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- Publisher
- Hindawi Publishing Corporation
- ISSN
- 2090-8865
- eISSN
- 2090-8873
- DOI
- 10.1155/2022/5297603
- Publisher site
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Abstract
Hindawi Journal of Analytical Methods in Chemistry Volume 2022, Article ID 5297603, 18 pages https://doi.org/10.1155/2022/5297603 Research Article Measurement of Pharmacokinetics and Tissue Distribution of Four Compounds from Nauclea officinalis in Rat Plasma and Tissues through HPLC-MS/MS 1 1 1 1 2 1,2 Yuhuang Wu, Liyan Li, Guxu Ming, Xinyue Ma, Changfu Liang, Yonghui Li , and Xiaoning He Hainan Provincial Key Lab of R&D on Tropic Herbs, School of Pharmacy, Hainan Medical University, Haikou 571199, China Te Second Afliated Hospital, Hainan Medical University, Haikou 571199, China Correspondence should be addressed to Yonghui Li; lyhssl@126.com and Xiaoning He; hexiaoningvv@aliyun.com Received 1 September 2022; Revised 29 November 2022; Accepted 6 December 2022; Published 21 December 2022 Academic Editor: Idaira Pacheco-Ferna´ndez Copyright © 2022 Yuhuang Wu et al. Tis is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. A rapid, sensitive, selective, and accurate HPLC–MS/MS method was developed and validated for the simultaneous determination of chlorogenic acid, naucleactonin C, khaephuoside A 3,4-dimethoxyphenyl-1-O-β-apiofuroseyl(1⟶ 2)-β-D-glucopyranoside in rat plasma and tissues after oral administration of Nauclea ofcinalis extracts. Chloramphenicol was used as an internal standard (IS). Te plasma and tissue samples were extracted by protein precipitation with methanol-ethyl acetate (1 :1, v/v) including 0.1% (v/v) formic acid. Te chromatographic separation was achieved by using an C18 column with gradient elution using mobile phase, which consisted of 0.1% formic acid water (A) and acetonitrile (B) and the fow rate of 0.8 mL/min. Mass spectrometric detection was performed in multiple reaction monitoring (MRM) mode utilizing electrospray ionization (ESI) in negative mode. Te developed method exhibited good linearity (determination coefcients, R ≥ 0.9849), and the lower limits of quantifcation were 2, 5, 5, and 25 ng/mL for chlorogenic acid, naucleactonin C, khaephuoside A, and 3,4-dimethoxyphenyl-1-O- β-apiofuroseyl(1⟶ 2)-β-D-glucopyranoside. Te intraday and interday precisions (relative standard deviation, RSD) were less than 12.65%, while the accuracy was ranged from 86.31 to 114.17%. Te recovery rate were 51.85–97.06%, 75.99–106.68%, 77.46–105.35%, and 68.36–103.75% for chlorogenic acid, naucleactonin C, khaephuoside A, and 3,4-dimethoxyphenyl-1-O- β-apiofuroseyl(1⟶ 2)-β-D-glucopyranoside the matrix efects were 50.17–116.62%, 86.75–115.99%, 45.79–87.44%, and 51.60–92.34% for chlorogenic acid, naucleactonin C, khaephuoside A, and 3,4-dimethoxyphenyl-1-O-β-apiofuroseyl(1⟶ 2)- β-D-glucopyranoside in diferent matrix. Te developed method was successfully applied to a pharmacokinetic study and tissue distribution of four compounds in rats after oral administration of Nauclea ofcinalis extracts. kinds of compounds have been isolated and identifed from 1. Introduction Nauclea ofcinalis, including alkaloids, phenolic acids, iri- Nauclea ofcinalis Pierre. ex Pitard, is one of the most doids, pentacyclic triterpenoids, and favonoids [8–12]. commonly used traditional medicines in China and is Previous studies showed that alkaloids were the main bio- mainly distributed in Hainan, Guangxi, Guangdong, and active compound in Nauclea ofcinalis [13–15]. Recent lit- other provinces in the south of China [1]. Modern phar- erature reported that phenolic compounds such as macological studies reported that Nauclea ofcinalis exhibits protocatechuic acid and chlorogenic acid possess antioxi- various biological properties such as antibacterial, anti-in- dant and antibacterial efects [16–18]. Terefore, the phe- fammatory, and analgesic activity [2–6]. As a traditional nolic compounds may be another kind of active ingredient Chinese medicine, the stems and twigs of Nauclea ofcinalis in Nauclea ofcinalis. are used for the treatment of colds, fever, acute tonsillitis, As a valuable work, the pharmacokinetics research could sore throat, and other diseases [1, 7]. A number of diferent not only explain the absorption, distribution, metabolism, 2 Journal of Analytical Methods in Chemistry and excretion of bioactive compounds but also reveal the (5.01–6.0 min). Te fow rate was 0.8 mL/min, and the in- mechanism of action and the cause of toxicity [19]. Several jection volume was 5μL. reports have been published on the pharmacokinetics of the Mass spectrometric detection was performed on an AB- alkaloids of Nauclea ofcinalis. For example, the pharma- SCIEX API 4000 mass spectrometer (AB SCIEX, Singapore) cokinetics of alkaloids of Nauclea ofcinalis extracts and with ESI in negative ion multiple reaction monitoring Danmu preparations in rat plasma [20–22] and the phar- (MRM) mode. Te optimized instrument parameters were macokinetics of Strictosamide in dog plasma [23]. However, as follows: nebulizer gas: 50 psi, heated by N gas: 55 psi, ion as one of the main active compounds of Nauclea ofcinalis, spray voltage: −4500 V and temperature: 550 C; nebulizer, the pharmacokinetics of phenolic acid compounds were blowback gas, and collision gas were nitrogen; declustering rarely researched. To our knowledge, the quantitative de- potential (DP), collision energy (CE), and collision cell exit tection of pharmacokinetics and tissue distribution of potential (CXP) of the four analytes and chloramphenicol chlorogenic acid, naucleactonin C, khaephuoside A, and 3,4- (IS) are shown in Table 1. dimethoxyphenyl-1-O-β-apiofuroseyl(1⟶ 2)-β-D-gluco- pyranoside of Nauclea ofcinalis extract have not been re- 2.3. Preparation of Nauclea ofcinalis Extract. Powdered ported in rats. Hence, it is necessary to establish rapid and Nauclea ofcinalis stems (1 kg) were accurately weighed and accurate approaches to investigate its pharmacokinetics and heated and refuxed by water (1 :10, w/v) for 2 h. Te ex- tissue distribution. tractions were combined and concentrated under reduced Te aim of the present investigation is to develop a re- pressure to obtain the 25 g of crude water extracts. liable and sensitive method based on HPLC-MS/MS to quantify the four compounds of Nauclea ofcinalis extract in the plasma and tissue distribution of rats. It is expected that 2.4. Preparations of Standard Solutions, IS, and Quality the results of this study can provide a useful reference for Control (QC) Samples. Te chlorogenic acid, naucleactonin understanding the mechanism of action, safety evaluation, C, khaephuoside A, and 3,4-dimethoxyphenyl-1-O- and clinical application of Nauclea ofcinalis. β-apiofuroseyl(1⟶ 2)-β-D-glucopyranoside and chlor- amphenicol (IS) were dissolved in methanol at a concen- tration of 1 mg/mL as the stock solution. A series of mixed 2. Material and Methods standard working solutions were prepared by diluting the primary mixed stock solution with methanol at appropriate 2.1.ChemicalsandReagents. Te stems of Nauclea ofcinalis ratios. All the solutions were kept away from light at 