Article Sidebar

Submitted
Jun 30, 2022
Accepted
Aug 21, 2022
Published
Sep 30, 2022
Download
pdf
Statistic
Read Counter : 465 Download : 358

Downloads

Download data is not yet available.

Main Article Content

Abstract

Soursop leaves contain alkaloids that have pharmacological effects. This study aims determine the total alkaloid content of soursop leaf ethanol extract using a validated HPLC method. Extraction was carried out by soxhletation followed by liquid-liquid extraction using chloroform. Optimizing HPLC conditions on the mobile phase, flow rate, concentration, and wavelength, further testing the validation of the analytical method. Optimizing HPLC conditions obtained optimal results at concentration of 500 ppm with a flow rate of 1.0 mL/min using acetonitrile: methanol: water (80:5:15) at wavelength of 272 nm. These results have met the validation requirements of analytical methods including System Suitability Test (UKS), selectivity, linearity with y = 2.01914x –219,97226, correlation coefficient value (r) = 0.9995, and Vx0 value 0.11201%. The limit of detection and quantization were 8,78525 ppm and 29,28418 ppm, respectively. The RSD percentage of the precision test is 1.0453%, and the accuracy test is obtained from the recovery of 98-102%. The result of determining the total alkaloid content  sample was 0.0862 ± 0.004 % (%w/w) of the entire alkaloid content (TALC) triple replication. This research concludes that the analytical method using the HPLC system for determining the total alkaloid content of the ethanolic extract of soursop leaves is validate

Keywords

Soursop leaf, HPLC, validation analytical method, total alkaloid content

Article Details

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

How to Cite
1.
Setyaningrum L, Purwaningtyas D, Puspaningtyas AR, Wigati D, Usman MR, Latifah MS. Development and Validation of RP-HPLC Analysis Method for Determination of Total Alkaloid Content of Soursop (Annona muricata L.) Leaf Extract. EKSAKTA [Internet]. 2022Sep.30 [cited 2024Nov.21];23(03):175-87. Available from: https://eksakta.ppj.unp.ac.id/index.php/eksakta/article/view/325

