between the two organs which can be attributed to the differences in the composition, permeability or accessibility of the sample matrix/organ tissues, rather than the chemical structures of the detected metabolites.

IDENTIFICATION, QUANTIFICATION AND VALIDATION OF STIGMASTEROL FROM ALPINIA CALCARATA USING HIGH-PERFORMANCE THIN LAYER CHROMATOGRAPHY METHOD

Objective: The present study is designed to develop a new simple, precise, rapid and selective high‐performance thin‐layer chromatographic (HPTLC) method for the determination of stigmasterol in methanolic rhizomes extract of Alpinia calcarata.Methods: As per International Conference on Harmonization (ICH) guidelines we have applied different concentrations of stigmasterol as standard on HPTLC plates for the quantification of stigmasterol from the Alpinia calcarata rhizomes. The concentration of standard stigmasterol is 1 mg/ml.Results: The retention factor of stigmasterol was 0.58. Linearity was obtained in the range of 50 ng‐250 ng for stigmasterol. The developed and validated HPTLC method was employed for stigmasterol in methanolic rhizomes extract of Alpinia calcarata for standardization of the content of the marker. The linear regression data for the calibration plots showed a good linear relationship with r=0.99977 for stigmasterol, respectively Satisfactory recoveries of 99.77 % were obtained for stigmasterol.Conclusion: The results obtained in validation assays indicate the accuracy and reliability of the developed HPTLC method for the quantification of stigmasterol in methanolic rhizomes extract of Alpinia calcarata

Concurrent analysis of the biologically active markers β-amyrin and β-sitosterol by applying a validated high-performance thin-layer chromatography method in the aerial parts of Tinospora cordifolia and Calotropis gigantia

In this study, we have developed a validated high-performance thin-layer chromatography (HPTLC) method for the concurrent estimation of the biomarkers β-amyrin and β-sitosterol in dichloromethane and ethanol extracts of the aerial parts of Tinospora cordifolia (TCDC and TCET) and Calotropis gigantia (CGDC and CGET). Chromatographic estimations were carried out on HPTLC (glass-backed silica gel 60 F254) plates with solvents hexane and ethyl acetate in the ratio of 7.5:2.5, v/v (as the mobile phase). Post development, the plate was derivatized with p-anisaldehyde reagent to furnish compact spots of β-amyrin and β-sitosterol and scanned at λmax = 530 nm. Well-resolved, compact as well as intense peaks of β-sitosterol (RF = 0.26 ± 0.001) and β-amyrin (RF = 0.39 ± 0.001) were found. The linear regression equation and the correlation coefficient square (r2) for β-amyrin (Y = 6.118x + 460.76 and 0.9959) and β-sitosterol (Y = 7.109x + 1069.1 and 0.9967) in the concentration range of 100–1400 ng spot−1 indicated g...

Development and validation of a high-performance thin-layer chromatographic—densitometric method for the quantification of guggulsterones E and Z in a polyherbal formulation containing Commiphora Mukul

The present article focuses on the quantitative analysis of a polyherbal formulation containing arjuna, guggul, and pushkarmoola in a combined dosage form which is used for its cardioprotective, anti-anginal, and hypolipidemic activities. Quantification of guggulsterones E and Z was done by developing and validating a high-performance thin-layer chromatography method after extraction by methanol. Densitometric evaluation was carried out on precoated silica gel G60 F254 high-performance thin-layer chromatography (HPTLC) plates using toluene—ethyl acetate (8:2, v/v) as the mobile phase. Based on the HPTLC results, the methanol extract of guggulsterone was chosen as it showed a better separation of spots as compared to another extract. After postderivatization of plates with anisaldehyde—sulfuric acid (10%) reagent, the methanol extracts gave different color spots (the arjuna extract gave a blue spot, the guggul extract gave orange and brown spot, and pushkarmoola gave dark blue spot). Scanning and densitome...

