http://informahealthcare.com/xen ISSN: 0049-8254 (print), 1366-5928 (electronic) Xenobiotica, 2014; 44(9): 855–860 ! 2014 Informa UK Ltd. DOI: 10.3109/00498254.2014.899407
RESEARCH ARTICLE
Comparative pharmacokinetic study of four major components after oral administration of pure compounds, herbs and Si–Ni–San to rats Jing Wen, Yanjuan Wang, Lina Yang, Weihua Zheng, Longshan Zhao, and Famei Li
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Department of Analytical Chemistry, Shenyang Pharmaceutical University, Shenyang, China
Abstract
Keywords
1. The pharmacokinetic differences of paeoniflorin, naringin, naringenin and glycyrrhetinic acid (GA) following oral administration of pure compounds, single herbs and Si–Ni–San (SNS) decoction to rats were studied. Blood samples were analyzed with a validated UPLC–MS/MS method. Student’s t-test was used for the statistical comparison. 2. The Cmax and AUC0–1 were 1470 ± 434 ng/mL and 4663 ± 916 ng h/mL for paeoniflorin, 64.29 ± 59.21 ng/mL and 311.8 ± 131.8 ng h/mL for naringin, 244.2 ± 138.8 ng/mL and 4761 ± 3167 ng h/mL for naringenin, and 1183 ± 294 ng/mL and 38 994 ± 14 377 ng h/mL for GA after oral administration of paeoniflorin, naringin and glycyrrhizic acid. The Cmax and AUC0–1 were 812.6 ± 259.6 ng/mL and 2489 ± 817 ng h/mL for paeoniflorin, 344.3 ± 234.9 ng/mL and 1479 ± 531 ng h/mL for naringin, 981.9 ± 465.4 ng/mL and 12 284 ± 6378 ng h/mL for naringenin, and 3164 ± 742 ng/mL and 78 817 ± 16 707 ng h/mL for GA after oral administration of SNS decoction. 3. There were significant differences between the pharmacokinetic behavior after oral administration of SNS decoction compared with pure components or herbs. The results indicated that some components in the other herbs of SNS had a pharmacokinetic interaction with paeoniflorin, naringin, naringenin and GA.
Glycyrrhetinic acid, naringin, naringenin, paeoniflorin, pharmacokinetics, Si–Ni–San, UPLC–MS/MS
Introduction The Chinese compound prescriptions are the main medications of traditional Chinese medicine (TCM) (Wu et al., 2009). Pharmacokinetic studies of active ingredients in herbs or prescriptions are useful to explain and predict the efficacy and toxicity of TCMs. Thus, it is valuable to perform pharmacokinetic studies for exploring the action mechanism and evaluating the compatibility of herbs or prescriptions. Si–Ni–San (SNS), one of the well-known TCM, is composed of Radix Bupleuri, Radix Paeoniae alba, Fructus Aurantii immaturus and Radix Glycyrrhiza with the ratio of 1:1:1:1. Pharmacological studies have shown that SNS had significant therapeutic effect on various experimental liver injury models (Jiang et al., 2003). It has been reported that the activity of SNS was stronger than those of single herbs, which was due to effective constituents in SNS acting on the each target of liver (Jiang & Xu, 2004). Paeoniflorin is one of the main active constituents in Radix Paeoniae, which has been used as a phytochemical marker for the quality control of Radix Paeoniae in Chinese Pharmacopoeia (Song et al., 2010). There are a few reports on the pharmacokinetics of
History Received 9 January 2014 Revised 23 February 2014 Accepted 25 February 2014 Published online 18 March 2014
paeoniflorin in the form of pure paeoniflorin, Paeonia lactiflora extract and prescriptions (Chen et al., 1999, 2002; Gan et al., 2012; Li et al., 2011). Naringin is a flavonoid present in many Citrus fruits and TCMs. Naringenin is the aglycone of naringin. The pharmacokinetics of naringin and naringenin have been reported after oral administration of pure naringin, herbs extract and prescriptions (Fang et al., 2006; Li et al., 2010; Yu et al., 2009). Glycyrrhizic acid (GL) is one of the bioactive constituents in Radix Glycyrrhiza. After orally administered, GL was converted to glycyrrhetinic acid (GA) by intestinal bacteria in rats (Takeda et al., 1996; Wang et al., 1994). The pharmacokinetic studies of GA after oral administration of GL have been reported (Gao et al., 2004; Zhang et al., 2010). Despite the number of pharmacokinetic studies of the four components, there is surprisingly no report on compared results of pharmacokinetics of the four components in pure compounds, herbs and SNS decoction. In this study, comparative pharmacokinetic study in rats after oral administration of pure compounds, single herbs and SNS decoction is researched.
Materials and methods Address for correspondence: Dr Jing Wen, Department of Analytical Chemistry, Shenyang Pharmaceutical University, 39 Mailbox, 103 Wenhua Road, Shenyang 110016, PR China. Tel: +86 24 2398 6290. Fax: +86 24 2398 6289. E-mail: [email protected]
Chemicals The reference standards of paeoniflorin, naringin, GA and lisinopril (internal standard, IS) (purity > 98%) were
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HPLC grade was obtained from Dikma (Richmond Hill, NY). All other reagents were of analytical grade. Instrumentations and analytical conditions The assay was performed on an ACQUITY UPLC–MS system (Waters Corp., Milford, MA) with an electrospray ionization (ESI) interface. An ACQUITY UPLCTM BEH C18 column (100 mm 2.1 mm, 1.7 lm) was employed. The column temperature was maintained at 35 C and a gradient elution with methanol (A)-water (2 mm ammonium acetate) (B) was used. The gradient program was as follow: 0–2.5 min, 25–90% A, 2.5–4.5 min, 90% A. The flow rate was set at 0.25 mL/min. The autosampler was conditioned at 4 C and the injection volume was 10 lL for analysis. The analytes and IS were all ionized by the ESI source in positive ionization mode. Quantification was performed using multiple reaction monitoring of the transitions of m/z 497.9 ! 178.9 for
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purchased from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China). Paeoniflorin (purity > 80%), naringin (purity > 98%) and GL (purity > 98%), which were used to administer to rats, and the reference standard of naringenin (purity > 98%) were purchased from Baozetang Medical Technology Co., Ltd. (Jiangsu, China). The structures of these compounds are shown in Figure 1. Radix Bupleuri was purchased from Northeast Big Pharmacy (Shenyang, China). Radix Paeoniae alba and Fructus Aurantii immaturus were purchased from Weikang Big Pharmacy (Shenyang, China). Radix Glycyrrhiza was purchased from Beijing Tongrentang Pharmacy (Shenyang, China). The above four medicinal materials were authenticated by Professor Jincai Lu (College of Traditional Chinese Materia Medica, Shenyang Pharmaceutical University). Methanol of HPLC grade was obtained from Tedia (Fairfield, OH). Ammonium acetate of
Xenobiotica, 2014; 44(9): 855–860
Figure 1. Chemical structures of paeoniflorin, naringin, naringenin, glycyrrhizic acid, glycyrrhetinic acid and lisinopril (IS).
