Article pubs.acs.org/JAFC
Cite This: J. Agric. Food Chem. 2018, 66, 3975−3982
Simultaneous Determination of Oleanolic Acid and Ursolic Acid by in Vivo Microdialysis via UHPLC-MS/MS Using Magnetic Dispersive Solid Phase Extraction Coupling with Microwave-Assisted Derivatization and Its Application to a Pharmacokinetic Study of Arctiumlappa L. Root Extract in Rats Zhenjia Zheng,† Xian-En Zhao,*,§ Shuyun Zhu,*,§ Jun Dang,‡ Xuguang Qiao,*,† Zhichang Qiu,† and Yanduo Tao‡ †
College of Food Science and Engineering, Shandong Agricultural University, 61 Daizong Street, Taian, Shandong 271018, P.R. China ‡ Qinghai Provincial Key Laboratory of Tibetan Medicine Research & Key Laboratory of Tibetan Medicine Research, Northwest Institute of Plateau Biology, Chinese Academy of Science, Xining, Qinghai 810001, P.R. China § College of Chemistry and Chemical Engineering, Qufu Normal University, Qufu, Shandong 273165, P.R. China S Supporting Information *
ABSTRACT: Simultaneous detection of oleanolic acid and ursolic acid in rat blood by in vivo microdialysis can provide important pharmacokinetics information. Microwave-assisted derivatization coupled with magnetic dispersive solid phase extraction was established for the determination of oleanolic acid and ursolic acid by liquid chromatography tandem mass spectrometry. 2′-Carbonyl-piperazine rhodamine B was first designed and synthesized as the derivatization reagent, which was easily adsorbed onto the surface of Fe3O4/graphene oxide. Simultaneous derivatization and extraction of oleanolic acid and ursolic acid were performed on Fe3O4/graphene oxide. The permanent positive charge of the derivatization reagent significantly improved the ionization efficiencies. The limits of detection were 0.025 and 0.020 ng/mL for oleanolic acid and ursolic acid, respectively. The validated method was shown to be promising for sensitive, accurate, and simultaneous determination of oleanolic acid and ursolic acid. It was used for their pharmacokinetics study in rat blood after oral administration of Arctiumlappa L. root extract. KEYWORDS: in vivo microdialysis, triterpenic acid, microwave-assisted derivatization, magnetic graphene oxide, pharmacokinetics, Arctiumlappa L. root
■
INTRODUCTION Oleanolic acid (OA) and ursolic acid (UA) belong to triterpenoid compounds that widely exist in herbs and fruits. They have been reported to have important pharmacological properties such as anti-inflammatory, hepatoprotective, antiulcer, antimicrobial, antitumor, anti-HIV, antihyperlipidemic activities, and so on.1,2 Therefore, a sensitive and rapid analytical method is necessary and helpful for their pharmacokinetic study to better understand the pharmacological activity of related foods and herbs. OA and UA are triterpine isomers with exactly the same chemical structures, with the only difference found in the position of a methyl group in the E ring (Figure 1); thus, they are difficult to separate and detect rapidly.3−5 A lot of methods have been reported for the determination of OA and UA, such as gas chromatography (GC),6 thin-layer chromatography (TLC),7 capillary electrophoresis (CE),8 high-performance liquid chromatography (HPLC) with UV,9 fluorescence detection,3 mass spectrometry (MS),4,5 or nuclear magnetic resonance (NMR).10 Each method has its own feature, but many of them show a limited enhancement on the sensitivity, accuracy and specificity. In the past decade, ultra high © 2018 American Chemical Society
performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS) in the multiple reaction monitoring mode (MRM) has aroused wide attention in rapid pharmaceutical analysis and bioanalysis in different biological matrixes.11 However, very low concentrations and strong matrix interferences in real samples usually make difficult the sensitive and accurate determination of compounds. Therefore, sensitive, accurate, rapid, and simultaneous determination of OA and UA is still a challenging task. However, OA and UA lack a chromophore and cannot easily gain a charge because of their carboxyl group; thus, they provide a very low sensitivity in relation to UV, fluorescence, and MS detection. Under these circumstances, sensitivity enhancement by chemical derivatization can solve problems.12,13 Some derivatization reagents have been reported for them by HPLC fluorescence detection14−18 and GC-MS.19 However, there are Received: Revised: Accepted: Published: 3975
December March 15, March 21, March 21,
21, 2017 2018 2018 2018 DOI: 10.1021/acs.jafc.7b06015 J. Agric. Food Chem. 2018, 66, 3975−3982
Article
Journal of Agricultural and Food Chemistry
Figure 1. Synthesized of CPR and the derivatization reaction scheme of CPR with OA and UA.