4 C were collected from Qiongzhong County, Hainan Province, until analysis. and identifed by Prof. Jianping Tian of Hainan Medical Te calibration standards were prepared by spiking 10 μL University. Standards of naucleactonin C, khaephuoside A, of the mixed standard solution and 10μL (500 ng/mL) IS and 3,4-dimethoxyphenyl-1-O-β-apiofuroseyl(1⟶ 2)- solution into 90μL rat blank matrixes at fnal concentrations β-D-glucopyranoside were isolated from Nauclea ofcinalis of 2–2400 ng/mL of chlorogenic acid, 5–2400 ng/mL of in our laboratory by silica gel column, semi-preparative high naucleactonin C, 5-2000 ng/mL of khaephuoside A, and performance liquid chromatography, which structure was 25–2400 ng/mL of 3,4-dimethoxyphenyl-1-O- identifed by NMR, and the HPLC purity was over than 98%. β-apiofuroseyl(1⟶ 2)-β-D-glucopyranoside. Te standards for chlorogenic acid were purchased from Te QC samples were prepared at concentrations of 2, 5, Chengdu Pufeide Biological Technology Co., Ltd (HPLC 100, and 2400 ng/mL for chlorogenic acid; 5, 8, 140, and >98%, Sichuan, China). Chloramphenicol was provided by 2400 ng/mL for naucleactonin C; 5, 8, 120, and 2000 ng/mL the National Institute for Food and Drug Control (Beijing, for khaephuoside A; 25, 50, 350, and 2400 ng/mL for 3,4- China). Methanol and acetonitrile were all chromato- dimethoxyphenyl-1-O-β-apiofuroseyl(1⟶ 2)-β-D-gluco- graphically pure and purchased from Fisher Scientifc (Fair pyranoside, respectively. Te IS concentration was 45 ng/mL Lawn, NJ, USA); ethyl acetate and formic acid were all in all calibration standards and QC. chromatographically pure and purchased from Aladdin Industrial Corporation (Shanghai, China); purifed water was prepared by a LabTower EDI system (Termo Scientifc, 2.5. Animals. Male Sprague–Dawley (SD) rats (250± 20 g USA). weight) were purchased from Tianqin Biotechnology Co. Ltd. (License No. 4307256220100013061), Changsha, China. Te animals were maintained at 22 ± 2 C and 60%± 10% 2.2. Instrument and Analytical Conditions. humidity with a 12 h light/dark cycle and allowed free access Chromatographic analysis was performed on the LC- to food and water. All animal experiments were performed 20ADXR System (Shimadzu, Japan). Te Phenomenex in accordance with the Institutional Animal Care and Use Kinete EVO C18 100 A (50 mm × 2.1 mm, 5μm) column was Committee at the Hainan Medical University (Haikou, used as the analytical column maintained at 40 C with (A) China). 0.1% formic acid aqueous solution and (B) acetonitrile as the mobile phase. Te gradient elution program was operated as follows: 2% B (0–0.5 min), 2–25% B (0.5–0.51 min), 25-55% 2.6. Preparation of Biological Samples. An aliquot of 100μL B (0.51–3.0 min), 55-95% B (3.0–3.01 min), 95% B of rat plasma and tissue homogenate were transferred to (3.01–5.0 min), 95–2% B (5.0–5.01 min), and 2% B a 2 mL tube. Subsequently, 10μL IS (500 ng/mL) were added Journal of Analytical Methods in Chemistry 3 Table 1: Optimized multiple reaction monitoring parameters for the detection of four analytes and IS. Retention time Precursor ion Quantitative ion DP CE CXP Compounds (min) (m/z) (m/z) (V) (V) (V) Chlorogenic acid 1.02 353.0 190.9 −51 −22.9 −10.5 Naucleactonin C 1.65 335.1 182.9 −110 −37 −9 Khaephuoside A 0.99 477.1 293.0 −98 −18 −6 3,4-Dimethoxyphenyl-1-O-β-apiofuroseyl(1⟶ 2)- 0.97 447.1 232.9 −84 −25 −14 β-D-glucopyranoside Chloramphenicol (IS) 1.16 320.8 175.9 −85 −22 −9 Abbreviations. DP, declustering potential; CE, collision energy; CXP, collision cell exit potential. followed by 1 mL of methanol-ethyl acetate (1 :1, v/v) accurately weighed and homogenized by using physiological including 0.1% (v/v) formic acid. Te sample was then saline at four times the tissue weight (w/v). Te homogenates vortexed for 1 min and centrifuged at 13,000 rpm for were stored at −80 C until analysis. 5 min. Tereafter, the supernatant was dried at 37 C with nitrogen, which the residue was reconstituted with 80 μL 3. Results and Discussion methanol, centrifuged at 13,000 rpm for 5 min, and 5 μL of 3.1.OptimizationofChromatographicandMassSpectrometric the sample were injected into the HPLC-MS/MS system Conditions. Optimization of chromatographic conditions: for analysis. by analyzing the separation efect and peak shape of four analytes and the IS on diferent columns, we found that the 2.7. Method Validation. Te method validation assays were Phenomenex Kinete EVO C18 column gave the best sepa- carried out according to the U.S. Food and Drug Admin- ration and peak shape. Subsequently, by exploring diferent istration (FDA) Bioanalytical Method Validation (Food and mobile phase systems, such as methanol-water and aceto- Drug Administration, 2018), including specifcity, linearity, nitrile-water, we found that the four analytes had better recovery, matrix efect, precision, accuracy, and stability response values in acetonitrile. Finally, by assessing the pH [24]. of the mobile phase (0.1, 0.2, 0.5, and 0.8% formic acid solution), the response values of four analytes were highest when the concentration of formic acid was 0.1%. Terefore, 2.8. Pharmacokinetic Study. Six male SD rats were tested in 0.1% formic acid water-acetonitrile was selected for use as pharmacokinetics studies. Te rats were housed at 22 ± 2 C the mobile phase. and fasted for 12 h with free access to water prior to dosing, Optimization of mass spectrometric conditions: the four which were orally administered Nauclea ofcinalis extracts analytes and the IS had strong [M-H] peaks in the negative at the dose of 2 g/kg (equivalent to 32.74 mg/kg chlorogenic ion mode, which can be easily broken and used for stable acid, 0.78 mg/kg naucleactonin C, 14.1 mg/kg khaephuoside fragment analysis and detection. Terefore, the ESI in A, and 10.39 mg/kg 3,4-dimethoxyphenyl-1-O- negative ion mode and Q1, MS2, and MRM scan modes were β-apiofuroseyl(1⟶ 2)-β-D-glucopyranoside). Te blood adopted. Te chemical structures, precursor ion, and samples (approximately 0.5 mL) were collected via the ve- product ion of four analytes and IS are shown in Figure 1. nous plexus of the eye socket at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, 1, 12, and 24 h under anesthesia. Next, each sample was im- 3.2. Optimization of the Sample Preparation. For sample mediately centrifuged at 4000 rpm for 10 min to acquire the processing, a simple protein precipitation method was frst plasma. Te plasma was transferred to new tubes and stored tried for methanol and acetonitrile, respectively, but the at −80 C until further use. Te pharmacokinetics parameters recoveries of chlorogenic acid and naucleactonin C were of the four compounds in rat plasma were calculated by the unsatisfactory. In addition, liquid-liquid extraction (LLE) noncompartment model using Drug and Statistics (DAS was tried for various solvents tested, including n-butanol, 3.3.0) software (Beijing, China). dichloromethane, isopropanol, methyl tert-butyl ether, and ethyl acetate, but no satisfactory recovery was obtained for 2.9. Tissue Distribution. Tirty rats were divided into fve khaephuoside A, and 3,4-dimethoxyphenyl-1-O- groups (n � 6) at random, which were orally administered β-apiofuroseyl(1⟶ 