[01] Koto-te-Nyiwa Ngbolua, Department of Biology, University of Kinshasa, Kinshasa, Democratic Republic of the Congo; Department of Biomedical Sciences, University of Gbadolite, Gbadolite, Democratic Republic of the Congo. [02] Benjamin Zoawe Gbolo, Department of Biology, University of Kinshasa, Kinshasa, Democratic Republic of the Congo. [03] Joseph Dipa Tshidibi, Department of Biology, University of Kinshasa, Kinshasa, Democratic Republic of the Congo. [04] Damien Sha Tshibey Tshibangu, Department of Chemistry, University of Kinshasa, Kinshasa, Democratic Republic of the Congo. [05] Patrick Bondo Memvanga, Faculty of Pharmaceutical Sciences, University of Kinshasa, Kinshasa, Democratic Republic of the Congo. [06] Pius Tshimankinda Mpiana, Department of Chemistry, University of Kinshasa, Kinshasa, Democratic Republic of the Congo.

Drepanoalpha^® is a poly-herbal formula (PHF) used in Democratic Republic of the Congo for the management of Sickle Cell Disease (SCD), a genetic disease due to the presence of hemoglobin S in the blood which could in hypoxia conduct to the formation of tactoids, the leading cause of erythrocyte sickling that plays a key role in the pathophysiology of SCD like vaso-occlusion (due to the loss of membrane elasticity) and hemolytic anemia. The aim of the present study was to evaluate the storage time effect on the bioactivity and the secondary metabolites (anthocyanis and flavonoids) and minerals (Fe, Mn, Zn and Cu) profiles of this PHF. Results revealed that flavonoids content of different batches is of the same order of magnitude: the percentage of ratio flavonoids/total polyphenols was 5.3% (batch A); 5.0% (batch B); 4.7% (batch C). However, the anthocyanins content of PHF varies according to the storage time. The percentage of ratio anthocyanis/total polyphenols was 3, 2% for batch A; 4, 7% for batch B; 8, 3% for batch C. Results revealed no significant difference between the average values of total ash (11.34 ± 0.35 84 g/100 g DM for A; 11.5 ± 0.12 84 g/100 g DM for B and 10 ± 0.84 84 g/100 g DM for C) of three batches (threshold: 5%). The different batches of Drepanoalpha^® contain Fe (0.535± 0.107 mg/100 g for A; 0.517 ± 0.103 mg/100 g for B; 0.54 ± 0.108 mg/100 g for C), Mn (4.366 ± 0.786 mg/100 g for A; 4.7 ± 0.799 mg/100 g for B; 4.318 ± 0.734 mg/100 g for C), and Zn (2.293 ± 0.321 mg/100 g for A; 2.389 ± 0.334 mg/100 g for B; 2.338 ± 0.327 mg/100 g for C) and are free of Cu. The micronutrient values of batches A, B and C are not significantly different (threshold: 5%) from each other. At 250 μg /mL, there is no significant difference (threshold: 5%) in anti-oxidative activity between the different batches. All samples displayed strong anti-sickling activity indicating thus at room temperature, the storage time (one or two years) has no effect on the bioactivity, flavonoids content and mineral profile of the PHF. It is desirable that Drepanoalpha^® could be formulated as syrup or capsule for a double blind, placebo-controlled and randomized cross-over clinical trials in SCD patients.