MS. 2′-Carbonyl-piperazine rhodamine B (CPR) was designed and synthesized as a derivatization reagent for the labeling of carboxyl group of OA and UA. This method was used for the simultaneous pharmacokinetic study of OA and UA in rat blood.
almost no synthesized derivatization reagents for the enhanced UHPLC-MS/MS determination of OA and UA. However, the UHPLC-MS/MS detection sensitivity is frequently compromised by the low contents of analytes and serious matrix effect from real samples and the derivatization procedure. Therefore, an efficient sample pretreatment procedure is necessary.20,21 Compared to the popular liquid− liquid extraction (LLE)11,12,22 and solid-phase extraction (SPE),23,24 dispersive solid-phase extraction (d-SPE) is time saving, easy to operate, and has low consumption of toxic organic solvents. Especially, the use of magnetic sorbents for magnetic dispersive solid phase extraction (MDSPE) has drawn significant attention because of their excellent dispersibility and ease of separation by an external magnetic field.25,26 Graphene oxide (GO), a carbon-based material with a high surface area to volume ratio, strong π−π stacking interactions, and high mechanical strength25,27,28 presents enormous advantages in the separation and determination of small amount of organic analytes depending on the hydrogen bonding, hydrophobic interactions, van der Waals forces, and electrostatic forces.29 In vivo microdialysis sampling is a preeminent technique for neuroscience, pharmacokinetics (PK), pharmacodynamics (PD), and clinical studies.30 It is commonly used for investigation of PK/PD because concentration of the unbound drug is more correlated to the pharmacological effects. By coupling with powerful analytical methods, in vivo microdialysis is able to overcome several disadvantages of conventional pharmacokinetic techniques which include continuous sampling in the same animal, minimizing the number of animals used, and also minimizing interanimal variation. However, drug quantification in microdialysis technique remains a major challenge because of the low concentration and the small volume of microdialysate. Therefore, a highly sensitive, accurate, and selective analytical method employing UHPLCMS/MS (MRM) is always recommended.31,32 In this study, a new method based on in vivo microdialysis by microwave-assisted derivatization coupling with MDSPE (MAD-MDSPE) in a single step was developed for the simultaneous determination of OA and UA by UHPLC-MS/
■
MATERIALS AND METHODS
Chemicals and Reagents. OA, UA, and the internal standard (IS) betulinic acid were purchased from National Institute for the control of pharmaceutical and biological products (Beijing, China). 1-Ethyl-3(3-(dimethylamino)propyl)-carbodiimide hydrochloride (EDC-HCl) and the HPLC grade formic acid were purchased from Sigma Co. (St. Louis, MO, United States). HPLC grade acetonitrile and methanol were purchased from Fisher Scientific Co. (Fair Lawn, NJ, United States). N,N-dimethylformamide (DMF) and pyridine were of analytical grade and obtained from Tianjin Guangcheng Chemical Reagent Co. (Tianjin, China). Pure water was obtained on a Millipore system (Bedford, MA, United States). All other reagents used were of HPLC grade or at least of analytical grade obtained commercially. Stock solutions of OA (10.0 μmol/L), UA (10.0 μmol/L), betulinic acid (IS, 10.0 μmol/L), and derivatization reagent CPR (100.0 μmol/ L) were prepared by HPLC grade acetonitrile. All working solutions with different concentrations were prepared by diluting corresponding stock solutions with acetonitrile. Solution of 0.10 mol/L coupling reagent EDC was prepared in HPLC acetonitrile. The quality control samples (QCs) containing OA (0.5, 5.0, 100 ng/mL) and UA (0.5, 5.0, 100 ng/mL) were prepared at 3 concentration levels by adding appropriate working standard solutions to drug-free rat microdialysates. When not in use, all the solutions were stored at 4 °C. Instrumentation. The UHPLC-MS/MS system consisted of an Agilent 1290 UHPLC system and an Agilent 6460 Triple Quadrupole MS/MS system (Agilent, United States). The chromatographic separation was realized on an Agilent SB C18 column (2.1 mm × 50 mm, 1.8 μm) at 30 °C column temperature with 2.0 μL injection volumes. The flow rate was constant at 0.2 mL/min. Eluent A was 5% acetonitrile/water (0.1% formic acid) and B was acetonitrile (0.1% formic acid). The linear binary gradient elution conditions were as follows: 65−82% B from 0 to 2 min; 82−98% B from 2 to 6 min; 98− 100% B from 6 to 8 min. During 0−5.5 min after injection, the flow was diverted into waste to protect the mass spectrometer from potential contaminations because there were no detectable analytes. The column was equilibrated using the initial mobile phase for 1.5 min for each injection. The mass spectrometer was run in positive ion 3976
DOI: 10.1021/acs.jafc.7b06015 J. Agric. Food Chem. 2018, 66, 3975−3982
Article
Journal of Agricultural and Food Chemistry
