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ObjectiveThe therapeutic effect of polysaccharides from Zanthoxyli Pericarpium on Alzheimer's disease（AD） was evaluated through establishing a mouse model of AD， and the structural characteristics of the polysaccharides was analyzed by sugar spectrum. MethodThe AD model of mice with rapid aging was established by intraperitoneal injection of D-galactose combined with gavage of aluminum trichloride， and the learning and memory ability of mice was evaluated by Morris water maze test， the histopathological status of brain and neuronal damage were observed by hematoxylin-eosin（HE） staining and Nissl staining. After hydrolysis of polysaccharides from Zanthoxyli Pericarpium with acid and different glycosidases， the characteristics of hydrolysates were analyzed by high performance thin layer chromatography（HPTLC） and fluorescence assisted carbohydrate gel electrophoresis（PACE）. HPTLC chromatography was performed on a silica gel 60 plate with sampling volume of 5 μL， developing solvent of ethyl acetate-glacial acetic acid-water（2∶2∶1）， developing twice， aniline-diphenylamine-phosphoric acid solution as chromogenic agent， and heating at 105 ℃ for 10 min， and then observed under sunlight. PACE experimental conditions were 34% separation gel and 8% concentration gel， electrophoresis buffer was 0.1 mol·L-1 tris（hydroxymethyl） aminomethane（Tris）-boric acid buffer（pH 8.2）. Electrophoresis was carried out at 0 ℃ and the loading amount was 3-6 μL. The sample ran to the front of the gel with a constant current of 15 mA， and imaged under ultraviolet 365 nm. ResultThe results of Morris water maze test showed that polysaccharides from Zanthoxyli Pericarpium significantly improved the learning and memory ability of AD model mice， shortened the escape latency， and significantly increased the number of crossing and the residence time in the target quadrant. The results of histopathological experiments showed that polysaccharides from Zanthoxyli Pericarpium could improve the pathological conditions and neuronal damage in the CA1 and CA3 regions of hippocampus of AD mice， and the number of Nissl corpuscles was significantly increased. The results of sugar spectrum analysis showed that the results of HPTLC and PACE analysis were basically consistent， polysaccharides from Zanthoxyli Pericarpium could be mainly hydrolyzed into small molecular sugars by cellulase and pectinase， indicating that they mainly contained β-1，4-glucosidic bond and α-1，4-galacturonic acid glycosidic bond， and could be slightly hydrolyzed by glucanase， β-galactosidase and β-mannase， indicating that they contained only a small amount of α-1，6-glucosidic bond， β-galactosidic bond， β-1，4-mannosidic bond. ConclusionPolysaccharides from Zanthoxyli Pericarpium has obvious therapeutic effect on AD mice， and its structure mainly contains β-1，4-glucosidic bond and α-1，4-galacturonic acid glycosidic bond， which can provide a reference for the structural analysis of traditional Chinese medicine polysaccharides.
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Sophorae Flavescentis Radix is the dried root of Sophora flavescens Ait. and Sophorae Tonkinensis Radix et Rhizoma is the dried root and rhizome of Sophora tonkinensis Gagnep. The two drugs are both from the same genus Sophora, having similar and different compositions and efficacies, however, their differences are not fully demonstrated in current standard. In this study, the high-performance thin-layer chromatography with multi-dimensional and multi-level features combined with electric spray mass spectrometry (HPTLC-ESI-MS) was used to discover and identify the characteristic zones in extracts of Sophorae Flavescentis Radix and Sophorae Tonkinensis Radix et Rhizoma, after optimizing the preparation method of the test solution and chromatographic parameters. As a result, 17 main characteristic zones were found on HPTLC chromatograms of Sophorae Flavescentis Radix and Sophorae Tonkinensis Radix et Rhizoma, among them, besides 3 known chemicals, another 12 unknown components were identified by HPTLC-ESI-MS, they are 1 alkaloid and 11 flavonoids. The identification results were verified by the reference standards partially and nuclear magnetic resonance spectra after guided-isolation. Finally, a unified HPTLC specific identification method with different markers was established to identify Sophorae Flavescentis Radix and Sophorae Tonkinensis Radix et Rhizoma simultaneously. Thanks to abundant chemical information provided when using diverse polarity mobile phases and derivatization reagents, the HPTLC technology offers a convenient strategy for discovery, quality evaluation, and identification of target chemicals when connecting with mass spectrometry.
