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Investigation of Nutraceutical Potential, in vitro Antioxidant and Free Radical Scavenging Activity of Indian Royal Jelly

Yıl 2023, Cilt: 27 Sayı: 3, 1289 - 1300, 28.06.2025

Rahul Dubey Lohidasan Sathiyanarayanan Laxmi Rao Sandeep Sankaran

Öz

Apis mellifera, a priceless bee species is known to produce various nutritional products. Royal Jelly (RJ) is one such bee product which has high nutritive value, functional and biological qualities. However factors such as bee species, environment, season, collection technique and larval age affect the composition of RJ at macro and micro levels thus there is a need to evaluate Indian Royal Jelly (IRJ) for its chemical properties. So in the present work IRJ samples collected from Southern, Central and Northern regions of India were evaluated for physicochemical parameters (nature, color, appearance, odor, and solubility,) residual content (moisture content, ash value and pesticide content), chemical attributes (total polyphenol content, total flavonoid content,), nutraceutical potential (total fat, protein, carbohydrate content, and energy), and antioxidant activity (DPPH assay, ABTS assay and reducing power,). The total flavonoid content and total phenolic content were in the range of 0.119-0.321 mg quercetin/g of IRJ and 25.2844-68.203 mg Gallic acid/g, respectively. High energy, protein and low fat value suggested IRJ as a suitable nutraceutical agent. Antioxidant activity (IC50) of IRJ samples was found to be in the order of IRJ II > IRJ IV > IRJ I > IRJ III. Overall, it was observed that the IRJ-II showed better nutritional efficacy, polyphenolic content, and antioxidant properties.