Figure 2. (A) Representative MRM chromatogram of CPR derivatives of internal standard (IS) betulinic acid, OA, and UA standards, and (B) product ion spectrum and the proposed fragmentation schematics of CPR-UA derivative. Preparation of Fe3O4/GO. The preparation of graphene oxide is described in Supporting Information according to the Hummers method with minor modifications.34 Fe3O4/GO was prepared based on chemical coprecipitation of Fe2+ and Fe3+ in alkaline media in the presence of GO as described in Supporting Information.35 MDSPE-MAD Procedure. Fe3O4/GO (8 mg) was put into a 1.5 mL vial, and then 30 μL of mixed standards or microdialysates, 30 μL of EDC (0.1 mol/L), and 200 μL of CPR reagent were added. The vial was sealed and immersed in the ultrasound bath for 2.0 min to form a homogeneous dispersed solution. This solution was radiated for 25 min in a microwave reactor (450 W) at 60 °C to achieve complete MDSPE-MAD procedure. The derivatization reaction scheme is shown in Figure 1. Then, the magnetic materials were separated rapidly from the derivatization solution by an external magnet. After that, CPR derivatives were eluted from the magnetic materials by 300 μL of methanol (containing 1.0% formic acid) under ultrasound for 1 min, and 2.0 μL of the solution was analyzed by UHPLC-MS/MS. Preparation of Arctiumlappa L. Root Extract. Dried Arctiumlappa L. root was purchased from China Beijing Tongrentang (Group) Co., Ltd. (Beijing). To 50 g of powdered raw herb (250 μm) in a 500 mL flask was added 300 mL of ethanol. The mixture was extracted 2 times by ethanol refluxing with 6 volume equivalents. The combined extracts were added with 20 mL of 30% β-cyclodextrin to increase the solubility, and then the solution was concentrated to 10 mL under vacuum to obtain herb solution for rats. The OA and UA contents in the extract were quantitatively determined by the developed method in this work. The contents of free OA and UA in the extract were 673.8 and 526.4 μg/g. After saponification reaction with 10% potassium hydroxide at 90 °C for 3 h, the conjugated OA
MRM mode of electrospray source. The optimal MS conditions were the same as those from our recent report in 2016.11 The fragmentor voltage (FV) and collision energy (CE) were also optimized for the target derivatives. Experimental conditions for the direct MRM detection of OA and UA were set according to the literature.4 Transmission electron microscope (TEM) images were obtained using the JEM-2100PLUS microscope (JEOL, Tokyo, Japan). In vivo microdialysis sampling was accomplished using in vivo microdialysis system from Sweden CMA Co., including a CMA 402 syringe pump (CMA, Solna, Sweden), a CMA 120 system (CMA, Solna, Sweden) for freely moving animals, and a microdialysis MAB6 probe (Stockholm, Sweden). ASI stereotaxic flat skull coordinates were purchased from ASI Instruments Inc. (MI, United States). The probe was perfused with Ringer’s solution (5 mmol/L) at a flow rate of 2.0 μL/min. Synthesis of CPR. The synthesis reaction schematic of CPR was shown in Figure 1. The synthesis of raw material N-hydroxysuccinimidyl rhodamine B ester (RB-S) was carried out as in our previous report.33 The synthesis procedure was as follows: RB-S (1.5 g) and 0.5 g piperazine were added into 50 mL of acetonitrile and 25 mL of sodium bicarbonate buffer (pH 8.5), and the solution was heated to 45 °C with continued agitation for 2 h. After reduced-pressure distillation of the solvent and recrystallization in dichloroethane/absolute ethanol (v/v, 1/1), CPR was obtained as dark red crystals with a yield of 55%. 1 HNMR (500 MHz, CDCl3/δ, ppm): 7.69 (t, J = 9.7 Hz, 2H), 7.61 (d, J = 13.5 Hz, 1H), 7.34 (d, J = 6.9 Hz, 1H), 7.19 (d, J = 9.5 Hz, 2H), 6.98 (d, J = 9.1 Hz, 2H), 6.75 (s, 2H), 3.82−3.55 (m, 12H), 3.03 (d, J = 34.4 Hz, 3H), 1.33 (t, J = 7.0 Hz, 11H). HRMS: [M + H]+ 511.30692. 3977
DOI: 10.1021/acs.jafc.7b06015 J. Agric. Food Chem. 2018, 66, 3975−3982
Article
Journal of Agricultural and Food Chemistry Table 1. MRM Parameters of OA, UA, and Betulinic Acid (IS) analytes
fragmentor (V)
quantitation transition (m/z)
collision energy (eV)
confirmation transition (m/z)
collision energy (eV)
OA UA betulinic acid (IS)
240 240 250
949.6 > 398.8 949.6 > 398.8 949.6 > 398.8
82 81 79
949.6 > 443.2 949.6 > 443.2 949.6 > 443.2
78 78 76
Figure 3. Optimization of MAD conditions (n = 5): (A) volumes of EDC, (B) volumes of CPR solution, (C) time (min), and (D) temperature (°C). and UA in the extract were determined, and their concentrations were 1438.5 and 1142.2 μg/g. Pharmacokinetics of OA and UA in Rats by in Vivo Microdialysis. Male Sprague−Dawley rats (200−220 g, n = 6) were purchased from Shandong Lukang Pharmaceutical Co. Ltd. The care and use of animals were in accordance with the related principles of China. Rats were narcotized with 20% urethane (1.2 g/kg, i.p.) before surgery and kept anesthetized during the whole surgery of MAB probe implantation. After recovering, rats were able to freely drink and eat for 24 h and were then fasted for 12 h with drinking water freely available before the pharmacokinetics test. Rats were orally administrated with extracts at a dose of 1.0 g/kg body weight. In vivo microdialysate samples from rat carotid artery were collected at 15, 30, 45, 60, 80, 100, 120, 180, 240, 360, 480, and 600 min after oral administration. Each time, 30 μL of microdialysates was used for MAD-MDSPE procedure and UHPLC-MS/MS analysis.