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Kokilakshadi Kashaya mentioned in Bhaishajya Ratnavali, Vataraktadhikara, it is a therapeutic formulation to treat Vatarakta. It is also used by Ayurvedic practitioners for treating hyperuricemia. The symptoms of hyperuricemia and gouty arthritis are similar to Vatarakta, a disease explained in classical Ayurvedic textbooks. Kokilakshadikwatha contains Kokilaksha and Guduchi and Pippalichurna given as Anupana of this formulation Physico chemical analysis of individual drug and formulation with modern parameters increase their scope and acceptance. The study was based on standard analytical parameters proposed by API. Method: Kokilakshadi Kwatha powder was evaluated for physico chemical analysis and phyto chemical screening. The analysis was done by using the parameters like Organoleptic features, loss on drying, acid soluble extractive, water soluble extractive. Results: Analytical parameters of individual drugs were done. All analytical parameter were within limit. Analytical parameter of Kokilakshadi Kwatha Churna like loss on drying 10.4%w/w, acid insoluble ash 0.79%, alcohol soluble extractive 11.2%w/w, water soluble extractive 7.8%w/w, pH 5.78 were obtained. High Performance Thin Layer Chromatography (HPTLC) profile of Kokilakshadikwatha powder showed 13 peaks at 254nm and 14 peaks at 366nm. Preliminary phytochemical screening test revealed the presence of steroids, alkaloids, phenols, flavonoids. Conclusion: The obtained data can be used for future comparative references
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Menopausal syndrome is a grouping of signs and symptoms associated with menopause. In Ayurveda, menopause is referred to as 'Rajonivrutti' (and menopausal syndrome as Rajonivruttianubandhaja vyadhies). Menopause's long-term risks include osteoporosis, cardiac problems, and Alzheimer's disease. Aims and objective: To study the Pharmacognostic, Phytochemical and HPTLC of Vayasthapana Gana Choorna and Vayasthapana Ghrita. Material and methods: Pharmacognostic, phytochemical and HPTLC of Vayasthapana Gana Choorna and Vayasthapana Ghrita have been carried out as per standard protocol. Result: Vayasthapana Gana Choorna showed the presence of mesocarp, asicular crystals, stone cells, scleroids, brown content, starch grains, colencyma cells, rhomboidal crystals, pitted vessels, parenchyma cells, simple trichome. Phytochemical parameters showed refractive index 1.3660, specific gravity 0.913, acid value 1.285, iodine value 212.1085 and in HPTLC, Methanol extract of Vayasthapana Ghrita at 254nm showed 6 spots and at 366nm 2 spots whereas in methanol extract of Vayasthapana Gana Choorna at 254nm 5 spots and in 366nm 4 spots were present. Conclusion: The applied pharmacognostic and HPTLC method has been shown to be selective, linear, precise and accurate. The method will be useful for quality control of the raw material and pharmaceutical preparations.