Comparative pharmacokinetic study
DOI: 10.3109/00498254.2014.899407
paeoniflorin, m/z 581.2 ! 273.0 for naringin, m/z 272.9 ! 152.8 for naringenin, m/z 470.9 ! 134.8 for GA and m/z 406.2 ! 246.1 for IS, respectively. The optimal MS parameters obtained were as follows: capillary voltage 3.0 kV; cone voltage for paeoniflorin, naringin, naringenin, GA and IS was 13, 15, 30, 20 and 30 kV, respectively; source temperature 100 C and desolvation temperature 450 C. Nitrogen was used as desolvation and cone gas with a flow rate of 500 and 30 L/h. Argon was used as the collision gas at a pressure of approximately 2.8 103 mbar. The optimized collision energy for paeoniflorin, naringin, naringenin, GA and IS was 20, 18, 25, 37 and 23 eV, respectively. All data collected in centroid mode were processed using MassLynxTM NT 4.1 software with a QuanLynxTM program (Waters Corp., Milford, MA).
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Preparation of Si–Ni–San decoction and single herbs The dried power of Radix Bupleuri (20 g), Radix Paeoniae alba (20 g), Fructus Aurantii immaturus (20 g) and Radix Glycyrrhiza (20 g) were mixed together. They were soaked in water for 0.5 h and then extracted three times with boiling water (1:12) each for 0.5 h, and the solution obtained by filtration was combined and concentrated to about 0.8 g/mL under vacuum at 60 C. The decoction was purified by alcohol precipitation with 95% alcohol (1:3), and then was left to stand overnight. The extract was concentrated under vacuum and then volatilized to dryness in water bath at 80 C. The concentrations of paeoniflorin, naringin and GL in the extract detected by HPLC, were 2.2, 21.5 and 5.8 mg/g (expressed in milligrams of pure chemical components per gram of Si–Ni–San), respectively. Oral administration of paeoniflorin, naringin and GL were given in the same doses. Radix Paeoniae alba, Fructus Aurantii immaturus and Radix Glycyrrhiza were prepared in the same method. Animals and treatments Forty-two male SD rats, weighing 200–240 g, were obtained from the Animal Center of Shenyang Pharmaceutical University (Shenyang, China). All protocols of animal experiments were approved in accordance with the Regulations of Experimental Animal Administration issued by the State Commission of Science and Technology of the People’s Republic of China. The rats were housed under controlled environmental conditions (temperature: 25 ± 2 C, relative humidity: 50 ± 10%, normal day and night cycle) with free access to food until 12 h prior to experiments. The animals had free access to water during the experiment. The rats were given orally with paeoniflorin (88 mg/kg), naringin (863 mg/kg), GL (235 mg/kg), Radix Paeoniae alba (10 g/kg), Fructus Aurantii immaturus (10 g/kg) and Radix Glycyrrhiza (10 g/kg) and SNS decoction (40 g/kg) (at a dose of containing paeoniflorin 88 mg/kg, naringin 863 mg/kg and GL 235 mg/kg). The residue was reconstituted in 5% CMC-Na to get a concentration equivalent to 4 g/mL of SNS. To calculate the administered dose, the contents of Radix Paeoniae alba, Fructus Aurantii immaturus, Radix Glycyrrhiza, paeoniflorin, naringin and GL in administration solution were 1, 1, 1, 0.0088, 0.0863 and 0.0235 g/mL, respectively.
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Animals were randomly divided into seven groups, with six rats in each. Blood samples (0.3 mL) were collected in 1.5 mL heparinized polythene tubes before dosing and 0.17, 0.33, 0.50, 0.75, 1.0, 2.0, 4.0, 6.0, 8.0, 10, 12, 14, 24, 30, 48, 60 and 72 h after dosing. The blood samples were immediately centrifuged at 13 000 rpm for 10 min and the plasma was stored at 20 C until analysis. Analytical method A UPLC–MS/MS method has been developed for the simultaneous determination of paeoniflorin, naringin, naringenin and GA in rat plasma (Wen et al., 2012). The calibration curves of the four analytes were linear in the ranges of 9.800–3920, 5.100–2040, 5.200–2080 and 10.60– 4240 ng/mL, respectively. The LLOQ of the four analytes were 9.800, 5.100, 5.200 and 10.60 ng/mL, respectively, with precision (relative standard deviation, RSD) below 20% and accuracy (relative error, RE) within ± 20%. The intra- and inter-day precision ranged 4.9–12% and 2.8–13%, respectively. The accuracy derived from QC samples was within 7.3–7.5% for three QC levels. The mean extraction recoveries were more than 84.5%. With regard to matrix effect, all the calculated values were between 90.93% and 109.8%. The concentrations of paeoniflorin, naringin, naringenin and GA measured in the stability study were between 89.5% and 102.9% of the initial values, indicating that the analytes in rat plasma were stable for 4 h at room, 15 d at 20 C, three freeze-thaw cycles and 12 h after pretreatment. In vivo pharmacokinetic study The above validated UPLC–MS/MS method was applied to comparative pharmacokinetic study of paeoniflorin, naringin, naringenin and GA in rats after oral administration of the pure compounds, singleherbs and SNS decoction. The maximum plasma concentrations (Cmax) and the time to reach the maximum concentrations (Tmax) were obtained directly from the observed data. The elimination rate constant (ke) was calculated on the slope of the linear regression of logtransformed concentration versus time using the last four measurable points. The elimination half-life (t1/2) was calculated using 0.693/ke. The area under plasma concentration–time curve (AUC0–t) to the last measurable plasma concentration (Ct) was estimated by using the linear trapezoidal rule. The area under the plasma concentration–time curve to time infinity (AUC0–1) was calculated as AUC0–1 ¼ AUC0–t + Ct/Ke. The mean residence time (MRT) was calculated as AUC0–1/AUC0–1. The plasma concentrations of paeoniflorin, naringin, naringenin and GA at different times were expressed in mean and standard deviation (SD), and the mean concentration–time curves were plotted. The significance of difference between each groups was assessed by unpaired Student’s t-test.
Results The mean plasma concentration–time curves of paeoniflorin, naringin, naringenin and GA after oral administration of pure compounds, single herbs and SNS decoction are presented in Figures 2–5. The corresponding pharmacokinetic parameters
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Figure 2. The mean ( ± SD, n ¼ 6) plasma concentration–time curves of paeoniflorin following oral administration of paeoniflorin, Radix Paeoniae alba and SNS decoction.
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Figure 5. The mean ( ± SD, n ¼ 6) plasma concentration–time curves of glycyrrhetinic acid following oral administration of glycyrrhizic acid, Radix Glycyrrhiza and SNS decoction.
Table 1. Pharmacokinetic parameters of paeoniflorin after oral administration of paeoniflorin, Radix Paeoniae alba and Si–Ni–San decoction to rats (mean ± SD, n ¼ 6).
Parameter
Paeoniflorin
Radix Paeoniae alba
Si–Ni–San decoction
Cmax (ng/mL) Tmax (h) t1/2 (h) AUC0–t (ng h/mL) AUC0–1 (ng h/mL) MRT0–1 (h)
1470 ± 434 0.75 ± 0.22 1.73 ± 0.29 4599 ± 929 4663 ± 916 2.96 ± 0.42
657.7 ± 139.7** 0.92 ± 0.13 2.59 ± 0.74* 2599 ± 469** 2719 ± 510** 3.98 ± 0.51**
812.6 ± 259.6** 0.70 ± 0.11m 3.19 ± 1.54 2282 ± 652** 2489 ± 817** 4.11 ± 2.34
*p50.05, **p50.01, versus oral administration of paeoniflorin; mp50.05, versus oral administration of Radix Paeoniae alba. Figure 3. The mean ( ± SD, n ¼ 6) plasma concentration–time curves of naringin following oral administration of naringin, Fructus Aurantii immaturus and SNS decoction.