best chromatographic separation and maximum MS sensitivity. The chromatographic conditions for optimization were similar to those in our previous study.11 The optimal conditions are described in the Materials and Methods section. The representative MRM chromatograms of CPR derivatives for standards and the internal standard are shown in Figure 2A. MS conditions were also optimized similar to our previous study.11 All the CPR derivatives showed intense [M + H]+ ions. They were set as precursor ions. The most abundant product ions for three CPR derivatives were m/z 398.8 and 443.2. In ESI-MS/MS conditions, these two specific product ions contained a permanent intramolecular positive charge. They brought enhanced sensitivity by increasing the ionization efficiency in the electrospray ionization. The proposed collision-induced dissociation pathways for the precursor ion of CPR-UA are shown in Figure 2B. In this study, m/z 398.8 was used for quantitative analysis, and m/z 443.2 was used for confirmation analysis. The optimal FVs and CEs for two
■
RESULTS AND DISCUSSION Optimization of Chromatography and MS Conditions. UHPLC and MS conditions were optimized for obtaining the 3978
DOI: 10.1021/acs.jafc.7b06015 J. Agric. Food Chem. 2018, 66, 3975−3982
Article
Journal of Agricultural and Food Chemistry transitions of OA, UA, and betulinic acid (IS) are shown in Table 1. Optimization of MAD-MDSPE. Optimization of the Volume of EDC Solution. Using EDC as the condensing agent, the carboxylic acid group of OA and UA can be selectively labeled. A great advantage of this derivatization reaction was that a small amount of water was allowed to be present in the EDC condensation derivatization system. Therefore, CPR coupling with EDC has the great advantage for the determination of triterpenic acids in aqueous sample (such as microdialyaste) and thus was selected in this work. The volumes of CPR solution were optimized in the range of 10−60 μL, as shown in Figure 3A. The peak area increased with the volumes of EDC from 10 to 30 μL and then decreased. Thus, 30 μL of EDC (0.1 mol/L) was selected. Optimization of the Volume of CPR Solution. To study the effect of CPR amount, the volume of CPR solution was optimized in the range of 50−350 μL, as shown in Figure 3B. No less than 200 μL of CPR solution was used, which ensured the thorough derivatization of analytes. The excess CPR was purified by the MDSPE procedure. Two-hundred microliters of CPR was chosen for the MAD-MDSPE procedure. Optimization of Derivatization Time and Temperature. The derivatization time was optimized from 10 to 40 min in the MAD-MDSPE procedure (microwave 450 W). As shown in Figure 3C, the derivatization reaction was completed rapidly under microwave assistance. There were no remarkable increases of peak areas when the derivatization reaction time was more than 25 min. Therefore, 25 min was used to perform the derivatization. The derivatization temperature was optimized from 40 to 70 °C while other conditions were kept constant, as shown in Figure 3D. Optimum peak areas were obtained when OA and UA standards were derivatized at 60 °C. Optimization of Sorbent Amount. To evaluate the effect of sorbent amount on extraction efficiency and derivatization efficiency of CPR derivatives, the amount of Fe3O4/GO was optimized in the range of 4−16 mg. The incremental amounts of sorbent up to 8 mg possibly helped the derivatization reaction by providing a sufficient surface for derivatives adsorption; but in higher amounts of it, lower extraction efficiency was obtained. Therefore, the MAD-MDSPE procedure was carried out with 8 mg of Fe3O4/GO. Optimization of Desorption Conditions. To achieve good desorption efficiency of CPR derivatives, many desorption solutions including acetonitrile, acetone, and methanol (each containing 1.0% formic acid) were estimated. The results indicated that methanol had the strongest desorption power of CPR derivatives (Figure 4A). Methanol was a stronger polar organic solvent than acetonitrile and acetone. Moreover, GO can be easily dispersed in polar solvents because of its polar groups on the surface. Therefore, methanol was selected to ensure sufficient desorption of the derivatives. The effect of the volumes of desorption solution on the desorption efficiency was also evaluated. When the methanol volume was increased to 300 μL, desorption efficiency was increased because of the high rate of Fe3O4/GO dispersion in methanol. The results showed that 300 μL of methanol (1.0% formic acid) was enough for the efficient desorption of the derivatives (Figure 4B). Desorption time was optimized in the range of 10−100 s (ultrasound 120 W, 40 kHz). Significantly increased peak areas of the CPR derivatives were detected in 10−60 s, and no
Figure 4. Optimization of desorption conditions (n = 5): (A) types of desorption solution and (B) volumes of desorption solution.
