A Sensitive Method for Determination of Platycodin D in Rat Plasma Using Liquid Chromatography/Tandem Mass Spectrometry and its Application to a Pharmacokinetic Study

Abstract

Platycodin D (PD), a major component isolated from the root of Platycodon grandiflorum, is widely used in traditional Chinese medicine. A sensitive rapid analytical method was established and validated to determine the PD in rat plasma. This method was further applied to assess the pharmacokinetics of PD in rats following administration of a single dose. Liquid chromatography tandem mass spectrometry (LC/MS/MS) in multiple reaction monitoring mode (MRM) was used in the method, and tubeimoside I was used as the internal standard (IS). A simple protein precipitation based on methanol (MeOH) was employed. The combination of a simple sample cleanup and short chromatographic running time (4 min) increased the throughput of the method substantially. The method was validated over the range of 0.5–1000 ng/mL with a correlation coefficient > 0.99. The lower limit of quantification was 0.5 ng/mL for PD in plasma. Intra- and inter-day accuracies for PD were 90–115 % and 96–108 %, respectively, and the inter-day precision was less than 15 %. After a single oral dose of 10 mg/kg of PD, its mean peak plasma concentration (C_max ) was 13.7 ± 4.5 ng/mL at 0.5 h. The area under the plasma concentration-time curve (AUC_0–24 h ) was 35.4 ± 16.1 h·ng/mL, and the elimination half-life (T_1/2 ) was 1.48 ± 0.13 h. In case of intravenous administration of PD at a dosage of 0.5 mg/kg, the area under the plasma concentration-time curve (AUC_0–24 h ) was 2203 ± 258 h · ng/mL, and the elimination half-life (T_1/2 ) was 6.57 ± 0.70 h. Based on the results, the oral bioavailability of PD in rats at 10 mg/kg is 0.079 %.

Key words

Platycodon grandiflorum A. DC - Campanulaceae - platycodin D - tubeimoside I - LC/MS/MS - pharmacokinetics