Anahtar Kelimeler

Indian Royal Jelly physicochemical nutraceuticals phenolic flavonoid antioxidant

Kaynakça

  • [1] Halliwell B, Gutteridge JMC. The antioxidant of human extracellular fluids. Arch Biochem Biophys. 1990;280: 1– 8. https://doi.org/10.1016/0003-9861(90)90510-6
  • [2] Nagai T, Sakai M, Inoue R, Inoue H, Suzuki N. Antioxidative activities of some commercially honeys, royal jelly, and propolis. Food Chem.2001; 75(2): 237–240. https://doi.org/10.1016/S0308-8146(01)00193-5.
  • [3] Ramadan MF, Al-Ghamdi A. Bioactive compounds and health-promoting properties of royal jelly: A review. J Funct Foods. 2012; 4(1):39-52. https://doi.org/10.1016/j.jff.2011.12.007.
  • [4] Ramanathan ANKG, Nair AJ, Sugunan VS. A review on royal jelly proteins and peptides. J Funct Foods. 2018; 44:255-264. https://doi.org/10.1016/j.jff.2018.03.008.
  • [5] Pavel CI, Mărghitaş LA, Bobiş O, Dezmirean DS, Şapcaliu A, Radoi I, Mădaş MN. Biological activities of royal jelly-review. Scient Pap Anim Sci Biotechnol. 2011; 44(2):108-118.
  • [6] Khazaei M, Ansarian A, Ghanbari E. New findings on biological actions and clinical applications of royal jelly: a review. J Diet Suppl. 2018; 15(5):757-775. https://doi.org/10.1080/19390211.2017.1363843
  • [7] Ahmad S, Campos MG, Fratini F, Altaye SZ, Li J. New insights into the biological and pharmaceutical properties of royal jelly. Int J Mol Sci. 2020; 21(2):382. https://doi.org/10.3390/ijms21020382.
  • [8] Sabatini A, Marcazzan GM, Caboni MF, Bogdanov S, de Almeida-Muradian LB. Quality and standardisation of royal jelly. J ApiProd ApiMed Sci 20091;(1): 1-6. http://dx.doi.org/10.3896/IBRA.4.01.1.04.
  • [9] Collazo N, Carpena M, Nuñez-Estevez B, Otero P, Simal-Gandara J, Prieto MA. Health promoting properties of bee royal jelly: Food of the queens. Nutrients. 2021; 13(2):543. https://doi.org/10.3390%2Fnu13020543.
  • [10] Melliou E, Chinou I. Chemistry and bioactivities of royal jelly. Stud Nat Prod Chem. 2014; 43:261-290. http://dx.doi.org/10.1016/B978-0-444-63430-6.00008-4.
  • [11] Bogdanov S. Royal jelly, bee brood: composition, health, medicine: a review. J Bee Prod Sci. 2011; 3(8):8-19.
  • [12] Vazhacharickal PJ. A review on health benefits and biological action of honey, propolis and royal jelly. J Med Plant. 2021; 9(5):01-13.
  • [13] Ajitha Nath KG, Adithya Krishna SB, Smrithi S, Jayakumaran Nair A, Sugunan VS. Biochemical characterization and biological evaluation of royal jelly from Apis cerana. J Food Tech Food Chem. 2019; 2:101.
  • [14] Hu FL, Bíliková K, Casabianca H, Daniele G, Espindola FS, Feng M, Guan C, Han B, Kraková TK, Li JK, Li L, Li XA, Simuth J, Wu LM, Wu YQ, Xue XF, Xue YB, Yamaguchi K, Zeng ZJ, Zheng HQ, Zhou JH. Standard methods for Apis mellifera royal jelly research. J Apicult Res. 2019; 58(2):1-68. https://doi.org/10.1080/00218839.2017.1286003.
  • [15] Balkanska R. Correlations of physicochemical parameters, antioxidant activity and total polyphenol content of fresh royal jelly samples. Int J Curr Microbiol Appl Sci. 2018; 7(4):3744-3750. https://doi.org/10.20546/ijcmas.2018.704.421.
  • [16] Nagai T, Inoue R, Suzuki N, Nagashima T. Antioxidant properties of enzymatic hydrolysates from royal jelly. J Med Food. 2006; 9(3):363-367. https://doi.org/10.1089/jmf.2006.9.363.
  • [17] Rajasekaran A, Sivagnanam G, Xavier R. Nutraceuticals as therapeutic agents: a review. Res J Pharm Tech. 2008; 1(4): 328–340.
  • [18] Kocot J, Kiełczykowska M, Luchowska-Kocot D, Kurzepa J, Musik I. Antioxidant potential of propolis, bee pollen, and royal jelly: Possible medical application. Oxid Med Cell Longev. 2018; 7074209. https://doi.org/10.1155%2F2018%2F7074209.
  • [19] Kanelis D, Tananaki C, Liolios V, Dimou M, Goras G, Rodopoulou MA, Karazafiris E, Thrasyvoulou A. A suggestion for royal jelly specifications. Arh Hig Rada Toksikol. 2015; 66(4): 275–284. https://doi.org/10.1515/aiht-2015-66-2651.
  • [20] Kolayli S, Sahin H, Can Z, Yildiz O, Malkoc M, Asadov A. A member of complementary medicinal food: anatolian royal jellies, their chemical compositions, and antioxidant properties. J Evid Based Complementary Altern Med. 2016; 21(4):NP43–48. https://doi.org/10.1177/2156587215618832.
  • [21] Oršolić N. Royal jelly: component efficiency, analysis, and standardisation. Arh Hig Rada Toksikol. 2013; 64(3): 445–461. https://doi.org/10.2478/10004-1254-64-2013-2332.
  • [22] Wytrychowski M, Daniele G, Casabianca H. Combination of sugar analysis and stable isotope ratio mass spectrometry to detect the use of artificial sugars in royal jelly production. Anal Bioanal Chem. 2012; 403(5): 1451–1456. https://doi.org/10.1007/s00216-012-5934-6.
  • [23] Liu JR, Yang YC, Shi LS, Peng CC. Antioxidant properties of royal jelly associated with larval age and time of harvest. J Agric Food Chem. 2008; 56(23): 11447–11452. https://doi.org/10.1021/jf802494e.
  • [24] El-Guendouz S, Machado AM, Aazza S, Lyoussi B, Miguel MG, Mateus MC, Figueiredo AC. Chemical characterization and biological properties of royal jelly samples from the Mediterranean area. Nat Prod Commun. 2020; 15(2): 1–13. https://doi.org/10.1177/1934578X20908080.
  • [25] Park MJ, Kim BY, Deng Y, Park HG, Choi YS, Lee KS, Jin BR. Antioxidant capacity of major royal jelly proteins of honeybee (Apis mellifera) royal jelly. J Asia-Pac Entomol 2020; 23(2):445-448. https://doi.org/10.1016/j.aspen.2020.03.007.
  • [26] Hindi NKK, Al-Charrakh AH, Naher HS, Abbas AS. Study of chemical analysis of Iraqi propolis and active component of propolis. J Sci. 2015; 5(11): 1095–1103.
  • [27] Pavel CI, Mărghitaş LA, Dezmirean DS, Tomoş LI, Bonta V, Şapcaliu A, Buttstedt A. Comparison between local and commercial royal jelly—use of antioxidant activity and 10-hydroxy-2-decenoic acid as quality parameter. J Apicult Res. 2014; 53(1):116-123. https://doi.org/10.3896/IBRA.1.53.1.12.
  • [28] Liming W, Jinhui Z, Xiaofeng X, Yi L, Jing Z. Fast determination of 26 amino acids and their content changes in royal jelly during storage using ultra-performance liquid chromatography. J Food Comp Anal. 2009; 22(3):242- 249. https://doi.org/10.1016/j.jfca.2008.10.022.
  • [29] Kokotou MG, Mantzourani C, Babaiti R, Kokotos G. Study of the royal jelly free fatty acids by liquid chromatography-high resolution mass spectrometry (LC-HRMS). Metabolites. 2020; 10(1):40. https://doi.org/10.3390/metabo10010040.
  • [30] Duong HA, Vu MT, Nguyen TD, Nguyen MH, Mai TD. Determination of 10-hydroxy-2-decenoic acid and free amino acids in royal jelly supplements with purpose-made capillary electrophoresis coupled with contactless conductivity detection. J Food Comp Anal. 2020; 87:103422. https://doi.org/10.1016/j.jfca.2020.103422.
  • [31] Matsui M. Analysis of 10-hydroxydecenoic acid in commercial royal jelly products by gas chromatography. Food Hyg Safe Sci. 1986; 27(3):212-217. https://doi.org/10.3358/shokueishi.27.212.
  • [32] Qi D, Ma C, Wang W, Zhang L, Hao J, Li J. Gas chromatography-mass spectrometry analysis reveals the differences in volatile components of royal jelly from different honeybee stocks. LWT. 2020; 124:109143. https://doi.org/10.1016/j.lwt.2020.109143.
  • [33] Sesta G. Determination of sugars in royal jelly by HPLC. Apidologie. 2006; 37(1):84-90. https://doi.org/10.1051/apido:2005061.
  • [34] Xue XF, Zhou JH, Wu LM, Fu LH, Zhao J. HPLC determination of adenosine in royal jelly. Food Chem. 2009; 115(2):715-719. https://doi.org/10.1016/j.foodchem.2008.12.003.
  • [35] Ciulu M, Floris I, Nurchi VM, Panzanelli A, Pilo MI, Spano N, Sanna G. HPLC determination of pantothenic acid in royal jelly. Anal Method. 2013; 5(23):6682-6685. https://doi.org/10.1039/C3AY41284A.
  • [36] Guo H, Ekusa A, Iwai K, Yonekura M, Takahata Y, Morimatsu F. Royal jelly peptides inhibit lipid peroxidation in vitro and in vivo. J Nutr Sci Vitaminol. 2008; 54(3):191-195. https://doi.org/10.3177/jnsv.54.191.
  • [37] Jamnik P, Goranovič D, Raspor P. Antioxidative action of royal jelly in the yeast cell. Exp Gerontol. 2007; 42(7):594-600. https://doi.org/10.1016/j.exger.2007.02.002. This is an open access article which is publicly available on our journal’s website under Institutional

