Development, Validation, and Application of HPLC Method for Quantification of Antihyperuricemic Compounds from Lippia nodiflora in Rat Plasma

Abstract

An HPLC method for simultaneous determination of arenarioside (1 ), verbascoside (2), 6-hydroxyluteolin (3), 6-hydroxyluteolin-7-O-glycoside (4), and nodifloretin (5) from Lippia nodiflora in rat plasma was developed and validated. The optimal chromatographic separation was achieved with a gradient mobile phase comprising 0.1% aqueous acetic acid and acetonitrile. The limit of detection was 78.1 ng/mL for 3 and 39.1 ng/mL for the other compounds (signal-to-noise ratio=3), whereas the limit of quantification was 312.5 ng/mL for 3 and 156.3 ng/mL for the other compounds (signal-to-noise ratio=12). The recovery values of compounds 1–5 ranged from 89.37–100.92%. Their accuracy values were between 96.48 and 105.81%, while their corresponding precision values were in the range of 0.75–9.06% for both intraday and inter-day analysis. The method was then applied in the first pharmacokinetic study of 1–5. Following intravenous administration, 1–5 were eliminated slowly from the body with a mean clearance value of 0.11, 0.13, 0.30, 0.09, and 0.23 L/kg h, respectively. Meanwhile, their peak plasma concentration upon oral administration was 8.97, 1.07, 1.06, 0.65, and 0.38 µg/mL, respectively. Compound 3 (5.97%) exhibited the highest absolute oral bioavailability value, followed by 1 (5.22%), 4 (3.13%), 2 (2.10%), and 5 (0.93%).

Key words

Lippia nodiflora - Verbenaceae - phenylethanoid glycosides - flavonoids - HPLC - pharmacokinetic

