Validation of High-Performance Liquid Chromatography (HPLC) Method for Quantification of Ethyl p-Methoxycinnamate in Kaempferia galanga Extract

http://www.doi.org/10.26538/tjnpr/v7i8.39

Authors

  • Oktavia R. Adianingsih Department of Pharmacy, Faculty of Medicine, Brawijaya University, Malang 65145, Indonesia
  • Bachtiar R. P. Ihsan Department of Pharmacy, Faculty of Medicine, Brawijaya University, Malang 65145, Indonesia
  • Oktavia E. Puspita Department of Pharmacy, Faculty of Medicine, Brawijaya University, Malang 65145, Indonesia
  • Kadek S. Maesayani Department of Pharmacy, Faculty of Medicine, Brawijaya University, Malang 65145, Indonesia

Keywords:

Method validation, High-performance liquid chromatography, Kaempferia galanga, Ethyl p-methoxycinnamate

Abstract

Kaempferia galanga has the potential to be developed as a raw material for traditional medicine because it has pharmacological activities such as analgesic and anti-inflammatory. Ethyl pmethoxycinnamate (EPMC) is one of the marker compounds for determining the dosage of traditional medicine. Determination of EPMC content in the Kaempferia galanga extract is important for the standardization of the extract. This study aims to validate the high-performance liquid chromatography (HPLC) method for determining EPMC content in Kaempferia galanga extract. Separation was performed on the C18 column (5 μm, 4.6 mm x 150 mm), using an isocratic mobile phase consisting of water: acetonitrile (40:60) at 30oC temperature. The flow rate was 1 mL/min with an injection volume of 10 μL. Detection was performed using a photodiode array (PDA) set at 308 nm. The method was validated as per the guidelines given by International Conference on Harmonization guidelines. The retention time of EPMC on standard EPMC and
Kaempferia galanga was 6.22 and 6.25 minutes, respectively. The linearity calibration curve was 10-60 µg/mL (r=0.9988) with the limit of detection (LOD) of 0.0011 µg/mL and limit of quantification (LOQ) of 0.0037 µg/mL. Adequate results were obtained for precision with a
relative standard deviation (RSD) of 1.19%-2.37% and accuracy with a recovery of 94.07%-113.82%. The average EPMC content in Kaempferia galanga was 54.13%. Therefore, this finding demonstrated that the method was proved selective for the quantification of EPMC in Kaempferia galanga extract. 

References

Kumar A. Phytochemistry, pharmacological activities and uses of traditional medicinal plant Kaempferia galanga L.–An overview. J. Ethnopharmacol. 2020; 253:112667. Doi: 10.1016/j.jep.2020.112667.

Khairullah AR, Solikhah TI, Ansori ANM, Hanisia RH, Puspitarani GA, Fadholly A, Ramandinianto SC. Medicinal importance of Kaempferia galanga L. (Zingiberaceae): A comprehensive review. J. HerbMed Pharmacol. 2021; 10(3):281-288. Doi: 10.34172/jhp.2021.32.

Dash PR, Mou KM, Erina IN, Ripa FA, Al Masud KN, Ali MS. Study of anthelmintic and insecticidal activities of different extracts of Kaempferia galanga . IJPSR. 2017; 8(2):729-733. Doi: 10.13040/IJPSR.0975-8232.

Wang SY, Zhao H, Xu HT, Han XD, Wu YS, Xu FF, Liu B. Kaempferia galanga L.: Progresses in phytochemistry, pharmacology, toxicology and ethnomedicinal uses. Front. Pharmacol. 2021; 12:675350. Doi: 10.3389/fphar.2021.675350.

Begum T, Gogoi R, Sarma N, Pandey SK, Lal M. Novel ethylp-methoxy cinnamate rich Kaempferia galanga (L.) essential oil and its pharmacological applications: special emphasis on anticholinesterase, anti-tyrosinase, α-amylase inhibitory, and genotoxic efficiencies. PeerJ. 2023; 11:14606. Doi: 10.7717/peerj.14606.

Zahara M, Hasanah M, Zalianda R. Identification of Zingiberaceae as medicinal plants in Gunung Cut Village, Aceh Barat Daya, Indonesia. JTHort. 2018; 1(1):24-28. Doi: 10.33089/jthort.v1i1.9.

Adianingsih OR, Widaryanto E, Saitama A, Zaini AH. Analysis of bioactive compounds present in Kaempferia galanga rhizome collected from different regions of East Java, Indonesia. IOP Conf. Ser.: Earth Environ. Sci. 2021; 913(1):012074. Doi: 10.1088/1755-1315/913/1/012074.

Septyanti C, Batubara I, Rafi M. HPLC fingerprint analysis combined with chemometrics for authentication of Kaempferia galanga from related species. Indones. J. Chem. 2016; 16(3):308-314. Doi: 10.22146/ijc.21146.