significant increase was obtained with the enhancement of desorption time in 60−100 s. In the end, 1.0 min was employed as the optimal desorption time. Method Validation. To investigate the applicability of the developed MAD-MDSPE coupled to UHPLC-MS/MS, linearity ranges, limits of detection (LODs), quantification (LOQs), repeatability, recovery, precision, accuracy, and matrix effect (ME) were determined. The linearity of this method was established using internal standard spiked calibration solutions at seven concentration levels. To evaluate the dynamic ranges of this method, six batches of calibration microdialysate samples were prepared and determined. The peak-area ratio between the analyte and IS of each spiked microdialysate sample was determined. The calibration curves were then constructed by plotting the peak-area ratio with the spiked concentrations using linear regression for each analyte, respectively. The regression equations were y = 9.836x + 0.082 (R = 0.991) for OA and y = 9.923x − 0.051 (R = 0.995) for UA, where y is the peak-area ratio of the analyte and IS and x is the concentration of analyte (ng/mL). The calibration curves covering the concentration range of 0.050−100 ng/mL showed good linearity with correlation coefficient R > 0.99. The LODs were 0.025 and 0.020 ng/mL for OA and UA (S/N > 3). The LOQs for OA and UA in microdialysates were 0.090 and 0.080 ng/mL (S/N > 10). 3979
DOI: 10.1021/acs.jafc.7b06015 J. Agric. Food Chem. 2018, 66, 3975−3982
Article
Journal of Agricultural and Food Chemistry Table 2. Results of Recovery, Matrix Effect, Repeatability, Precision, and Accuracy for OA and UA (n = 6) repeatability (RSD, %) analyte OA
UA
spiked levels (ng/mL) 0.50 5.0 50.0 0.50 5.0 50.0
recovery (%) 98.4 97.6 104.1 98.7 109.8 103.5
± ± ± ± ± ±
4.2 3.1 5.1 5.1 4.3 3.6
matrix effect (%) 98.6 94.6 103.3 111.3 103.7 92.6
± ± ± ± ± ±
6.5 6.1 7.0 6.5 6.9 4.5
intraday precision (RSD, %)
interday precision (RSD, %)
accuracy (%)
peak area
retention time
peak area
retention time
peak area
retention time
intraday
interday
7.35 6.62 4.23 4.75 5.52 6.26
3.14 2.34 4.50 3.70 2.88 2.70
3.59 5.81 3.87 4.41 5.27 5.70
3.35 2.57 2.90 4.20 3.77 3.29
4.80 3.45 2.33 5.58 6.01 5.12
3.50 3.21 3.89 2.20 5.74 3.10
93.0 99.1 101.8 101.4 98.0 96.4
92.8 112.3 87.2 85.1 103.4 110.5
As shown in Table 2, the precision was in the range of 2.20− 6.01%, and the accuracy was in the range of 85.1−112.3% from the actual QCs. The intra- and interday accuracy and precision were all within 15% by FDA. The matrix effect of the analytes ranged from 92.6 to 111.3% at 3 concentration levels. The repeatability of the method was in the range of 2.34−7.35%. The recoveries of the OA and UA were in the range of 97.6− 109.8% at 3 concentration levels. These results indicated that the developed method could be used well for the sensitive, specific, and accurate determination of OA and UA in rat microdialysate samples. Pharmacokinetics of OA and UA from in Vivo Rat Blood Microdialysates. The developed MAD-MDSPE coupled to UHPLC-MS/MS method has been used to analyze microdialysate samples from rat blood after oral administration of Arctiumlappa L. root extract. Typical MRM chromatograms of internal standard (IS) betulinic acid, OA, and UA derivatives in a rat blood microdialysate sample are shown in Figure 5. The Figure 6. Mean concentration−time curves of OA and UA in rat plasma microdialysates after oral administration of Arctiumlappa L. root extract. Each point represents the mean ± standard error (n = 6).