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Nagabala -Arjunadi Yoga, is the combination of Nagabala and Arjuna Churna mentioned in Chakradatta, Hridroga Chikitsa, is prepared by giving Bhavana of Rasonadi Kwatha. Hridroga (cardiovascular disorders) are the most common health concern of the present era. It is the leading cause of death worldwide. Ancient Samhitas contain many formulations in the context of Hridroga, whose applicability is unexplored. Churna and Kwatha are the main dosage forms used in clinical practice. But compared to Churna and Kwatha, tablets are more patient compatible in terms of palatability and possess increased shelf life. Hence, Nagabala-Arjunadi Yoga, a tablet dosage form is developed using Nagabala- Arjuna Churna and Rasonadi Kwatha. No scientific evaluation data for this drug is available to date. The present study was done to evaluate the pharmacognostical and pharmaceutical profile of Nagabala-Arjunadi Yoga. The microscopic examination of the Nagabala- Arjunadi Yoga showed the presence of rosette crystals, rhomboidal crystals, simple fibres, oil globules and stones cells. The physicochemical analysis showed that pH value, hardness, loss on drying, ash value, water extractive value and methanol extractive value was 5.8, 3.5kg/cm2, 7.949%, 3.03%, 17.43%, 16.14% respectively. The HPTLC densitograms at UV 254 nm and UV 366nm using Toluene and Ethyl acetate in the ratio 9:1 showed maximum peak height in 3rd peak corresponding to the Rf value 0.18 and 0.17 respectively. The finding observed in the present study can be used as reference for future quality control.
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Marketing of Guggulu, an exudate obtained from the plant Commiphora mukul and its preparations is a great concern of different ayurvedic pharmaceutical houses due to its adulteration, substitution and non-availability of genuine samples. Standardization of raw drugs and formulations with modern analytical tools increase their scope, acceptance and scientific validity. In the present study, an attempt was made for the physicochemical analysis of Kanchanara guggulu tablets (having Guggulu as the major ingredient) prepared as per references in Sharangadhara Samhita and Bhaishajya Ratnavali to develop an analytical profile of the formulation. The study was based on the standard analytical parameters proposed by API and PLIM. Six samples of tablets were prepared as per two references (three for each reference) which were available in market following the same manufacturing procedures. The study comprised of three stages- pharmacognosy of raw drugs, pharmaceutical work and analytical study. The analysis was done using the parameters like organoleptic evaluation, weight variation, hardness, friability, disintegration time, ash values, extractive values, loss on drying, pH, HPTLC and microbial contamination and an analytical profile was developed as per both references.
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Aims: Hypericum perforatum L., known as “Hofarighun” is a widely used herbal drug in Traditional Persian Medicine (TPM). Detection of non-relevant plants, instead of this species, in the herbal market encourages the need for the establishment of it’s chemical authentication and standardization, through implying rapid and efficient phytochemical techniques. Study Design: Twelve Hypericum samples were acquired from traditional medicine markets of different regions of Iran (Tehran, Sanandaj, Mashhad, Kerman, Bandar Abbas, Ahvaz, Yazd, Babol, Yasuj, Shiraz (Chehel Giah), Shiraz (Kazerun Gate), and Shiraz (Adloo Zerehi), based on microscopic characterization. Positive control was taken in the form of cultivated specimen of H. perforatum. Place and Duration of Study: Study was performed in Medicinal plants processing Research Center, SUMS, Shiraz in the months between February to December 2021. Methodology: Essential oil samples were injected into a gas chromatograph (GC) and compounds were identified as per the spectra obtained. Total phenol, flavonoid and HPTLC analysis of samples were also done. Results: ?-pinene was found in highest proportion in majority of samples i.e. 35.55-63.69%. However other compounds such as 1-dodecanol (10.82%), caryophyllene (15.87%) and ?-cubebene (15.14%) were also analyzed in samples and the cultivated sample respectively. Total phenol and flavonoid content among the Hypericum extracts were found to be between 50.31±3.22 to 262.76±8.12 mg Gallic Acid Equivalent (GAE)/g of Ext. and 13.47±1.68 to 79.26±5.78 mg Quercetin Equivalent (QE)/g of Ext., respectively. Conclusion: The noticeable findings of present study can be used as a framework for authentication of Hypericum perforatum samples. The methods used were found to be feasible and efficient in detection of adultrations and may contribute to minimize the safety and efficacy concerns over the samples available in the traditional herbal pharmacies.