2)-β-D-glucopyranoside. Te extrac- Nauclea ofcinalis extracts at a dose of 2 g/kg for tissue tion of chlorogenic acid should be in an acidic condition. distribution conducted at 0.5, 1, 2, 4, and 6 h. During the Terefore, the extraction results of chlorogenic acid at collection, the heart, liver, spleen, lung, kidney, stomach, diferent pH are investigated. small intestine, and brain were rinsed with physiological Finally, the combination method using mixture solution saline solution to get rid of the blood or content and blotted of methanol-ethyl acetate (1 :1, v/v) including 0.1% (v/v) on flter paper and then weighed. Each tissue sample was formic acid was selected. Consequently, satisfactory and 4 Journal of Analytical Methods in Chemistry 100 293.0 190.9 100 OMe OMe HO OH N O HO OH H H H O O OMe O HO HO O HO O OH O Naucleactonin C HO HO 182.9 HO OH 335.1 477.1 Chlorogenic acid Khaephuoside A 148.0 168.9 289.0 233.0 249.0 277.0 236.9 262.9 133.9 141.8 0 0 0 100 200 300 400 100 200 300 400 200 300 400 500 m (s) m (s) m (s) OMe 89.0 151.9 100 100 OH NO O HO O OMe HO HO HO O 3,4-dimethoxyphenyl-1-O-β- OH Cl apiofuroseyl(1→2)-β-D- glucopyranoside Cl HO OH Chloramphenicol (IS) 124.9 232.9 447.1 175.9 193.9 256.9 98.9 71.0 137.9 150.8 320.8 149.0 112.9 130.9 59.0 161.0 190.9 249.0 85.0 126.8 148.0 81.2 292.7 120.9 218.8 110.8 126.8 173.0 0 0 100 200 300 400 500 50 150 250 350 m (s) m (s) Figure 1: Chemical structures, precursor ion, and product ion mass spectra of four analytes and IS. 5.3e2 1.02 1.01 1.8e3 2.7e4 0 6 06 0 6 1.65 1.65 8.5e2 2.0e2 3.9e4 0 6 0 6 0.99 2.5e2 1.01 1.7e3 2.1e4 3 3 0 6 0 6 0 6 0.97 1.0e2 0.99 1.2e3 3.4e3 4 4 0 6 06 0 6 1.16 2.0e2 1.16 5.7e4 3.1e4 5 5 0 6 0 6 Time (min) Time (min) Time (min) (a) (b) (c) Figure 2: Typical chromatograms of four analyte and IS in rat plasma: (a) blank plasma samples, (b) blank plasma samples spiked with four analytes (LLOQ) and IS, (c) rat plasma samples at 0.5 h after oral administration the Nauclea ofcinalis extracts spiked with IS; (1) chlorogenic acid, (2) naucleactonin C, (3) khaephuoside A, (4) 3,4-dimethoxyphenyl-1-O-β-apiofuroseyl(1⟶ 2)-β-D-glucopyranoside, (5) chloramphenicol (IS). Intensity (%) Intensity (%) Intensity (%) Intensity (%) Intensity (%) Journal of Analytical Methods in Chemistry 5 consistent recovery from plasma and tissues samples was Naucleactonin C, Khaephuoside A, 3,4-dimethoxyphenyl-1- achieved for four analytes and IS. O-β-apiofuroseyl(1⟶ 2)-β-D-glucopyranoside, and IS were eluted approximately at retention time of 1.02 min, 1.65 min, 0.99 min, 0.97 min, and 1.16 min, respectively. No 3.3. Method Validation interfering endogenous substances were found at the re- spective retention times. 3.3.1. Selectivity. Te selectivity was assessed by analyzing chromatograms of blank rat plasma and tissue homogenates from diferent batches, blank plasma and tissue homoge- 3.3.2. Linearity of Calibration Curve and Lower Limit of nates spiked with analytes and IS, and plasma, tissue samples Quantifcation. Te calibration curves were constructed by obtained from rats after oral administration of Nauclea plotting the peak areas ratios of chlorogenic acid/IS, Nau- ofcinalis extracts (n � 6). Figure 2 shows typical chro- cleactonin C/IS, Khaephuoside A/IS, and 3,4-dimethox- matograms of four analytes and IS in rat plasma (Figures S1 yphenyl-1-O-β-apiofuroseyl(1⟶ 2)-β-D-glucopyranoside/ and S2 of the Supporting Material show typical chro- IS versus theoretical concentrations. Tis method exhibited matograms of four analytes and IS in liver tissue and kidney a good linear response for the range of concentrations from 2 tissue). Under the given condition, Chlorogenic acid, to 2500 ng/mL in plasma and from 2 to 2500 ng/mL in Table 2: Summary of standard curves, correlation coefcients, LLOQ, and linear ranges of four analytes in plasma and tissue samples. Linear regression LLOQ Compounds Matrix R Linear range (ng/mL) equation (ng/mL) Plasma y � 0.0288x + 0.00193 0.9994 2 2–2500 Heart y � 0.0063x − 0.00771 0.9849 2 2–2500 Liver y � 0.0267x − 0.0323 0.9980 2 2–2500 Spleen y � 0.0392x + 0.0277 0.9886 2 2–2500 Lung y � 0.0442x + 0.0239 0.9958 2 2–2500 Chlorogenic acid Kidney y � 0.0217x − 0.0302 0.9950 2 2–2500 Small y � 0.0334x − 0.0401 0.9964 2 2–2500 intestine Stomach y � 0.0402x − 0.0129 0.9952 2 2–2500 Brain y � 0.0499x − 0.0562 0.9960 2 2–2500 Plasma y � 0.00535x + 0.0207 0.9964 5 5–2500 Heart y � 0.00313x − 0.00116 0.9926 5 5–2500 Liver y � 0.00395x − 0.0101 0.9964 5 5–2500 Spleen y � 0.00511x − 0.00526 0.9950 5 5–2500 Lung y � 0.00534x − 0.00263 0.9980 5 5–2500 Naucleactonin C Kidney y � 0.00363x − 0.0098 0.9960 5 5–2500 Small y � 0.00224x − 0.00527 0.9968 5 5–2500 intestine Stomach y � 0.00465x − 0.0138 0.9972 5 5–2500 Brain y � 0.00552x − 0.0163 0.9932 5 5–2500 Plasma y � 0.00316x + 0.0153 0.9954 5 5–2000 Heart y � 0.00673x − 0.005 0.9946 5 5–2000 Liver y � 0.0063x − 0.00502 0.9962 5 5–2000 Spleen y � 0.011x − 0.0054 0.9980 5 5–2000 Lung y � 0.0099x − 0.00166 0.9978 5 5–2000 Khaephuoside A Kidney y � 0.00531x − 0.0047 0.9954 5 5–2000 Small y � 0.0058x − 0.0156 0.9906 5 5–2000 intestine Stomach y � 0.00745x − 0.00837 0.9920 5 5–2000 Brain y � 0.0126x + 0.000377 0.9938 5 5–2000 Plasma y � 0.000845x + 0.00396 0.9984 25 25–2500 Heart y � 0.000804x + 0.00159 0.9849 25 25–2500 Liver y � 0.000873x − 0.00515 0.9968 25 25–2500 Spleen y � 0.00134x + 0.00947 0.9894 25 25–2500 3,4-Dimethoxyphenyl-1-O-β-apiofuroseyl(1⟶ 2)- Lung y � 0.00138x + 0.00626 0.9964 25 25–2500 β-D-glucopyranoside Kidney y � 0.000633x − 0.000538 0.9930 25 25–2500 Small y � 0.00109x + 0.000966 0.9962 25 25–2500 intestine Stomach y � 0.00119x + 0.00549 0.9964 25 25–2500 Brain y � 0.00163x − 0.0149 0.9952 25 25–2500 Abbreviations. LLOQ, lower limit of quantifcation; R , determination coefcients. 6 Journal of Analytical Methods in Chemistry tissues; all determination coefcients (R ) were greater than accurate for the determination of four analytes in rat plasma 0.9849. Te lower limit of quantifcation (LLOQ) was de- and tissue samples. fned as the lowest concentration of a signal-noise (S/N) ratios of 10 :1, respectively, the precision was less than 15% and the accuracy was within±20%. Te limits were adequate 3.3.4. Extraction Recovery and Matrix Efect. Six batches of blank plasma from independent sources were used to obtain for studies of pharmacokinetics and tissue distribution by oral administration of Nauclea ofcinalis extracts. Data from extracted samples, postextracted spiked samples, and unextracted samples at high, medium, low, and LLOQ the determination are shown in Table 2. concentrations. Te peak areas of the three types of samples were recorded as A, B, and C, respectively. Extraction re- 3.3.3. Accuracy and Precision. Accuracy and precision were covery was evaluated by A/B. Matrix efects were calculated evaluated by analyzing QC samples at high, medium, low, by comparing the A/C of analytes. Te extraction, recovery, and LLOQ concentrations (n � 6) on the same day and three and matrix efect of four analytes in rat plasma and tissues consecutive days using the standard curve, respectively. Te are shown in Table 4. Te recovery rate were 