Planta Med 2016; 82(01/02): 113-120
DOI: 10.1055/s-0035-1558115
Pharmacokinetic Investigations
Original Papers
Georg Thieme Verlag KG Stuttgart · New York

A Simple and Sensitive LC-MS/MS Method for Determination of Four Major Active Diterpenoids from Andrographis paniculata in Human Plasma and Its Application to a Pilot Study

Nanthanit Pholphana
Laboratory of Pharmacology, Chulabhorn Research Institute, Bangkok, Thailand
,
Duangchit Panomvana
Translational Research Unit (TRU), Chulabhorn Research Institute, Bangkok, Thailand
,
Nuchanart Rangkadilok
Laboratory of Pharmacology, Chulabhorn Research Institute, Bangkok, Thailand
Environmental Toxicology Program, Chulabhorn Graduate Institute, Bangkok, Thailand
,
Tawit Suriyo
Laboratory of Pharmacology, Chulabhorn Research Institute, Bangkok, Thailand
,
Teerapat Ungtrakul
Chulabhorn Hospital, Bangkok, Thailand
,
Wanwisa Pongpun
Chulabhorn Hospital, Bangkok, Thailand
,
Saichit Thaeopattha
Translational Research Unit (TRU), Chulabhorn Research Institute, Bangkok, Thailand
,
Jutamaad Satayavivad
Laboratory of Pharmacology, Chulabhorn Research Institute, Bangkok, Thailand
Environmental Toxicology Program, Chulabhorn Graduate Institute, Bangkok, Thailand
Center of Excellence on Environmental Health and Toxicology, Office of Higher Education Commission, Ministry of Education, Bangkok, Thailand
› Author Affiliations
Further Information

Publication History

received 03 June 2015
revised 21 August 2015

accepted 29 August 2015

Publication Date:
17 November 2015 (eFirst)

Abstract

Andrographis paniculata contains four major active diterpenoids, including andrographolide (1), 14-deoxy-11, 12-didehydroandrographolide (2), neoandrographolide (3), and 14-deoxyandrographolide (4), which exhibit differences in types and/or degrees of their pharmacological activity. Previous pharmacokinetic studies in humans reported only the parameters of compound 1 and its analytical method in human plasma. The purpose of this study was to develop a simple, sensitive, and selective liquid chromatography tandem-mass spectrometry technique for the simultaneous determination of all four major active diterpenoids in the A. paniculata product in human plasma. These four diterpenoids in plasma samples were extracted by a simple protein precipitation method with methanol and separated on a Kinetex C18 column using a gradient system with a mobile phase of acetonitrile and water. The liquid chromatography tandem-mass spectrometry was performed in the negative mode, and the multiple reaction monitoring mode was used for the quantitation. The method showed a good linearity over a wide concentration range of 2.50–500 ng/mL for 1 and over the range of 1.00–500 ng/mL for the other diterpenoids with a correlation coefficient R2 > 0.995. The lower limit of quantification of 1 was found to be 2.50 ng/mL, while those of the other diterpenoids were 1.00 ng/mL. The intraday and interday accuracy (relative error) ranged from 0.03 % to 10.03 %, and the intraday and interday precisions (relative standard deviation) were in the range of 2.05–9.67 %. The extraction recovery (86.54–111.56 %) with a relative standard deviation of 2.78–8.61 % and the matrix effect (85.15–112.36 %) were within the acceptance criteria. Moreover, these four major active diterpenoids were stable in plasma samples at the studied storage conditions with a relative error ≤–9.79 % and a relative standard deviation ≤ 9.26 %. Hence, this present method was successfully validated and used in the pilot study to determine the pharmacokinetic parameters of all four major active diterpenoids in human plasma after multiple oral doses of the A. paniculata product were administered to a healthy, Thai female volunteer.