Butterfly pea (Clitoria ternatea L.) flowers infusion causes very mild damage to histological structure of albino rats’ (Rattus norvegicus Berkehnout, 1769) liver and kidney
Year 2026,
Volume: 30 Issue: 1, 208 - 224, 11.01.2026
Nur Ainun Oktavia Pusparini
,
Tia Apriliyani
,
Ardaning Nuriliani
Abstract
Butterfly pea flowers (BPF) are widely used in Indonesia as a natural food and beverage colorants. BPF is also traditionally used to relieve diseases such as diabetes and cardiovascular diseases. However, few research has been conducted on the potential toxic effects of BPF infusion. Therefore, further research is necessary to evaluate its safe consumption dosage. The objective of this study was to investigate acute and subchronic toxicity of BPF infusion on histological structure of the kidney and liver in male albino rat. Acute toxicity treatment, BPF infusion was orally administered at the doses of 2500 and 5000 mg kg-1 BW for 14 days. For the subchronic toxicity treatment, BPF infusion was orally administered at the doses of 250, 500, and 1000 mg kg-1 BW. Histological preparations were performed using the paraffin method, followed by hematoxylin-eosin (HE) staining. The histological structures of the livers and kidney were evaluated using an ordinal scoring method. Data were analyzed using the Mann-Whitney test for acute treatments and the Kruskal-Wallis test for subchronic treatments (p ≤ 0.05). The results indicated very mild liver damage in the acute treatment groups, characterized by cellular degeneration. In the subchronic treatment groups, very mild damage was observed, including cellular degeneration and vacuolization. Additionally, very mild kidney damage, such as leukocyte infiltration and tubular hemorrhage, was detected in both acute and subchronic treatment groups. These findings suggest that BPF infusion is relatively safe for consumption, provided it is not consumed over an extended period.
Ethical Statement
The Ethics Committee of the Faculty of Veterinary Medicine, Universitas Gadjah Mada, authorized all methods related to the use and care of test animals in this study. The approval was granted under letter number: 127/ECFKH/Eks./2023
Supporting Institution
Faculty of Biology, Universitas Gadjah Mada
Thanks
This research was funded by Hibah Rekognisi Tugas Akhir (RTA Grand), Universitas Gadjah Mada 2024 (Number: 4971/UN1.P1/PT.01.01/2024).
References
-
[1] Kementerian Perdagangan Republik Indonesia. Penganugerahan UKM pangan award 2024. https://www.kemendag.go.id/berita/foto/penganugerahan-ukm-pangan-award-2024 (accesed on 21 November 2024)
-
[2] Indonesian Food and Drug Authority’s. Bahan tambahan pangan. https://jdih.pom.go.id/download/flip/1251/11/2021 (accesed on 15 November 2024)
-
[3] Fikkra KL and Holinesti R. The effect of substituting butterfly pea extract as a natural dye in mochi production. Jurnal Pendidikan Tata Boga dan Teknologi. 2024; 5(1): 89-95 https://doi.org/10.24036/jptbt.v5i1.12919
-
[4] Jaiswal AS, Kadam KA, Gangurde H, Mouraiya MM, Dhokane PA. Isolation and evaluation of food dye from butterfly peas. International journal of creative research thoughts. Int J Creat Res Thoughts. 2023; 11(8): f570-f583.
-
[5] Widowati W, Darsono L, Utomo HS, Sabrina AHN, Natariza MR, Valentinus Tarigan AC, Waluyo NW, Gleyriena AM, Siahaan BH, Oktaviani R. Antidiabetic and hepatoprotection effect of butterfly pea flower (Clitoria ternatea L.) through antioxidant, anti-inflammatory, lower LDH, ACP, AST, and ALT on diabetes mellitus and dyslipidemia rat. Heliyon. 2024;10(8):e29812. https://doi.org/10.1016/j.heliyon.2024.e29812
-
[6] Wang Y, Liu T, Xie Y, Na L, Liu Y, Wen J, Zhang M, Feng W, Huang J, Guo Y, Junior T, Wang D, Granato D. Clitoria ternatea blue petal extract protects against obesity, oxidative stress, and inflammation induced by a high-fat, high-fructose diet in C57BL/6 mice. Food Res Int. 2022; 162: 1-12. https://doi.org/10.1016/j.foodres.2022.112008
-
[7] Al-Shamrani SM, Safhi FA, Mobasher MA, Saleem RM, Alharthi A, Alshaya DS, Awad NS. Antiproliferative effect of Clitoria ternatea ethanolic extract against colorectal, breast, and medullary thyroid cancer cell lines. Separations. 2022; 9: 311. https://doi.org/10.3390/separations9110331
-
[8] Pebiansyah A, Rahayuningsih N, Yeni Aprilia A, Nuryadin Zain D. Aktivitas hepatoprotektif ekstrak etanol bunga telang (Clitoria ternatea L.) pada tikus putih yang diinduksi parasetamol. Jurnal Ilmiah Manuntung. 2022; 8(1):100-105.