In vitro antioxidant assessment and a rapid HPTLC bioautographic method for the detection of anticholinesterase inhibitory activity of Geophila repens

ObjectiveGeophila repens (L.) I. M. Johnst. (Rubiaceae), a small, creeping, perennial herb, is claimed to have memory-enhancing property. The goal of this study was to assess its antioxidant and anticholinesterase activity and conduct a rapid bioautographic enzyme assay for screening acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibition of G. repens extracts. MethodsAntioxidant activity of G. repens extracts was assessed by performing 1,1-diphenyl-2-picrylhydrazyl (DPPH), nitric oxide (NO), superoxide (SOD), hydroxyl (OH) and total antioxidant capacity (TAC) assays. Anticholinesterase activity was investigated by quantifying the AChE and BChE inhibitory activities of chloroform (CGR), ethyl acetate (EGR) and methanol (MGR) extract fractions from G. repens leaves. A rapid high-performance thin-layer chromatography (HPTLC) bioautographic method for the detection of AChE and BChE inhibition was performed. ResultsAmong all extract fractions, EGR exhibited the highest half maximal inhibitory concentration (IC50) in DPPH, SOD, NO, OH and TAC assays, with IC50 of (38.33 ± 3.21), (45.14 ± 1.78), (59.81 ± 1.32), (39.45 ± 0.79) and (43.76 ± 0.81) µg/mL respectively. EGR displayed competitive, reversible inhibition of AChE and BChE activities with IC50 of (68.63 ± 0.45) and (59.45 ± 0.45) µg/mL, respectively. Total phenolic and flavonoids contents of EGR were found to be 360.42 mg gallic acid equivalents and 257.31 mg quercetin equivalents per gram of extract. Phytoconstituents of the EGR extract that were inhibitors of cholinesterase produced white spots on the yellow background of HPTLC plates in the bioautographic test. ConclusionThe results of this study revealed that phenols and flavonoids could be responsible for the antioxidant, anticholinesterase activities of G. repens.

A new and highly sensitive TLC method to measure hypericin using chemiluminescence

ABSTRACTHypericin is a polyphenolic compound belonging to the group of polyphenols and is the active constituents of Hypericum perforatum (Saint John’s wort). We present a new high-performance thin-layer chromatography (HPTLC) method to measure a large number of hypericin extracts using chemiluminescence. On a 10 × 10 cm HPTLC plate (LiChrospher® Merck, 1.05586), more than 40 tracks can be simultaneously quantified using a piezoelectric application system (pipeJet) which can apply 56 nL of a methanolic hypericin extract contactless with high precision. For separation, a solvent mixture of ethyl acetate, water, formic acid, methyl tert-butyl ether, and cyclohexane (180 + 14 + 14 + 80 + 30, v/v) was used. The Rf-value of hypericin is 0.27. The method presented is specific for hypericin and offers a limit of quantification of 690 pg hypericin per band.

Simultaneous HPTLC Determination of Rhein and Aloe-emodin in Senna Alata Leaves from Thailand and their Commercial Products

A high performance thin layer chromatographic (HPTLC) method was developed and validated for simultaneous determination of rhein and aloe-emodin, major anthraquinone constituents, in S. alata leaves. The separation was performed on a silica gel 60 F₂₅₄ HPTLC plate using ethyl acetate: methanol: water (100:17:10, v/v/v) as the development system. Validation of the analytical method for rhein and aloe-emodin promoted acceptable parameters. Good linearity in the range of 40-480 ng/band was obtained while intra-day and inter-day precisions were shown to be precise with relative standard deviations of less than 5%. The average percentage recoveries of rhein and aloe-emodin were 98.8% and 98.9%, suggesting acceptable accuracy. The content of rhein and aloe-emodin in S. alata leaves collected from 5 different provinces in Thailand analyzed by the validated HPTLC method were in the ranges of 0.098 ± 0.017 - 0.30±0.02%, w/w, and 0.081 ± 0.0006 - 0.34 ? 0.0009 %, w/w, respectively. Five commercial products of S. alata tea available in the market were purchased and analyzed for rhein and aloe-emodin contents. The contents of rhein and aloe-emodin in the tea samples were in the ranges of 0.085 ± 0.004 - 0:23 ± 0.04%, w/w, and 0.096 ±0.006 - 0.30 ± 0.01 %, w/w, respectively. The HPTLC method is rapid, reliable, sensitive and economical for routine analysis of rhein and aloe-emodin contents in-S. alata leaf raw materials and its commercial products.