Table 3. Plasma Pharmacokinetic Parameters after Oral Administration of Arctiumlappa L. Root Extract to Rats parameter t1/2 Tmax Cmax AUC0−t AUC0−∞ MRT Vz/F CL/F
unit min min ng/mL ng/mL·min ng/mL·min min (ng)/(ng/mL) (ng)/(ng/mL)/ min
UA
OA
294.67 ± 83.66 50 ± 7.75 40.55 ± 4.11 6897.44 ± 428.33 9491.76 ± 953.31 457.42 ± 87.94 36.60 ± 7.35 0.09 ± 0.009
333.89 ± 107.88 45 ± 9.49 10.51 ± 3.26 1249.10 ± 245.91 1749.03 ± 470.50 473.65 ± 110.73 228.31 ± 40.28 0.50 ± 0.12
were quite different. OA appeared to be absorbed slightly quicker into the plasma with a Tmax of 45 min, while UA absorbed relatively slowly with a Tmax of 50 min. However, the absolute bioavailability of UA was obviously better than that of OA with Cmax of 40 vs 10 ng/mL. This finding is consistent with a previous study,5 which will be helpful for future pharmacology, pharmacodynamics, and drug development. In conclusion, we developed a rapid, selective, and sensitive strategy based on MAD-MDSPE coupled to UHPLC-MS/MS (MRM) for the simultaneous determination of OA and UA in rat blood microdialysates. The derivatization, extraction, and purification of OA and UA occurred on the surface of Fe3O4/ GO and were integrated into one step. Furthermore, the validated method was successfully used in the pharmacokinetics study.
Figure 5. Typical MRM chromatograms of internal standard (IS) betulinic acid, OA, and UA derivatives in a rat blood microdialysate sample.
mean plasma drug concentration−time profiles are shown in Figure 6. Table 3 presented the pharmacokinetic parameters, including the maximum plasma concentration (Cmax), the time for reaching the maximum concentration (Tmax), terminal halflife (t1/2), the area under the concentration−time curve (AUC), mean residence time (MRT), the apparent volume of distribution (Vz/F) and time-averaged total body clearance (CL/F). After oral administration, both OA and UA could be absorbed into the blood. However, their systemic exposures 3980
DOI: 10.1021/acs.jafc.7b06015 J. Agric. Food Chem. 2018, 66, 3975−3982
Article
Journal of Agricultural and Food Chemistry
■
(9) Sowa, I.; Wojciak-Kosior, M.; Rokicka, K.; Kocjan, R.; Szymczak, G. Application of solid phase extraction with the use of silica modified with polyaniline film for pretreatment of samples from plant material before HPLC determination of triterpenicacids. Talanta 2014, 122, 51−57. (10) Dais, P.; Plessel, R.; Williamson, K.; Hatzakis, E. Complete H-1 and C-13 NMR assignment and P-31 NMR determination of pentacyclic triterpenic acids. Anal. Methods 2017, 9, 949−957. (11) He, Y. R.; Zhao, X. E.; Wang, R. J.; Wei, N.; Sun, J.; Dang, J.; Chen, G.; Liu, Z. Q.; Zhu, S. Y.; You, J. M. Simultaneous Determination of food-related biogenic amines and precursor amino acids using in situ derivatization ultrasound-assisted dispersive liquidliquid microextraction by ultra-high-performance liquid chromatography tandem mass spectrometry. J. Agric. Food Chem. 2016, 64, 8225−8234. (12) Zhao, X.-E.; Lv, T.; Zhu, S. Y.; Qu, F.; Chen, G.; He, Y. R.; Wei, N.; Li, G. L.; Xia, L.; Sun, Z. W.; Zhang, S. J.; You, J. M.; Liu, S.; Liu, Z. G.; Sun, J.; Liu, S. Y. Dual ultrasonic-assisted dispersive liquid-liquid microextraction coupled with microwave-assisted derivatization for simultaneous determination of 20(S)-protopanaxadiol and 20(S)protopanaxatriol by ultra high performance liquid chromatographytandem mass spectrometry. J. Chromatogr. A 2016, 1437, 49−57. (13) Mung, D.; Li, L. Development of chemical isotope labeling LCMS for milk metabolomics: comprehensive and quantitative profiling of the amine/phenol submetabolome. Anal. Chem. 2017, 89, 4435− 4443. (14) Li, G. L.; You, J. M.; Song, C. H.; Xia, L. A.; Zheng, J.; Suo, Y. R. Development of a new HPLC method with precolumn fluorescent derivatization for rapid, selective and sensitive detection of triterpenic acids in fruits. J. Agric. Food Chem. 2011, 59, 2972−2979. (15) Chen, G.; Li, J.; Song, C. H.; Suo, Y. R.; You, J. M. A sensitive and efficient method for simultaneous trace detection and identification of triterpene