Table 2. Pharmacokinetic parameters of naringin after oral administration of naringin, Fructus Aurantii immaturus and Si–Ni–San decoction to rats (mean ± SD, n ¼ 6).
Parameter Cmax (ng/mL) Tmax (h) t1/2 (h) AUC0–t (ng h/mL) AUC0–1 (ng h/mL) MRT0–1 (h)
Naringin 64.29 ± 59.21 0.71 ± 0.24 7.95 ± 2.05 230.3 ± 94.6 311.8 ± 131.8 10.26 ± 3.22
Fructus Aurantii immaturus
Si–Ni–San decoction
53.01 ± 25.04 344.3 ± 234.9* m 0.80 ± 0.21 0.95 ± 0.59 10.95 ± 3.00 5.76 ± 2.12m 276.6 ± 40.2 1423 ± 550** mm 465.9 ± 130.9 1479 ± 531** m 15.37 ± 4.42 10.02 ± 1.62m
*p50.05, **p50.01, versus oral administration of naringin; m p50.05, mmp50.01, versus oral administration of Fructus Aurantii immaturus.
Figure 4. The mean ( ± SD, n ¼ 6) plasma concentration–time curves of naringenin following oral administration of naringin, Fructus Aurantii immaturus and SNS decoction.
are shown in Tables 1–4. The Cmax and AUC0–1 of paeoniflorin were 1470 ± 434 ng/mL and 4663 ± 916 ng h/ mL after oral administration of paeoniflorin, and were 812.6 ± 259.6 ng/mL and 2489 ± 817 ng h/mL after oral administration of SNS decoction, respectively. The Cmax and AUC0–1 of naringin were 64.29 ± 59.21 ng/mL and 311.8 ± 131.8 ng h/mL after oral administration of naringin and were 344.3 ± 234.9 ng/mL and 1479 ± 531 ng h/mL after
Table 3. Pharmacokinetic parameters of naringenin after oral administration of naringin, Fructus Aurantii immaturus and Si–Ni–San decoction to rats (mean ± SD, n ¼ 6).
Parameter Cmax (ng/mL) Tmax (h) t1/2 (h) AUC0–t (ng h/mL) AUC0–1 (ng h/mL) MRT0–1 (h)
Naringin
Fructus Aurantii immaturus
Si–Ni–San decoction
244.2 ± 138.8 730.1 ± 190.3** 981.9 ± 465.4** 19.00 ± 5.48 13.60 ± 0.89 13.6 ± 0.89 5.58 ± 1.81 4.91 ± 1.23 5.00 ± 1.46 4695 ± 3152 11 408 ± 2355** 12 242 ± 6381* 4761 ± 3167 11 479 ± 2372** 12 284 ± 6378* 21.25 ± 1.47 17.49 ± 2.34** 16.25 ± 1.64**
*p50.05, **p50.01, versus oral administration of naringin.
Comparative pharmacokinetic study
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oral administration of SNS decoction, respectively. The Cmax and AUC0–1 of naringenin were 244.2 ± 138.8 ng/mL and 4761 ± 3167 ng h/mL after oral administration of naringin, and were 981.9 ± 465.4 ng/mL and 12 284 ± 6378 ng h/mL after oral administration of SNS decoction, respectively. The Cmax and AUC0–1 of GA were 1183 ± 294 ng/mL and 38 994 ± 14 377 ng h/mL after oral administration of GL, and were 3164 ± 742 ng/mL and 78 817 ± 16 707 ng h/mL after oral administration of SNS decoction, respectively.
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Discussion It was reported that the bioavailability of paeoniflorin in rats after oral administration was very low (Takeda et al., 1995). The extremely low bioavailability was resulted from its poor absorption in the intestine (Chen et al., 2011). Its aglycones are connected with sugar moieties, and some literatures have reported that the presence of sugar moieties could affect solubility and permeability, which might lead to the low intrinsic permeability of compounds in the intestine (Chen et al., 2008). As shown in Table 1, the Cmax and AUC of paeoniflorin were lower after oral administration of Radix Paeoniae alba or SNS decoction than those for same amount of paeoniflorin. These data indicated that the total systemic exposure level of paeoniflorin was lower in the Radix Paeoniae alba group or SNS decoction group than those in the Paeoniflorin group. These results implied that some components in Radix Paeoniae alba or in the other herb of SNS might have pharmacokinetic interactions with paeoniflorin to decrease the total systemic exposure level in vivo. It was reported that GA can induce the functions of p-glycoprotein (p-gp) and CYP3A (Hou et al., 2012), and p-gp-mediated efflux inhibits the intestinal absorption of paeoniflorin (Liu et al., 2006). The major part of paeoniflorin is metabolized or transformed to paeoniflorgenin by intestinal bacterial or b-glucosidase (Xu et al., 2013). The transformation of paeoniflorin to its metabolites might be increased by other components in Radix Paeoniae alba or SNS decoction. In our results, naringin was quickly absorbed and also quickly eliminated in the body. And the plasma profile displayed bimodal phenomenon after oral administration SNS decoction (Figure 3). It was confirmed that the ability of rapid absorption of naringin resulted in the appearance of the first peak. Bimodal phenomenon of naringin was due to the enterohepatic circulation in rats. Naringin could not be detected 14 h after oral administration of pure naringin or Fructus Aurantii immaturus. The Cmax and AUC of naringin were higher after oral administration of SNS decoction than those of naringin or Fructus Aurantii immaturus. These results implied that some components in the other herbs of SNS, but not in Fructus Aurantii immaturus, might reinforce the absorption of naringin. First, it has been reported that some surfactants such as Tween-80 can open the tight junction of intestinal epithelial cells to promote the absorption of drugs (Bittner et al., 2002). There were different kinds of saponins, such as saikosaponin, which has surfactant activity in SNS. It seems that saponins might promote the absorption of naringin. Second, poor permeation, p-gp-mediated efflux and
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Table 4. Pharmacokinetic parameters of glycyrrhetinic acid after oral administration of glycyrrhizic acid, Radix Glycyrrhiza and Si–Ni–San decoction to rats (mean ± SD, n ¼ 6).