Simultaneous quantification of picrosides I and II from Picrorhiza kurroa, Picrolax capsules, and Picrolax suspension: High-performance thin-layer chromatographic—densitometric method developed with Kutkin

Kutkin, a mixture of two iridoid glycosides, viz., picrosides I and II, is a major active constituent of Picrorhiza kurroa (family: Scrophulariaceae). P. kurroa is reported to be effective in many ailments, viz., liver disorders, cancer, asthma, and inflammation. Various herbal and Ayurvedic preparations containing P. kurroa are available on the market. In these formulations, quality is not maintained in the absence of quick, efficient, and economic quantification methods for active markers. In the present study, a high-performance thin-layer chromatographic (HPTLC)—densitometric method was developed for the simultaneous quantification of picrosides I and II using kutkin from P. kurroa crude drug, P. kurroa extract, and its marketed formulations, viz., Picrolax capsules and Picrolax suspension. The method was validated as per the International Conference on Harmonization (ICH) guidelines. The samples were prepared and applied in triplicate on HPTLC plates. The plates were developed using the mobile phase ...

DEVELOPMENT OF A DENSITOMETRIC HIGH-PERFORMANCE THIN-LAYER CHROMATOGRAPHIC METHOD FOR THE QUANTITATIVE ANALYSIS OF URSOLIC ACID IN THE LEAVES OF SPECIES OF GENUS TECOMA AND TABEBUIA OF BIGNONIACEAE FAMILY

<p><strong>Objective: </strong>A simple and sensitive high-performance thin–layer chromatographic (HPTLC) method was developed for the quantification of ursolic acid in the leaves of three species of genus <em>Tecoma</em> (<em>Tecoma gaudichaudi</em> DC, <em>Tecoma capensis</em> (Thunb.) Lindl, <em>Tecoma stans</em> (L.) Juss. ex Kunth) and genus <em>Tabebuia</em> (<em>Tabebuia rosea</em> Bertol) belong to family Bignoniaceae.</p><p><strong>Methods: </strong>Chromatography was performed on Silica gel 60 F254 precoated HPTLC Plates with optimised mobile phase pet ether: ethyl acetate: formic acid (7:3:0.5, v/v/v). The plate was derivatized with p-anisaldehyde reagent and scanned at 540 nm. The developed method was found to give a compact spot for ursolic acid at R<sub>f </sub>value 0.43±0.01. The method was validated using International Council for Harmonization (ICH) guidelines, including linearity, precision, accuracy, and robustness.</p><p><strong>Results: </strong>Ursolic acid was found to be present in four species, i.e., <em>Tecoma gaudichaudi</em> DC (1.48%w/w), <em>Tecoma capensis</em> (Thunb.) Lindl. (0.79%w/w), <em>Tecoma stans</em> (L.) Juss. Ex Kunth (1.11%w/w), <em>Tabebuia rosea</em> (Bertol.) (1.13%w/w). A good linearity relationship was found to be (200-1400ng band−1) with a correlation coefficient (r<sup>2</sup>) value of 0.9946 with ursolic acid. Limit of detection and limit of quantification was considered to be 40.66, 123.21 ng per band respectively. The developed method was found to be accurate and precise with 1.32%, 1.19% (%RSD) for interday and intraday precision. The accuracy of the method was performed by recovery studies at three different concentration levels, and the average percentage recovery was found to be 98.05% for ursolic acid.</p><p><strong>Conclusion</strong>:<strong> </strong>The proposed method for the quantitation of ursolic acid was found to be reproducible and simple.</p>

Determination of oleanolic acid, ursolic acid, lupeol, and stigmasterol by high-performance thin-layer chromatographic method in Oldenlandia corymbosa Linn.