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OBJECTIVE To optimize the existing t hin layer chromatography （TLC）identification and content determination methods of Jizhi syrup. METHODS High performance thin-layer chromatography （HPTLC）was used to identify five medicinal materials in Jizhi syrup ，such as Ilex chinensis ，Houttuynia cordata ，Peucedanum praeruptorum ，Citrus aurantium ，Glycyrrhiza uralensis. High performance liquid chromatography （HPLC）was used to determine the contents of procatechuic acid ，ephedrine hydrochloride and naringin in Jizhi syrup. RESULTS HPTLC results showed that the identification spots of pedunculoside ， praeruptorin A ，naringin，and liquiritin were clearly displayed ，and the retention factors were in the range of 0.2 to 0.8. After validation，the method had been proved to be strongly specific ，robust and repeatable. HPLC results showed that the linear ranges of protocatechuic acid ，ephedrine hydrochloride and naringin were 4.32-431.67，1.14-114.17 and 7.02-702.33 μg/mL（all r＞ 0.996），respectively. The average recoveries were 100.61％，100.40％ and 99.22％，and RSDs were all less than 2.00％. RSDs of precision（n＝6），stability（24 h，n＝7）and repeatability （n＝6）were all less than 2.00％. The average contents of the three components in 10 batches were 623.3，152.1，1 213.9 μg/mL（RSD＜10.00％），respectively. CONCLUSIONS In this study ， HPTLC method of one-plate multi-drug is established for the identification of Jizhi syrup. One sample pretreatment method and two TLC conditions are used to realize the rapid identification of five kinds of medicinal materials. An HPLC method is established to determine the content of Jizhi syrup ，which realizes the fast quantification of three active components in Jizhi syrup ，and can be used to optimize the identification and content determination items in the existing legal quality standards of Jizhi syrup.
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A rapid and feasible method of HPTLC is standardized for quantification of anethole in essential oil’s extract and from herbal formulations of fennel seed. The developeddensitometric HPTLC method was performed to estimate the existence of anethole in the essential oil, extract and herbal formulations of fennel with the optimized concentration of hexane: Ethyl acetate (8:2%, v/v, mobile phase) on glass coated silica gel 60 F254plates (20 × 10 cm) scanned with the absorbance of λ260nm under densitometric condition. The Linearity of regressions revealed a satisfactory relationship between peak area and concentration of anethole in between the range of 100-600 ng/spot. This reliable method was validated as per the ICH guidelines to fulfill the necessary parameters such as accuracy and robustness. The amount of anethole in essential oil (0.098 ± 0.002%),extract (0.101 ± 0.004%)and three herbal formulations A (0.024 ± 0.004%), C (0.019 ± 0.002%) whileanethole is not detected in B formulations from fennel seed was completely estimated by the developed method. The standardized methods and its validation gave new insights of HPTLC based detection and quantification of anethole in other aromatic plants as well as in other pharmacological formulations
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Background: Cyclea peltata is one of the herbs mentioned in ancient scriptures of Ayurveda and is used indifferent types of Ayurvedic gritham preparations. Moreover, in traditional/tribal medicine C. peltata isused as digestive, anti-inflammatory, diuretic and to treat jaundice, digestive disorders, etc.Objective: Activity guided fractionation of C. peltata and in correlation with the levels of bioactivecompound tetrandrine.Materials and methods: Preliminary phytochemical screening, estimation of total alkaloid content,preparation of different extracts of C. peltata (crude extract CP, hexane extract HCP, chloroform extractCCP, methanol extract MCP, alkaloid fraction ACP). In vitro anti-inflammatory studies using RAW264.7 cells and in vitro antioxidant assays of the different extracts of C. peltata. HPTLC estimation oftetrandrine (TET) was carried out using solvent system toluene: ethyl acetate: diethylamine (7.2: 2: 0.8)and isolation of TET from ACP.Results: Preliminary phytochemical studies of C. peltata showed the presence of alkaloid content in allextracts. Whereas, saponins, steroids and terpenoids were detected in CP and CCP. ACP and TET showedsignificant in vitro anti-inflammatory and antioxidant activity when compared to other extracts. ACP andTET (100 mg/ml) treatment significantly inhibited the mRNA expression of iNOS, COX-2, TNF-a in LPStreated RAW 264.7 cells. HPTLC estimation of bioactive compound tetrandrine was highest in ACP228.4 mg/mg followed by CP-29.62 mg/mg, CCP-23.46 mg/mg, MCP-18.82 mg/mg and HCP-1.25 mg/mg. TEThas been isolated from ACP.Conclusion: The results of the present in vitro assays revealed that the alkaloid fraction (ACP) is the mostactive fraction when compared to other extracts and has a positive correlation with the levels of bioactivecompound tetrandrine.