-
[9] Zain DN, Pebiansyah A, Aprilia Y. Aktivitas nefroprotektif ekstrak etanol bunga telang (Clitoria ternatea L.) terhadap tikus yang diinduksi paracetamol. Pharmacoscript. 2021; 4(2):173-180.
-
[10] Dewi S, Astuti KI, Rusida ER. Uji toksisitas akut ekstrak etanol 70% bunga telang (Clitoria ternatea L.) terhadap tikus betina galur wistar dengan metode OECD 425. Jurnal Ilmiah Farmasi Farmasyifa 2023; 6(1): 60–66. https://doi.org/10.29313/jiff.v6i1.10420
-
[11] Khatib A, Tofrizal, Arsanty D. Toxicity effect of Clitoria ternatea l. extract in liver and kidney histopathological examination in mus musculus. IIUM Med J Malaysia. 2024; 23(1). https://doi.org/10.31436/imjm.v23i01.2318
-
[12] De Wet H, Ngubane SC. Traditional Herbal remedies used by women in a rural community in Northern Maputaland (South Africa) for the treatment of gynaecology and obstetric complaints. South Afr J Bot. 2014; 94: 129–139. https://doi.org/10.1016/j.sajb.2014.06.009
-
[13] Treuting PM, Dintzis SM, Montine KS. Comparative anatomy and histology a mouse, rat, and human atlas, second ed., Academic Press, United States 2018.
-
[14] Indrianingsih AW, Wulanjati MP, Windarsih A, Bhattacharjya DK, Suzuki T, Katayama T. In vitro studies of antioxidant, antidiabetic, and antibacterial activities of Theobroma cacao, Anonna muricata and Clitoria ternatea. Biocatal Agric Biotechnol 2021;33:101995. https://doi.org/10.1016/j.bcab.2021.101995.
-
[15] Sopyan MAP. Undergraduated Thesis. Aktivitas antibakteri staphylococcus aureus pada produk fermentasi teh kombucha bunga telang (Clitoria ternatea L.). Department of Biology, Faculty of Biology, Gadjah Mada University, Yogyakarta, Indonesia, 2024.
-
[16] Baker HJ, Lindsey R, Wesibroth SH. The Laboratory Rat: Biology and Diseases. vol. 1. New York: Academic Press; 1979.
-
[17] Taek AY, Ndaong NA, Gaina CA. Gambaran histopatologi hepar tikus putih (Rattus norvegicus) jantan pasca pemberian ekstrak ınfusa buah pare (Momordica charantica L.) lokal NTT. Journal Veteriner Nusantara 2020; 3(2): 89–96.