Parameter
Glycyrrhizic acid
Radix Glycyrrhiza
Si–Ni–San decoction
Cmax (ng/mL) 1183 ± 294 3118 ± 730** 3164 ± 742** Tmax (h) 11.67 ± 0.81 24.00 ± 0.00** 27.60 ± 3.28** m t1/2 (h) 10.78 ± 2.05 12.07 ± 2.63 9.40 ± 1.15 AUC0–t 37 947 ± 13 894 66 029 ± 18 608* 77 480 ± 16 674** (ng h/mL) 38 994 ± 14 377 67 407 ± 19 555* 78 817 ± 16 707** AUC0–1 (ng h/mL) 29.94 ± 2.45 25.57 ± 3.15* 32.85 ± 4.07m MRT0–1 (h) *p50.05, **p50.01, versus oral administration of glycyrrhizic acid; m p50.05, versus oral administration of Radix Glycyrrhiza.
hydrolysis via a glucosidase contributed to the poor bioavailability of naringin and inhibitors of p-gp can enhance the oral bioavailability and intestinal permeability. The inhibitors in SNS could result in the increase of plasma concentration of naringin. Naringenin must be absorbed as the aglycone after cleavage of naringin by intestinal bacteria, thus naringenin was produced slowly in the body (Figure 4). Naringin, after absorption, must be hydrolyzed to naringenin or other naringin disaccharides. Then, naringenin could be conjugated with glucuronic acid after oral administration of SNS decoction (Qiao et al., 2012). The Cmax and AUC of naringenin were higher after oral administration of Fructus Aurantii immaturus or SNS decoction than those of naringin. This phenomenon indicated that other components in Fructus Aurantii immaturus or SNS decoction may increase the transformation of naringin to its metabolite naringenin. GL generated GA by taking off two molecules from glucuronic acid. GL was transformed to GA by intestinal bacteria. The Cmax, Tmax and AUC of GA were higher after oral administration of Radix Glycyrrhiza or SNS decoction than those of GL (Table 4). This phenomenon indicated that other components in Radix Glycyrrhiza increased the transformation of GL to GA, and the other components in the other herbs of SNS extended the onset time.
Conclusions In conclusion, some herbs or components in TCM may be substrates, inhibitors or inducers of cytochrome P450 and thus may affect the pharmacokinetic of each other. Interactions occurred in the prescription resulting in different pharmacokinetics of pure compounds, herbs and Si–Ni–San. There were statistically significant differences in pharmacokinetic parameters of paeoniflorin, naringin, naringenin and GA following oral administration of pure compounds, single herbs and SNS decoction. The main explanation for these differences seems to be that the interactions occurred in the prescription. It is demonstrated for the first time that these four components have different pharmacokinetic behaviors after oral administration of pure compounds, single herbs and SNS decoction, which reveals the potential compatibility mechanism of TCMs.
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Declaration of interest The authors declare no conflict of interest. This work is supported by grant from the National Foundation of Natural Sciences of China (no. 81102786).
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References Bittner B, Guenzi A, Fullhardt P, et al. (2002). Improvement of the bioavailability of colchicine in rats by co-administration of d-alphatocopherol polyethylene glycol 1000 succinate and a polyethoxylated derivative of 12-hydroxy-stearic acid. Arzneimittelforschung 52: 684–8. Chen LC, Lee MH, Chou MH, et al. (1999). Pharmacokinetic study of paeoniflorin in mice after oral administration of Paeoniae Radix extract. J Chromatogr B 735:33–40. Chen LC, Chou MH, Lin MF, Yang LL. (2002). Pharmacokinetics of paeoniflorin after oral administration of Shao-yao Gan-chao Tang in mice. Jpn J Pharmacol 88:250–5. Chen Y, Wang JY, Yuan L, et al. (2011). Interaction of the main components from the traditional Chinese drug pair Chaihu-Shaoyao based on rat intestinal absorption. Molecules 16:9600–10. Chen Y, Zhao YH, Jia XB, Hu M. (2008). Intestinal absorption mechanisms of prenylated flavonoids present in the heat-processed Epimedium koreanum Nakai (Yin Yanghuo). Pharm Res 25:2190–9. Fang T, Wang Y, Ma Y, et al. (2006). A rapid LC/MS/MS quantitation assay for naringin and its two metabolites in rats plasma. J Pharm Biomed Anal 40:454–9. Gan PP, Zhong MZ, Huang X, et al. (2012). Pharmacokinetic comparisons of albiflorin and paeoniflorin after oral administration of Shaoyao-Gancao-Tang and single herb Paeony decoction to rats. Planta Med 78:237–43. Gao QT, Chen XH, Bi KS. (2004). Comparative pharmacokinetic behavior of glycyrrhetic acid after oral administration of glycyrrhizic acid and Gancao-Fuzi-Tang. Biol Pharm Bull 27:226–8. Hou YC, Lin SP, Chao PDL. (2012). Liquorice reduced cyclosporine bioavailability by activating p-glycoprotein and CYP3A. Food Chem 135:2307–12. Jiang JY, Zhou C, Xu Q. (2003). Alleviating effects of Si–Ni–San, a traditional Chinese prescription, on experimental liver injury and its mechanisms. Biol Pharm Bull 26:1089–94. Jiang JY, Xu Q. (2004). Effect of Si–Ni–San and its individual component drugs on experimental liver injury. Chin J Nat Med 2: 45–9.
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Li YF, Wang M, Wang XY, et al. (2011). Pharmacokinetic properties of albiflorin and paeoniflorin after oral administration of pure compound, Radix Paeoniae alba extract and Danggui-Shaoyao-San extract to rats. J Asian Nat Prod Res 13:117–27. Li XH, Xiong ZL, Lu S, et al. (2010). Pharmacokinetics of naringin and its metabolite naringenin in rats after oral administration of Rhizoma Drynariae extract assayed by UPLC–MS/MS. Chin J Nat Med 8:40–6. Liu Z, Jiang Z, Liu L, Hu M. (2006). Mechanisms responsible for poor oral bioavailability of paeoniflorin: role of intestinal disposition and interactions with sinomenine. Pharm Res 23:2768–80. Qiao Y, Wen J, Song Y, et al. (2012). Study on in vitro and in vivo material base of Si–Ni–San by UPLC–PDA–MS/MS. Chin J Chin Mater Med 37:1672–6. Song J, Wang XD, Zhao JN, et al. (2010). Influence on experimental liver injury of extraction from Peony and Licorice decoction. Pharmacol Clin Chin Mater Med 26:40–2. Takeda S, Ishihara K, Wakui Y, et al. (1996). Bioavailability study of glycyrrhetic acid after oral administration of glycyrrhizin in rats: relevance to the intestinal bacterial hydrolysis. J Pharm Pharmacol 48: 902–5. Takeda S, Isono T, Wakui Y, et al. (1995). Absorption and excretion of paeoniflorin in rats. J Pharm Pharmacol 47:1036–40. Wang Z, Kurosaki Y, Nakayama T, Kimura T. (1994). Mechanism of gastrointestinal absorption of glycyrrhizin in rats. Biol Pharm Bull 17: 1399–403. Wen J, Qiao Y, Yang J, et al. (2012). UPLC–MS/MS determination of paeoniflorin, naringin, naringenin and glycyrrhetinic acid in rat plasma and its application to a pharmacokinetic study after oral administration of Si–Ni–San decoction. J Pharm Biomed Anal 66: 271–7. Wu H, Zhu ZY, Zhang GQ, et al. (2009). Comparative pharmacokinetic study of paeoniflorin after oral administration of pure paeoniflorin, extract of Cortex Moutan and Shuang-Dan prescription to rats. J Ethnopharmacol 125:444–9. Xu CH, Wang P, Wang Y, et al. (2013). Pharmacokinetic comparisons of two different combinations of Shaoyao-Gancao decoction in rats: competing mechanisms between paeoniflorin and glycyrrhetinic acid. J Ethnopharmacol 149:443–52. Yu Q, Xiang J, Tang W, et al. (2009). Simultaneous determination of the 10 major components of Da-Cheng-Qi decoction in dog plasma by liquid chromatography tandem mass spectrometry. J Chromatogr B 877:2025–31. Zhang QF, Chen XH, Yi LX, et al. (2010). Pharmacokinetics of 2 epimers of glycyrrhetic acid in rats. Cent South Pharm 8:350–3.