acids and its application to pharmacokinetic study. Talanta 2012, 98, 101−111. (16) Wang, Y. W.; Suo, Y. R.; Sun, Y. N.; You, J. M. Determination of triterpene acids from 37 different varieties of raspberry using precolumn derivatization and HPLC fluorescence detection. Chromatographia 2016, 79, 1515−1525. (17) Wozniak, L.; Marszalek, K.; Skapska, S.; Jedrzejczak, R. Novel method for hplc analysis of triterpenic acids using 9-anthryldiazomethane derivatization and fluorescence detection. Chromatographia 2017, 80, 1527−1533. (18) You, J. M.; Wu, D.; Zhao, M.; Li, G. L.; Gong, P. W.; Wu, Y. Y.; Guo, Y.; Chen, G.; Zhao, X. E.; Sun, Z. W.; Xia, L.; Wu, Y. N. Development of a facile and sensitive HPLC-FLD method via fluorescence labeling for triterpenic acid bioavailability investigation. Biomed. Chromatogr. 2017, 31, e3894. (19) Jemmali, Z.; Chartier, A.; Dufresne, C.; Elfakir, C. Optimization of the derivatization protocol of pentacyclic triterpenes prior to their gas chromatography-mass spectrometry analysis in plant extracts. Talanta 2016, 147, 35−43. (20) Tang, S.; Zhang, H.; Lee, H. K. Advances in sample extraction. Anal. Chem. 2016, 88, 228−249. (21) Xu, L. N.; Qi, X. Y.; Li, X. J.; Bai, Y.; Liu, H. W. Recent advances in applications of nanomaterials for sample preparation. Talanta 2016, 146, 714−726. (22) Zhao, X. E.; He, Y. R.; Li, M.; Chen, G.; Wei, N.; Wang, X.; Sun, J.; Zhu, S. Y.; You, J. M. Analysis of amino acid and monoamine neurotransmitters and their metabolites in rat urine of Alzheimer’s disease using in situ ultrasound-assisted derivatization dispersive liquid-liquid microextraction with UHPLC−MS/MS. J. Pharm. Biomed. Anal. 2017, 135, 186−198. (23) Cao, Z. Y.; Ma, Y. N.; Sun, L. H.; Mou, R. X.; Zhu, Z. W.; Chen, M. X. Direct determination of six cytokinin nucleotide monophosphates in coconut flesh by reversed-phase liquid chromatography-tandem mass spectrometry. J. Agric. Food Chem. 2017, 65, 9909−9915. (24) Xiang, L.; Chen, L.; Xiao, T.; Mo, C. H.; Li, Y. W.; Cai, Q. Y.; Li, H.; Zhou, D. M.; Wong, M. H. Determination of trace perfluoroalkyl
ASSOCIATED CONTENT
S Supporting Information *
The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acs.jafc.7b06015.
■
Preparation of Fe3O4/GO (PDF)
AUTHOR INFORMATION
Corresponding Authors
*E-mail:
[email protected]; Tel: +86-537-4456301; Fax: +86-537-4456305. *E-mail:
[email protected]. *E-mail:
[email protected]. ORCID
Xian-En Zhao: 0000-0003-3500-9518 Shuyun Zhu: 0000-0002-9632-8187 Funding
This work was supported by the National Natural Science Foundation of China (Grants 21775088, 21405094, and 81303179), the Special Fund for Agro-scientific Research in the Public Interest (Grant 201503142), the Innovation Platform for the Development and Construction of Special Project of Key Laboratory of Tibetan Medicine Research of Qinghai Province (Grant 2017-ZJ-Y11), and the Open Projects Program of the Key Laboratory of Tibetan Medicine Research of Chinese Academy of Sciences. Notes
The authors declare no competing financial interest.
■
REFERENCES
(1) Gautam, R.; Jachak, S. M. Recent developments in antiinflammatory natural products. Med. Res. Rev. 2009, 29, 767−820. (2) Yin, M. C.; Lin, M. C.; Mong, M. C.; Lin, C. Y. Bioavailability, distribution, and antioxidative effects of selected triterpenes in mice. J. Agric. Food Chem. 2012, 60, 7697−7701. (3) Wu, H. L.; Li, G. L.; Liu, S. C.; Liu, D.; Chen, G.; Hu, N.; Suo, Y. R. Simultaneous determination of six triterpenic acids in some Chinese medicinal herbs using ultrasound-assisted dispersive liquid-liquid microextraction and high-performance liquid chromatography with fluorescence detection. J. Pharm. Biomed. Anal. 2015, 107, 98−107. (4) Sanchez-Avila, N.; Priego-Capote, F.; Ruiz-Jimenez, J.; de Castro, M. D. L. Fast and selective determination of triterpenic compounds in olive leaves by liquid chromatography-tandem mass spectrometry with multiple reaction monitoring after microwave-assisted extraction. Talanta 2009, 78, 40−48. (5) Zhao, L.; Li, W. H.; Li, Y. Y.; Xu, X.; Lv, L.; Wang, X. X.; Chai, Y. F.; Zhang, G. Q. Simultaneous