HPLC method development and validation of cytotoxic agent phenyl-heptatriyne in Bidens pilosa with ultrasonic-assisted cloud point extraction and preconcentration.
Extraction and pre-concentration of a bioactive marker compound, phenyl-1,3,5-heptatriyne from Bidens pilosa, prior to HPLC has been demonstrated using both organic and ecofriendly solvents. Non-ionic surfactants, viz. Triton X-100, Triton X-114 and Genapol X-80, were used for extraction. No back-extraction or liquid chromatographic steps were required to remove the target phytochemical from the surfactant-rich extractant phase. The optimized cloud point extraction procedure has been shown to be a potentially useful methodology for the preconcentration of the target analyte, with a preconcentration factor of 4-99. Moreover, the method is simple, sensitive, rapid and consumes lesser solvent than traditional methods. An isocratic chromatographic separation and quantitation was accomplished on a C(18) column with acetonitrile-acidified aqueous as mobile phase at a flow rate of 1.0 mL/min, UV detection at 254 nm and specificity with photo diode-array detector (PDA) and MS. Under the optimum experimental conditions recovery was satisfactory (99.18-100.33%) without interference from the surfactant. The method seems to be reliable with intraday precision and interday precision below 2.0%. Good linearity was obtained in the working range from 7.5 to 30 µg/mL with correlation coefficient >0.99. The limits of detection and quantitation were 1.84 and 6.13 µg/mL, respectively. The method was validated following international guidelines and successfully applied for quantitative assays of cytotoxic compound phenyl-1,3,5-heptatriyne in Bidens pilosa.
Trivedi
Priyanka
P
Analytical Chemistry Department, Central Institute of Medicinal and Aromatic Plants, Council of Scientific and Industrial Research, PO-CIMAP, Lucknow, India.
Kumar
Jonnala Kotesh
JK
Negi
Arvind Singh
AS
Shanker
Karuna
K
D000480:Alkynes / Q000032:analysis* / Q000737:chemistry*; D000704:Analysis of Variance; D031176:Bidens / Q000737:chemistry*; D005591:Chemical Fractionation / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D016018:Least-Squares Analysis; D016014:Linear Models; D013058:Mass Spectrometry; D008823:Micelles; D017830:Octoxynol; D010936:Plant Extracts / Q000737:chemistry*; D018515:Plant Leaves / Q000737:chemistry; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013010:Sonication
C560575:1-phenyl-1,3,5-heptatriyne; D000480:Alkynes; D008823:Micelles; D010936:Plant Extracts; C019518:phenylheptatriyne; D017830:Octoxynol