51.85–97.06%, acceptable limits of accuracy were required to be within 75.99–106.68%, 77.46–105.35%, and 68.36–103.75% for ±15% of the actual value except when at LLOQ, and the chlorogenic acid, naucleactonin C, khaephuoside A, and 3,4- intra- and interday precision and accuracy data of the de- dimethoxyphenyl-1-O-β-apiofuroseyl(1⟶ 2)-β-D-gluco- termination were shown in Table 3. In our study, the in- pyranoside and the matrix efects were 50.17–116.62%, traday precision ranged from 0.62 to 12.65%, and intraday 86.75–115.99%, 45.79–87.44%, and 51.60–92.34% for accuracy ranged from 86.48 to 114.17%. Te interday pre- chlorogenic acid, naucleactonin C, khaephuoside A, and 3,4- cision ranged from 2.42 to 12.61%, and the interday accuracy dimethoxyphenyl-1-O-β-apiofuroseyl(1⟶ 2)-β-D-gluco- ranged from 86.31 to 113.99%. Te results showed the ac- pyranoside in diferent matrices, indicating some ion inhibitory curacy and precision were within the acceptable limits, efects for four compounds in the biological samples, the RSD (relative standard deviation) of recovery and matrix efect were which proved the method was reproducible, reliable, and Table 3: Summary of intraday and interday precisions and accuracies of four analytes in plasma and tissue samples. Intraday (n � 6) Interday (n � 18) Compounds Matrix Concentration (ng/mL) Accuracy RSD (%) Accuracy RSD (%) 2 104.02± 11.91 11.45 100.64± 11.67 11.60 4 97.72± 9.23 9.45 96.09± 7.08 7.36 Plasma 100 104.42± 6.23 5.97 104.29± 10.6 10.16 2400 101.22± 3.20 3.16 96.59± 8.11 8.40 2 101.90± 12.14 11.92 109.34± 11.74 10.74 4 99.35± 3.89 3.91 93.67± 6.13 6.54 Heart 100 96.57± 4.81 4.98 94.46± 7.80 8.26 2400 109.00± 5.90 5.41 102.88± 6.85 6.66 2 101.65± 2.82 2.78 100.88± 5.38 5.33 4 101.45± 2.98 2.93 98.59± 6.72 6.82 Liver 100 100.30± 2.89 2.88 97.78± 4.89 5.00 2400 95.07± 5.74 6.04 97.11± 5.58 5.75 2 96.40± 7.63 7.92 102.14± 8.90 8.71 4 95.40± 4.20 4.40 101.28± 9.61 9.49 Spleen 100 102.40± 2.00 1.95 111.52± 7.00 6.27 2400 94.93± 4.81 5.07 108.26± 10.41 9.62 2 101.53± 6.40 6.30 101.22± 7.45 7.36 4 99.75± 6.09 6.10 93.33± 11.17 11.97 Chlorogenic acid Lung 100 108.33± 5.47 5.04 111.33± 6.49 5.83 2400 100.60± 3.35 3.33 101.63± 5.07 4.99 2 98.12± 5.36 5.47 96.23± 5.33 5.54 4 100.13± 2.17 2.17 92.09± 7.21 7.83 Kidney 100 99.37± 5.20 5.23 96.10± 6.36 6.62 2400 104.47± 5.79 5.54 99.47± 6.06 6.10 2 93.27± 4.13 4.43 96.03± 7.86 8.19 4 86.48± 5.09 5.89 93.40± 10.75 11.51 Small intestine 100 89.75± 7.96 8.87 96.61± 10.06 10.41 2400 92.70± 6.63 7.16 93.01± 5.27 5.66 2 105.87± 7.87 7.43 107.84± 7.45 6.90 4 93.17± 7.02 7.53 98.76± 6.44 6.52 Stomach 100 106.00± 3.95 3.73 106.35± 4.46 4.19 2400 100.63± 4.18 4.15 99.41± 4.81 4.84 2 109.27± 6.74 6.17 104.39± 9.47 9.07 4 101.00± 3.81 3.77 101.29± 5.42 5.35 Brain 100 106.00± 4.94 4.66 106.77± 7.16 6.70 2400 101.27± 2.25 2.22 105.62± 7.04 6.67 Journal of Analytical Methods in Chemistry 7 Table 3: Continued. Intraday (n � 6) Interday (n � 18) Compounds Matrix Concentration (ng/mL) Accuracy RSD (%) Accuracy RSD (%) 5 95.62± 12.10 12.65 98.24± 10.30 10.48 8 100.53± 11.78 11.71 96.43± 12.16 12.61 Plasma 140 99.95± 6.49 6.50 95.76± 7.18 7.50 2400 97.05± 4.80 4.95 95.50± 6.23 6.52 5 111.27± 7.32 6.58 107.46± 10.58 9.85 8 99.28± 5.48 5.52 101.15± 11.46 11.33 Heart 140 93.45± 4.34 4.65 93.19± 5.33 5.72 2400 105.92± 9.54 9.01 104.46± 6.66 6.38 5 100.90± 5.40 5.36 108.30± 7.82 7.22 8 99.47± 2.82 2.84 100.52± 5.00 4.98 Liver 140 88.60± 2.60 2.93 93.53± 6.33 6.77 2400 102.50± 1.52 1.48 101.42± 2.46 2.42 5 111.00± 6.99 6.29 104.42± 9.31 8.91 8 98.90± 3.55 3.59 100.74± 6.77 6.72 Spleen 140 96.07± 3.95 4.11 93.72± 5.30 5.66 2400 100.02± 3.39 3.39 103.13± 5.06 4.91 5 113.83± 4.07 3.58 113.44± 5.37 4.73 8 103.28± 10.46 10.13 102.03± 10.16 9.96 Naucleactonin C Lung 140 104.17± 3.76 3.61 103.83± 4.05 3.90 2400 107.83± 4.07 3.77 107.06± 5.02 4.69 5 100.70± 4.28 4.25 103.81± 6.61 6.36 8 98.38± 5.31 5.40 99.18± 4.36 4.39 Kidney 140 95.37± 4.19 4.39 100.44± 7.07 7.04 2400 106.80± 9.74 9.12 104.87± 6.46 6.16 5 109.98± 10.97 9.97 105.41± 10.19 9.67 8 99.30± 8.94 9.01 100.44± 9.18 9.14 Small intestine 140 88.73± 8.96 10.10 86.31± 7.66 8.88 2400 103.30± 6.82 6.60 98.26± 7.60 7.73 5 109.83± 7.05 6.42 102.67± 10.09 9.83 8 101.58± 2.96 2.92 95.73± 8.07 8.43 Stomach 140 94.88± 2.74 2.88 92.09± 5.17 5.62 2400 113.17± 4.88 4.31 108.96± 7.21 6.62 5 105.75± 10.81 10.22 105.82± 8.77 8.29 8 104.78± 7.46 7.12 100.57± 8.11 8.06 Brain 140 101.22± 4.59 4.53 98.93± 5.73 5.80 2400 113.83± 3.31 2.91 111.39± 5.54 4.98 5 101.63± 9.93 9.77 105.66± 10.50 9.94 8 101.02± 9.64 9.55 101.78± 7.85 7.71 Plasma 120 112.33± 4.76 4.24 106.16± 6.11 5.76 1800 104.08± 4.39 4.21 102.52± 4.41 4.31 5 113.50± 9.18 8.09 103.33± 10.92 10.57 8 100.82± 6.19 6.14 98.68± 7.06 7.16 Heart 120 96.73± 2.92 3.02 89.72± 8.01 8.93 1800 109.12± 9.45 8.66 102.43± 8.13 7.94 5 99.97± 7.06 7.07 101.51± 8.49 8.36 8 104.10± 5.07 4.87 101.81± 4.02 3.95 Liver 120 99.33± 0.62 0.62 100.43± 4.55 4.53 1800 108.00± 6.10 5.65 103.94± 5.60 5.39 5 98.73± 11.21 11.35 103.62± 9.47 9.14 8 100.98± 6.25 6.19 103.08± 6.75 6.55 Spleen 120 99.35± 2.52 2.54 109.89± 8.84 8.04 1800 109.00± 4.60 4.22 113.06± 5.36 4.74 5 108.17± 10.19 9.42 109.93± 7.6 6.92 8 109.90± 11.25 10.24 106.91± 9.95 9.31 Khaephuoside A Lung 120 90.32± 6.12 6.78 95.54± 6.53 6.83 1800 100.73± 2.02 2.00 103.65± 6.91 6.66 5 106.82± 7.91 7.40 109.94± 6.29 5.72 8 113.67± 2.94 2.59 111.40± 6.12 5.49 Kidney 120 105.30± 6.14 5.83 107.78± 6.91 6.41 1800 105.02± 9.81 9.34 108.84± 7.60 6.99 5 106.50± 8.53 8.01 104.08± 10.11 9.71 8 99.98± 9.43 9.13 101.89± 11.58 11.36 Small intestine 120 89.90± 6.84 7.61 93.74± 7.80 8.32 1800 104.45± 6.22 5.96 105.44± 5.15 4.89 5 96.63± 9.15 9.47 95.98± 10.26 10.69 8 90.92± 7.96 8.76 97.66± 8.61 8.82 Stomach 120 104.00± 6.13 5.90 103.24± 5.31 5.15 1800 109.83± 4.83 4.40 110.50± 4.54 4.11 5 108.90± 9.74 8.95 107.11± 7.94 7.41 8 108.55± 9.90 9.12 107.23± 8.01 7.47 Brain 120 113.17± 6.71 5.93 110.13± 6.09 5.53 1800 100.50± 3.99 3.97 108.44± 7.63 7.03 8 Journal of Analytical Methods in Chemistry Table 3: Continued. Intraday (n � 6) Interday (n � 18) Compounds Matrix Concentration (ng/mL) Accuracy RSD (%) Accuracy RSD (%) 25 109.00± 4.05 3.72 101.34± 9.10 8.98 50 105.82± 10.95 10.35 102.61± 7.89 7.69 Plasma 350 109.83± 5.23 4.76 104.49± 8.60 8.23 2400 96.82± 2.76 2.85 100.71± 7.50 7.45 25 99.12± 5.06 5.10 96.73± 8.08 8.35 50 96.45± 6.75 7.00 98.20± 8.28 8.43 Heart 350 89.15± 4.83 5.42 90.19± 6.82 7.56 2400 99.80± 10.89 10.91 99.33± 7.66 7.71 25 105.07± 6.62 6.30 103.78± 6.39 6.16 50 108.48± 7.39 6.82 108.22± 7.38 6.82 Liver 350 112.15± 6.89 6.14 113.99± 4.83 4.24 2400 98.28± 3.67 3.73 100.28± 3.24 3.23 25 102.03± 10.68 10.47 104.78± 8.63 8.23 50 99.90± 520 5.21 103.82± 7.77 7.48 Spleen 350 97.70± 4.12 4.21 109.12± 9.07 8.31 2400 100.65± 5.46 5.43 110.22± 8.94 8.11 25 96.17± 7.59 7.89 99.80± 10.99 11.01 3,4-Dimethoxyphenyl-1-O-β- 50 106.78± 7.08 6.63 107.49± 8.16 7.59 apiofuroseyl(1⟶ 2)-β- Lung 350 96.42± 4.49 4.66 100.05± 5.50 5.49 D-glucopyranoside 2400 99.67± 6.29 6.31 99.32± 5.68 5.72 25 97.02± 9.75 10.05 96.23± 9.38 9.74 50 108.00± 5.66 5.24 106.69± 7.00 6.56 Kidney 350 99.63± 5.83 5.85 100.46± 6.78 6.75 2400 96.50± 7.41 7.68 97.74± 6.31 