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In this research work to evaluate in vitro antioxidant activity and HPTLC finger printing analysis of Physalis peruviana fruits. The chemical fingerprinting was carried out by high performance thin layer chromatography. It was carried out by the CAMAG HPTLC system equipped with Linomat V sample applicator, twin through plate development chamber, TLC scanner III and integration software WIN CATS-4.02. Physalis peruviana fruit extract was tested for phytochemical screening and in vitro anti-oxidant enzymes like 2, 2-diphenyl-1-picryl hydrazyl radical (DPPH), total antioxidant activity and reducing ability. Physalis peruviana fruit extract effectively scavenged free radicals at all different concentrations and showed its potent antioxidant activity. Phytochemical analysis revealed the presence of various major phytoconstituents like alkaloids, flavonoids, saponins, phenolics, tannins and anthraquinones. The HPTLC fingerprint qualitatively revealed predominant amount of quercetin. Physalis peruviana fruit extract will be subjected to further extensive studies to isolate and identify their active constituents which are useful for understanding their mechanism of action as antioxidants.
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Alocasia indica is perennial herb growing widely and used as traditional medicine in India, China and Bangladesh. The divine herb has potent medicinal values for the treatment of different type of illnesses. The HPTLC techniques were used to separate active components from ethanolic extract of tuber part of A. indica. This examination was intended to designed a HPTLC fingerprint profile of crude extract of the plant in ethanol. A HPTLC method for the isolation of various active constituents in A. indica ethanolic extract have been developed and solvent system for quercetin the mobile phase used was toluene: ethyl acetate: formic acid (5:2:1) and for analysis of β-sitosterol the mobile phase used was chloroform: ethyl acetate: formic acid (6:4:1) . In the present investigation, HPTLC fingerprint of extract of dried tuber part of A. indica have been performed and the results demonstrated that important information for standardization. The HPTLC system for routine quality control of present species can be used for ethanolic extract and serve in qualitative, quantitative and was appropriate for standardization of the plant.
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Background: Increasing popularity of Mathan Tailam (mattan tailam, pachai ennai) for the treatment ofdiabetic foot ulcer necessitated standardization and quality control of this medicated oil both in largescale production and quality check in marketed drug.Objective: Present study aims to develop standard operating procedure for the preparation of MathanTailam as per Siddha Formulary of India and its standardization using suitable analytical techniques.Materials and methods: Mathan Tailam was prepared as per Siddha Formulary of India. Physicochemicalparameters and preliminary phytochemical screening were carried out using standard methods. The inhouse prepared sample underwent physico-chemical analysis, qualitative phytochemical analysis, Gaschromatography-mass spectrum (GC-MS) analysis, High performance thin layer chromatographic(HPTLC) fingerprinting profile and inductively coupled plasma-optical emission spectroscopic (ICP-OES)analysis.Results: Physico-chemical parameters of the prepared formulation were comparable to that of coconutoil. Aqueous methonolic extract of this drug was found to be positive for alkaloid, saponin, coumarin,steroid, triterpinoid, quinine and furan. The GC-MS values were comparable to that of the base used i.e.,the coconut oil. HPTLC fingerprinting profile revealed the presence of phytochemicals in the medicatedoil derived from both coconut oil and Datura metel. ICP-OES addressed the mineral portion of theformulation and its safety in heavy metal aspect.Conclusion: All these parameters can be utilized for the overall quality check over its preparation andformulation.© 2017 Transdisciplinary University, Bangalore and World Ayurveda Foundation. Publishing Services byElsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