Oldenlandia corymbosa Linn. (Rubiaceae) is an important herb traditionally used as a febrifuge and liver tonic. In this study, a high-performance thin-layer chromatography (HPTLC) method has been established for the quantification of four bioactive markers, oleanolic acid (OA), ursolic acid (UA), lupeol (LU), and stigmasterol (ST), in the whole plant of O. corymbosa. Separation was achieved on silica gel 60 F254 HPTLC plates using hexane-ethyl acetate-methanol (8.2:1.8:0.5, v/v) for oleanolic acid and ursolic acid; and toluene-methanol (9.4:0.6, v/v) for lupeol and stigmasterol as the mobile phases. The quantitation of the four markers was carried out using the densitometric scanning at 540 nm after derivatization using sulfuric acid reagent. The linear regression analysis data for the calibration plots showed a good linear relationship (r2 = 0.9831–0.9979) in the concentration range of 1200–4200 ng for oleanolic acid, 400–1400 ng for ursolic acid, 100–500 ng for lupeol, and 500–2500 ng per spot for stigm...

Two selective HPTLC methods for determination of some angiotensin II receptor antagonists in tablets and biological fluids.

Two simple, selective, precise and highly sensitive high-performance thin-layer chromatography (HPTLC) methods have been developed and validated for analysis of five angiotensin II receptor antagonists, namely losartan, irbesartan valsartan, candesartan and olmesartan, which are widely used in clinical practice. HPTLC of the drugs was performed on pre-coated silica gel HPTLC plates 60 F254 by development using a mobile phase composed of chloroform-acetone-glacial acetic acid (7.8:1.5:0.7m v/v/v), which was suitable for all of the studied drugs. The first method depended on utilizing reflectance/fluorescence mode for detection while the second method depended on using 2,3,5,6-tetrachloro-1,4-benzoquinone as spraying reagent for the first time to form orange spots scanned at 460 nm. A good linear relationship was obtained over the concentration ranges of 1.2-60 and 360-3000 ng/band while detection and quantification limits were in the ranges of 0.07-0.43, 45.2-140.49 and 0.21-1.29, 137.05-425.74 ng/band for reflectance/fluorescence and reflectance/absorbance methods respectively. The developed methods were applied successfully for their determination in tablets and spiked human plasma for reflectance/fluorescence method with good accuracy and precision, and so can be applied in the pharmacokinetic and bioavailability studies.

Thin-Layer Chromatography/Desorption Atmospheric Pressure Photoionization Orbitrap Mass Spectrometry of Lipids.

Desorption atmospheric pressure photoionization (DAPPI) allows surface analysis in the open atmosphere and is thus an appropriate method for the direct coupling of thin-layer chromatography (TLC) and mass spectrometry (MS). Here, the capability of DAPPI-MS for ionizing and detecting lipids, namely, cholesterol, triacylglycerols, 1,2-diol diesters, wax esters, cholesteryl esters, and hydrocarbons, from TLC and high-performance thin-layer chromatography (HPTLC) plates in MS and MS2 modes was tested. Limits of detection for lipid standards separated using normal-phase (NP)-TLC and NP-HPTLC were established. TLC/DAPPI-MS was applied for lipids of vernix caseosa, a white creamy proteolipid biofilm that progressively coats the fetus during the last trimester of the pregnancy, and plant oils including caraway, parsley, safflower, and jojoba oils. Various lipids were identified by means of high resolution/accurate mass measurement of Orbitrap and comparison of the retardation factors with standards. Lipid class separation was carried out on the NP-HPTLC plates, whereas individual triacylglycerol and wax ester species were separated on the reversed-phase HPTLC plates. DAPPI-MS was found to be a simple, rapid, and efficient approach for detecting lipids separated by TLC.