RESEARCH ARTICLE
Comparative pharmacokinetic study of four major components after oral administration of pure compounds, herbs and Si–Ni–San to rats Jing Wen, Yanjuan Wang, Lina Yang, Weihua Zheng, Longshan Zhao, and Famei Li
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Department of Analytical Chemistry, Shenyang Pharmaceutical University, Shenyang, China
Abstract
Keywords
1. The pharmacokinetic differences of paeoniflorin, naringin, naringenin and glycyrrhetinic acid (GA) following oral administration of pure compounds, single herbs and Si–Ni–San (SNS) decoction to rats were studied. Blood samples were analyzed with a validated UPLC–MS/MS method. Student’s t-test was used for the statistical comparison. 2. The Cmax and AUC0–1 were 1470 ± 434 ng/mL and 4663 ± 916 ng h/mL for paeoniflorin, 64.29 ± 59.21 ng/mL and 311.8 ± 131.8 ng h/mL for naringin, 244.2 ± 138.8 ng/mL and 4761 ± 3167 ng h/mL for naringenin, and 1183 ± 294 ng/mL and 38 994 ± 14 377 ng h/mL for GA after oral administration of paeoniflorin, naringin and glycyrrhizic acid. The Cmax and AUC0–1 were 812.6 ± 259.6 ng/mL and 2489 ± 817 ng h/mL for paeoniflorin, 344.3 ± 234.9 ng/mL and 1479 ± 531 ng h/mL for naringin, 981.9 ± 465.4 ng/mL and 12 284 ± 6378 ng h/mL for naringenin, and 3164 ± 742 ng/mL and 78 817 ± 16 707 ng h/mL for GA after oral administration of SNS decoction. 3. There were significant differences between the pharmacokinetic behavior after oral administration of SNS decoction compared with pure components or herbs. The results indicated that some components in the other herbs of SNS had a pharmacokinetic interaction with paeoniflorin, naringin, naringenin and GA.
Glycyrrhetinic acid, naringin, naringenin, paeoniflorin, pharmacokinetics, Si–Ni–San, UPLC–MS/MS
Introduction The Chinese compound prescriptions are the main medications of traditional Chinese medicine (TCM) (Wu et al., 2009). Pharmacokinetic studies of active ingredients in herbs or prescriptions are useful to explain and predict the efficacy and toxicity of TCMs. Thus, it is valuable to perform pharmacokinetic studies for exploring the action mechanism and evaluating the compatibility of herbs or prescriptions. Si–Ni–San (SNS), one of the well-known TCM, is composed of Radix Bupleuri, Radix Paeoniae alba, Fructus Aurantii immaturus and Radix Glycyrrhiza with the ratio of 1:1:1:1. Pharmacological studies have shown that SNS had significant therapeutic effect on various experimental liver injury models (Jiang et al., 2003). It has been reported that the activity of SNS was stronger than those of single herbs, which was due to effective constituents in SNS acting on the each target of liver (Jiang & Xu, 2004). Paeoniflorin is one of the main active constituents in Radix Paeoniae, which has been used as a phytochemical marker for the quality control of Radix Paeoniae in Chinese Pharmacopoeia (Song et al., 2010). There are a few reports on the pharmacokinetics of
History Received 9 January 2014 Revised 23 February 2014 Accepted 25 February 2014 Published online 18 March 2014
paeoniflorin in the form of pure paeoniflorin, Paeonia lactiflora extract and prescriptions (Chen et al., 1999, 2002; Gan et al., 2012; Li et al., 2011). Naringin is a flavonoid present in many Citrus fruits and TCMs. Naringenin is the aglycone of naringin. The pharmacokinetics of naringin and naringenin have been reported after oral administration of pure naringin, herbs extract and prescriptions (Fang et al., 2006; Li et al., 2010; Yu et al., 2009). Glycyrrhizic acid (GL) is one of the bioactive constituents in Radix Glycyrrhiza. After orally administered, GL was converted to glycyrrhetinic acid (GA) by intestinal bacteria in rats (Takeda et al., 1996; Wang et al., 1994). The pharmacokinetic studies of GA after oral administration of GL have been reported (Gao et al., 2004; Zhang et al., 2010). Despite the number of pharmacokinetic studies of the four components, there is surprisingly no report on compared results of pharmacokinetics of the four components in pure compounds, herbs and SNS decoction. In this study, comparative pharmacokinetic study in rats after oral administration of pure compounds, single herbs and SNS decoction is researched.
Materials and methods Address for correspondence: Dr Jing Wen, Department of Analytical Chemistry, Shenyang Pharmaceutical University, 39 Mailbox, 103 Wenhua Road, Shenyang 110016, PR China. Tel: +86 24 2398 6290. Fax: +86 24 2398 6289. E-mail: [email protected]
Chemicals The reference standards of paeoniflorin, naringin, GA and lisinopril (internal standard, IS) (purity > 98%) were
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HPLC grade was obtained from Dikma (Richmond Hill, NY). All other reagents were of analytical grade. Instrumentations and analytical conditions The assay was performed on an ACQUITY UPLC–MS system (Waters Corp., Milford, MA) with an electrospray ionization (ESI) interface. An ACQUITY UPLCTM BEH C18 column (100 mm 2.1 mm, 1.7 lm) was employed. The column temperature was maintained at 35 C and a gradient elution with methanol (A)-water (2 mm ammonium acetate) (B) was used. The gradient program was as follow: 0–2.5 min, 25–90% A, 2.5–4.5 min, 90% A. The flow rate was set at 0.25 mL/min. The autosampler was conditioned at 4 C and the injection volume was 10 lL for analysis. The analytes and IS were all ionized by the ESI source in positive ionization mode. Quantification was performed using multiple reaction monitoring of the transitions of m/z 497.9 ! 178.9 for
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purchased from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China). Paeoniflorin (purity > 80%), naringin (purity > 98%) and GL (purity > 98%), which were used to administer to rats, and the reference standard of naringenin (purity > 98%) were purchased from Baozetang Medical Technology Co., Ltd. (Jiangsu, China). The structures of these compounds are shown in Figure 1. Radix Bupleuri was purchased from Northeast Big Pharmacy (Shenyang, China). Radix Paeoniae alba and Fructus Aurantii immaturus were purchased from Weikang Big Pharmacy (Shenyang, China). Radix Glycyrrhiza was purchased from Beijing Tongrentang Pharmacy (Shenyang, China). The above four medicinal materials were authenticated by Professor Jincai Lu (College of Traditional Chinese Materia Medica, Shenyang Pharmaceutical University). Methanol of HPLC grade was obtained from Tedia (Fairfield, OH). Ammonium acetate of
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Figure 1. Chemical structures of paeoniflorin, naringin, naringenin, glycyrrhizic acid, glycyrrhetinic acid and lisinopril (IS).
