Asian Journal of Research in Chemistry

Open Access Open Access  Restricted Access Subscription Access
Open Access Open Access Open Access  Restricted Access Restricted Access Subscription Access

Development and Validation of Stability Indicating HPTLC Method for Estimation of Ursolic Acid


     

   Subscribe/Renew Journal


A stability indicating high performance thin layer chromatographic (HPTLC) method was developed and validated for estimation of ursolic acid. Study was performed on pre-coated silica gel TLC plates using toluene: ethyl acetate: formic acid (7:3:0.1, v/v/v) as the mobile phase. After development, plates were derivatized with Anisaldehyde- Sulphuric acid reagent. Detection and quantification were performed by densitometric scanning at 530 nm. Method was validated according to ICH Q2(R<SUB>1</SUB>)guideline. The linear regression analysis for the calibration plot showed good linear relationship over the concentration range of 200-600 ng/spot. The correlation coefficient of calibration curve was found to be 0.9986. The intra and inter day precision was found to be less than 2%. Ursolic acid was also subjected to acid, base, oxidation, heat, humidity and photo-degradation studies. The degradation products obtained were well resolved from the pure drug with significantly different Rf values. As the method could effectively separate the drugs from its degradation products, it can be used for stability-indicating analysis.

Keywords

Ursolic Acid, Stability Indicating Method, High Performance Thin Layer Chromatography (HPTLC), Method Validation
Subscription Login to verify subscription
User
Notifications
Font Size


  • http://www.85xianji.com/xiangxi.asp?id=170 (assessed on 09/02/2013)

  • http://suite101.com/article/ursolic-acid-a92128#.UWLDtJP-G3s (assessed 10/02/2013)

  • Amin P, Nayak B and Patel K. Extraction and estimation of different marker compounds by HPTLC from Alstonia scholaris leaves and flowers. International Journal of Pharmaceutical Science. 2012: 2731-2738.

  • Shetty PG, et al. Pharmacokinetic analysis of ursolic acid in Alstonia scholaris R. Br. by high-performance thin-layer chromatography. Journal of Planer Chromatography. 20(2); 2007: 117-120.

  • Rout K, et al. Thin-layer chromatographic separation and validated HPTLC method for quantification of ursolic acid in various Ocimum Species. Journal of Food and Drug Analysis. 20(4); 2012: 865-871.

  • Gupta M, et al. Determination of ursolic acid a biomarker in different Swertia species through high performance thin layer chromatography. Chinese Medicine. 2; 2011: 121-124.

  • Mucaji P and Nagy M. Contribution to the TLC separation of ursolic and oleanolic acid mixture. Acta Facultatis Pharmceuticae Universitatis Comenianae. 58; 2011: 56 - 61.

  • Almeida M, et al. Determination of ursolic acid from polymeric nanoparticles using high-performance liquid chromatography method. Latin American Journal of Pharmacy. 31(8); 2012: 1217-1221.

  • Liu Ch and Liu Y. Determination of ursolic acid in herbs of Verbena officinalis by HPLC. Zhongguo Zhong Yao Za Zhi. 27(12); 2002: 916-918.

  • Taralkar SV and Chattopadhyay S. A HPLC method for determination of ursolic acid and betulinic acids from their methanolic extracts of Vitex Negundo Linn. Journal of Analytical and Bioanalytical Techniques. 3(3); 2012: 134.

  • Maurya A and Srivastava S. Determination of ursolic acid and ursolic acid lactone in the leaves of Eucalyptus tereticornis by HPLC. Journal of Brazilian Chemical Society. 23(3); 2012: 468-472.

  • http://en.cnki.com.cn/Article_en/CJFDTOTALSZGY200711007. htm (assessed on 14/02/2013)

  • Zacchigna M, et al. Rapid HPLC analysis for quantitative determination of the two isomeric triterpenic acids, oleanolic acid and ursolic acid, in Plantago Major. Scientia Pharmaceutica. 77; 2009: 79-86.

