Rapid RP-HPLC technique for the determination of phyllanthin as bulk and its quantification in Phyllanthus amarus extract
Keywords:
Phyllanthus amarus, Phyllanthin, HPLC and Plant extractAbstract
A simple, specific and precise high performance liquid chromatographic method has been developed and validated for the estimation of Phyllanthin, an important bitter lignan present in Phyllanthus amarus. Furthermore, the developed method was used to successfully quantify the Phyllanthin in the plant extract also. The mobile phase optimized for the RP-HPLC was ethanol-water 66:34 (%v/v) and that was very simple and cost effective. The detection was carried out using variable wavelength UV–VIS detector set at 229 nm. Linearity for the developed method was found over the concentration range 1–50 μg/ml with a correlation coefficient of 0.999. Method was validated according to ICH guidelines and found to be suitable for rapid quantification of Phyllanthin in bulk and in plant extracts.
References
Thagarajan SP, Thiruneelakantan K,
Subramaniam S, Sundaravelu T. Invitro
inactivation of HBsAg by Eclipta alba and
Phyllanthus niruri Linn. Ind. J. Med. Res.
;76:124-130.
Thagarajan SP, Subramaniam S,
Thirunalasundari T. Effect of Phyllanthusamarus on chronic carriers of Hepatitis B
virus. The Lancet. 1988;11:764-766.
Venkateswaran PS, Millman I, Blumberg
BS. Effect of an extract from Phyllanthus
niruri on Hepatitis B and Woodchuck
Hepatitis viruses: Invitro and invivo
studies, Proc. Natl. Acad. Sci. USA.
;84:274-278.
Mehrotra R, Rawat S, Kulshreshtha DK.
Invitro effect of Phyllanthus amarus on
Hepatitis B virus. Ind. J Med. Res.
;93:71-73.
Jayaram S, Thyagarajan SP. Inhibition of
HBsAg secrection from Alexander cell
line by Phyllanthus amarus. Indian
J.Pathol. Microbiol. 1996;39:211-215.
Yeh SF, Hong CY, Huang YL, Liu TY,
Choo KB, Chou CK. Effect of an extract
from Phyllanthus amarus on Hepatitis B
surface antigen gene expression in human
hepatoma cells. Antiviral Res.
;20:185-192.
Lee CD, Ott M, Thayagarajan SP, Shafritz
DA, Burk RD, Gupta S. Phyllanthus
amarus down regulates Hepatitis B virus
mRNA transcription and replication. Eur.
J. Clin. Invest. 1996;26:1069-1076.
Ott M, Thyagarajan SP, Gupta S.
Phyllanthus amarus suppresses Hepatitis
B virus by interrupting interaction
between HBV enhancer I and cellular
transcription factors. Eur. J. Clin. Invest.
;27:908-915.
Ramakrishnan PN, Murugesan R,
Palanichamy S, Murugesh N. Oral
hypoglycaemic effect of Phyllanthus
niruri Linn. Leaves. Ind.J.Pharmaceu. Sci.
;44:10-12.
Moshi MJ, Uiso FC, Mahunnah R.L. A
study of the effect of Phyllanthus amarus
extract on blood glucose in rabbits. Int J.
Pharmacog. 1997;35:167-173.
Srividya N, Periwal S. Diuretic,
hypotensive and hypoglycaemic effect of
Phyllanthus amarus. Ind. J. Exp. Biol.
;33:861-864.
Sivaprakasam K, Yasodha R, Sivanandam
G, Veluchamy G. Clinical evaluation of
Phyllanthus amarus Schum & Thonn. in
Diabetes Mellitus. Seminar on research in
Ayurveda and Siddha, CCRAS, New
Delhi. 1995;20-22: p17.
Tripathi SC, Patnaik GK, Visen PKS.
Evaluation of hepatoprotective activity of
Phyllanthus amarus against
experimentally induced liver damage in
rats. Proc. 25th Ind. Pharmacol. Soc.
Conf., Muzaffarpur, Bihar, India.
;December 5th - 8th:82.
Sane RT, Kuber VV, Chalissery MS.,
Menon S. Hepatoprotection by
Phyllanthus amarus and Phyllanthus
debili in CCL4 induced liver dysfunction.
Current Science. 1995;68:1243-1246.
Venkateswaran PS, Millman I, Blumberg
BS. Effects of an extract from Phyllanthus
niruri on Hepatitis B and Woodchuck
hepatitis viruses Invitro and Invivo
studies Proc. Natl. Acad. Sci. USA.
;84:274-278.
Calixto JB, Santos AR, Cechinel Filho V,
Yunes RA. A review of the plants of the
genus Phyllanthus: their chemistry.
Pharmacology and therapeutic potential.
Med Res Rev. 1998;18:225-258.
Shakila A, Rajeswari R. Effect ofGA3,
certain chemicals and media on
germination, growth and yield of
Phyllanthus niruri. Plant Arch.
;6:121–125.
ICH Haromonised Tripartite Guideline,
International Conference on
Harmonisation on Technical
Requirements for Registration of
Pharmaceuticals for Human Use, Q2A,
Text on Validation of Analytical
Procedures, Step 4 of the ICH process,
and Q2B, Validation of Analytical
Procedures: Methodology, Step 4 of the
ICH process, 1996, ICH Steering
Committee.