Comparative pharmacokinetic study
DOI: 10.3109/00498254.2014.899407
paeoniflorin, m/z 581.2 ! 273.0 for naringin, m/z 272.9 ! 152.8 for naringenin, m/z 470.9 ! 134.8 for GA and m/z 406.2 ! 246.1 for IS, respectively. The optimal MS parameters obtained were as follows: capillary voltage 3.0 kV; cone voltage for paeoniflorin, naringin, naringenin, GA and IS was 13, 15, 30, 20 and 30 kV, respectively; source temperature 100 C and desolvation temperature 450 C. Nitrogen was used as desolvation and cone gas with a flow rate of 500 and 30 L/h. Argon was used as the collision gas at a pressure of approximately 2.8 103 mbar. The optimized collision energy for paeoniflorin, naringin, naringenin, GA and IS was 20, 18, 25, 37 and 23 eV, respectively. All data collected in centroid mode were processed using MassLynxTM NT 4.1 software with a QuanLynxTM program (Waters Corp., Milford, MA).
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Preparation of Si–Ni–San decoction and single herbs The dried power of Radix Bupleuri (20 g), Radix Paeoniae alba (20 g), Fructus Aurantii immaturus (20 g) and Radix Glycyrrhiza (20 g) were mixed together. They were soaked in water for 0.5 h and then extracted three times with boiling water (1:12) each for 0.5 h, and the solution obtained by filtration was combined and concentrated to about 0.8 g/mL under vacuum at 60 C. The decoction was purified by alcohol precipitation with 95% alcohol (1:3), and then was left to stand overnight. The extract was concentrated under vacuum and then volatilized to dryness in water bath at 80 C. The concentrations of paeoniflorin, naringin and GL in the extract detected by HPLC, were 2.2, 21.5 and 5.8 mg/g (expressed in milligrams of pure chemical components per gram of Si–Ni–San), respectively. Oral administration of paeoniflorin, naringin and GL were given in the same doses. Radix Paeoniae alba, Fructus Aurantii immaturus and Radix Glycyrrhiza were prepared in the same method. Animals and treatments Forty-two male SD rats, weighing 200–240 g, were obtained from the Animal Center of Shenyang Pharmaceutical University (Shenyang, China). All protocols of animal experiments were approved in accordance with the Regulations of Experimental Animal Administration issued by the State Commission of Science and Technology of the People’s Republic of China. The rats were housed under controlled environmental conditions (temperature: 25 ± 2 C, relative humidity: 50 ± 10%, normal day and night cycle) with free access to food until 12 h prior to experiments. The animals had free access to water during the experiment. The rats were given orally with paeoniflorin (88 mg/kg), naringin (863 mg/kg), GL (235 mg/kg), Radix Paeoniae alba (10 g/kg), Fructus Aurantii immaturus (10 g/kg) and Radix Glycyrrhiza (10 g/kg) and SNS decoction (40 g/kg) (at a dose of containing paeoniflorin 88 mg/kg, naringin 863 mg/kg and GL 235 mg/kg). The residue was reconstituted in 5% CMC-Na to get a concentration equivalent to 4 g/mL of SNS. To calculate the administered dose, the contents of Radix Paeoniae alba, Fructus Aurantii immaturus, Radix Glycyrrhiza, paeoniflorin, naringin and GL in administration solution were 1, 1, 1, 0.0088, 0.0863 and 0.0235 g/mL, respectively.
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Animals were randomly divided into seven groups, with six rats in each. Blood samples (0.3 mL) were collected in 1.5 mL heparinized polythene tubes before dosing and 0.17, 0.33, 0.50, 0.75, 1.0, 2.0, 4.0, 6.0, 8.0, 10, 12, 14, 24, 30, 48, 60 and 72 h after dosing. The blood samples were immediately centrifuged at 13 000 rpm for 10 min and the plasma was stored at 20 C until analysis. Analytical method A UPLC–MS/MS method has been developed for the simultaneous determination of paeoniflorin, naringin, naringenin and GA in rat plasma (Wen et al., 2012). The calibration curves of the four analytes were linear in the ranges of 9.800–3920, 5.100–2040, 5.200–2080 and 10.60– 4240 ng/mL, respectively. The LLOQ of the four analytes were 9.800, 5.100, 5.200 and 10.60 ng/mL, respectively, with precision (relative standard deviation, RSD) below 20% and accuracy (relative error, RE) within ± 20%. The intra- and inter-day precision ranged 4.9–12% and 2.8–13%, respectively. The accuracy derived from QC samples was within 7.3–7.5% for three QC levels. The mean extraction recoveries were more than 84.5%. With regard to matrix effect, all the calculated values were between 90.93% and 109.8%. The concentrations of paeoniflorin, naringin, naringenin and GA measured in the stability study were between 89.5% and 102.9% of the initial values, indicating that the analytes in rat plasma were stable for 4 h at room, 15 d at 20 C, three freeze-thaw cycles and 12 h after pretreatment. In vivo pharmacokinetic study The above validated UPLC–MS/MS method was applied to comparative pharmacokinetic study of paeoniflorin, naringin, naringenin and GA in rats after oral administration of the pure compounds, singleherbs and SNS decoction. The maximum plasma concentrations (Cmax) and the time to reach the maximum concentrations (Tmax) were obtained directly from the observed data. The elimination rate constant (ke) was calculated on the slope of the linear regression of logtransformed concentration versus time using the last four measurable points. The elimination half-life (t1/2) was calculated using 0.693/ke. The area under plasma concentration–time curve (AUC0–t) to the last measurable plasma concentration (Ct) was estimated by using the linear trapezoidal rule. The area under the plasma concentration–time curve to time infinity (AUC0–1) was calculated as AUC0–1 ¼ AUC0–t + Ct/Ke. The mean residence time (MRT) was calculated as AUC0–1/AUC0–1. The plasma concentrations of paeoniflorin, naringin, naringenin and GA at different times were expressed in mean and standard deviation (SD), and the mean concentration–time curves were plotted. The significance of difference between each groups was assessed by unpaired Student’s t-test.
Results The mean plasma concentration–time curves of paeoniflorin, naringin, naringenin and GA after oral administration of pure compounds, single herbs and SNS decoction are presented in Figures 2–5. The corresponding pharmacokinetic parameters
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Figure 2. The mean ( ± SD, n ¼ 6) plasma concentration–time curves of paeoniflorin following oral administration of paeoniflorin, Radix Paeoniae alba and SNS decoction.
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Figure 5. The mean ( ± SD, n ¼ 6) plasma concentration–time curves of glycyrrhetinic acid following oral administration of glycyrrhizic acid, Radix Glycyrrhiza and SNS decoction.
Table 1. Pharmacokinetic parameters of paeoniflorin after oral administration of paeoniflorin, Radix Paeoniae alba and Si–Ni–San decoction to rats (mean ± SD, n ¼ 6).
Parameter
Paeoniflorin
Radix Paeoniae alba
Si–Ni–San decoction
Cmax (ng/mL) Tmax (h) t1/2 (h) AUC0–t (ng h/mL) AUC0–1 (ng h/mL) MRT0–1 (h)
1470 ± 434 0.75 ± 0.22 1.73 ± 0.29 4599 ± 929 4663 ± 916 2.96 ± 0.42
657.7 ± 139.7** 0.92 ± 0.13 2.59 ± 0.74* 2599 ± 469** 2719 ± 510** 3.98 ± 0.51**
812.6 ± 259.6** 0.70 ± 0.11m 3.19 ± 1.54 2282 ± 652** 2489 ± 817** 4.11 ± 2.34
*p50.05, **p50.01, versus oral administration of paeoniflorin; mp50.05, versus oral administration of Radix Paeoniae alba. Figure 3. The mean ( ± SD, n ¼ 6) plasma concentration–time curves of naringin following oral administration of naringin, Fructus Aurantii immaturus and SNS decoction.
