DEVELOPMENT, VALIDATION OF HPTLC METHOD FOR SIMULTANEOUS QUANTITATION OF LUTEOLIN, APIGENIN FROM CARDIOSPERMUM HALICACABUM LINN. AND HYDNOCARPUS PENTANDRA (BUCH.-HAM.) OKEN

Objectives: To develop and validate High Performance Thin Layer Chromatographic (HPTLC) method for simultaneous quantitation of luteolin and apigenin from dried leaf powder of Cardiospermum halicacabum Linn. and dried seed hull powder of Hydnocarpus pentandra (Buch.-Ham.) Oken. Methods: Chromatography was performed using methanolic extracts of dried leaf powder of Cardiospermum halicacabum Linn. and dried seed hull powder of Hydnocarpus pentandra (Buch.-Ham.) Oken. Separation of luteolin and apigenin from methanolic extracts of both the plant materials was achieved on Silica gel 60F254 TLC plates using a suitable solvent system. Detection and quantitation of luetolin and apigenin was done by densitometric scanning at λ=349 nm. The developed HPTLC method has been validated using International Conference on Harmonization (ICH) guidelines. Results: The validated HPTLC method was used for simultaneous quantitation of luteolin and apigenin from methanolic extracts of dried leaf powder of Cardiospermum halicacabum Linn. and dried seed hull powder of Hydnocarpus pentandra (Buch.-Ham.) Oken using their respective calibration curves. Amounts of luteolin and apigenin present in dried leaf powder of Cardiospermum halicacabum Linn. are 0.2119mg/g and 0.9089mg/g respectively. Amounts of luteolin and apigenin present in dried seed hull powder of Hydnocarpus pentandra (Buch.-Ham.) Oken are 0.0696mg/g and 0.0320mg/g. Conclusion: The developed method is simple, precise and accurate and can also be used for routine quality control analysis and for the quantitation of luteolin and apigenin in herbal raw materials as well as in their formulations

HPLC METHOD DEVELOPMENT AND VALIDATION FOR QUANTITATION OF TRIGONELLINE FROM MIRABILIS JALAPA LINN. LEAVES AND ENHANCEMENT IN EXTRACTION YIELD FROM ULTRA FINE POWDER

Objective: Development and validation of a simple and reliable HPLC method for determination of an alkaloid, trigonelline, in the methanolic extract of Mirabilis jalapa Linn. leaves and comparing the extraction yields of trigonelline from micro powder and ultrafine powder.Methods: The quantitation of trigonelline was carried out on a Phenomenex (Luna 5 U RP C8 (2) column, 25 cm x 4.6 mm, i.d. 5 µm), using mobile phase comprising of distilled water containing HCl (pH adjusted to 3.5) and methanol in the volume ratio of 70:30, which was delivered at the flow rate of 0.5 ml per min, at 35 °C column temperature. The detection and quantitation of trigonelline were carried out using PDA detector at the wavelength λ=264 nm.Ultra-fine powder of Mirabilis jalapa Linn. was prepared using simple stepwise powdering method. The dried leaves of Mirabilis jalapa Linn. were ground using ice jacketed domestic mixer. This powder was sieved through a BSS 85 mesh sieve and considered as a micro powder. Further fine grinding was done by jet milling, followed by ball milling. This powder was considered as an ultra-fine powder.Results: The proposed HPLC method for quantitation of trigonelline from dried leaf powder of Mirabilis jalapa Linn. is rapid, simple, accurate and precise.Conclusion: The amount of trigonelline obtained using methanolic extracts of Mirabilis jalapa Linn. Ultra-fine powder and the micro powder was found to be 1.1103 mg/g and 0.7258 mg/g respectively

Detection and Estimation of alpha-Amyrin, beta-Sitosterol, Lupeol, and n-Triacontane in Two Medicinal Plants by High Performance Thin Layer Chromatography

A normal phase high performance thin layer chromatography (HPTLC) method has been developed and validated for simultaneous estimation of four components, namely, alpha-amyrin, beta-sitosterol, lupeol, and n-triacontane from two medicinally important plants, Leptadenia reticulata Wight & Arn. and Pluchea lanceolata (DC.) CB. Clarke. In Ayurveda, both plants have been reported to possess immunomodulatory activity. Chromatographic separation of the four components from the methanolic extracts of whole plant powders of Leptadenia reticulata Wight & Arn. and Pluchea lanceolata (DC.) CB. Clarke. was performed on TLC aluminium plates precoated with silica gel 60F254 using a suitable mobile phase. The densitometric scanning was done after derivatization at λ = 580 nm for α-amyrin, β-sitosterol, and lupeol, and at 366 nm for n-triacontane. The developed HPTLC method has been validated and used for simultaneous quantitation of the four components from the methanolic extracts of whole plant powders of Leptadenia reticulata Wight & Arn. and Pluchea lanceolata (DC.) CB. Clarke. The developed HPTLC method is simple, rapid, and precise and can be used for routine quality control

HPLC METHOD DEVELOPMENT AND VALIDATION FOR QUANTITATION OF TRIGONELLINE FROM MIRABILIS JALAPA LINN. LEAVES AND ENHANCEMENT IN EXTRACTION YIELD FROM ULTRA FINE POWDER

Objective: Development and validation of a simple and reliable HPLC method for determination of an alkaloid, trigonelline, in the methanolic extract of Mirabilis jalapa Linn. leaves and comparing the extraction yields of trigonelline from micro powder and ultrafine powder.Methods: The quantitation of trigonelline was carried out on a Phenomenex (Luna 5 U RP C8 (2) column, 25 cm x 4.6 mm, i.d. 5 µm), using mobile phase comprising of distilled water containing HCl (pH adjusted to 3.5) and methanol in the volume ratio of 70:30, which was delivered at the flow rate of 0.5 ml per min, at 35 °C column temperature. The detection and quantitation of trigonelline were carried out using PDA detector at the wavelength λ=264 nm.Ultra-fine powder of Mirabilis jalapa Linn. was prepared using simple stepwise powdering method. The dried leaves of Mirabilis jalapa Linn. were ground using ice jacketed domestic mixer. This powder was sieved through a BSS 85 mesh sieve and considered as a micro powder. Further fine grinding was done by jet milling, followed by ball milling. This powder was considered as an ultra-fine powder.Results: The proposed HPLC method for quantitation of trigonelline from dried leaf powder of Mirabilis jalapa Linn. is rapid, simple, accurate and precise.Conclusion: The amount of trigonelline obtained using methanolic extracts of Mirabilis jalapa Linn. Ultra-fine powder and the micro powder was found to be 1.1103 mg/g and 0.7258 mg/g respectively.</jats:p