Published28 Sep Abstract Heracleum candicans Wall. Apiaceae is widely used in Indian system of medicines as an aphrodisiac, nerve tonic and also in the treatment of skin diseases with reports of being rich in furocoumarins which are extensively used in pharmaceutical industry for their photosensitizing activity on human skin. A simple high performance thin layer chromatography HPTLC method has been developed for the simultaneous determination of psoralen 1 and heraclenol 2 in the fruits of H. The analytes were separated on silica gel F plates with toluene : diethy lether : acetic acid 6 : 4 : 1 and scanned densitometrically at nm. The method was validated in terms of precision, repeatability, and accuracy.
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Published28 Sep Abstract Heracleum candicans Wall. Apiaceae is widely used in Indian system of medicines as an aphrodisiac, nerve tonic and also in the treatment of skin diseases with reports of being rich in furocoumarins which are extensively used in pharmaceutical industry for their photosensitizing activity on human skin. A simple high performance thin layer chromatography HPTLC method has been developed for the simultaneous determination of psoralen 1 and heraclenol 2 in the fruits of H.
The analytes were separated on silica gel F plates with toluene : diethy lether : acetic acid 6 : 4 : 1 and scanned densitometrically at nm. The method was validated in terms of precision, repeatability, and accuracy. The two compounds were quantified in fruits of H. The method was found to be very simple, accurate, precise, and economical and can be used for routine quality control. Introduction Heracleum candicans Wall. In Indian system of medicines, it is used as an aphrodisiac, nerve tonic and also used in the treatment of skin diseases [ 2 ].
The plant has been shown to possess potent stimulatory effect on melanogenesis with significant enhancement of cell proliferation [ 3 ]. Dichloromethane extract of shoots of H.
Chemical investigation of the plant has revealed the presence of furocoumarins, namely, bergapten [ 2 ], heraclenol, xanthotoxin, phellopterin, angelicin, imperatorin, xanthotoxol [ 5 ], heraclenin [ 6 ], candibirin [ 7 ], and 8-gernoxypsoralen [ 8 ]. Activity-guided isolation has also shown heraclenin to be the anti-inflammatory principle present in H. Furocoumarins are polyphenolic compounds, synthesized from L-phenylalanine, which may occur in a linear form with the furan ring attached to the 6, 7 position of the benzopyrene nucleus.
These are biologically active natural compounds found in many plants particularly in members of the Umbelliferae and Rutaceae families [ 9 , 10 ]. They are typically phototoxic compounds leading to photodermatitis upon exposure to UV light [ 11 ].
In humans and experimental animals, chronic furocoumarin treatment, in combination with UV light, is used in the therapy of psoriasis [ 12 ]. On the molecular level furocoumarins bind to cellular constituents such as proteins and lipids, can damage lysosomes, lead to the formation of reactive oxygen species, and can contribute to the formation of novel antigens by covalent modification of proteins [ 13 ].
Furocoumarins of H. There has already been an attempt made at the spectrophotometric estimation of the total furocoumarin bitter principles from roots of H. HPLC as a tool for the quantitative estimation of furocoumarins in fruits of H. Thus keeping in view the importance of furocoumarins, quantitative estimation of these compounds in H.
We have made an attempt to qualitatively evaluate H. An attempt has also been made to validate the developed method in terms of precision, repeatability, LOD, LOQ, and accuracy.
Experimental 2. Plant Material and Reagents Fruits of H. The samples were authenticated and herbarium of the same was deposited in the NBRI, Lucknow, India voucher specimen no. Precoated silica gel 60 F cm, 0. Solutions of sample and standard compounds 1, 2, and 3 of known concentrations were applied on TLC F in band form as 6 mm wide and 10 mm from the bottom.
The scanner conditions used were slit width mm, wavelength mm, and absorption reflection scan mode. These parameters were kept constant throughout. Preparation of Standard Solutions of Psoralen and Heraclenol Stock solutions of psoralen 1 and heraclenol 2 1.
The combined extracts were concentrated under vacuum. The extracts were obtained in a yield of After development, the plates were air dried and scanned at nm. The peak areas were recorded. Calibration curves were prepared by plotting peak areas versus concentration. The plate was developed and scanned as described above.
The amount of 1 and 2 in the sample was calculated using the calibration curves. Peak identification was achieved in comparison of both and UV absorption spectrum with those obtained for standards. Method Validation Precision was checked by repeated scanning of the same spots of 1 3. Variability of the method was studied by analyzing aliquots of standard solutions of 1 0.
The accuracy of the method was determined by recovery studies. Plant samples spiked with three different amounts of 1 1. Different dilutions of standard solutions of 1 and 2 in methanol were applied.
Limit of detection LOD and limit of quantification LOQ were determined on the basis of-signal to-noise ratios of 3 : 1 and 10 : 1. Results and Discussion Earlier a simple TLC method has been reported for the simultaneous determination of only two compounds from the roots of Heracleum candicans [ 18 ]. The quantitative estimation of furocoumarins in fruits was also reported by HPLC only [ 17 ].
In systems other than toluene: Diethyl ether: Acetic acid either the resolution of the spots was not very clear or there was interference from the matrix. Best resolution and reproducible peaks were obtained only by using toluene : diethyl ether : acetic acid 6 : 4 : 1 as mobile phase 1: , The specificity of the method was ascertained by analyzing standards and the samples.
The spots for psoralen 1 and heraclenol 2 in the sample were confirmed by comparing the values and the spectra of the spots with those of the standards Figure 1. The overlay spectra of identified peaks in the standards as well as the corresponding peak in the extracted samples showed total superimposition at peak start, peak maximum, and peak end, thus confirming the purity of the peaks in the extracted solutions.
The method was validated in terms of precision, repeatability, and accuracy Table 1. These correlation coefficients indicated good linearity between the concentration and peak area in the applied concentration range. The intraday and interday precision values for 1 and 2 indicated that the method was precise and reproducible. Psoralen 1 and heraclenol 2 contents were determined quantitatively in the fruits of H.
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Author Seema and Devkant Sangwan. Heracleum candicans Wall. Seema and Devkant Sangwan. Flowers are yellowish often tinged with brown.
Heracleum candicans Wall.
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