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International Journal of Farm Sciences
Year : 2016, Volume : 6, Issue : 3
First page : ( 70) Last page : ( 76)
Print ISSN : 2229-3744. Online ISSN : 2250-0499.

Chemical profiling assisted quality assessment of podophyllotoxin from Podophyllum hexandrum by LC-APCI-3Q-MS/MS

Sharma DK, Kumar Anil*, Kumar Jitender1

Department of Chemistry, Himachal Pradesh University, Shimla-171005, HP, India

1School of Biotechnology, Shoolini University, Bhajol-173223, HP, India

*Email forcorrespondence: anilthakur.iitd@gmail.com

Online published on 25 July, 2018.

Abstract

A fast and sensitive HPLC coupled with APCI-3Q mass spectrometer was utilized to assist the quality assessment of podophyllotoxin from Podophyllum hexandrum. More than ten peaks were separated and detected on a SeQuant TM ZIC®-HILIC LC column along with an Oasis MCX 96well solid-phase extraction cartridge with 35 min. P hexandrum phytoextract was monitored by a APCI-3Q 5500 mass spectrometer with both positive and negative APCI ionization modes. Multiple reaction monitoring was used for quantification using the precursor to product ion pairs of m/z 414.4 to 437.5 adduct with Sodium, m/z 414.4 to 432.4 adduct with water and m/z 414.4 to 828.8 dimer. The assay had a calibration range from 0.2 to 5 μg/ml and a lower limit of quantification of 0.68 μg/ml. The inter- and intra-day precisions were <0.4 per cent and the accuracies were within the range of 99.9–100.1 per cent. The mean recovery ranged from 82.0 to 98.7 per cent and internal standard normalized matrix effect from 0.813 to 0.899. The analyte was stable under all tested conditions. This ICH Q2b validated method can successfully be employed in the absence of reference standards for the marker and is particularly useful in view of the scarcity of the chemical standard of podophyllotoxin.

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Keywords

Podophyllotoxin (PPT), Podophyllum hexandrun, ICH, APCI-3Q, LC-MS/MS.

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