References

  1. M. K. Simo, M. D. Nguepi, M. L. Sameza, R. K. Toghueo, F. B. Fekam, and G. Froldi (2018) Cameroonian medicinal plants belonging to Annonaceae family: radical scavenging and antifungal activities. Natural Product Research, vol. 32, no. 17, pp. 2092–2095.
  2. T. Abdel-Rahman, A. S. Hussein, S. Beshir, A. R. Hamed, E. Ali, and S. S. El-Tanany. (2019). Antimicrobial activity of terpenoids extracted from Annona muricata seeds and its endophytic Aspergillus Niger strain SH3 either singly or in combination. Open Access Macedonian Journal of Medical Sciences, vol. 7, no. 19, pp. 3127–3131
  3. C. Z. Cagnini et al., (2021). Antimicrobial activity of Annona muricata leaf oleoresin. Natural Product Research, pp. 1–7.
  4. D. Neglo et al., (2021). Evaluation of the Modulatory Effect of Annona muricata Extracts on the Activity of Some Selected Antibiotics against Biofilm-Forming MRSA. Evidence-based Complementary and Alternative Medicine
  5. Quantitative Assessment of the Relative Antineoplastic Potential of the n-butanolic Leaf Extract of Annona Muricata Linn in Normal and Immortalized Human Cell Lines, (2012), Asian Pacific Journal of Cancer Prevention, vol. 13, no. 2, pp. 699–704 [Online]. Available: http://journal.waocp.org/article_26296.html
  6. K. Utari, E. Nursafitri, R. Sari, A. K. Winda, and A. S. Harti. (2013). Kegunaan daun sirsak (Annona muricata L) untuk membunuh sel kanker dan pengganti kemoterapi. Jurnal Kesehatan Kusuma Husada.
  7. Minarni et al., (2017). Anticancer activity test of ethyl acetate extract of endophytic fungi isolated from soursop leaf (Annona muricata L.). Asian Pacific Journal of Tropical Medicine, vol. 10, no. 6, pp. 566–571.
  8. I. Rady et al., (2018). Anticancer Properties of Graviola (Annona muricata): A Comprehensive Mechanistic Review. Oxidative Medicine and Cellular Longevity.
  9. R. M. K. Toghueo et al., (2019). Antiplasmodial potential and GC-MS fingerprint of endophytic fungal extracts derived from Cameroonian Annona muricata. Journal of Ethnopharmacology, vol. 235, pp. 111–121.
  10. E. Soekaryo. (2019). Uji Inhibisi Enzim Siklooksigenase-2 (Cox-2) Dari Ekstrak Daun Sirsak (Annona Muricata Linn.) sebagai Antiinflamasi
  11. V. S. Rahayu, C. Miyarso, and N. Z. W. Kiromah, (2021). Test Of Analgesic Effect Of The Aquadest Extract Of Sousoup Leaf (Annona Muricata L)) On Mice (Mus Musculus. Proceeding of The URECOL, pp. 932–941.
  12. S. Fofana et al., (2012). Alkaloids From Leaves Of Annona muricata.
  13. A. S. S. C. Lúcio, J. R. G. da S. Almeida, E. V. L. da-Cunha, J. F. Tavares, and J. M. Barbosa Filho, (2015). Alkaloids of the Annonaceae: Occurrence and a Compilation of Their Biological Activities. Alkaloids: Chemistry and Biology, vol. 74, pp. 233–409.
  14. S. Z. Moghadamtousi, M. Fadaeinasab, S. Nikzad, G. Mohan, H. M. Ali, and H. A. Kadir, (2015). Annona muricata (Annonaceae): A review of its traditional uses, isolated acetogenins and biological activities. International Journal of Molecular Sciences, vol. 16, no. 7. MDPI AG, pp. 15625–15658.
  15. H. Rivai, W. R. Susandri, and Z. Azizah. (2019). Analisis Kualitatif dan Analisis Kuantitatif dari Ekstrak Heksan, Aseton, Etanol, dan Air dari Daun Sirsak (Annona muricata L.).
  16. A. P. Ratu, S. Setyahadi, and P. Simanjuntak. (2017). Aktivitas Fraksi etanol daun sirsak (Annona muricata Linn.) Sebagai Penghambat asetilkolinesterase. Parapemikir: Jurnal Ilmiah Farmasi, vol. 6, no. 2.
  17. A. S. Nugraha, R. Haritakun, J. M. Lambert, C. T. Dillon, and P. A. Keller. (2021). Alkaloids from the root of Indonesian Annona muricata L. Natural Product Research, vol. 35, no. 3, pp. 481–489.
  18. H. Parbuntari, Y. Prestica, R. Gunawan, M. N. Nurman, F. Adella, and N. Padang. (2018). Preliminary Phytochemical Screening (Qualitative Analysis) of Cacao Leaves (Theobroma Cacao L.). EKSAKTA Berkala Ilmiah Bidang MIPA 19(2):40-45
  19. C. Z. Lin, S. M. Xie, C. C. Zhu, Z. D. Bairu, S. Q. Kangsa, and D. Zhu. (2013). Simultaneous determination of five diterpenoid alkaloids in Herba Delphinii by HPLC/ELSD. Journal of Pharmaceutical Analysis, vol. 3, no. 6, pp. 447–451.
  20. I. Ivanov, S. Berkov, and A. Pavlov. (2009). Improved hplc metod for the determination of amaryllidaceae alkaloids. Biotechnology and Biotechnological Equipment, vol. 23, pp. 809–813
  21. L. Li, W. Long, X. Wan, Q. Ding, F. Zhang, and D. Wan. (2015). Studies on quantitative determination of total alkaloids and berberine in five origins of crude medicine ‘sankezhen’. Journal of Chromatographic Science, vol. 53, no. 2, pp. 307–311
  22. N. Aini. (2016). Penentuan kadar alkaloid pada ekstrak daun tanaman menggunakan metode nir dan kemometrik. Skripsi. Universitas Jember, Jember, Jawa Timur
  23. J. Xiao et al., (2019). High performance liquid chromatography determination and optimization of the extraction process for the total alkaloids from traditional herb stephania cepharantha hayata. Molecules, vol. 24, no. 3