Choironi NA, Pudyastuti B, Gumelar G, Fareza MS, Wijaya TH, Setyono J. Formula Optimization of the SelfNanoemulsifying Drug Delivery System (SNEDDS) of Ethyl-p-methoxycinnamate (EPMC). ALCHEMY J. Pen. Kim. 2022; 18(2):205-213. Doi: 1.20961/alchemy.18.2.56847.205-213.

Alam G, Mishra AK. Traditional and modern approaches for standardization of herbal drugs: a review. Acta Biomedica Scientia. 2017; 4(1):40-55. Doi: 10.21276/abs.2017.4.1.9.

Indrayanto G. The importance of method validation in herbal drug research. J. Pharm. Biomed. Anal. 2022; 214:114735. Doi: 10.1016/j.jpba.2022.114735.

Ihsan BRP, Maysaroh I, Nurhayati IP. Method validation of ultra high performance chromatography-double mass spectrometry (UHPLC-MS/MS) for analysis of curcumin in ethanol extract of turmeric (Curcuma longa) with various comparisons. PJI. 2018; 4(1):29-34.

Ihsan BRP, Hafid AF, Primaharinastiti R, Widyawaruyanti A, Yuwono M. Development and validation of HPLC method for determination of andrographolide in raw material and tablet of ethyl acetate fractions of Andrographis paniculata. RJPT. 2020; 13(9):4291-4296. Doi: 10.5958/0974-360X.2020.00758.1.

Ihsan BRP, Delina AP, Shalas AF. Determination of quercetin in extracts and herbal products of Phyllanthus niruri by TLC densitometry method. PLANAR. 2022; 2:31-37. Doi: 10.18860/planar.v2i0.1855.

Mukherjee PK, Bahadur S, Chaudhary SK, Kar A, Mukherjee K. Quality related safety issue-evidence-based validation of herbal medicine farm to pharma. In: Mukherjee PK. Evidence-Based Validation of Herbal Medicine. Amsterdam: Elsevier Inc; 2015. 1-28 p. Doi: 10.1016/B978- 0-12-800874-4.00001-5.

Ministry of Health of the Republic of Indonesia. 2017. Indonesian Herbal Pharmacopoeia. Jakarta: Ministry of Health of the Republic of Indonesia.

Khuntia S, Sahoo D, Kar B, Sahoo S. Metabolic profiling of Kaempferia galanga leaf and rhizome extract using GCMS. J. Appl. Adv. Res. 2022; 7:35-41. Doi: 10.21839/jaar.2022.v7.7704.

Tsizin S, Bokka R, Keshet U, Alon T, Fialkov AB, Tal N, Amirav A. Comparison of electrospray LC–MS, LC–MS with Cold EI and GC–MS with Cold EI for sample identification. Int J Mass Spectrom. 2017; 422:119-125. Doi: 10.1016/j.ijms.2017.09.006.

Tetali SD, Acharya S, Ankari AB, Nanakram V, Raghavendra AS. Metabolomics of Withania somnifera (L.) Dunal: Advances and applications. J. Ethnopharmacol. 2021; 267:113469. Doi: 10.1016/j.jep.2020.113469.

Zhang K, Wu W, Tian S. Comparative study on the determination of ethyl p-methoxycinnamate in Kaempferia galanga rhizome by HPTLCS and HPLC. JPC–J PLANAR CHROMAT. 2020; 33(1):51-57. Doi: 10.1007/s00764-019-00011-1.

Hussain A, Pu H, Sun DW. Measurements of lycopene contents in fruit: A review of recent developments in conventional and novel techniques. Crit. Rev. Food Sci. Nutr. 2019; 59(5):758-769. Doi: 10.1080/10408398.2018.1518896.

Zhao X, Liu Z, Sun F, Yao L, Yang G, Wang K. Bile acid detection techniques and bile acid-related diseases. Front. Physiol. 2022; 13:826740. Doi: 10.3389/fphys.2022.826740.

Mane S. Racemic drug resolution: a comprehensive guide. Anal. Methods. 2016; 8(42):7567-7586. Doi:10.1039/c6ay02015a.

International Conference on Harmonization (ICH). Q2 (R1) Validation of analytical procedures: Text and Methodology. 1995.

Phattanaphakdee W, Ditipaeng C, Uttayarat P, Thongnopkoon T, Athikomkulchai S, Chittasupho C. Development and validation of HPLC method for determination of quercetin in hydrogel transdermal patches loaded with red onion peel extract. Trop. J. Nat. Prod. Res. 2022; 6(9):1210-1214. Doi: 10.26538/tjnpr/v6i8.8.