6.46 25 98.25± 8.92 9.08 97.96± 7.73 7.90 50 101.58± 6.41 6.31 101.89± 7.13 7.00 Small intestine 350 89.58± 4.27 4.76 98.17± 8.39 8.55 2400 100.18± 4.77 4.76 101.32± 4.83 4.77 25 103.63± 5.21 5.02 99.07± 9.64 9.73 50 101.90± 9.32 9.14 97.96± 9.33 9.52 Stomach 350 114.17± 3.19 2.79 109.34± 6.14 5.62 2400 100.62± 3.42 3.40 99.42± 4.66 4.69 25 106.95± 12.13 11.34 99.13± 11.26 11.36 50 105.78± 7.63 7.22 107.10± 7.86 7.34 Brain 350 110.00± 9.01 8.19 112.56± 6.93 6.16 2400 99.42± 2.90 2.91 101.13± 5.44 5.38 less than 15%. Tere were no signifcant diferences in re- four analytes are shown in Figure 3, and the main phar- coveries and matrix efects between the four compounds at macokinetic parameters were estimated of four analytes in diferent concentrations in the same matrix. Table 6. Tese compounds were detected at 5 min after oral administration of Nauclea ofcinalis extracts, indicating four 3.3.5. Stability. Te stability was evaluated by analyzing QC compounds were rapidly absorbed, exhibiting a therapeutic samples at high, medium, low, and LLOQ concentrations efect. Chlorogenic acid, khaephuoside A, and 3,4-dime- (n � 6) under diferent storage conditions, including room thoxyphenyl-1-O-β-apiofuroseyl(1⟶ 2)-β-D-glucopyr- temperature for 4 h, auto-sampler (4 C) for 8 h, three freeze- anoside were rapidly absorbed and peaked at approximately thaw cycles (freeze cycle was at −20 C while thaw cycle was 0.5 h, while naucleactonin C peaked at 1.66 h, indicating that done at room temperature) and −80 C for 30 days. Te naucleactonin C was absorbed relatively slowly. Te content results are shown in Table 5. It was evident from these data of naucleactonin C was much lower than that of the other that four storage conditions have no signifcant efect on the three compounds in the Nauclea ofcinalis extracts, but sample. Te results indicated that four analytes were stable naucleactonin C was present in a much larger amount than under these four storage conditions. other three compounds for AUC (area under the plasma 0−t concentration–time curve) and AUC , indicating that 0−∞ 3.4.PharmacokineticStudy. Te validated analytical method naucleactonin C may had excellent bioavailability. However, was successfully applied to the quantitative estimation of the bioavailability of chlorogenic acid and khaephuoside A chlorogenic acid, naucleactonin C, khaephuoside A, and 3,4- may not be satisfactory, the CL of chlorogenic acid and dimethoxyphenyl-1-O-β-apiofuroseyl(1⟶ 2)-β-D-gluco- khaephuoside A were 62.93± 15.996 and 54.40± 6.04 L/h/kg pyranoside in the plasma samples following oral adminis- after oral administration of Nauclea ofcinalis extract, re- tration of Nauclea ofcinalis extract at a dosage of 2 g/kg. spectively. Excessive clearance rate will afect the residence Pharmacokinetic parameters of four compounds were cal- time of the drug in the body, thereby reducing the efcacy. In culated by noncompartmental model using DAS 3.3.0 addition, literature reported that the prototype chlorogenic software. Te mean plasma concentration–time-curve of acid absorbed into the blood by oral chlorogenic acid Journal of Analytical Methods in Chemistry 9 Table 4: Summary of extraction recovery and matrix efect of four analytes in plasma and tissue samples (n � 6). Compounds Matrix Concentration (ng/mL) Matrix efect (%) RSD (%) Extraction recovery (%) RSD (%) 2 97.26± 5.35 5.50 80.45± 4.36 5.42 4 109.75± 4.40 4.01 80.04± 3.08 3.85 Plasma 100 82.80± 6.51 7.86 69.49± 4.05 5.84 2400 91.66± 1.53 1.67 72.26± 1.16 1.61 2 116.54± 3.03 2.60 80.53± 9.62 11.95 4 98.55± 2.71 2.75 76.82± 4.77 6.21 Heart 100 97.78± 1.76 1.80 62.02± 2.24 3.61 2400 93.25± 0.76 0.81 63.32± 0.58 0.91 2 83.69± 4.24 5.06 97.06± 3.75 3.86 4 79.96± 4.79 5.94 79.07± 2.90 3.66 Liver 100 63.82± 2.51 3.94 66.44± 0.24 0.37 2400 80.01± 0.90 1.12 69.83± 1.55 2.22 2 99.89± 2.61 2.61 80.09± 1.22 1.52 4 100.56± 1.99 1.98 86.82± 2.88 3.32 Spleen 100 92.05± 1.01 1.10 67.29± 1.81 2.69 2400 91.66± 0.81 0.88 73.12± 1.61 2.20 2 104.73± 1.78 1.70 81.80± 2.52 3.08 4 99.34± 2.07 2.09 88.65± 1.55 1.75 Chlorogenic acid Lung 100 98.65± 1.24 1.25 80.24± 2.11 2.63 2400 96.68± 1.81 1.88 76.89± 1.69 2.20 2 90.49± 5.66 6.26 78.34± 4.02 5.13 4 98.52± 4.21 4.27 86.92± 3.00 3.46 Kidney 100 95.03± 5.94 6.25 70.73± 4.90 6.92 2400 89.55± 2.38 3.16 72.70± 1.50 2.07 2 56.96± 6.16 10.81 56.20± 3.36 5.98 4 63.67± 2.46 4.14 51.85± 1.94 3.73 Small intestine 100 53.67± 1.06 1.97 61.26± 1.87 3.05 2400 50.17± 1.72 3.42 62.66± 1.06 1.70 2 114.40± 5.01 4.46 85.33± 5.20 6.10 4 116.62± 8.80 7.55 91.66± 6.11 6.67 Stomach 100 102.87± 3.24 3.15 66.48± 3.94 5.93 2400 98.38± 1.63 1.65 69.41± 1.56 2.25 2 97.89± 6.42 6.56 82.33± 4.65 5.65 4 105.09± 2.99 2.84 75.80± 3.59 4.74 Brain 100 90.02± 5.46 6.06 71.37± 1.17 1.63 2400 90.56± 2.16 2.39 86.06± 1.68 1.95 5 104.57± 6.94 6.64 100.30± 11.77 11.74 8 102.54± 2.55 2.48 106.68± 6.31 5.91 Plasma 140 104.07± 2.88 2.77 96.41± 1.66 1.72 2400 102.64± 2.09 2.04 88.27± 1.05 1.19 5 98.86± 2.67 2.70 97.71± 1.95 2.00 8 99.69± 3.96 3.97 96.33± 7.19 7.46 Heart 140 106.24± 4.87 4.58 85.37± 2.90 3.40 2400 99.81± 1.48 1.48 86.76± 1.61 1.86 5 101.81± 10.88 10.69 92.06± 9.87 10.72 8 111.16± 3.12 2.81 85.03± 4.91 5.77 Liver 140 97.92± 5.09 5.20 84.88± 4.52 5.33 2400 97.02± 1.11 1.14 80.54± 0.83 1.03 5 90.83± 3.65 4.02 100.21± 9.21 9.19 8 93.78± 6.07 6.47 99.90± 5.49 5.50 Spleen 140 97.28± 2.71 2.79 84.00± 1.54 1.83 2400 100.67± 0.79 0.78 84.60± 1.40 1.65 5 98.17± 7.63 7.77 105.63± 3.87 3.66 8 115.99± 7.97 5.01 99.21± 12.43 12.53 Naucleactonin C Lung 140 98.73± 1.39 1.41 95.48± 2.88 3.01 2400 99.21± 0.91 0.92 84.40± 1.82 2.15 5 99.90± 5.10 5.10 94.08± 8.11 8.62 8 100.35± 5.14 5.12 105.08± 5.22 4.97 Kidney 140 101.18± 0.70 0.69 86.02± 4.39 5.10 2400 102.49± 1.87 1.83 86.64± 2.26 2.60 5 98.00± 5.51 5.62 98.55± 9.61 9.75 8 99.37± 8.48 8.90 82.00± 5.03 6.14 Small intestine 140 95.42± 5.52 5.79 91.73± 1.67 1.82 2400 86.75± 2.24 2.58 86.61± 3.15 3.63 5 101.53± 3.55 3.50 97.93± 4.19 4.28 8 101.21± 3.87 3.82 99.25± 3.17 3.20 Stomach 140 99.62± 1.19 1.20 82.46± 1.88 2.28 2400 101.37± 1.35 1.33 75.99± 2.77 3.64 5 97.50± 6.50 6.67 88.08± 3.12 3.54 8 105.47± 5.14 4.87 88.38± 7.32 8.29 Brain 140 102.02± 0.85 0.83 97.63± 4.35 4.46 2400 101.24± 2.44 2.41 99.47± 1.42 1.43 10 Journal of Analytical Methods in Chemistry Table 4: Continued. Compounds Matrix Concentration (ng/mL) Matrix efect (%) RSD (%) Extraction recovery (%) RSD (%) 5 64.06± 5.66 8.84 88.84± 5.10 5.75 8 65.86± 5.65 8.57 92.35± 8.73 9.45 Plasma 120 55.55± 4.05 7.29 85.88± 2.06 2.40 1800 58.01± 1.93 3.33 85.24± 3.30 3.87 5 70.92± 8.09 11.41 96.07± 5.37 5.59 8 77.13± 3.56 4.61 82.29± 3.95 4.80 Heart 120 54.79± 1.14 2.08 92.18± 1.64 1.78 1800 59.93± 4.64 7.75 86.98± 4.87 5.59 5 73.90± 6.84 9.26 105.35± 11.02 10.46 8 53.20± 1.59 2.98 103.34± 3.75 3.63 Liver 120 58.17± 1.12 1.93 77.46± 2.17 2.80 1800 61.29± 2.80 4.58 84.49± 1.70 2.01 5 68.33± 1.41 2.07 90.99± 7.89 8.67 8 60.36± 2.58 4.28 96.38± 2.77 2.88 Spleen 120 58.12± 0.42 0.73 90.31± 2.80 3.10 1800 61.44± 1.13 1.84 86.75± 0.98 1.13 5 84.39± 4.20 4.97 77.71± 3.73 4.81 8 72.06± 6.89 9.56 87.06± 7.30 8.38 Khaephuoside A Lung 120 59.00± 2.55 4.33 86.34± 3.48 4.03 1800 61.35± 4.41 7.19 