Quantitative analysis of phytochemicals fromInula capparoots

Inula cappa (family Compositae) is used in the Ayurvedic medicinal system for the treatment of bronchitis, diabetes, fever, hypertension, and rheumatism. The proposed high-performance thin-layer chromatography (HPTLC) study offers coherent evaluation of isoalantolactone, germacranolide, β-sitosterol, and lupeol from I. cappa root. Methanolic solutions of isoalantolactone, germacranolide, β-sitosterol, and lupeol were applied on an HPTLC plate and they were scanned at 525 nm. The mobile phase toluene—methanol (9.4:0.6, v/v) was used for all the phytochemicals. After development, all the plates were air-dried at room temperature, derivatized with anisaldehyde–sulfuric acid reagent and heated at 105°C. This study aids the identification of these compounds and provides an easy and simple method for the simultaneous estimation of these markers in the I. cappa roots. The method would serve as an expedient tool in routine analyses to corroborate the drug through good constancy.

Chemical fingerprinting and quantification of biomarker wedelolactone in different sources of “Bhringaraja” by high-performance thin-layer chromatography and method validation

Two different botanical sources, Eclipta alba and Wedelia calendulacea are used as “Bhringaraja” in the Ayurvedic s ystem o f medicine. In the present study, an effort has been made to evaluate different sources by using high-performance thin-layer chromatography (HPTLC) as an analytical tool. Wedelolactone, one of the primary constituents of these plants, was taken as the marker compound for the evaluation. An HPTLC method was developed and validated for the evaluation of different sources of “Bhringaraja”. The chromatographic system was developed using silica gel 60 F254 HPTLC plates with the mobile phase toluene–chloroform–ethyl alcohol–formic acid (5:4:1:0.5, v/v). Linearity was found between the concentration ranges of 80 to 280 ng spot−1 (R2: 0.9994), and limit of detection (LOD) and limit of quantification (LOQ) were found to be 0.36 ng spot−1 and 1.09 ng spot−1. The study has shown the presence of wedelolactone at the concentration of 0.26% w/w and 0.05% w/w in E. alba and W. calendulacea, respect...

Fingerprint Study of Curcuma xanthorrhiza Rhizome by High Performance Thin Layer Chromatography (HPTLC)

The rhizome of Curcuma xanthorrhiza Roxb is intensively used in Indonesia as traditional medicine. It is widely used for hepatoprotective and anti inflammatory activities. To ensure the quality of its extract, we have studied the fingerprint or phytochemical analysis. This research was aimed to produce a chromatogram profile of the rhizome by HPTLC. The HPTLC fingerprint chromatogram of C. xanthorrhiza rhizome was performed using HPTLC plate of silica gel 60 F254 as the stationary phase and chloroform-methanol (97:3) as the mobile phase. Spot detection was carried out by TLC photo documentary system at 254 and 366 nm and TLC scanner at 427 nm. The developed method was validated according to ICH guidelines by determination of specificity and precision. We found that the specifity and precission of the method were met the acceptance criteria. In conclusion, the developed method is valid and could be used for quality control and standardization of herbal medicine containing C. xanthorrhiza rhizome.