Sickle Cell Disease, Traditional Medicine, Medicinal Foods, Anti-sickling Activity, Polyphenols

[01] Girot R, Bégué P, Galacteros F. La drépanocytose. John Libbey Eurotext Paris, 2003. [02] Bardakjian MJ, Dhont JL, Du Crocq R. Bonnes pratiques de l’étude de l’hémoglobine Ann. Biol. Clin. 2003; 61: 401-09. [03] Mpiana PT, Ngbolua KN, Atibu EK, Kasonga TM, Bokota MT, Mudogo V. In vitro effects of anthocyanins extracts from Justicia secunda Vahl on the solubility of hemoglobin S and membrane stability of sickle erythrocytes. Blood Transfusion. 2010; 8 (4): 248-54. [04] Mpiana PT, Mudogo V, Tshibangu DST, Kitwa EK, Kanangila AB, Lumbu JBS, Ngbolua KN, Atibu EK, Kakule MK. Antisickling activity of anthocyanins from Bombax pentadrum, Ficus capensis and Ziziphus mucronata: Photodegradation effect. J. Ethnopharmacology. 2008; 120: 413-418. [05] Mpiana PT, Makelele LK, Oleko RW, Bokota MT, Tshibangu DST, Ngbolua KN, Mbala MB, Atibu EK, Nsimba SM. Antisickling activity of medicinal plants used in the management of Sickle cell Disease in Tshopo District, D. R. Congo, Australian Journal of Medical Herbalism. 2010; 22 (4): 132-137. [06] Ngbolua KN, Mpiana PT, Tshibangu DST, Gbolo BZ. Bioactivity of medicinal plants traditionally used for the management of sickle cell disease in Democratic Republic of the Congo: State of the art and future directions in: Sickle cell disease: Genetics, management and prognosis. New York. Nova Science Publishers, 2015. [07] De Montalembert M, Tshilolo L. Les progrès thérapeutiques dans la prise en charge.de la drépanocytose sont-ils applicables en Afrique subsaharienne. Med. Trop. 2007; 67: 612-616. [08] Ngbolua KN, Mpiana PT, Akoundze BJ, Mwanza BF, Tshibangu DST, Masengo AC, Liesse JM, Takaisi K. Anti-sickling and bacterial inhibitory effects of two medicinal foods from the Congo River basin: Gnetum africanum Welw. (Gnetaceae) and Grewia coriacea Mast. (Malvaceae). Current Traditional Medicine. 2016; 2 (1): 34-41. [09] Mpiana PT, Ngbolua KN, Tshibangu DST. Les Alicaments et la drepanocytose: Une mini revue. Comptes Rendus Chimie. 2016; 1 (6): 884-89. French. [10] Tshilanda DD, Onyamboko DNV, Babady BP, Mutwale KP, Tsalu VP, Tshibangu DST, Ngombe KN, Ngbolua KN, Mpiana PT. Chemical fingerprint and anti-sickling activity of Rosmarinic acid and Methanolic extracts from three Species of Ocimum from DR Congo. Journal of Biosciences and Medicines. 2016; 4: 59-68. http://dx.doi.org/10.4236/jbm.2016.41008 [11] Ngbolua KN, Rafatro H, Rakotoarimanana H, Mudogo V, Mpiana PT, Tshibangu DST, Tshilanda DD. In vitro anti-erythrocyte sickling effect of lunularic acid of natural origin. International Blood Research & Reviews 4 (3): 1-6, 2015. [12] Tshibangu DST, Shode FO, Koorbanally N, Mudogo V, Mpiana PT, Ngbolua KN. Antisickling triterpenoids from Callistemon viminalis, Meulaleuca bracteata var. Revolution Gold, Syzygium guineense and Syzygium cordatum. The 14th NAPRECA Symposium and AAMPS Ethnoverterinary Medicine Symposium 8th-12th August, 2011. International Centre for Insect Physiology and Ecology (ICIPE): Kasarani, Nairobi, Kenya, pp. 296-300 (YS 27). [13] Tshilanda DD, Onyamboko DNV, Babady PB, Ngbolua KN, Tshibangu DST, Dibwe EF, Mpiana PT. Anti-sickling Activity of Ursolic Acid Isolated from the Leaves of Ocimum gratissimum L. (Lamiaceae). Nat. Prod. Bioprospect. 