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OBJECTIVE: To carry out HPTLC and HPLC fingerprint analysis of 18 batches of Ganoderma samples using two kinds of reference substance of Ganoderma extract, G. lucidum Extract Reference Substance(CZERS) and G. sinense Extract Reference Substance(ZZERS). METHODS: HPTLC Fingerprint was used to analyze triterpene acids and sterols in Ganoderma with chloroform-acetonitrile-methanol-formic acid (13∶2∶0.5∶0.5, develop 3 times) and cyclohexane-ethyl acetate-methanol-formic acid (15∶5∶0.5∶0.5, develop 2 times) respectively. HPLC Fingerprint analysis was conducted using Kromasil 100-5 C18 column (4.6 mm×250 mm, 5 μm) kept at 25 ℃. Mobile phase A was acetonitrile and B was 0.02% phosphoric acid; gradient elution procedure was as follows: 0-40 min, 29%→33% A; 40-70 min, 33%→65%A; 70-105 min, 65%→100%A; 105-120 min, 100% A; flow rate was 1.0 mL•min-1. DAD detector was adopted with detection wavelength set at 244 nm. The injection volume was 10 μL. RESULTS: By using ERS and fingerprint analysis, G. lucidum, G. sessile and G. lucidum could be distinguished. The components of G. lucidum in different species and growth patterns were different. CONCLUSION: There are many varieties of G. lucidum, which can be divided into wild and artificial cultures, and the culture media of artificial culture are different, which leads to the difference of individual components of different G. lucidum. Fingerprint analysis based on ERS of specific varieties are more suitable for the overall quality control of G. lucidum.
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Objective: The study was conducted to develop and validate a high performance thin-layer chromatography (HPTLC)-densitometric method for the quantitative analysis of morin in Maclura cochinchinensis heartwood collected from different locations in Thailand. Methods: HPTLC analysis was performed on an aluminium sheet of silica gel 60 F254 using toluene: ethyl acetate: formic acid (36:12:7, volume percent) as a mobile phase. The densitometric scanning was performed at the wavelength 410 nm. HPTLC method was validated according to ICH guideline. Results: The proposed HPTLC method showed acceptable validation parameters. The content of morin in M. cochinchinensis heartwood collected from eight different provinces in Thailand were in the ranges of 1.53%−2.73%. Conclusion: The simple and sensitive HPTLC method was successfully developed and validated for determination of morin in M. cochinchinensis heartwood. The proposed HPTLC method was found to be simple, fast and inexpensive, and can be used for the routine quality control of raw materials.
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Background: Trikatu, Sitopaladi, Hingavastaka, Avipattikara, Sringyadi and Talisadya are very popular Ayurvedic (churna) medicines practiced in India; however, unfortunately, they possess several qualitycontrol issues.Objective: The aim of this study was to find out a simple, accurate and sensitive HPTLC method for thedetection and quantification of marker molecule, piperine (alkaloid) on these Ayurvedic formulations forstandardization.Materials and methods: Methanolic extraction (reflux) was performed from the above six churnas as wellas three single ingredients Piper longum (pipul), Piper nigrum (marich) and Piper chaba (chai). HPTLC wasdone using piperine as a standard. The mobile phase was a mixture of toluene-ethyl acetate (7:3, v/v) anddetection at 342l.Results: The Rf was detected at 0.39. Piperine was quantified in all samples. P. nigrum showed higherpiperine than P. longum and P. chaba. The maximum piperine was noted in Hingavastaka churna andfollowed by Sringyadi churna, Sitopaladi churna, Talisadya churna, Trikatu churna and Avipattikara churna.Conclusion: This method can be successfully employed for standardization and quantitative analysis ofpiperine in Ayurvedic formulations (churnas) and also be helpful to clinicians and pharmacists to drawsignificant role of piperine present in all these samples.© 2017 Transdisciplinary University, Bangalore and World Ayurveda Foundation. Publishing Services byElsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