Table 2. Pharmacokinetic parameters of naringin after oral administration of naringin, Fructus Aurantii immaturus and Si–Ni–San decoction to rats (mean ± SD, n ¼ 6).
Parameter Cmax (ng/mL) Tmax (h) t1/2 (h) AUC0–t (ng h/mL) AUC0–1 (ng h/mL) MRT0–1 (h)
Naringin 64.29 ± 59.21 0.71 ± 0.24 7.95 ± 2.05 230.3 ± 94.6 311.8 ± 131.8 10.26 ± 3.22
Fructus Aurantii immaturus
Si–Ni–San decoction
53.01 ± 25.04 344.3 ± 234.9* m 0.80 ± 0.21 0.95 ± 0.59 10.95 ± 3.00 5.76 ± 2.12m 276.6 ± 40.2 1423 ± 550** mm 465.9 ± 130.9 1479 ± 531** m 15.37 ± 4.42 10.02 ± 1.62m
*p50.05, **p50.01, versus oral administration of naringin; m p50.05, mmp50.01, versus oral administration of Fructus Aurantii immaturus.
Figure 4. The mean ( ± SD, n ¼ 6) plasma concentration–time curves of naringenin following oral administration of naringin, Fructus Aurantii immaturus and SNS decoction.
are shown in Tables 1–4. The Cmax and AUC0–1 of paeoniflorin were 1470 ± 434 ng/mL and 4663 ± 916 ng h/ mL after oral administration of paeoniflorin, and were 812.6 ± 259.6 ng/mL and 2489 ± 817 ng h/mL after oral administration of SNS decoction, respectively. The Cmax and AUC0–1 of naringin were 64.29 ± 59.21 ng/mL and 311.8 ± 131.8 ng h/mL after oral administration of naringin and were 344.3 ± 234.9 ng/mL and 1479 ± 531 ng h/mL after
Table 3. Pharmacokinetic parameters of naringenin after oral administration of naringin, Fructus Aurantii immaturus and Si–Ni–San decoction to rats (mean ± SD, n ¼ 6).
Parameter Cmax (ng/mL) Tmax (h) t1/2 (h) AUC0–t (ng h/mL) AUC0–1 (ng h/mL) MRT0–1 (h)
Naringin
Fructus Aurantii immaturus
Si–Ni–San decoction
244.2 ± 138.8 730.1 ± 190.3** 981.9 ± 465.4** 19.00 ± 5.48 13.60 ± 0.89 13.6 ± 0.89 5.58 ± 1.81 4.91 ± 1.23 5.00 ± 1.46 4695 ± 3152 11 408 ± 2355** 12 242 ± 6381* 4761 ± 3167 11 479 ± 2372** 12 284 ± 6378* 21.25 ± 1.47 17.49 ± 2.34** 16.25 ± 1.64**
*p50.05, **p50.01, versus oral administration of naringin.
Comparative pharmacokinetic study
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oral administration of SNS decoction, respectively. The Cmax and AUC0–1 of naringenin were 244.2 ± 138.8 ng/mL and 4761 ± 3167 ng h/mL after oral administration of naringin, and were 981.9 ± 465.4 ng/mL and 12 284 ± 6378 ng h/mL after oral administration of SNS decoction, respectively. The Cmax and AUC0–1 of GA were 1183 ± 294 ng/mL and 38 994 ± 14 377 ng h/mL after oral administration of GL, and were 3164 ± 742 ng/mL and 78 817 ± 16 707 ng h/mL after oral administration of SNS decoction, respectively.
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Discussion It was reported that the bioavailability of paeoniflorin in rats after oral administration was very low (Takeda et al., 1995). The extremely low bioavailability was resulted from its poor absorption in the intestine (Chen et al., 2011). Its aglycones are connected with sugar moieties, and some literatures have reported that the presence of sugar moieties could affect solubility and permeability, which might lead to the low intrinsic permeability of compounds in the intestine (Chen et al., 2008). As shown in Table 1, the Cmax and AUC of paeoniflorin were lower after oral administration of Radix Paeoniae alba or SNS decoction than those for same amount of paeoniflorin. These data indicated that the total systemic exposure level of paeoniflorin was lower in the Radix Paeoniae alba group or SNS decoction group than those in the Paeoniflorin group. These results implied that some components in Radix Paeoniae alba or in the other herb of SNS might have pharmacokinetic interactions with paeoniflorin to decrease the total systemic exposure level in vivo. It was reported that GA can induce the functions of p-glycoprotein (p-gp) and CYP3A (Hou et al., 2012), and p-gp-mediated efflux inhibits the intestinal absorption of paeoniflorin (Liu et al., 2006). The major part of paeoniflorin is metabolized or transformed to paeoniflorgenin by intestinal bacterial or b-glucosidase (Xu et al., 2013). The transformation of paeoniflorin to its metabolites might be increased by other components in Radix Paeoniae alba or SNS decoction. In our results, naringin was quickly absorbed and also quickly eliminated in the body. And the plasma profile displayed bimodal phenomenon after oral administration SNS decoction (Figure 3). It was confirmed that the ability of rapid absorption of naringin resulted in the appearance of the first peak. Bimodal phenomenon of naringin was due to the enterohepatic circulation in rats. Naringin could not be detected 14 h after oral administration of pure naringin or Fructus Aurantii immaturus. The Cmax and AUC of naringin were higher after oral administration of SNS decoction than those of naringin or Fructus Aurantii immaturus. These results implied that some components in the other herbs of SNS, but not in Fructus Aurantii immaturus, might reinforce the absorption of naringin. First, it has been reported that some surfactants such as Tween-80 can open the tight junction of intestinal epithelial cells to promote the absorption of drugs (Bittner et al., 2002). There were different kinds of saponins, such as saikosaponin, which has surfactant activity in SNS. It seems that saponins might promote the absorption of naringin. Second, poor permeation, p-gp-mediated efflux and
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Table 4. Pharmacokinetic parameters of glycyrrhetinic acid after oral administration of glycyrrhizic acid, Radix Glycyrrhiza and Si–Ni–San decoction to rats (mean ± SD, n ¼ 6).