Yunarto N, Calvin CC, Sulistyowati I, Oktoberia IS, Reswandaru UN, Elya B, Mihardja LK. Development and validation of a high-performance liquid chromatographybased method for catechin isolated from the leaves of gambir (Uncaria gambir Roxb). Trop. J. Nat. Prod. Res. 2023;

(3):2569-2573. Doi: 10.26538/tjnpr/v7i3.16.

Ghanjaoui ME, Mandil A, Ait Sidi Mou A, Slimani R. High performance liquid chromatography quality control. Int. J. Adv. Chem. 2020; 8:160-169. Doi: 10.14419/ijac.v8i1.30723.

Srivastava N, Mishra S, Iqbal H, Chanda D, Shanker K. Standardization of Kaempferia galanga L. rhizome and vasorelaxation effect of its key metabolite ethyl pmethoxycinnamate. J. Ethnopharmacol. 2021; 271:113911. Doi: 10.1016/j.jep.2021.113911.

Winingsih W, Husein SG, Ramdhani RPN. Analysis of ethyl p-methoxycinnamate from Kaempferia galanga L. extract by high performance liquid chromatography. J. Trop. Pharm. 2021; 5(4):353-358. Doi: 10.25026/jtpc.v5i4.3

United States Pharmacopoeia 43 National Formulary 38 (USP43-NF38). United States Pharmacopoeial Convention Inc., USA. 2020.

Patil MS, Patil RR, Chalikwar SS, Surana SJ, Firke SD. Analytical Method Development And Validation: A Review. IJPBA. 2019; 7(3):70-81. Doi: 10.32553/ijpba.v7i3.126.

AOAC International. 2016. Appendix K: Guidelines for Dietary Supplements and Botanicals. AOAC Official Method of Analysis. AOAC International.

Reddy PS, Jagarlapudi VSK, Sekaran CB. Determination of edoxaban in bulk and in tablet dosage form by stabilityindicating high-performance liquid chromatography. Pharm. Sci. 2016; 22(1):35-41. Doi: 10.15171/PS.2016.07.

Srivastava N, Singh S, Gupta AC, Shanker K, Bawankule DU, Luqman S. Aromatic ginger (Kaempferia galanga L.) extracts with ameliorative and protective potential as a functional food, beyond its flavor and nutritional benefits. Toxicol. Rep. 2019; 6:521-528. Doi: 10.1016/j.toxrep.2019.05.014.

Mukkasombut N, Pipatrattanaseree W, Itharat A. Validation of HPLC method for the determination of anti-allergic compounds in ethanolic extract of Prasaprohyai remedy, a Thai traditional medicine. TMJ. 2020; 20(1):74-83.

Dwita LP, Hikmawanti NPE, Yeni, Supandi. Extract, fractions, and ethyl-p-methoxycinnamate isolate from Kaempferia galanga Elicit anti-inflammatory activity by limiting leukotriene B4 (LTB4) production. J Tradit Complement Med. 2021; 11(6):563-569. Doi: 10.1016/j.jtcme.2021.06.004.

Lallo S, Hardianti B, Sartini S, Ismail I, Laela D, Hayakawa Y. Ethyl p-methoxycinnamate: an active anti-metastasis agent and chemosensitizer targeting NFκB from Kaempferia galanga for melanoma cells. Life. 2022; 12(3):337. Doi: 10.3390/life12030337.

Chittasupho C, Ditsri S, Singh S, Kanlayavattanakul M, Duangnin N, Ruksiriwanich W, Athikomkulchai S. Ultraviolet radiation protective and anti-inflammatory effects of Kaempferia galanga L. rhizome oil and microemulsion: formulation, characterization, and hydrogel preparation. Gels. 2022; 8: 639. Doi: 10.3390/gels8100639

Chasanah U, Yulian F, Yulianty R, Irianto A, Radjaram A. The Effects of binder on the physical characteristic of squeezed rhizome of Kaempferia galanga L lozenges. HSIC. 2017; 2: 264-270.

Pratama G, Yanuarti R, Ilhamdy AF, Suhana, MP.Formulation of sunscreen cream from Eucheuma cottonii and Kaempferia galanga (zingiberaceae). IOP Conf.Ser.: Earth Environ. Sci. 2019; 278: 012062.

Published

2023-08-31

How to Cite

Adianingsih, O. R., Ihsan, B. R. P., Puspita, O. E., & Maesayani, K. S. (2023). Validation of High-Performance Liquid Chromatography (HPLC) Method for Quantification of Ethyl p-Methoxycinnamate in Kaempferia galanga Extract: http://www.doi.org/10.26538/tjnpr/v7i8.39. Tropical Journal of Natural Product Research (TJNPR), 7(8), 3829–3835. Retrieved from https://tjnpr.org/index.php/home/article/view/2459