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Sahadevi (Cyanthillium cinereum (L.) H.Rob.) (Family Asteraceae) commonly known as Purple Fleabane in English, Sahadei in Hindi and Poovankurunthila in Malayalam, an erect annual branched herb with pubescent cylindrical stem found as a weed throughout India is extensively used in folkore medicine. The present paper highlights the pharmacognostical and phytochemical characters of the plant to give clear standards for identification of the drug. Microscopic evaluation of root, stem and leaf as well powder microscopy of the plant were carried out. Physicochemical parameters like moisture content, total ash, water insoluble ash, acid insoluble ash, volatile oil content, sugar content, fibre content, alcohol soluble extractive and water soluble extractive were studied. Preliminary phytochemical analysis of the plant Sahadevi [Cyanthillium cinereum (L.) H.Rob.] showed the presence of steroid, flavonoid, glycoside, saponins and tannin. The present study signifies the use of TLC and HPTLC fingerprint profiles of aqueous and alcoholic extracts of the drug for determining the identity, purity of the drug and also for developing standards. The findings drawn from the study substantiates the genuineness of the drug Sahadevi [Cyanthillium cinereum (L.) H.Rob.], which is at par with the descriptions available in the authentic books.
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Objective: To develop a new simple, selective and precise high-performance thin layer chromatographic method for determination of Dapagliflozin (DAPA) in bulk and tablet dosage form Methods: The present study describes development and validation of High performance thin layer chromatographic method for DAPA. The chromatographic separation was carried out on Merck precoated silica gel aluminium plate 60 F254 using Chloroform: Methanol (9:1v/v) as mobile phase. Quantitative determination of drug was carried out by densitometric scanning of plates at 223 nm using Camag TLC Scanner. Results: The chromatographic condition shows compact band with the retention factor for dapaglifloxin as 0.21 ± 0.004. The method was validated as per ICH guidelines for linearity, accuracy, precision and robustness. Response was found to be linear in the concentration range of 400 ng/ band to 1200 ng/band with linear regression value of 0.9953 with respect to peak area and concentration value The LOD and LOQ was found to be 1.2083 ng/band and 3.6616ng/ band. The percentage assay was found to be 100±0.05. Conclusion: This method under statistical analysis proved a selective, repeatable and accurate analysis of the drug. This method can be used for quantitative analysis of dapaglifloxin in the bulk drug and in tablet.
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Amrtottara kvatha is a decoction which is used primarily in the management of hyperpyrexia (jwara). Fresh stem of Tinospora cordifolia (Willd.) Miers (Guduchi), dried fruit rind of Terminalia chebula Retz (Haritaki), and dried rhizome of Zingiber offficinale Roscoe (Shunti) in the ratio 3:2:1 are its ingredients. From the point of view of drug design, it has the peculiarity that, its ingredients are specified to be added in a particular ratio, rather than in equal amounts as is the case in most of the compound formulations in Ayurveda. But, the rationale behind the drug design are not detailed enough to give much information regarding the effect of alteration of the drug ratio etc. Thus, in the present study an exploration was made with respect to its phytochemistry, by comparing with the decoction prepared in the general ratio of 1:1:1. The compounds Gallic acid and Ellagic acid were used for quantitative evaluation and comparison using HPTLC method. Estimation of Gallic acid in the samples showed that the amount of Gallic acid in Amrtottara kvatha prepared in classical ratio of 3:2:1 is significantly higher than that prepared in the altered ratio of 1:1:1 with a p value <0.05, though the amount by weight of Haritaki (source of Gallic acid) was unaltered in both, thus indicating the possibility of some complex phytochemical interactions among the constituents. With respect to Ellagic acid, there was no statistically significant difference in its quantity in the two decoctions compared. The method developed for HPTLC analysis in this study can be used as a technique for standardization of Amrtottara kvatha.