Fingerprint Study of Curcuma xanthorrhiza Rhizome by High Performance Thin Layer Chromatography (HPTLC)

The rhizome of Curcuma xanthorrhiza Roxb is intensively used in Indonesia as traditional medicine. It is widely used for hepatoprotective and anti inflammatory activities. To ensure the quality of its extract, we have studied the fingerprint or phytochemical analysis. This research was aimed to produce a chromatogram profile of the rhizome by HPTLC. The HPTLC fingerprint chromatogram of C. xanthorrhiza rhizome was performed using HPTLC plate of silica gel 60 F254 as the stationary phase and chloroform-methanol (97:3) as the mobile phase. Spot detection was carried out by TLC photo documentary system at 254 and 366 nm and TLC scanner at 427 nm. The developed method was validated according to ICH guidelines by determination of specificity and precision. We found that the specifity and precission of the method were met the acceptance criteria. In conclusion, the developed method is valid and could be used for quality control and standardization of herbal medicine containing C. xanthorrhiza rhizome.

Assessment of antioxidant activity in Victorian marine algal extracts using high performance thin-layer chromatography and multivariate analysis

The aim of this study was to develop and validate a rapid and simple high performance thin layer chromatographic (HPTLC) method to screen for antioxidant activity in algal samples. 16 algal species were collected from local Victorian beaches. Fucoxanthin, one of the most abundant marine carotenoids was quantified directly from the HPTLC plates before derivatization, while derivatization either with 2,2-diphenyl-1-picrylhydrazyl (DPPH) or ferric chloride (FeCl3) was used to analyze antioxidants in marine algae, based on their ability to scavenge non biological stable free radical (DPPH) or to chelate iron ions. Principal component analysis of obtained HPTLC fingerprints has classified algae species into 5 groups according to their chemical/antioxidant profiles. The investigated brown algae samples were found to be rich in non-and moderate-polar compounds and phenolic compounds with antioxidant activity. Most of the phenolic iron chelators also have shown free radical scavenging activity. Strong positive and significant correlations between total phenolic content and DPPH radical scavenging activity showed that, phenolic compounds, including flavonoids are the main contributors of antioxidant activity in these species. The results suggest that certain brown algae possess significantly higher antioxidant potential when compared to red or green algae and could be considered for future applications in medicine, dietary supplements, cosmetics or food industries. Cystophora monilifera extract was found to have the highest antioxidant concentration, followed by Zonaria angustata, Cystophora pectinate, Codium fragile, and Cystophora pectinata. Fucoxanthin was found mainly in the brown algae species. The proposed methods provide an edge in terms of screening for antioxidants and quantification of antioxidant constituents in complex mixtures. The current application also demonstrates flexibility and versatility of a standard HPTLC system in the drug discovery. Proposed methods could be used for the bioassay-guided isolation of unknown natural antioxidants and subsequent identification if combined with spectroscopic identification.

Quantitative determination of alliin in dried garlic cloves and products by high-performance thin-layer chromatography

Purpose: To standardize the garlic samples and its products for alliin contents. Methods: A direct high-performance thin-layer chromatographic (HPTLC) method was developed to determine alliin in Chinese (CG) and Indian garlic (IG) and two other marketed products from USA and UK, respectively. Scanning and quantification were performed at 205 nm. Furthermore, chromatography was performed on pre-coated HPTLC plates with the solvent mixture, n – hexane:ethyl acetate (29:1 v/v), as the mobile phase. In addition, the parameters suggested by International Conference on Harmonization for analytical procedures were considered to validate the proposed method. Results: The system gave a compact spot of alliin at RF = 0.19 ± 0.01 The linear regression data for the calibration plots showed a good linearity (r 2 = 0.996) in the concentration range of 200 - 1600 ng. Linear regression equation was represented by Y = 1.792x + 182.855, while LOD and LOQ values were 40.42 ngband -1 and 111.72 ng.band -1 , respectively. The method showed excellent accuracy with recovery of 98.20 – 99.10 % and good precision with RSD of 1 - 2.65 %. Conclusion: The proposed method is selective, sensitive and accurate for the determination of alliin in garlic and its products. It is also evident from the results obtained that raw Indian garlic has higher alliin content than Chinese garlic. Keywords: Garlic, HPTLC, Alliin, Hypercholesteremia, Quantification