  • Chun-Hua Zhou, et al. Oleanolic and ursolic acid in the fruit of Eriobotrya japonica Lindl. Journal of Medicinal Plants Research. 5(9); 2011: 1735-1740.

  • Sundaram RS, et al. Quantification of bioactive principles in Indian traditional herb Ocimum Sanctum Linn. (Holy Basil) Leaves by high performance liquid chromatography. Asian Journal of Biomedical and Pharmaceutical Sciences. 1(3); 2011: 35-41.

  • Srivastava P and Chaturvedi R. Simultaneous determination and quantification of three pentacyclic triterpenoids—betulinic acid, oleanolic acid, and ursolic acid—in cell cultures of Lantana camara L. In Vitro Cellular and Developmental Biology Plant. 46; 2010: 549–557.

  • Xia Y, et al. Quantitation of ursolic acid in human plasma by ultra performance liquid chromatography tandem mass spectrometry and its pharmacokinetic study. Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences. 879(2); 2011: 219-224.

  • Qiongfeng Liao, et al. LC-MS determination and pharmacokinetic studies of ursolic acid in rat plasma after administration of the traditional chinese medicinal preparation Lu-Ying extract. Yakugaku Zassi. 125(6); 2005: 509-515.

  • Sundaram RS,et al. LC-MS quantification of rosmarinic acid and ursolic acid in the Ocimum Sanctum Linn. leaf extract (Holy Basil, Tulsi), Journal of Liquid Chromatography & Related Technologies. 35(5); 2012: 634-650.

  • Zhi-Rong Tan, et al. LC–MS–MS quantitative determination of ursolic acid in human plasma and its application to pharmacokinetic studies. Chromatographia. 72; 2010: 1107- 1113.

  • Lacikova L, et al. A rapid tandem mass spectrometric assay for determination of ursolic acid –application to analysis of ursolic acid in four species of Staphylea L. and leaves of Staphylea Pinnata L. gathered during ontogenesis. Chromatographia. 63; 2006: 117-122.

  • Janicsak G,et al. Gas Chromatographic method for routine determination of oleanolic and ursolic acids in medicinal plants. Chromatographia. 58; 2003: 295-299.

  • Razborsek MA, et al. Determination of oleanolic, betulinic and ursolic acid in lamiaceae and mass spectral fragmentation of their trimethylsilylated derivatives. Chromatographia. 67; 2008: 433- 440.

  • ICH Harmonized Tripartite Guideline Q2 (R1): Validation Of Analytical Procedure: Text and Methodology. London; 2005.

  • Bakshi M and Singh S. Development of validated stability indicating assay-methods – critical review. Journal of Pharmaceutical and Biomedical Analysis. 28; 2002:1011- 1040.


Abstract Views: 413

PDF Views: 2




  • Development and Validation of Stability Indicating HPTLC Method for Estimation of Ursolic Acid

Abstract Views: 413  |  PDF Views: 2

Authors

Abstract


A stability indicating high performance thin layer chromatographic (HPTLC) method was developed and validated for estimation of ursolic acid. Study was performed on pre-coated silica gel TLC plates using toluene: ethyl acetate: formic acid (7:3:0.1, v/v/v) as the mobile phase. After development, plates were derivatized with Anisaldehyde- Sulphuric acid reagent. Detection and quantification were performed by densitometric scanning at 530 nm. Method was validated according to ICH Q2(R<SUB>1</SUB>)guideline. The linear regression analysis for the calibration plot showed good linear relationship over the concentration range of 200-600 ng/spot. The correlation coefficient of calibration curve was found to be 0.9986. The intra and inter day precision was found to be less than 2%. Ursolic acid was also subjected to acid, base, oxidation, heat, humidity and photo-degradation studies. The degradation products obtained were well resolved from the pure drug with significantly different Rf values. As the method could effectively separate the drugs from its degradation products, it can be used for stability-indicating analysis.

Keywords


Ursolic Acid, Stability Indicating Method, High Performance Thin Layer Chromatography (HPTLC), Method Validation

References