  24. S. Ahmed, A. Kadhim Alanbari, and H. R. R. Al-Taai. (2019). HPLC profiles of some bioactive compounds in Annona muricata L. and Garcinia mangostana L. peels. International Conference on Food, Nutrition and Agriculture (ICFNA19). Istanbul, Turkey.
  25. S. Wahyuni and P. Marpaung. (2020). Determination of Total Alkaloid Levels Extracts of Akar Kuning (Fibraurea chloroleuca Miers) Based on The Differences of Ethanol Concentrations by Spectrofotometry UV-Vis Method. Dalton: Jurnal Pendidikan Kimia dan Ilmu Kimia, Volume 3 Nomor 2.
  26. R. Ayuningtyas, R. Primaharinastiti, M. Yuwono. (2020). Optimasi dan Validasi Metode KCKT untuk Identifikasi dan Penetapan Kadar Metabolit Nitrofuran dalam Bakso Udang the HPLC Methods for Identificaton and Determination of Nitrofuran Metabolite Levels in Shrimps Meatballs. Jurnal Farmasi Dan Ilmu Kefarmasian Indonesia, vol. 8, no. 2, p. 117
  27. D. Wulandaria, G. Gusrizala, and T. A. Zaharaha. (2020). Optimasi dan Validasi Metode Penentuan Kadar Asam Glikolat dan Asam Laktat Dalam Krim Menggunakan Kromatografi Cair Kinerja Tinggi, ALCHEMY Jurnal Penelitian Kimia, vol. 16, no. 1, pp. 10–24
  28. S. Sudjarwo, F. -Annuryanti, M. Nur Alfi, and F. Annuryanti. (2019). Validation and Development of Tlc-Densitometry Method For Standardization Of Soursop Leaf Extract (Annona Muricata Linn.) With Quercetin The Effectiveness And Toxicity Of Glucomannan In Porang And Konjac Powder For Niddm In Mus Musculus View Project Validation And Development Of Tlc-Densitometry Method For Standardization Of Soursop Leaf Extract (Annona Muricata Linn.) With Quercetin. International Journal of Pharmaceutical Sciences and Research, vol. 10, no. 2, p. 686
  29. N. S. D. Brahmana. (2020). Kemampuan Daya Hambat Beberapa Produk Sabun Cair Antibakteri Terhadap Bakteri Staphylococcus aureus. Skripsi. Universitas Medan Area. Medan
  30. R. K. Patel, J. B. Patel, and P. D. Trivedi. (2015). Spectrophotometric method for the estimation of total alkaloids in the Tinospora cordifolia M. and its herbal formulations. Int J Pharm Pharm Sci, vol. 7, no. 10, pp. 249–251
  31. P. N. Patil (2012). Caffeine in various samples and their analysis with HPLC-A review. International Journal of Pharmaceutical Sciences Review and Research 16(2):76-83.
  32. S. Chowdhury, M. Maleque, and M. H. Shihan. (2012). Development and Validation of a Simple RP-HPLC Method for Determination of Caffeine in Pharmaceutical Dosage Forms. Asian Journal of Pharmaceutical Analysis, vol. 2, pp. 1–4
  33. A. A. Rahim, S. Nofrizal, and B. Saad. (2014). Rapid tea catechins and caffeine determination by HPLC using microwave-assisted extraction and silica monolithic column. Food Chemistry, vol. 147, pp. 262–268.
  34. T. S. Ningtyas and I. Fajriati. (2020). Analisis Pemanis buatan Natrium Siklamat pada minuman ringan dengan Kromatografi cair kinerja tinggi. Indonesian Journal of Halal Science, vol. 1, no. 1, pp. 30–35
  35. I. G. Gandjar and A. Rohman. (2007). Kimia farmasi analisis. Pustaka Pelajar, Yogyakarta
  36. M. Yuwono and G. Indrayanto. (2005). Validation of chromatographic methods of analysis. Profiles of drug substances, excipients and related methodology, vol. 32, pp. 243–259.
  37. Association of Official Analytical Chemists. (2013). AOAC Appendix K : Guidelines for Dietary Supplements and Botanicals. pp. 1–32
  38. A.C. Moffat M.D. Osselton B. Widdop J. Watts. (2011). Review of: Clarke’s Analysis of Drugs and Poisons, 4th edn. Journal of Analytical Toxicology, Volume 36, Issue 5
  39. N. H. Fajriana and I. Fajriati. (2018). Analisis Kadar Kafein Kopi Arabika (Coffea arabica L.) pada Variasi Temperatur Sangrai secara Spektrofotometri Ultra Violet. Analit: Analytical and Environmental Chemistry, vol. 3, no. 2
  40. L. R. Snyder, J. J. Kirkland, and J. W. Dolan. (2011). Introduction to modern liquid chromatography. John Wiley & Sons
  41. L. Dhooghe et al.. (2008). Development and validation of an HPLC-method for the determination of alkaloids in the stem bark extract of Nauclea pobeguinii, Talanta, vol. 76, no. 2, pp. 462–468
  42. M. Cao et al., (2020). HPLC-based qualitative and quantitative analyses of alkaloids in chewable areca products from different geographic regions. J AOAC Int, vol. 103, no. 5, pp. 1400–1405
  43. P. Naveen, H. Lingaraju, M. Deepak, B. Medhini, and K. Prasad. (2018). Method Development and Validation for the Determination of Caffeine: An Alkaloid from Coffea arabica by High-performance Liquid Chromatography Method. Pharmacognosy Research, vol. 10, no. 1, pp. 88–91
  44. R. Saad and H. Bustan. (2016). Validation and Determination of Caffeine Contents in Energy Drinks Available on the Iraqi Market by Using High Performance liquid chromatography (HPLC). International Journal of Pharmacy &Pharmaceutical Reasearch. Vol 6, issue 1
  45. D. Sylvi, F. Azima, and S. Anggini. (2021). The Effect of Additional Fruit Eggplant (Solanum betaceum cav.) Juice on the Characteristics of Black Tea (Camelia sinensis) Beverage. EKSAKTA :Berkala Ilmiah Bidang MIPA. vol. 22, pp. 270–283