Abstract
A quantitative method using silica gel 60F254 high performance thin layer chromatography plates, automated bandwise sample application, and automated visible mode densitometric method has been developed for the determination of 24β-ethylcholesta-5,22E,25-triene-3β-ol (ECTO) in the aerial part of Clerodendrum phlomidis. ECTO was used as a chemical marker for the standardization of C. phlomidis plant extracts. The separation was performed on silica gel 60F254 TLC plates using chloroform-methanol (98.5: 1.5, v/v) as mobile phase. The quantitation of ECTO was carried out using the densitometric reflection/absorption mode at 650 nm after post chromatographic derivatization with anisaldehyde reagent. A precise and accurate quantification can be performed in the linear working concentration range of 150–400 ng band−1 with good correlation (r 2 = 0.996). The method was validated for peak purities, precision, robustness, limit of detection (LOD) and quantitation (LOQ), etc. as per ICH guidelines.



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References
Chopra RN, Nayar SL, Chopra IC (1956) Glossary of Indian medicinal plants. CSIR, New Delhi, p 71
Nadkarni AK (1976) Indian materia medica. Popular Prakashan, Bombay, India
Krishnamurthy KH, Masilamoney P, Govindraj N (1972) J Res Ind Med 7:27–36
Rajasekaran A, Ponnusamy K (2006) Turk J Biol 30:139–142
Garg SC, Siddiqui N (1992) J Res Edu Indian Med 11:1–3
Kole RK, Chowdhary A (1994) Pesticide Res J 6:26–33
Bhattacharya SK, Bajpai HS (1975) J Res Indian Med 10:1–6
Rani S, Ahmed N, Rajaram S, Sluja R, Thenmozhi S, Murugesan T (1999) J Ethnopharmacol 68:315–319
Pandey, R (2006) Chemical investigation of some indigenous plant. Ph.D. Thesis, Dr. Ram Manohar. Lohia Avadh University, Faizabad, India
Akihisa T, Ghosh P, Thakur S, Oshikiri S, Tamura T, Matsumoto T (1988) Phytochemistry 27:241–244
Akihisa T, Ghosh P, Thakur S, Nagata H, Tamura T, Matsumoto T (1990) Phytochemistry 29:1639–1641
Akihisa T, Tamura T, Matsumoto T, Kokke WCMC, Ghosh P, Thakur S (1990) J Chem Soc Perkin Transcription India 2213–2218
Pandey R, Verma RK, Gupta MM (2007) J Sep Sci (in press)
Abidi SL (2001) J Chromatogr A 935:173–201
Volin P (2001) J Chromatogr A 935:125–140
Sánchez-Machado DI, López-Hernández J, Paseiro-Losada P, López-Cervantes J (2004) Biomed Chromatogr 18:183–190
Larsen T, Axelsen J, Ravn HW (2004) J Chromatogr A 1026:301–304
Reich E, Schibli A (2007) A high-performance thin-layer chromatography for the analysis of medicinal plants. Thieme Medical Publishers, New York, p 13
Subramanian SS, Nair AGR, Vedantham TNC (1973) Phytochemistry 12:2078–2079
Rehman AU, Begum S, Saied S, Choudhary MI, Akhtar F (1997) Phytochemistry 45:1721–1722
ICH (Q2A) (1994) Note for guidance on validation of analytical methods: definitions and terminology. International Conference on Harmonisation, IFPMA, Geneva
ICH (Q2B) (1996) Note for guidance on validation of analytical procedure: methodology. International Conferences on Harmonisation, IFPMA, Geneva
Acknowledgments
We are thankful to the Director, CIMAP for his keen interest during the course of work and Council of Scientific and Industrial Research (CSIR) and Department of AYUSH, New Delhi for providing financial supports in the form of Golden Triangle Project.
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Shanker, K., Singh, S.C., Pant, S. et al. Quantitative TLC Analysis of Sterol (24β-Ethylcholesta-5,22E,25-triene-3β-ol) in Agnimantha (Clerodendrum phlomidis Linn). Chroma 67, 269–274 (2008). https://doi.org/10.1365/s10337-007-0502-5
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DOI: https://doi.org/10.1365/s10337-007-0502-5