References

• 1 Choi Y H, Yoo D S, Cha M R, Choi C W, Kim Y S, Choi S U, Lee K R, Ryu S Y. Antiproliferative effects of saponins from the roots of Platycodon grandiflorum on cultured human tumor cells.  J Nat Prod. 2010;  73 1863-1867
• 2 Choi Y H, Yoo D S, Choi C W, Cha M R, Kim Y S, Lee H S, Lee K R, Ryu S Y. Platyconic acid A, a genuine triterpenoid saponin from the roots of Platycodon grandiflorum.  Molecules. 2008;  13 2871-2879
• 3 Fu W W, Dou D Q, Zhao C J, Shimizu N, Pei Y P, Pei Y H, Chen Y J, Takeda T. Triterpenoid saponins from Platycodon grandiflorum.  J Asian Nat Prod Res. 2007;  9 35-40
• 4 Fu W W, Shimizu N, Dou D Q, Takeda T, Fu R, Pei Y H, Chen Y J. Five new triterpenoid saponins from the roots of Platycodon grandiflorum.  Chem Pharm Bull (Tokyo). 2006;  54 557-560
• 5 Kim Y S, Kim J S, Choi S U, Kim J S, Lee H S, Roh S H, Jeong Y C, Kim Y K, Ryu S Y. Isolation of a new saponin and cytotoxic effect of saponins from the root of Platycodon grandiflorum on human tumor cell lines.  Planta Med. 2005;  71 566-568
• 6 Fu W W, Shimizu N, Takeda T, Dou D Q, Chen B, Pei Y H, Chen Y J. New A-ring lactone triterpenoid saponins from the roots of Platycodon grandiflorum.  Chem Pharm Bull (Tokyo). 2006;  54 1285-1287
• 7 Saeki T, Koike K, Nikaido T. A comparative study on commercial, botanical gardens and wild samples of the roots of Platycodon grandiflorum by HPLC analysis.  Planta Med. 1999;  65 428-431
• 8 Arai I, Komatsu Y, Hirai Y, Shingu K, Ida Y, Yamaura H, Yamamoto T, Kuroiwa Y, Sasaki K, Taguchi S. Stimulative effects of saponin from kikyo-to, a Japanese herbal medicine, on pancreatic exocrine secretion of conscious rats.  Planta Med. 1997;  63 419-424
• 9 Li W, Xiang L, Zhang J, Zheng Y N, Han L K, Saito M. A new triterpenoid saponin from the roots of Platycodon grandiflorum.  Chin Chem Lett. 2007;  18 306-308
• 10 Shin C Y, Lee W J, Lee E B, Choi E Y, Ko K H. Platycodin D and D3 increase airway mucin release in vivo and in vitro in rats and hamsters.  Planta Med. 2002;  68 221-225
• 11 Kim Y P, Lee E B, Kim S Y, Li D, Ban H S, Lim S S, Shin K H, Ohuchi K. Inhibition of prostaglandin E2 production by platycodin D isolated from the root of Platycodon grandiflorum.  Planta Med. 2001;  67 362-364
• 12 Lee H, Kang R, Kim Y S, Chung S I, Yoon Y. Platycodin D inhibits adipogenesis of 3T3-L1 cells by modulating Kruppel-like factor 2 and peroxisome proliferator-activated receptor gamma.  Phytother Res. 2009;  24 S161-S167
• 13 Zheng J, He J, Ji B, Li Y, Zhang X. Antihyperglycemic effects of Platycodon grandiflorum (Jacq.) A. DC. extract on streptozotocin-induced diabetic mice.  Plant Foods Hum Nutr. 2007;  62 7-11
• 14 Kim J Y, Park K W, Moon K D, Lee M K, Choi J, Yee S T, Shim K H, Seo K I. Induction of apoptosis in HT-29 colon cancer cells by crude saponin from Platycodi Radix.  Food Chem Toxicol. 2008;  46 3753-3758
• 15 Ahn K S, Noh E J, Zhao H L, Jung S H, Kang S S, Kim Y S. Inhibition of inducible nitric oxide synthase and cyclooxygenase II by Platycodon grandiflorum saponins via suppression of nuclear factor-kappaB activation in RAW 264.7 cells.  Life Sci. 2005;  76 2315-2328
• 16 Kim J Y, Hwang Y P, Kim D H, Han E H, Chung Y C, Roh S H, Jeong H G. Inhibitory effect of the saponins derived from roots of Platycodon grandiflorum on carrageenan-induced inflammation.  Biosci Biotechnol Biochem. 2006;  70 858-864
• 17 Zhao H L, Sim J S, Shim S H, Ha Y W, Kang S S, Kim Y S. Antiobese and hypolipidemic effects of platycodin saponins in diet-induced obese rats: evidences for lipase inhibition and calorie intake restriction.  Int J Obes (Lond). 2005;  29 983-990
• 18 Zhao H L, Harding S V, Marinangeli C P, Kim Y S, Jones P J. Hypocholesterolemic and anti-obesity effects of saponins from Platycodon grandiflorum in hamsters fed atherogenic diets.  J Food Sci. 2008;  73 H195-H200
• 19 Lee K J, Choi C Y, Chung Y C, Kim Y S, Ryu S Y, Roh S H, Jeong H G. Protective effect of saponins derived from roots of Platycodon grandiflorum on tert-butyl hydroperoxide-induced oxidative hepatotoxicity.  Toxicol Lett. 2004;  147 271-282
• 20 Kim M O, Moon D O, Choi Y H, Lee J D, Kim N D, Kim G Y. Platycodin D induces mitotic arrest in vitro, leading to endoreduplication, inhibition of proliferation and apoptosis in leukemia cells.  Int J Cancer. 2008;  122 2674-2681
• 21 Kim M O, Moon D O, Choi Y H, Shin D Y, Kang H S, Choi B T, Lee J D, Li W, Kim G Y. Platycodin D induces apoptosis and decreases telomerase activity in human leukemia cells.  Cancer Lett. 2008;  261 98-107
• 22 Ahn K S, Hahn B S, Kwack K, Lee E B, Kim Y S. Platycodin D-induced apoptosis through nuclear factor-kappaB activation in immortalized keratinocytes.  Eur J Pharmacol. 2006;  537 1-11
• 23 Huang J H, Huang X H, Chen Z Y, Zheng Q S, Sun R Y. Dose conversion among different animals and healthy volunteers in pharmacological study.  Chin J Clin Pharmacol Ther. 2004;  9 1069-1072
• 24 Ye J, Xiao M T, Tang X C, Huang Y Y. HPLC-ELSD for determining the content of three platycodins in Radix Platycodoi.  J Xi'an Jiaotong Univ (Med Sci). 2010;  31 640-642
• 25 Xie Y, Ye Y P, Sun H X, Li D. Contribution of the glycidic moieties to the haemolytic and adjuvant activity of platycodigenin-type saponins from the root of Platycodon grandiflorum.  Vaccine. 2008;  26 3452-3460
• 26 Liu H, Yang J, Du F, Gao X, Ma X, Huang Y, Xu F, Niu W, Wang F, Mao Y, Sun Y, Lu T, Liu C, Zhang B, Li C. Absorption and disposition of ginsenosides after oral administration of Panax notoginseng extract to rats.  Drug Metab Dispos. 2009;  37 2290-2298
• 27 Dai J Y, Yang J L, Li C. Transport and metabolism of flavonoids from Chinese herbal remedy Xiaochaihu- tang across human intestinal Caco-2 cell monolayers.  Acta Pharmacol Sin. 2008;  29 1086-1093
• 28 Xie Y, Deng W, Sun H, Li D. Platycodin D2 is a potential less hemolytic saponin adjuvant eliciting Th1 and Th2 immune responses.  Int Immunopharmacol. 2008;  8 1143-1150
• 29 Xie Y, He S W, Sun H X, Li D. Platycodin D2 improves specific cellular and humoral responses to hepatitis B surface antigen in mice.  Chem Biodivers. 2010;  7 178-185
• 30 Chung J W, Noh E J, Zhao H L, Sim J S, Ha Y W, Shin E M, Lee E B, Cheong C S, Kim Y S. Anti-inflammatory activity of prosapogenin methyl ester of platycodin D via nuclear factor-kappaB pathway inhibition.  Biol Pharm Bull. 2008;  31 2114-2120