-
[18] Haerani H, Sijid StA, Zulkarnain Z. Pengaruh pemberian cuka aren terhadap kadar gula darah dan histopatologi pankreas mencit (Mus musculus) ICR jantan. Teknosains: Media Informasi Sains Dan Teknologi. 2023; 17(2): 210–219. https://doi.org/10.24252/teknosains.v17i2.36424
-
[19] Ringgi TN, Sitompul YY, Amalo FA, Winarso A. Gambaran patologi anatomi dan histopatologi organ visceral kambing kacang (Capra aegagrus hircus) yang terinfeksi sistiserkus. Jurnal Veteriner Nusantara. 2023; 6(2): 293–305. https://doi.org/10.35508/jvn.v6i2.8538
-
[20] Sari W, Okavia IW, Ceriana R, Sunarti S. Struktur mikroskopis hati ıkan seurukan (Osteochilus vittatus) dari Sungai Krueng Sabee Kabupaten Aceh Jaya yang tercemar limbah penggilingan bijih emas. BIOTIK: Jurnal Ilmiah Biologi Teknologi dan Kependidikan 2017; 4(1) :33-40. https://doi.org/10.22373/biotik.v4i1.1068
-
[21] Simangunsong MVE, Febriani Y, Saputri M, Arisa D, Zarqa Afifah G. Effectiveness of butterfly pea ethanol extract on decreasing blood glucose levels of male mice. Jambura J Health Sci Res. 2023; 5(2): 707-721. https://doi.org/10.35971/jjhsr.v5i2.18789
-
[22] Waruwu IS, Rawar EA, Kristiyani A. Penetapan kadar flavonoid total dan fenolik total serta uji penghambatan denaturasi protein dalam seduhan teh bunga telang (Clitoria ternatea L.). Majalah Farmasi dan Farmakologi. 2023; 27(2): 47–51. https://doi.org/10.20956/mff.v27i2.26250
-
[23] Rahayu S, Vifta R, Susilo J. Uji aktivitas antioksidan ekstrak etanol bunga telang (Clitoria ternatea L.) dari Kabupaten Lombok Utara dan Wonosobo menggunakan metode FRAP. Generics: J Res Pharm. 2021; 1(2): 1–9. https://doi.org/10.14710/genres.v1i2.9836
-
[24] Sotler R. Prooxidant Activities of antioxidants and their ımpact on health. Acta Clin Croat. 2019; 58(4): 726-736. https://doi.org/10.20471/acc.2019.58.04.20
-
[25] Prochazkova D, Bousova I, Wilhelmova N. Antioxidant and prooxidant properties of flavonoids. Fitoterapia. 2011; 82: 513–523. https://doi.org/10.1016/j.fitote.2011.01.018
-
[26] Purwaningsih S, Handharyan E, Lestari IR. Pengujian toksisitas sub akut ekstrak hipokotil bakau hitam pada tikus galur sprague dawley. Jurnal Akutika. 2015; 6(1): 30–40.
-
[27] Neuman MG, Cohen L, Opris M, Nanau RM, Jeong H. Hepatotoxicity of pyrrolizidine alkaloids. J Pharm Pharm Sci. 2015; 18(4): 825-843. https://doi.org/10.18433/J3BG7J
-
[28] Atere TG, Ajao AT. Toxicological implications of crude alkaloidal fraction from Cnestis ferruginea D.C root on liver function indices of male Wistar rats. Int J Biomed Health Sci. 2009;5:145–156.
-
[29] Qin Y, Wu X, Huang W, Gong G, Li D, He Y, Zhao Y. Acute toxicity and sub-chronic toxicity of steroidal saponins from Dioscorea zingiberensis C.H.Wright in rodents. J Ethnopharmacol. 2009; 126 (3): 543–550. https://doi.org/10.1016/j.jep.2009.08.047
-
[30] Chung KT, Wong TY, Wei C-I, Huang Y-W, Lin Y. Tannins and human health: A Review. Crit Rev Food Sci Nutr. 1998; 38(6): 421–464. https://doi.org/10.1080/10408699891274273
-
[31] Nourshargh S and Alon R. Leukocyte migration into flamed tissue. Immunity. 2014; 41(5):694-707. https://doi.org/10.1016/j.immuni.2014.10.008
-
[32] Jutila MA 5.14 Leukocyte trafficking. In: Comprehensive toxicology (Second edition). Elsevier, Oxford, 2010, pp.271-284. https://doi.org/10.1016/B978-0-08-046884-6.00615-1
-
[33] Schelling JR. Tubular atrophy in the pathogenesis of chronic kidney disease progression. Pediatr Nephrol. 2016; 31(5): 693-706. https://doi.org/10.1007/s00467-015-3169-4
-
[34] Kanagasundaram N. Pathophysiology of acute kidney injury. Ann Clin Biochem. 2015; 52(2): 193-205. https://doi.org/10.1177/0004563214556820
-
[35] Rodríguez MAL, Abraham SMV, Vázquez VMY, de León Garza B. Histopathological findings in renal biopsies in Anderson–Fabry disease. Case series. Revista Médica Del Hospital General de México. 2018; 81 (4): 243–247. https://doi.org/10.1016/j.hgmx.2016.08.010