Parameter
Glycyrrhizic acid
Radix Glycyrrhiza
Si–Ni–San decoction
Cmax (ng/mL) 1183 ± 294 3118 ± 730** 3164 ± 742** Tmax (h) 11.67 ± 0.81 24.00 ± 0.00** 27.60 ± 3.28** m t1/2 (h) 10.78 ± 2.05 12.07 ± 2.63 9.40 ± 1.15 AUC0–t 37 947 ± 13 894 66 029 ± 18 608* 77 480 ± 16 674** (ng h/mL) 38 994 ± 14 377 67 407 ± 19 555* 78 817 ± 16 707** AUC0–1 (ng h/mL) 29.94 ± 2.45 25.57 ± 3.15* 32.85 ± 4.07m MRT0–1 (h) *p50.05, **p50.01, versus oral administration of glycyrrhizic acid; m p50.05, versus oral administration of Radix Glycyrrhiza.
hydrolysis via a glucosidase contributed to the poor bioavailability of naringin and inhibitors of p-gp can enhance the oral bioavailability and intestinal permeability. The inhibitors in SNS could result in the increase of plasma concentration of naringin. Naringenin must be absorbed as the aglycone after cleavage of naringin by intestinal bacteria, thus naringenin was produced slowly in the body (Figure 4). Naringin, after absorption, must be hydrolyzed to naringenin or other naringin disaccharides. Then, naringenin could be conjugated with glucuronic acid after oral administration of SNS decoction (Qiao et al., 2012). The Cmax and AUC of naringenin were higher after oral administration of Fructus Aurantii immaturus or SNS decoction than those of naringin. This phenomenon indicated that other components in Fructus Aurantii immaturus or SNS decoction may increase the transformation of naringin to its metabolite naringenin. GL generated GA by taking off two molecules from glucuronic acid. GL was transformed to GA by intestinal bacteria. The Cmax, Tmax and AUC of GA were higher after oral administration of Radix Glycyrrhiza or SNS decoction than those of GL (Table 4). This phenomenon indicated that other components in Radix Glycyrrhiza increased the transformation of GL to GA, and the other components in the other herbs of SNS extended the onset time.
Conclusions In conclusion, some herbs or components in TCM may be substrates, inhibitors or inducers of cytochrome P450 and thus may affect the pharmacokinetic of each other. Interactions occurred in the prescription resulting in different pharmacokinetics of pure compounds, herbs and Si–Ni–San. There were statistically significant differences in pharmacokinetic parameters of paeoniflorin, naringin, naringenin and GA following oral administration of pure compounds, single herbs and SNS decoction. The main explanation for these differences seems to be that the interactions occurred in the prescription. It is demonstrated for the first time that these four components have different pharmacokinetic behaviors after oral administration of pure compounds, single herbs and SNS decoction, which reveals the potential compatibility mechanism of TCMs.
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Declaration of interest The authors declare no conflict of interest. This work is supported by grant from the National Foundation of Natural Sciences of China (no. 81102786).
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References Bittner B, Guenzi A, Fullhardt P, et al. (2002). Improvement of the bioavailability of colchicine in rats by co-administration of d-alphatocopherol polyethylene glycol 1000 succinate and a polyethoxylated derivative of 12-hydroxy-stearic acid. Arzneimittelforschung 52: 684–8. Chen LC, Lee MH, Chou MH, et al. (1999). Pharmacokinetic study of paeoniflorin in mice after oral administration of Paeoniae Radix extract. J Chromatogr B 735:33–40. Chen LC, Chou MH, Lin MF, Yang LL. (2002). Pharmacokinetics of paeoniflorin after oral administration of Shao-yao Gan-chao Tang in mice. Jpn J Pharmacol 88:250–5. Chen Y, Wang JY, Yuan L, et al. (2011). Interaction of the main components from the traditional Chinese drug pair Chaihu-Shaoyao based on rat intestinal absorption. Molecules 16:9600–10. Chen Y, Zhao YH, Jia XB, Hu M. (2008). Intestinal absorption mechanisms of prenylated flavonoids present in the heat-processed Epimedium koreanum Nakai (Yin Yanghuo). Pharm Res 25:2190–9. Fang T, Wang Y, Ma Y, et al. (2006). A rapid LC/MS/MS quantitation assay for naringin and its two metabolites in rats plasma. J Pharm Biomed Anal 40:454–9. Gan PP, Zhong MZ, Huang X, et al. (2012). Pharmacokinetic comparisons of albiflorin and paeoniflorin after oral administration of Shaoyao-Gancao-Tang and single herb Paeony decoction to rats. Planta Med 78:237–43. Gao QT, Chen XH, Bi KS. (2004). Comparative pharmacokinetic behavior of glycyrrhetic acid after oral administration of glycyrrhizic acid and Gancao-Fuzi-Tang. Biol Pharm Bull 27:226–8. Hou YC, Lin SP, Chao PDL. (2012). Liquorice reduced cyclosporine bioavailability by activating p-glycoprotein and CYP3A. Food Chem 135:2307–12. Jiang JY, Zhou C, Xu Q. (2003). Alleviating effects of Si–Ni–San, a traditional Chinese prescription, on experimental liver injury and its mechanisms. Biol Pharm Bull 26:1089–94. Jiang JY, Xu Q. (2004). Effect of Si–Ni–San and its individual component drugs on experimental liver injury. Chin J Nat Med 2: 45–9.
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Li YF, Wang M, Wang XY, et al. (2011). Pharmacokinetic properties of albiflorin and paeoniflorin after oral administration of pure compound, Radix Paeoniae alba extract and Danggui-Shaoyao-San extract to rats. J Asian Nat Prod Res 13:117–27. Li XH, Xiong ZL, Lu S, et al. (2010). Pharmacokinetics of naringin and its metabolite naringenin in rats after oral administration of Rhizoma Drynariae extract assayed by UPLC–MS/MS. Chin J Nat Med 8:40–6. Liu Z, Jiang Z, Liu L, Hu M. (2006). Mechanisms responsible for poor oral bioavailability of paeoniflorin: role of intestinal disposition and interactions with sinomenine. Pharm Res 23:2768–80. Qiao Y, Wen J, Song Y, et al. (2012). Study on in vitro and in vivo material base of Si–Ni–San by UPLC–PDA–MS/MS. Chin J Chin Mater Med 37:1672–6. Song J, Wang XD, Zhao JN, et al. (2010). Influence on experimental liver injury of extraction from Peony and Licorice decoction. Pharmacol Clin Chin Mater Med 26:40–2. Takeda S, Ishihara K, Wakui Y, et al. (1996). Bioavailability study of glycyrrhetic acid after oral administration of glycyrrhizin in rats: relevance to the intestinal bacterial hydrolysis. J Pharm Pharmacol 48: 902–5. Takeda S, Isono T, Wakui Y, et al. (1995). Absorption and excretion of paeoniflorin in rats. J Pharm Pharmacol 47:1036–40. Wang Z, Kurosaki Y, Nakayama T, Kimura T. (1994). Mechanism of gastrointestinal absorption of glycyrrhizin in rats. Biol Pharm Bull 17: 1399–403. Wen J, Qiao Y, Yang J, et al. (2012). UPLC–MS/MS determination of paeoniflorin, naringin, naringenin and glycyrrhetinic acid in rat plasma and its application to a pharmacokinetic study after oral administration of Si–Ni–San decoction. J Pharm Biomed Anal 66: 271–7. Wu H, Zhu ZY, Zhang GQ, et al. (2009). Comparative pharmacokinetic study of paeoniflorin after oral administration of pure paeoniflorin, extract of Cortex Moutan and Shuang-Dan prescription to rats. J Ethnopharmacol 125:444–9. Xu CH, Wang P, Wang Y, et al. (2013). Pharmacokinetic comparisons of two different combinations of Shaoyao-Gancao decoction in rats: competing mechanisms between paeoniflorin and glycyrrhetinic acid. J Ethnopharmacol 149:443–52. Yu Q, Xiang J, Tang W, et al. (2009). Simultaneous determination of the 10 major components of Da-Cheng-Qi decoction in dog plasma by liquid chromatography tandem mass spectrometry. J Chromatogr B 877:2025–31. Zhang QF, Chen XH, Yi LX, et al. (2010). Pharmacokinetics of 2 epimers of glycyrrhetic acid in rats. Cent South Pharm 8:350–3.
