Method Development and Validation for the Estimation of Valerenic Acid Content in Valerian Tablets Using RP-HPLC Method

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M. Sahiti
B. M. Gurupadayya


Valerian (Valeriana officinalis) has been used from so long as a traditional medicine since it has therapeutic effects such as enhances sleep, acts as an anxiolytic for nervous unrest etc. For determining the dose of herbal formulations and to estimate the quality of dosage form, we need to develop an effective reproducible validated method for the determination of the bioactive molecules essential for the sedative activity of the valerian tablets. So, the primary objective of the present work was to develop an efficient and sensitive method for the determination of valerenic acid content in valerian tablets using reverse-phase high-performance liquid chromatography technique. Since it is RP-HPLC method the column used was Phenomenex C18 Luna® 5 µm (250 x 4.6mm) column. The LC system used was Shimadzu UFLC, Nexera series utilizing a Photo-diode Array detector (SPD-M20A) detector and the software used was LC Solutions. The mobile phase used is acetonitrile: Ortho-phosphoric -acid in the ratio of 97:3 v/v. Ortho-phosphoric acid (gradient mode) employed was of 5% solution with a pH of 3.5. The chromatographic conditions employed are 1ml/min flow rate and absorbances were observed at 220 nm. The selected chromatographic conditions were found to give effective separation of Valerenic acid at 4.4±0.2 min. Validation was carried out for linearity, system suitability, accuracy, precision (intra-day and inter-day), and robustness, the limit of detection and limit of quantification which were found to be within acceptable limits according to ICH Guidelines. Thus, the method proposed was found to be accurate, precise, reproducible and analyte-specific. In this work, based on the observations made from the literature review a relatively best suitable method was developed for the estimation of valerenic acid in valerian tablets with shortened retention time along with better resolution thus, reducing the wastage of solvents and the time of analysis.

RP-HPLC, Valeriana officinalis, valerenic acid, validation.

Article Details

How to Cite
Sahiti, M., & Gurupadayya, B. M. (2019). Method Development and Validation for the Estimation of Valerenic Acid Content in Valerian Tablets Using RP-HPLC Method. South Asian Research Journal of Natural Products, 2(3), 1-9. Retrieved from
Original Research Article


Mills S, Bone K. Principles and practice of phytotherapy. Modern Herbal Medicine. Churchill Livingstone; 2000.

Rotblatt M. American herbal pharma-copoeia and therapeutic compendia: analytical, quality control and therapeutic monographs. The Journal of Alternative and Complementary Medicine. 2000;6(1): 95-6.

Pizzorno JE, Murray MT. textbook of natural medicine-e-book. Elsevier Health Sciences; 2012.

Dietz BM, Mahady GB, Pauli GF, Farnsworth NR. Valerian extract and valerenic acid are partial agonists of the 5-HT5a receptor in vitro. Molecular Brain Research. 2005;138(2):191-7.

Taibi DM, Landis CA. Valerian and other CAM botanicals in treatment of sleep disturbances. In Complementary and Alternative Therapies and the Aging Population. Academic Press. 2009;57- 81.

Robinson L. Herbal Monograph: Valeriana officinalis; 2012.

Singh N, Gupta AP, Singh B, Kaul VK. Quantification of valerenic acid in Valeriana jatamansi and Valeriana officinalis by HPTLC. Chromatographia. 2006;63(3-4):209-13.

Pavlovic M, Kovacevic N, Tzakou O, Couladis M. The essential oil of Valeriana officinalis L. sl growing wild in Western Serbia. Journal of Essential Oil Research. 2004;16(5):397-9.

Mikell JR, Ganzera M, Khan IA. Analysis of sesquiterpenes in Valeriana officinalis by capillary electrophoresis. Die Pharmazie. 2001;56(12):946-8.

Ghafari S, Esmaeili S, Aref H, Naghibi F, Mosaddegh M. Qualitative and quantitative analysis of some brands of valerian pharmaceutical products. Studies on Ethno-Medicine. 2009;3(1):61-4.

Safaralie A, Fatemi S, Sefidkon F. Essential oil composition of Valeriana officinalis L. roots cultivated in Iran: Comparative analysis between super-critical CO2 extraction and hydro-distillation. Journal of Chromatography A. 2008;1180(1-2):159-64.

Srivastava A, Tiwari SS, Srivastava S, Rawat AK. HPTLC method for quantification of valerenic acid in Ayurvedic drug Jatamansi and its substitutes. Journal of liquid chromatography & related technologies. 2010;33(18):1679-88.

Ebrahimzadeh H, rajabian T, Ekhteraei TS, Bashiri SZ, Niknam V, Zareei M. Quantification of valerenic acid and its derivatives in some species of Valeriana l. and Centranthus longiflorus stev.

Hassan E, Tayebeh R, Samaneh ET, Zeinalabedin BS, Vahid N, Mehdi Z. Quantification of Valerenic acid and its derivatives in some species of Valeriana L and Centranthus longiflorus Stev. Asian Journal of Plant Sciences. 2008;7(2):195-200.

Goppel M, Franz G. Stability control of valerian ground material and extracts: A new HPLC-method for the routine quantification of valerenic acids and lignans. Die Pharmazie-An International Journal of Pharmaceutical Sciences. 2004; 59(6):446-52.

Asadollahi-Baboli M. Comprehensive analysis of Valeriana officinalis L. essential oil using GC-MS coupled with integrated chemometric resolution techniques. International Journal of Food Properties. 2015;18(3):597-607.

Boyadzhiev L, Kancheva D, Gourdon C, Metcheva D. Extraction of valerenic acids from valerian (Valeriana officinalis L.) rhizomes. Die Pharmazie-An International Journal of Pharmaceutical Sciences. 2004; 59(9):727-8.

Torrado JJ. In vitro release of valerenic and hydroxyvalerenic acids from valerian tablets. Die Pharmazie. An International Journal of Pharmaceutical Sciences. 2003; 58(9):636-8.

Shohet D, Wills RB, Stuart DL. Valepotriates and valerenic acids in commercial preparations of valerian available in Australia. Die Pharmazie. 2001;56(11):860-3.

Kopp S, Schweizer WB, Altmann KH. Total synthesis of valerenic acid. Synlett. 2009; 11:1769-72.