• References

• 1 Khare CP. Indian medicinal plants: An illustrated dictionary. New York, USA: Springer-Verlag; 2007: 480-481
• 2 Umberto Quattrocchi FLS CRC world dictionary of medicinal and poisonous plants. common names, scientific names, eponyms, synonyms, and etymology. Boca Raton, Florida, USA: Taylor and Francis Group, LLC; 2012: 2895-2896
• 3 Sharma RA, Singh R. A review on Phyla nodiflora Linn.: A wild wetland medicinal herb. Int J Pharm Sci Rev Res 2013; 20: 57-63
• 4 Narendra ND, Bhupesh CN, Rameswara RG, Bastipati S. Evaluation of hepatoprotective activity of Phyla nodiflora L. against CCl₄ induced hepatotoxicity. Int J Innov Drug Disc 2012; 2: 16-21
• 5 Yang XR, Chen AM, Ma YF, Gao Y, Gao ZM, Fu BY, Sun F, Qiao JL, Li Q, Wan SQ, Werner H, Chuang YF, Zhu XS. Traditional Chinese medicine: A manual from A-Z: Symptoms, therapy, and herbal remedies. Heidelberg, New York, USA: Springer-Verlag; Berlin: 2003. 340
• 6 Barua AK, Chakrabarti P, Sanyal PK. Structure of nodifloretin - new flavone from Lippia nodiflora. Trans Bose Res Inst (Calcutta) 1971; 33-34: 5-8
• 7 Barnabas C, Gunasingh G, Nagarajan S. Flavonoids from the flowers of Phyla nodiflora Linn. Indian J Chem, Sect B 1980; 19B: 822
• 8 Sudha A, Srinivasan P. Bioassay-guided isolation and antioxidant evaluation of flavonoid compound from aerial parts of Lippia nodiflora . BioMed Res Int 2014; 2014; 1-10
• 9 Tomás-Barberán FA, Harborne JB, Self R. Twelve 6-oxygenated flavone sulphates from Lippia nodiflora and L. canescens . Phytochemistry 1987; 26: 2281-2284
• 10 Khalil AT, Lahloub MF, Salama OM. Phenolic compounds from Lippia nodiflora . J Pharm Sci 1995; 11: 256-265
• 11 Siddiqui BS, Ahmad F, Sattar F, Begum S. Chemical constituents from the aerial parts of Lippia nodiflora Linn. Arch Pharm Res 2007; 30: 1507-1510
• 12 Siddiqui BS, Ahmed F, Ali SK, Perwaiz S, Begum S. Steroidal constituents from the aerial parts of Lippia nodiflora Linn. Nat Prod Res 2009; 23: 436-441
• 13 Dodoala S, Diviti R, Koganti B, Prasad KVSRG. Effect of ethanolic extract of Phyla nodiflora (Linn.) Greene against calculi producing diet induced urolithiasis. Indian J Nat Prod Resour 2010; 1: 314-321
• 14 Gadhvi R, Mishra GJ, Reddy MN, Nivserkar M. Antihypertensive efficacy of Lippia nodiflora - whole plant on uninephrectomized doca - salt hypertensive rats. IOSR J Pharm 2012; 2: 24-28
• 15 Shukla S, Saluja AK, Pandya SS. In-vitro antioxidant activity of aerial parts of Lippia nodiflora Rich. Pharmacologyonline 2009; 2: 450-459
• 16 Durairaj A, Mazumder UK, Gupta M, Selvan VT. Effect on inhibition of proliferation and antioxidant enzyme level of Lippia nodiflora in EAC cell line treated mice. J Complement. Integr Med 2009; 6: 713-714
• 17 Balakrishnan G, Janakarajan L, Balakrishnan A, Lakshmi BS. Molecular basis of the anti-inflammatory property exhibited by cyclopentano phenanthrenol isolated from Lippia nodiflora . Immunol Invest 2010; 39: 713-739
• 18 Cheng LC, Murugaiyah V, Chan KL. In vitro xanthine oxidase inhibitory studies of Lippia nodiflora and isolated flavonoids and phenylethanoid glycosides as potential uric acid-lowering agents. Nat Prod Commun 2015; 10: 945-948
• 19 Cheng LC, Murugaiyah V, Chan KL. Flavonoids and phenylethanoid glycosides from Lippia nodiflora as promising antihyperuricemic agents and elucidation of their mechanism of action. J Ethnopharmacol 2015; 176: 485-493
• 20 Li Y, Gan L, Li GQ, Deng L, Zhang X, Deng YL. Pharmacokinetics of plantamajoside and acteoside from Plantago asiatica in rats by liquid chromatography-mass spectrometry. J Pharm Biomed Anal 2014; 89: 251-256
• 21 Wu YT, Lin LC, Sung JS, Tsai TH. Determination of acteoside in Cistanche deserticola and Boschniakia rossica and its pharmacokinetics in freely-moving rats using LC-MS/MS. J Chromatogr B 2006; 844: 89-95
• 22 Zhang W, Huo SX, Wen YL, Xing H, Zhang Q, Li N, Zhao D, Sun XL, Xu J, Yan M, Chen XJ. Pharmacokinetics of acteoside following single dose intragastric and intravenous administrations in dogs. Chin J Nat Med 2015; 13: 634-640
• 23 Wu YT, Tsai TR, Lin LC, Tsai TH. Liquid chromatographic method with amperometric detection to determine acteoside in rat blood and brain microdialysates and its application to pharmacokinetic study. J Chromatogr B 2007; 853: 281-286
• 24 Cheng LC, Murugaiyah V, Chan KL. Developing a validated HPLC method for the phytochemical analysis of antihyperuricemic phenylethanoid glycosides and flavonoids in Lippia nodiflora . Nat Prod Commun 2017; 12: 1681-1684
• 25 Chen HJ, Inbaraj BS, Chen BH. Determination of phenolic acids and flavonoids in Taraxacum formosanum Kitam by liquid chromatography tandem mass spectrometry coupled with a post-column derivatization technique. Int J Mol Sci 2012; 13: 260-285
• 26 Kao TH, Huang SC, Inbaraj BS, Chen BH. Determination of flavonoids and saponins in Gynostemma pentaphyllum (Thunb.) Makino by liquid chromatography mass spectrometry. Anal Chim Acta 2008; 626: 200-211
• 27 Inbaraj BS, Lu H, Kao TH, Chen BH. Simultaneous determination of phenolic acids and flavonoids in Lycium barbarum Linnaeus by HPLC-DAD-ESI-MS. J Pharm Biomed Anal 2010; 51: 549-556
• 28 Tuszyńska M. Validation of the analytical method for the determination of flavonoids in broccoli. J Hort Res 2014; 22: 131-140
• 29 Zu YG, Li CY, Fu YJ, Zhao CJ. Simultaneous determination of catechin, rutin, quercetin kaempferol and isorhamnetin in the extract of sea buckthorn (Hippophae rhamnoides L.) leaves by RP-HPLC with DAD. J Pharm Biomed Anal 2006; 41: 714-719
• 30 Thilakarathna SH, Vasantha Rupasinghe HP. Flavonoid bioavailability and attempts for bioavailability enhancement. Nutrients 2013; 5: 3367-3387
• 31 Murugaiyah V, Chan KL. Analysis of lignans from Phyllanthus niruri L. in plasma using a simple HPLC method with fluorescence detection and its application in a pharmacokinetic study. J Chromatogr B 2007; 852: 138-144
• 32 Sun DQ, Xue AY, Wu J, Zhang B, Yu JL, Li Q, Sun C. Simultaneous determination of acetylpuerarin and puerarin in rat plasma by liquid chromatography-tandem mass spectrometry: Application to a pharmacokinetic study following intravenous and oral administration. J Chromatogr B 2015; 995-996: 24-30
• 33 ICH Harmonised Tripartite Guidelines. Validation of analytical procedures: Text and methodology Q2(R1), International Conference on Harmonisation (ICH) of technical requirements for registration of pharmaceuticals for human use, Current Step 4 version, Parent guideline dated 27 October 1994 (Complementary guidelines on methodology dated 6 November 1996 incorporated in November 2005); Available at https://www.ich.org/fileadmin/Public_Web_Site/ICH_Products/Guidelines/Quality/Q2_R1_Guideline.pdf Acessed 7 Feb 2019)
• 34 National Research Council (NRC). Guide for the care and use of laboratory animals. seventh ed., Institute of Laboratory Animal Resources, Commission on Life Sciences. Washington, D.C: National Academy Press; 1996: 1-125
• 35 Public Health Service (PHS). Human Health Research Act. 1985 revised Public health service policy on humane care and use of laboratory animals. Bethesda, Maryland, US: US: Department of Health and Human Services, National Institute of Health; 1996: 99-158