79.39± 6.48 8.17 5 57.46± 1.80 3.13 97.31± 4.38 4.50 8 51.41± 2.91 5.66 99.78± 3.41 3.42 Kidney 120 45.79± 1.30 2.84 99.65± 1.23 1.23 1800 47.23± 1.38 2.93 101.56± 2.17 2.14 5 56.14± 1.47 2.61 102.93± 10.44 10.15 8 67.00± 5.43 8.10 94.63± 6.27 6.63 Small intestine 120 62.69± 2.06 3.29 100.39± 5.00 4.98 1800 71.51± 2.08 2.91 99.78± 1.06 1.06 5 56.84± 3.21 5.64 95.49± 6.60 6.91 8 63.57± 5.59 8.80 83.81± 6.89 8.22 Stomach 120 54.53± 2.44 4.48 77.49± 4.08 5.27 1800 55.57± 1.07 1.92 79.96± 0.87 1.09 5 76.07± 1.69 2.22 93.48± 8.70 9.30 8 76.50± 6.23 8.15 102.39± 6.80 6.64 Brain 120 87.44± 3.23 3.70 90.50± 0.99 1.09 1800 79.81± 2.15 2.70 100.48± 1.42 1.41 25 64.80± 4.95 7.64 88.62± 8.56 9.66 50 65.25± 0.81 1.24 89.23± 4.24 4.76 Plasma 350 60.35± 3.67 6.08 85.65± 1.17 1.37 2400 55.98± 2.34 4.18 84.82± 3.16 3.72 25 79.81± 2.39 2.99 80.56± 4.05 5.03 50 73.97± 1.84 2.49 89.91± 2.89 3.21 Heart 350 69.51± 4.04 5.81 85.34± 4.20 4.92 2400 64.96± 2.28 3.51 85.00± 3.15 3.71 25 55.02± 4.02 7.31 88.70± 8.38 9.45 50 53.00± 1.63 3.08 94.52± 4.76 5.04 Liver 350 52.69± 2.12 4.03 83.51± 2.63 3.15 2400 51.60± 1.44 2.80 87.94± 2.88 3.27 25 81.22± 3.27 4.02 98.19± 2.87 2.92 50 87.52± 4.51 5.16 92.02± 3.15 3.42 Spleen 350 81.89± 1.03 1.26 89.66± 2.42 2.70 2400 80.06± 3.07 3.83 80.48± 2.15 2.68 25 91.11± 4.01 4.40 68.36± 4.42 6.47 3,4-Dimethoxyphenyl-1-O- 50 92.34± 3.46 3.75 77.05± 2.14 2.78 β-apiofuroseyl(1⟶ 2)-β- Lung 350 77.28± 4.36 5.65 83.53± 3.42 4.09 D-glucopyranoside 2400 76.69± 2.73 3.56 75.92± 0.73 0.97 25 63.39± 1.54 2.43 86.96± 4.51 5.19 50 64.43± 1.53 2.38 99.12± 2.65 2.67 Kidney 350 53.76± 3.07 5.72 101.00± 1.81 1.79 2400 60.75± 1.53 2.51 103.75± 3.17 3.05 25 64.61± 4.23 6.55 98.39± 11.15 11.33 50 68.08± 3.68 5.40 90.4± 3.02 3.34 Small intestine 350 70.33± 6.13 8.71 101.86± 5.70 5.60 2400 64.51± 2.35 3.64 99.20± 1.01 1.02 25 68.84± 1.18 1.72 92.61± 3.13 3.38 50 62.45± 2.20 3.52 88.53± 3.44 3.89 Stomach 350 65.27± 1.67 2.56 77.56± 1.83 2.36 2400 62.11± 0.87 1.40 83.08± 2.16 2.60 25 71.85± 1.85 2.57 90.54± 1.48 1.64 50 78.40± 2.29 2.92 86.20± 4.87 5.65 Brain 350 85.96± 2.91 3.39 93.34± 1.88 2.02 2400 75.94± 1.52 2.00 101.87± 1.82 1.78 Journal of Analytical Methods in Chemistry 11 Table 5: Summary of stability of four analytes in plasma and tissue samples (n � 6). Ambient temperature Autosampler at 4 C for Tree freeze-thraw −80 C for 30 days for 4 h 8 h cycles Concentration Compounds Matrix (ng/mL) RSD RSD RSD RSD Accuracy Accuracy Accuracy Accuracy (%) (%) (%) (%) 2 101.68± 8.39 8.25 98.25± 7.83 7.97 101.23± 12.44 12.29 93.35± 10.78 11.55 4 86.78± 6.16 7.09 97.90± 11.88 12.14 95.02± 5.39 5.67 98.92± 12.32 12.46 Plasma 100 112.33± 3.72 3.31 111.33± 4.37 3.92 100.68± 6.22 6.18 113.17± 4.96 4.38 2400 98.40± 5.56 5.65 99.47± 7.59 7.63 91.98± 4.75 5.17 112.67± 7.58 6.73 2 102.73± 12.98 12.64 101.70± 10.91 10.72 91.72± 6.33 6.90 88.98± 5.73 6.43 4 96.88± 11.06 11.42 99.50± 6.21 6.24 99.03± 10.17 10.27 102.27± 10.30 10.07 Heart 100 112.83± 9.11 8.07 111.93± 11.77 10.52 111.00± 7.72 6.96 112.62± 8.15 7.24 2400 108.60± 8.41 7.75 101.43± 10.34 10.20 110.55± 8.11 7.33 104.85± 10.69 10.19 2 88.47± 6.93 7.83 87.82± 6.45 7.35 88.47± 7.22 8.16 87.43± 7.74 8.86 4 86.27± 7.32 8.48 87.33± 5.74 6.57 86.00± 6.23 7.24 87.30± 4.35 4.98 Liver 100 99.82± 2.89 2.89 106.33± 2.88 2.70 102.27± 4.97 4.86 87.13± 5.77 6.62 2400 104.17± 2.14 2.05 100.92± 3.08 3.05 99.98± 1.86 1.86 87.02± 5.24 6.03 2 97.62± 8.11 8.30 109.83± 9.95 9.06 105.55± 7.56 7.17 109.70± 13.05 11.90 4 105.82± 9.91 9.37 100.15± 9.69 9.67 103.68± 9.71 9.36 98.88± 4.16 4.20 Spleen 100 112.00± 7.82 6.98 111.12± 7.31 6.58 106.82± 6.88 6.44 111.17± 7.17 6.45 2400 101.52± 7.73 7.61 104.00± 7.45 7.17 106.67± 5.43 5.09 104.73± 7.04 6.72 2 103.27± 8.01 7.75 111.67± 5.43 4.86 112.93± 9.57 8.48 113.77± 9.20 8.08 4 101.83± 6.41 6.30 100.55± 8.58 8.53 100.07± 10.39 10.38 98.77± 6.15 6.23 Chlorogenic acid Lung 100 111.50± 5.47 4.90 110.17± 10.93 9.92 110.17± 7.14 6.48 110.03± 9.33 8.46 2400 100.67± 10.17 10.11 98.78± 7.01 7.10 104.88± 8.23 7.85 110.17± 4.54 4.12 2 110.07± 7.34 6.67 88.18± 4.98 5.64 97.22± 7.67 7.89 89.87± 9.23 10.27 4 87.63± 7.76 8.86 88.40± 6.19 7.00 87.60± 9.26 10.58 89.60± 9.05 10.10 Kidney 100 112.00± 7.01 6.26 111.92± 8.86 7.91 111.17± 7.44 6.69 103.22± 11.32 10.97 2400 100.55± 5.11 5.09 98.10± 4.87 4.97 100.45± 3.96 3.94 101.23± 4.69 4.63 2 105.00± 8.20 7.81 102.63± 3.52 3.43 103.90± 4.49 4.32 101.42± 7.82 7.71 Small 4 91.73± 4.49 4.89 99.47± 5.10 5.13 99.78± 7.70 7.72 106.70± 5.43 5.09 intestine 100 95.90± 12.31 12.84 108.67± 12.04 11.08 100.93± 12.33 12.22 99.07± 12.41 12.52 2400 112.00± 5.37 4.79 111.67± 7.26 6.50 111.67± 6.12 5.48 110.83± 6.68 6.02 2 113.08± 11.06 9.78 111.67± 6.25 5.60 113.58± 8.22 7.24 109.08± 9.97 9.14 4 102.70± 10.17 9.90 97.82± 9.05 9.25 98.87± 9.82 9.94 89.87± 7.04 7.84 Stomach 100 93.75± 8.65 9.22 94.82± 5.05 5.33 93.47± 6.76 7.23 107.33± 5.89 5.49 2400 107.17± 8.68 8.10 96.60± 3.60 3.72 100.23± 4.63 4.62 99.73± 10.09 10.12 2 110.33± 7.55 6.85 106.15± 12.36 11.65 105.65± 10.92 10.34 90.73± 10.25 11.30 4 89.12± 9.00 10.10 87.57± 8.47 9.67 90.68± 10.51 11.59 105.32± 9.72 9.23 Brain 100 91.93± 9.93 10.80 109.17± 7.44 6.82 108.13± 9.91 9.17 105.48± 6.92 6.56 2400 102.93± 5.79 5.62 106.72± 9.83 9.21 96.57± 10.12 10.48 97.98± 9.56 9.75 12 Journal of Analytical Methods in Chemistry Table 5: Continued. Ambient temperature Autosampler at 4 C for Tree freeze-thraw −80 C for 30 days for 4 h 8 h cycles Concentration Compounds Matrix (ng/mL) RSD RSD RSD RSD Accuracy Accuracy Accuracy Accuracy (%) (%) (%) (%) 5 101.95± 10.33 10.14 106.67± 10.88 10.20 91.87± 8.63 9.39 109.57± 12.66 11.55 8 108.88± 1.87 1.72 105.01± 6.32 6.02 105.88± 1.87 1.77 113.33± 5.32 4.69 Plasma 140 87.67± 9.20 10.50 87.88± 4.14 4.71 111.88± 1.87 1.67 111.57± 9.60 8.60 2400 88.93± 6.01 6.75 89.25± 7.58 8.49 89.17± 7.49 8.40 113.83± 7.14 6.27 5 111.97± 11.63 10.39 100.78± 9.63 9.56 107.98± 9.45 8.75 107.42± 10.24 9.53 8 110.50± 4.42 4.00 107.25± 9.25 8.62 109.08± 8.25 7.56 110.87± 9.04 8.15 Heart 140 102.78± 6.89 6.71 104.88± 3.80 3.62 100.40± 9.13 9.09 98.28± 5.70 5.80 2400 112.33± 10.41 9.26 111.50± 5.68 5.10 112.33± 5.72 5.09 110.50± 4.81 4.35 5 101.78± 11.83 11.63 107.70± 5.36 4.98 98.32± 6.66 6.78 98.38± 12.07 12.27 8 87.60± 5.90 6.73 86.47± 4.52 5.23 90.60± 4.91 5.42 86.00± 8.60 10.00 Liver 140 88.57± 2.44 2.76 95.00± 4.15 4.37 96.28± 4.36 4.53 97.32± 7.17 7.36 2400 98.93± 3.81 3.85 91.37± 2.51 2.75 89.35± 3.70 4.15 89.97± 7.32 8.14 5 110.33± 8.71 7.89 112.92± 8.72 7.72 110.67± 8.57 7.75 109.68± 12.57 11.46 8 99.28± 13.31 13.41 93.42± 8.00 8.57 96.13± 11.98 12.46 101.90± 10.25 10.06 Spleen 140 93.13± 6.10 6.55 95.27± 7.08 7.43 89.18± 8.23 9.23 92.35± 2.70 2.92 2400 103.33± 8.37 8.10 105.67± 8.50 8.05 107.73± 8.10 7.51 109.83± 4.96 4.51 5 109.50± 11.47 10.48 100.40± 10.26 10.22 101.05± 5.33 5.28 95.98± 10.47 10.91 8 94.73± 8.92 9.41 91.40± 9.71 10.63 101.90± 10.45 10.25 99.03± 10.49 10.59 Naucleactonin C Lung 140 102.38± 4.12 4.03 94.45± 5.92 6.27 93.75± 8.53 9.09 92.58± 3.60 3.89 2400 111.37± 9.27 8.33 111.85± 9.53 8.52 112.125± 9.66 8.61 111.67± 3.39 3.03 5 109.33± 7.55 6.91 106.08± 8.96 8.44 98.92± 9.91 10.02 97.18± 10.46 10.76 8 99.10± 4.24 4.27 97.38± 