-
[36] Kiss N, Hamar P. Histopathological Evaluation of Contrast-Induced Acute Kidney Injury Rodent Models. Biomed Res Int. 2016; 2016: 1–15. https://doi.org/10.1155/2016/3763250
-
[37] Suzuki N, Kojima K, Malvica S, Yamasaki K, Chikamatsu Y, Oe Y, Nagasawa T, Kondo E, Sanada S, Aiba S, Sato H, Miyazaki M, Ito S, Sato M, Tanaka T, Kinoshita K, Asano Y, Rosenberg AZ, Okamoto K, Shido K. Deep learning-based histopathological assessment of tubulo-interstitial injury in chronic kidney diseases. Commun Med. 2025; 5(1): 1-15. https://doi.org/10.1038/s43856-024-00708-3
-
[38] Dikkala PK, Kakarlapudi J, Rokalla P, Vedantam SK, Kaur A, Kaur K, Sharma M, Sridhar K. Computational screening of phytochemicals for anti-diabetic drug discovery. InPhytochemistry, Computational Tools and Databases in Drug Discovery 2023 Jan 1 (pp. 285-311). Elsevier. https://doi.org/10.1016/B978-0-323-90593-0.00009-5
-
[39] Shi J, Arunasalam K, Yeung D, Kakuda Y, Mittal G, Jiang Y. Saponins from edible legumes: chemistry, processing, and health benefits. J Med Food. 2004; 7(1): 67–78. https://doi.org/10.1089/109662004322984734
-
[40] Diwan FH, Abdel-Hassan A, Mohammed ST. Effect of saponin on mortality and histopathological changes in mice. East Mediterr Health J. 2000; 6: 345–351.
-
[41] Brandao-Costa R, Batistaa J, Nascimento T, Porto A. Renal function effects of FDS, a saponin ısolated from Filicium decipiens seeds. Biochemical and Histopathological Studies. J Plant Sci Phytopathol. 2019; 3: 107–110. https://doi.org/10.29328/journal.jpsp.1001040
-
[42] Karta J, Pandjaitan M, Rahminiwati M. Evaluation of acute oral toxicity of butterfly pea root extract on experimental mice. 3rd International Conference on Instrumentation, Communications, Information Technology, and Biomedical Engineering (ICICI-BME), 2013. 317–323. http://dx.doi.org/10.1109/ICICI-BME.2013.6698516.
-
[43] Seely JC. A Brief review of kidney dvelopment, maturation, developmental abnormalities, and drug toxicity: juvenile animal relevancy. J Toxicol Pathol. 2017; 30(2): 125-133. https://doi.org/10.1293/tox.2017-0006.
-
[44] Djohari M, Efendi TT, Rahmawati N, Husnawati H, Aryani F. Uji toksisitas akut dan tertunda fraksi butanol daun akar kaik-kaik (Uncaria cordata (Lour) Merr.) pada mencit putih (Mus musculus L.). Jurnal Ilmiah Ibnu Sina (JIIS). 2022; 7(1): 203–211. https://doi.org/10.36387/jiis.v7i1.874
-
[45] Nurma S, Safika S, Hamdika YP, Andriyanto A. Toksisitas akut ınfusa daun pepaya calina jantan (Carica papaya L.) pada mencit betina. Curr Biomed. 2023; 1(1): 46–53. https://doi.org/10.29244/currbiomed.1.1.46-53
-
[46] Mandal R, Loeffler AG, Salamat S, Fritsch MK. Organ weight changes associated with body mass index determined from a medical autopsy population. Am J Forens Med Pathol. 2012;33(4):382-389. https://doi.org/10.1097/paf.0b013e3182518e5f
-
[47] Yu Q, Yu F, Li Q, Zhang J, Peng Y, Wang X, Li T, Yin N, Sun G, Ouyang H, Chen Y, Mine Y, Tsao R, Zhang H. Anthocyanin-rich butterfly pea flower extract ameliorating low-grade inflammation in a high-fat-diet and lipopolysaccharide-induced mouse model. J Agric Food Chem. 2023;71: 11941–11956. https://doi.org/10.1021/acs.jafc.3c02696
-
[48] Wang Y, Liu T, Xie Y, Li N, Liu Y, Wen J, Zhang M, Feng W, Huang J, Guo Y, Kabbas Junior T, Wang D, Granato D. Clitoria ternatea blue petal extract protects against obesity, oxidative stress, and inflammation induced by a high-fat, high-fructose diet in C57BL/6 mice. Food Res Int. 2022;162: 112008. https://doi.org/10.1016/j.foodres.2022.112008
-
[49] Purwanto UMS, Aprilia K, Sulistiyani. Antioxidant activity of telang (Clitoria ternatea L.) Extract in ınhibiting lipid peroxidation. Curr Biochem. 2022; 9(9): 26–37.