Determination of oleanolic and ursolic acids in rat plasma by HPLC−MS: application to a pharmacokinetic study after oral administration of different combinations of qinggansanjie decoction extracts. J. Chromatogr. Sci. 2015, 53, 1185− 1192. (6) Avila, N. S.; Capote, F. P.; de Castro, M. D. L. Ultrasoundassisted extraction and silylation prior to gas chromatography−mass spectrometry for the characterization of the triterpenic fraction in olive leaves. J. Chromatogr. A 2007, 1165, 158−165. (7) Banerjee, A.; Sane, R.; Mangaonkar, K.; Shailajan, S.; Deshpande, A.; Gundi, G. Quantitation of oleanolic acid in Oldenlandia corymbosa L. whole-plant powder by high-performance thin-layer chromatography. J. Planar Chromatogr.–Mod. TLC 2006, 19, 68−72. (8) Gao, R. B.; Wang, L. T.; Yang, Y.; Ni, J. M.; Zhao, L.; Dong, S. Q.; Guo, M. Simultaneous determination of oleanolic acid, ursolic acid, quercetin and apigenin in Swertia mussotii Franch by capillary zone electrophoresis with running buffer modifier. Biomed. Chromatogr. 2015, 29, 402−409. 3981
DOI: 10.1021/acs.jafc.7b06015 J. Agric. Food Chem. 2018, 66, 3975−3982
Article
Journal of Agricultural and Food Chemistry carboxylic acids in edible crop matrices: matrix effect and method development. J. Agric. Food Chem. 2017, 65, 8763−8772. (25) Xu, J.; Wu, P.; Ye, E. C.; Yuan, B. F.; Feng, Y. Q. Metal oxides in sample pretreatment. TrAC, Trends Anal. Chem. 2016, 80, 41−56. (26) Wang, A. X.; Lu, H. Z.; Xu, S. F. Preparation of magnetic hollow molecularly imprinted polymers for detection of triazines in food samples. J. Agric. Food Chem. 2016, 64, 5110−5116. (27) Wen, Y. Y.; Chen, L.; Li, J. H.; Liu, D. Y.; Chen, L. X. Recent advances in solid-phase sorbents for sample preparation prior to chromatographic analysis. TrAC, Trends Anal. Chem. 2014, 59, 26−41. (28) Pei, H. M.; Zhu, S. Y.; Yang, M. H.; Kong, R. M.; Zheng, Y. Q.; Qu, F. L. Graphene oxide quantum dots@silver core-shell nanocrystals as turn-on fluorescent nanoprobe for ultrasensitive detection of prostate specific antigen. Biosens. Bioelectron. 2015, 74, 909−914. (29) Ren, H.; Kulkarni, D. D.; Kodiyath, R.; Xu, W. N.; Choi, I.; Tsukruk, V. V. Competitive adsorption of dopamine and rhodamine 6g on the surface of graphene oxide. ACS Appl. Mater. Interfaces 2014, 6, 2459−2470. (30) Shannon, R. J.; Carpenter, K. L. H.; Guilfoyle, M. R.; Helmy, A.; Hutchinson, P. J. Cerebral microdialysis in clinical studies of drugs: pharmacokinetic applications. J. Pharmacokinet. Pharmacodyn. 2013, 40, 343−358. (31) Zhao, X. E.; He, Y. R.; Yan, P.; Wei, N.; Wang, R. J.; Sun, J.; Zheng, L. F.; Zhu, S. Y.; You, J. M. Sensitive and accurate determination of neurotransmitters from in vivo rat brain microdialysate of Parkinson’s disease using in situ ultrasound-assisted derivatization dispersive liquid−liquid microextraction by UHPLCMS/MS. RSC Adv. 2016, 6, 108635−108644. (32) Kong, W. M.; Mohamed, Z.; Alshawsh, M. A.; Chik, Z. Evaluation of pharmacokinetics and blood-brain barrier permeabilityof mitragynine using in vivo microdialysis technique. J. Pharm. Biomed. Anal. 2017, 143, 43−47. (33) Zheng, L. F.; Zhao, X. E.; Ji, W. H.; Wang, X.; Tao, Y. D.; Sun, J.; Xu, Y. Q.; Wang, X.; Zhu, S. Y.; You, J. M. Core-shell magnetic molecularly imprinted polymers used rhodamine B hydroxyproline derivate as template combined with in situ derivatization for the specific measurement of L-hydroxyproline. J. Chromatogr. A 2018, 1532, 30−39. (34) Anastassiades, M.; Lehotay, S. J.; Štajnbaher, D.; Schenck, F. J. Fast and easy multiresidue method employing acetonitrile extraction/ partitioning and dispersive solid-phase extraction for the determination of pesticide residues in produce. J. AOAC Int. 2003, 86, 412−431. (35) Sun, J. P.; Liang, Q. L.; Han, Q.; Zhang, X. Q.; Ding, M. Y. Onestep synthesis of magnetic graphene oxide nanocomposite and its application in magnetic solid phase extraction of heavy metal ions from biological samples. Talanta 2015, 132, 557−563.
3982
DOI: 10.1021/acs.jafc.7b06015 J. Agric. Food Chem. 2018, 66, 3975−3982