7.31 7.51 91.10± 9.20 10.10 94.77± 7.62 8.04 Kidney 140 96.08± 7.36 7.66 95.43± 7.99 8.37 101.10± 4.98 4.92 100.25± 8.25 8.23 2400 108.00± 5.62 5.20 103.87± 4.68 4.51 104.57± 6.09 5.82 102.30± 5.90 5.77 5 111.90± 12.07 10.79 111.67± 8.14 7.29 111.17± 8.33 7.49 112.67± 8.78 7.79 Small 8 103.33± 3.39 3.28 100.03± 11.46 11.46 100.60± 10.24 10.18 104.25± 9.49 9.10 intestine 140 88.60± 9.48 10.70 89.80± 8.30 9.24 87.88± 5.57 6.34 90.17± 7.26 8.05 2400 107.48± 8.23 7.66 103.75± 6.68 6.44 108.43± 8.65 7.98 102.32± 8.65 8.45 5 111.00± 7.90 7.12 112.17± 7.25 6.46 113.12± 9.97 8.81 113.83± 2.99 2.63 8 107.32± 12.96 12.08 98.37± 10.75 10.93 105.90± 9.13 8.62 103.55± 9.49 9.16 Stomach 140 88.15± 7.91 8.98 86.38± 8.38 9.70 87.12± 5.24 6.1 88.28± 6.31 7.15 2400 100.53± 7.29 7.25 99.52± 5.85 5.88 104.90± 5.53 5.28 102.32± 8.22 8.03 5 106.35± 11.82 11.11 109.83± 6.05 5.51 98.92± 10.65 10.77 104.10± 10.88 10.45 8 88.87± 10.43 11.74 90.65± 6.37 7.03 89.95± 9.23 10.26 88.82± 10.57 11.90 Brain 140 88.33± 6.74 7.63 87.92± 8.15 9.27 87.20± 4.98 5.71 87.93± 5.81 6.61 2400 99.43± 6.38 6.41 112.67± 6.62 5.88 109.67± 6.44 5.87 109.00± 5.22 4.78 Journal of Analytical Methods in Chemistry 13 Table 5: Continued. Ambient temperature Autosampler at 4 C for Tree freeze-thraw −80 C for 30 days for 4 h 8 h cycles Concentration Compounds Matrix (ng/mL) RSD RSD RSD RSD Accuracy Accuracy Accuracy Accuracy (%) (%) (%) (%) 5 99.10± 9.25 9.33 90.97± 9.01 9.90 93.93± 11.26 11.98 101.18± 3.53 3.49 8 91.38± 12.76 13.96 94.88± 12.31 12.97 90.67± 9.29 10.25 108.17± 9.20 8.50 Plasma 120 101.35± 4.33 4.28 101.32± 3.40 3.36 96.75± 4.55 4.70 107.30± 11.48 10.70 1800 99.35± 6.04 6.08 101.93± 6.90 6.77 96.13± 4.03 4.19 112.67± 7.66 6.80 5 107.60± 11.23 10.44 104.15± 9.92 9.52 110.67± 10.82 9.78 107.45± 9.35 8.70 8 112.67± 7.23 6.42 111.83± 8.23 7.36 111.33± 5.92 5.32 101.00± 12.05 11.93 Heart 120 111.67± 6.22 5.57 112.67± 5.89 5.23 108.58± 11.51 10.60 113.50± 7.20 6.35 1800 110.83± 2.79 2.51 106.58± 8.77 8.22 113.67± 4.27 3.76 101.90± 12.30 12.07 5 93.87± 9.31 9.92 106.17± 4.88 4.59 95.62± 9.82 10.27 100.87± 10.27 10.19 8 108.95± 10.77 9.88 101.97± 7.93 7.78 105.20± 10.21 9.70 91.12± 10.79 11.84 Liver 120 107.67± 6.22 5.78 111.50± 7.26 6.51 101.62± 9.62 9.47 104.23± 8.68 8.33 1800 108.83± 2.32 2.13 109.83± 5.38 4.90 103.20± 7.52 7.29 108.93± 9.52 8.74 5 110.30± 12.31 11.16 112.17± 7.94 7.07 111.00± 10.26 9.24 111.50± 8.41 7.54 8 112.17± 5.64 5.02 111.67± 6.19 5.54 111.38± 7.36 6.58 110.67± 7.17 6.48 Spleen 120 107.85± 7.83 7.26 105.45± 7.95 7.54 98.83± 7.51 7.60 105.42± 10.79 10.24 1800 109.77± 8.61 7.85 111.17± 8.18 7.36 113.17± 3.31 2.93 111.83± 7.57 6.77 5 113.33± 8.71 7.69 106.65± 12.43 11.65 98.87± 9.02 9.12 102.98± 9.50 9.23 8 105.32± 7.81 7.41 103.18± 10.97 10.63 105.03± 9.86 9.38 104.00± 11.73 11.28 Khaephuoside A Lung 120 101.27± 3.53 3.49 99.07± 11.01 11.11 102.02± 6.36 6.23 110.83± 5.19 4.69 1800 110.83± 8.23 7.43 112.80± 7.46 6.61 110.00± 8.88 8.07 112.50± 6.80 6.05 5 96.42± 6.24 6.47 99.65± 10.53 10.57 101.62± 11.37 11.19 112.78± 11.43 10.13 8 113.17± 6.46 5.71 110.83± 7.88 7.11 113.00± 6.99 6.18 113.17± 7.19 6.36 Kidney 120 111.67± 5.32 4.76 111.83± 5.31 4.75 112.00± 5.93 5.30 112.00± 5.62 5.02 1800 102.03± 4.44 4.35 97.37± 6.84 7.02 100.50± 5.53 5.50 101.12± 6.56 6.48 5 110.02± 8.19 7.44 112.83± 3.43 3.04 100.88± 9.39 9.30 94.98± 9.42 9.91 Small 8 105.62± 9.76 9.24 109.53± 10.14 9.25 101.00± 10.42 10.31 107.70± 11.47 10.65 intestine 120 103.30± 7.75 7.50 106.00± 7.29 6.88 102.80± 6.09 5.93 101.58± 6.26 6.16 1800 113.00± 6.13 5.43 110.83± 8.13 7.34 113.67± 8.19 7.20 113.12± 12.47 11.02 5 113.68± 8.42 7.41 112.13± 10.23 9.12 111.12± 10.43 9.38 110.67± 6.09 5.50 8 111.67± 6.59 5.90 111.00± 5.22 4.70 109.67± 3.33 3.03 104.50± 4.71 4.51 Stomach 120 113.17± 3.66 3.23 111.50± 5.86 5.25 112.17± 6.15 5.48 110.92± 8.36 7.53 1800 109.83± 7.36 6.70 111.92± 10.99 9.82 110.50± 6.28 5.69 113.83± 5.56 4.89 5 109.82± 10.26 9.34 106.90± 12.09 11.31 103.05± 9.12 8.85 112.50± 5.58 4.96 8 111.00± 7.42 6.52 112.50± 7.06 6.28 113.33± 3.83 3.38 111.33± 6.09 5.47 Brain 120 112.50± 5.13 4.56 112.33± 6.47 5.76 113.17± 5.31 4.69 110.17± 8.28 7.52 1800 105.82± 8.31 7.86 112.83± 7.68 6.81 110.83± 7.52 6.79 113.83± 4.49 3.95 14 Journal of Analytical Methods in Chemistry Table 5: Continued. Ambient temperature Autosampler at 4 C for Tree freeze-thraw −80 C for 30 days for 4 h 8 h cycles Concentration Compounds Matrix (ng/mL) RSD RSD RSD RSD Accuracy Accuracy Accuracy Accuracy (%) (%) (%) (%) 25 86.25± 5.40 6.26 94.95± 9.80 10.32 102.87± 10.37 10.08 105.60± 10.19 9.65 50 87.20± 7.91 9.07 98.98± 11.30 11.30 94.53± 9.84 10.41 107.88± 14.96 13.87 Plasma 350 112.83± 4.58 4.06 112.67± 6.62 5.88 108.33± 5.13 4.73 109.07± 1.87 1.72 2400 89.98± 4.76 5.29 89.43± 7.42 8.30 86.07± 3.39 3.93 111.74± 3.76 3.37 25 98.10± 9.43 9.61 100.45± 10.89 10.85 97.55± 9.01 9.24 88.38± 5.69 6.44 50 94.60± 11.14 7.80 102.70± 7.98 7.77 97.05± 8.42 8.68 99.12± 9.56 9.65 Heart 350 102.57± 4.92 4.80 112.63± 98.93 7.93 110.33± 5.82 5.27 108.38± 7.34 6.77 2400 102.23± 6.64 6.49 92.98± 8.87 9.54 101.57± 6.73 6.63 99.57± 9.46 9.50 25 91.10± 10.37 11.39 102.88± 10.85 10.54 97.32± 10.44 10.73 104.70± 11.97 11.43 50 105.60± 9.04 8.56 112.50± 5.65 5.02 107.48± 10.54 9.80 95.25± 11.29 11.86 Liver 350 97.50± 6.63 6.80 113.00± 4.00 3.54 103.12± 5.26 5.10 108.67± 8.98 8.27 2400 104.32± 10.18 9.76 100.42± 6.85 6.83 96.25± 5.74 5.96 107.68± 7.73 7.18 25 88.72± 9.93 11.19 104.47± 9.00 8.62 90.93± 9.15 10.07 112.83± 3.60 3.19 50 113.17± 10.59 9.36 108.35± 10.58 9.77 95.62± 10.90 11.40 107.68± 10.39 9.65 Spleen 350 111.33± 7.63 6.86 111.68± 9.21 8.25 112.17± 7.99 7.12 112.17± 8.40 7.49 2400 102.27± 11.50 11.24 112.00± 8.67 7.74 105.32± 6.59 6.26 111.00± 6.03 5.44 25 90.52± 10.77 11.90 96.80± 11.29 11.67 97.07± 10.87 11.20 97.92± 11.34 11.59 3,4-Dimethoxyphenyl-1-O- 50 112.67± 7.84 6.96 111.02± 7.81 7.04 100.02± 12.46 12.46 109.25± 11.46 10.49 Lung β-apiofuroseyl(1⟶ 2)-β-D-glucopyranoside 350 111.33± 6.89 6.19 109.47± 11.10 10.14 103.68± 11.62 11.21 112.33± 6.92 6.16 2400 98.85± 9.83 9.94 101.82± 8.67 8.52 100.62± 7.71 7.66 109.12± 10.69 9.79 25 100.05± 11.92 11.92 101.42± 8.63 8.51 96.23± 9.14 9.50 110.83± 6.24 5.63 50 98.37± 7.53 7.65 102.72± 10.29 10.02 106.75± 9.15 8.57 99.88± 9.65 9.66 Kidney 350 90.05± 8.47 9.40 102.03± 4.61 4.52 111.83± 7.03 6.28 105.40± 10.28 9.75 2400 104.72± 8.96 8.56 106.27± 10.19 9.59 107.85± 9.16 8.49 104.82± 8.59 8.20 25 87.47± 7.17 8.20 96.42± 9.13 9.47 94.10± 8.05 8.55 99.25± 6.06 6.11 Small 50 109.83± 2.56 2.33 100.75± 6.11 6.07 106.83± 5.34 5.00 101.80± 8.78 8.62 intestine 350 111.17± 7.73 6.95 113.67± 8.76 7.70 113.00± 3.79 3.36 106.33± 5.39 5.07 2400 98.50± 5.49 5.57 104.65± 7.85 7.50 99.92± 5.94 5.94 94.90± 8.62 9.08 25 111.83± 5.78 5.17 110.50± 4.42 4.00 113.00± 6.32 5.60 106.32± 9.48 8.92 50 112.75± 10.97 9.73 112.67± 3.67 3.26 112.83± 5.60 4.96 109.83± 8.66 7.88 Stomach 350 113.17± 3.87 3.42 108.17± 9.68 8.95 111.00± 6.20 5.58 111.83± 5.98 5.35 2400 101.27± 7.89 7.80 94.40± 6.00 6.35 96.82± 8.08 8.34 103.08± 9.60 9.32 25 99.82± 9.82 9.84 107.80± 12.79 11.86 100.68± 9.80 9.73 105.37± 7.31 6.93 50 109.35± 12.15 11.11 112.83± 4.26 3.78 113.77± 10.36 9.11 113.83± 3.87 3.40 Brain 350 111.50± 8.64 7.75 110.83± 7.47 6.74 111.17± 7.63 6.27 112.83± 7.78 6.90 2400 98.95± 3.97 4.01 100.10± 9.43 9.42 98.23± 5.39 5.49 101.50± 9.94 9.80 Journal of Analytical Methods in Chemistry 15 Chlorogenic acid Naucleactonin C 100 400 50 200 0 0 0.083 0.25 0.5 1 2468 10 12 24 0.083 0.25 0.5 1 2 4 6 8 10 12 24 Time (h) Time (h) Khaephuoside A 3,4-dimethoxyphenyl-1-O-β-apiofuroseyl (1→2)-β-D- glucopyranoside 0 0 0.083 0.25 0.5 1 2 4 6 8 10 0.083 0.25 0.5 1 2 4 Time (h) Time (h) Figure 3: Concentration-time profles of four analytes in rat plasma following oral administration of Nauclea ofcinalis extracts at a dose of 2 g/kg. Table 6: Summary of primary pharmacokinetic parameters of four analytes (n � 6). 