-
[50] Arsianti A, Pasha Mahindra S, Nur Azizah N, Megawati Fajrin A, Dameria Nadapdap L. Phytochemical analysis, antioxidant and anticancer effects of clitoria ternatae extract on breast T47D cancer cells. Indones J Med Chem Bioinformatics. 2022; 1(1):3. https://doi.org/10.7454/ijmcb.v1i1.1003
-
[51] Fang J, Zeng L, He Y, Liu X, Zhang T, Wang Q. Effects of dietary tannic acid on obesity and gut microbiota in C57BL/6J mice fed with high-fat diet. Foods. 2022; 11(21): 1-15. https://doi.org/10.3390/foods11213325
-
[52] Manzoor F, Nisa MU, Hussain HA, Khan MK, Ahmad RS, Ahmad N, Imran M, Umbreen H. Effect of hydrolysable tannin on nutrient ıntake obesity and other associated metabolic risk factors in polycystic rats. Transl Med Commun. 2021; 6: 1-10. https://doi.org/10.1186/s41231-021-00089-y
-
[53] Kusumawati D, Puspitasari R. Evaluasi sifat fisik sediaan sabun cair ekstrak bunga telang (Clitoria ternatea). Indonesia J Med Pharm Sci. 2023; 2(2): 48–53. https://doi.org/10.30659/ijmps.v2i2.147
-
[54] Surasa NJ, Utami NR, Isnaeni W. Struktur mikroanatomi hati dan kadar kolesterol total plasma darah tikus putih strain wistar pasca suplementasi minyak lemuru dan minyak sawit. Biosaintifika: J Biol Biol Educ. 2014; 6(2): 141-151. https://doi.org/10.15294/biosaintifika.v6i2.3778
-
[55] Merlani V, Febrina D, Sunarti. Studi toksisitas akut ekstrak etanol bunga telang (Clitoria ternatea L.) pada tikus putih. Jurnal Ilmiah Farmasi Terapan & Kesehatan 2024;2:17–27.
-
[56] Yuliana F. Undergraduated Thesis. Toksisitas akut ekstrak air bunga telang (Clitoria ternatea) pada tikus betina. Department of Pharmacy, Faculty of Mathematics and Natural Science, Pakuan University, Bogor, Jawa Barat, Indonesia, 2023.
-
[57] Indonesian Food and Drug Authority. Acuan sediaan herbal. vol 5. 1st ed. Indonesia: Badan POM Ri;2010.
-
[58] Bryda EC. The mighty mouse: the impact of rodents on advances in biomedical research. Missouri Med. 2013; 110:207–211.
-
[59] Gad SC. Rodents model for toxicity testing and biomarkers. Biomarkers in Toxicology, Elsevier; 2014, p. 7–69. https://doi.org/10.1016/B978-0-12-404630-6.00002-6.
-
[60] Arifin W and Zahiruddin W. Sample size calculation in animal studies using resource equation approach. Malay J Med Sci. 2017; 24(5):101-105. https://doi.org/10.21315/mjms2017.24.5.11
-
[61] Indonesian Food and Drug Authority. Pedoman Uji Toksisitas Praklinik secara in vivo. Indonesia: Badan POM RI; 2022.
-
[62] Cook DJ. Cellular pathology. Oxford: Reed Educational and Profesional Publishing Ltd.; 2003.
-
[63] Firdiana VA, Gunanegara RF, Sunarti, Nuriliani A. Bambara groundnut ameliorates kidney histology in female mice with protein deficiency. Med J Indonesia. 2023;32(3): 161–167. https://doi.org/10.13181/mji.oa.237030
-
[64] Hammad FT, Al-Salam S, Hammad WF, Yasin J, Lubbad L. Despite initial recovery of GFR, long-term renal functions deteriorate following short periods of unilateral ureteral obstruction. Am J Physiol Renal Physiol. 2020;319: 523-533. https://doi.org/10.1152/ajprenal.00096.2020