2015; 5: 215-221. doi 10.1007/s13659-015-0070-6. [14] Ngbolua KN, Mpiana PT, Tshibangu DST, Mazasa PP, Gbolo BZ, Atibu EK, Kadima JN and Kasali FM. In vitro antisickling and radical scavenging activities of a poly-herbal formula (Drepanoalpha^®) in sickle cell eruthrocyte and acute toxicity study in Wistar albinos rats. European Journal of Medicinal Plants. 2014; 4 (10): 1251-67. [15] Ngbolua KN, Mpiana PT. The possible role of a congolese polyherbal formula (Drepanoalpha^®) as source of epigenetic modulators in sickle cell disease: A hypothesis. J. of Advancement in Medical and Life Sciences. 2014; 2 (1): 1-3. [16] Mpiana PT, Kasali FM, Bwirhonde F, Gbolo BZ, Tshibangu DST, Ngbolua KN, et al. Acute and sub-acute oral toxicity studies of Drepanoalpha^® (a poly-herbal formula used in the management of sickle cell disease) in guinea-pig. British Journal of Pharmaceutical Research. 2016; 10 (5): 1-8. [17] Ngbolua KN, Tshidibi JD, Tshibangu DST, Memvanga PB, Gbolo BZ, Tshilanda DD, Mpiana PT. Drepanoalpha^®: An Overview on the Quality Control Process and Standardization Feature of an Antisickling Herbal Drug from Democratic Republic of the Congo. J. of Modern Drug Discovery and Drug Delivery Research. 2016; V4I1. DOI: 10.15297/JMDDR.V4I1.01. [18] Gbolo BZ, Asamboa LS, Bongo GN, Tshibangu DST, Kasali FM, Memvanga PB, et al. Bioactivity and chemical analysis of Drepanoalpha: An anti-sickle cell anemia poly-herbal formula from Congo-Kinshasa American Journal of Phytomedicine and Clinical Theurapetics. 2017; 5 (1): 1-5. [19] Gbolo BZ, Tshibangu DST, Asamboa LT, Bongo NG, Kasali MF, Feza BV, Ngbolua KN, Mpiana PT. Sickle cell anemia therapeutic approach based on Drepanoalpha^®: About 34 cases. Journal of Complementary and Alternative Medical Research 2017; 4 (2): 1-8. [20] Ngbolua KN, Tshilanda DD, Djolu RD, Falanga MC, Masengo AC, Tshibangu DST, Iteku BJ, Mudogo V, Mpiana PT. Anti-Sickle Cell Anemia and Bacterial Inhibitory Effects of Uvariodendron molundense (Diels) R. E. Fr. (Annonaceae) from Ubangi River Basin, DR Congo. Journal of Biosciences and Medicines. 2017; 5: 71-84. [21] Tshibangu DST, Ngbolua KN, Lengbiye EM, Tshilanda DD, Mvingu KB, Iteku BJ, Mbala MB, Mudogo V, Mpiana PT. Chemical composition and bioactivity of Canarium schweinfurthii stem bark extracts from DR Congo against Sickle cell disease and associated bacteria. Journal of Pharmacognosy and Phytochemistry. 2016; 5 (4): 181-187. [22] Giusti MM, Wrolstad RE. Characterization and measurement of anthocyanins by UV-visible spectroscopy. In Current Protocols in Food Analytical Chemistry; Wrolstad, R. E., Ed.; John Wiley & Sons: New York, NY, USA, pp. 7–47, 2001. [23] Ruch RJ, Cheng S-J, Klaunig JE. Prevention of cytotoxicity and inhibition of intercellular communication by antioxidant catechins isolated from chinese green tea. Carcinogenesis. 1989; 10 (6): 1003-1008. [24] Prieto P, Pineda M, Aguillar M. Spectrophotometric quantitation of antioxidant capacity through the formation of phosphomolybdenum complex: specific application to determination of vitamin E. Anal Biochem. 1999; 269: 337-341. [25] Laurent L. Eléments minéraux. In: Techniques d’analyses et de contrôle dans les industries agroalimentaires. Volume 4. Lavoisier (Ed). Paris, 1999. [26] Mbemba F. Aliments et denrées alimentaires traditionnels du Bandundu en RD Congo: Répertoire et composition en nutriments, Ed L’Harmattan, Paris, 2013. [27] Prasad AS. Clinical, Immunological, Anti-inflammatory and Antioxidant roles of Zinc. Exp Gerentology. 2008; 45 (5): 370-377.