3,4-dimethoxyphenyl-1-O- Chlorogenic acid Naucleactonin C Khaephuoside A Parameters Unit β-apiofuroseyl(1⟶ 2)-β-D-glucopyranoside (mean± SD) (mean± SD) (mean± SD) (mean± SD) μg/ AUC (0 −t) 482.76± 124.95 5281.36± 440.02 238.70± 46.64 212.99± 49.29 L∗h AUC μg/ 554.37± 166.14 5405.02± 416.99 262.16± 32.24 440.17± 216.99 (0 −∞) L∗h MRT (0 −t) h 5.33± 0.65 4.76± 1.57 3.19± 0.24 1.6± 0.14 t1/2 h 10.64± 6.21 4.59± 1.65 3.12± 1.70 3.99± 2.87 T h 0.25± 0.00 1.67± 0.52 0.29± 0.10 0.33± 0.13 max L/h/ CL 62.94± 16.00 0.15± 0.01 54.40± 6.04 28.63± 13.64 kg C μg/L 242.42± 84.67 1008.58± 368.84 106.18± 23.30 144.13± 35.59 max Vz/F L/kg 915.76± 451.75 0.966± 0.359 253.289± 158.11 127.43± 65.93 Abbreviations. AUC, area under the plasma concentration–time curve; MRT, mean residence time; t1/2, elimination half-life; T , time to Cmax; CL, max clearance rate; C , maximum plasma concentration; Vz/F, apparent distribution volume. max accounts for only 30% [25], which greatly afected the 1-O-β-apiofuroseyl(1⟶ 2)-β-D-glucopyranoside in rats. Te pharmacological activity of chlorogenic acid from Nauclea result is shown in Figure 4 that naucleactonin C and khae- ofcinalis. Terefore, considering the clinical application of phuoside A could be detected in all studied tissue. However, Nauclea ofcinalis [26], we could design Nauclea ofcinalis chlorogenic acid and 3,4-dimethoxyphenyl-1-O- to diferent dosage forms with advanced technology in order β-apiofuroseyl(1⟶ 2)-β-D-glucopyranoside were not de- to improve the bioavailability and fnally increase the tected in the spleen. Te concentration orders in eight diferent efcacy. tissues were ranked as 3,4-dimethoxyphenyl-1-O- β-apiofuroseyl(1⟶ 2)-β-D-glucopyranoside>Khaephuoside A>Naucleactonin C>Chlorogenic acid in the small intestine, 3.5. Tissue Distribution Study. Tis method was also applied stomach, heart, liver, spleen, lung, and brain tissues, and to investigate the tissue distribution of chlorogenic acid, naucleactonin C>3,4-dimethoxyphenyl-1-O-β-apiofuroseyl naucleactonin C, khaephuoside A, and 3,4-dimethoxyphenyl- (1⟶ 2)-β-D-glucopyranoside>Khaephuoside A>Chloro Concentration (ng/mL) Concentration (ng/mL) Concentration (ng/mL) Concentration (ng/mL) 16 Journal of Analytical Methods in Chemistry Chlorogenic acid Naucleactonin C 0 0 heart liver spleen lung kidney small stomach brain heart liver spleen lung kidney small stomach brain intestine intestine 0.5 h 4 h 0.5 h 4 h 1 h 6 h 1 h 6 h 2 h 2 h Khaephuoside A 3,4-dimethoxyphenyl-1-O-β-apiofuroseyl (1→2)-β-D- glucopyranoside 0 0 heart liver spleen lung kidney small stomach brain heart liver spleen lung kidney small stomach brain intestine intestine 0.5h 4 h 0.5 h 4 h 1 h 6 h 1 h 6 h 2 h 2 h Figure 4: Mean concentration of four analytes in the heart, liver, spleen, lung, kidney, small intestine, stomach, and brain at 0.5, 1, 2, 4, and 6 h after oral administration of Nauclea ofcinalis extracts at a dose of 2 g/kg. genic acid in the kidney. Te contents of the four compounds frst-pass efect of oral administration might be the main in the liver, kidney, and lung were relatively high within cause of high distribution in the liver. after oral administration of Nauclea ofcinalis extrac, indicating that the liver, kidney, and lung may be the main 4. Conclusion target organs for the pharmacological efects of Nauclea ofcinalis. Tis result is consistent with existing research We report the development and validation of a sensitive, on the inhibition of bronchitis by Nauclea ofcinalis rapid, and reliable HPLC–MS/MS analytical method for the [27]. Te results show that all compounds except simultaneous determination and quantifcation of chloro- chlorogenic acid were more abundant in the small in- genic acid, naucleactonin C, khaephuoside A, and 3,4- testine than in the stomach, indicating that most of dimethoxyphenyl-1-O-β-apiofuroseyl(1⟶ 2)-β-D-gluco- naucleactonin C, khaephuoside A, and 3,4-dimethox- pyranoside in the plasma and tissues of rats. Tis validated yphenyl-1-O-β-apiofuroseyl(1⟶2)-β-D-glucopyrano- analytical method was assessed on the basis of the FDA side possibly absorbed by the small intestine after oral guidelines for bioanalytical method validation and applied administration of Nauclea ofcinalis extract for enter the the study of pharmacokinetics and tissue distribution of oral systemic circulation system. With the exception of administration of Nauclea ofcinalis extract in SD rats. To chlorogenic acid, the peak times and concentrations of the best of our knowledge, this is the frst study that de- the other three compounds in most tissues are nearly termined the pharmacokinetic and tissue distribution of identical to those in plasma, which means that the naucleactonin C, khaephuoside A, and 3,4-dimethox- distribution of the other three compounds depends on yphenyl-1-O-β-apiofuroseyl(1⟶ 2)-β-D-glucopyranoside. the blood fow or perfusion rate of the organ [28]. Te In addition, the pharmacokinetics and tissue distribution of peak concentration of chlorogenic acid in most tissues is chlorogenic acid after oral administration of Nauclea of- much lower than in plasma and may be related to the cinalis extract in rats was also reported for the frst time. absorption of the compound in the tissue. Drugs enter tissues that receive high blood fow frst, followed by Data Availability those that receive low blood fow [29]. Our study demonstrated that the four compounds are mainly Te data used to support the fndings of this study are in- distributed in organs with relatively large blood fows, cluded within the article and the supplementary information such as the liver and kidneys. In addition to this, the fles. Concentration (ng/g) Concentration (ng/g) Concentration (ng/g) Concentration (ng/g) Journal of Analytical Methods in Chemistry 17 Chinese Traditional and Herbal Drugs, vol. 48, no. 1, Conflicts of Interest pp. 52–57, 2017. [12] N. X. Li, J. J. Zhang, Y. Zhang et al., “Chromatographic Te authors declares that there are no conficts of interest. fngerprints analysis and determination of seven components in Danmu preparations by HPLC-DAD/QTOF-MS,” Chinese Acknowledgments Medicine, vol. 15, no. 1, p. 19, 2020. [13] S. Y. Mai, Y. H. Li, X. G. Zhang, Y. R. Wang, J. Q. Zhang, and Tis work was supported by the Hainan Provincial Natural A. 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Journal
Journal of Analytical Methods in Chemistry – Hindawi Publishing Corporation
Published: Dec 21, 2022