Yıl 2023, Cilt: 27 Sayı: 3, 1289 - 1300, 28.06.2025

Rahul Dubey Lohidasan Sathiyanarayanan Laxmi Rao Sandeep Sankaran

Öz

Kaynakça

  • [1] Halliwell B, Gutteridge JMC. The antioxidant of human extracellular fluids. Arch Biochem Biophys. 1990;280: 1– 8. https://doi.org/10.1016/0003-9861(90)90510-6
  • [2] Nagai T, Sakai M, Inoue R, Inoue H, Suzuki N. Antioxidative activities of some commercially honeys, royal jelly, and propolis. Food Chem.2001; 75(2): 237–240. https://doi.org/10.1016/S0308-8146(01)00193-5.
  • [3] Ramadan MF, Al-Ghamdi A. Bioactive compounds and health-promoting properties of royal jelly: A review. J Funct Foods. 2012; 4(1):39-52. https://doi.org/10.1016/j.jff.2011.12.007.
  • [4] Ramanathan ANKG, Nair AJ, Sugunan VS. A review on royal jelly proteins and peptides. J Funct Foods. 2018; 44:255-264. https://doi.org/10.1016/j.jff.2018.03.008.
  • [5] Pavel CI, Mărghitaş LA, Bobiş O, Dezmirean DS, Şapcaliu A, Radoi I, Mădaş MN. Biological activities of royal jelly-review. Scient Pap Anim Sci Biotechnol. 2011; 44(2):108-118.
  • [6] Khazaei M, Ansarian A, Ghanbari E. New findings on biological actions and clinical applications of royal jelly: a review. J Diet Suppl. 2018; 15(5):757-775. https://doi.org/10.1080/19390211.2017.1363843
  • [7] Ahmad S, Campos MG, Fratini F, Altaye SZ, Li J. New insights into the biological and pharmaceutical properties of royal jelly. Int J Mol Sci. 2020; 21(2):382. https://doi.org/10.3390/ijms21020382.
  • [8] Sabatini A, Marcazzan GM, Caboni MF, Bogdanov S, de Almeida-Muradian LB. Quality and standardisation of royal jelly. J ApiProd ApiMed Sci 20091;(1): 1-6. http://dx.doi.org/10.3896/IBRA.4.01.1.04.
  • [9] Collazo N, Carpena M, Nuñez-Estevez B, Otero P, Simal-Gandara J, Prieto MA. Health promoting properties of bee royal jelly: Food of the queens. Nutrients. 2021; 13(2):543. https://doi.org/10.3390%2Fnu13020543.
  • [10] Melliou E, Chinou I. Chemistry and bioactivities of royal jelly. Stud Nat Prod Chem. 2014; 43:261-290. http://dx.doi.org/10.1016/B978-0-444-63430-6.00008-4.
  • [11] Bogdanov S. Royal jelly, bee brood: composition, health, medicine: a review. J Bee Prod Sci. 2011; 3(8):8-19.
  • [12] Vazhacharickal PJ. A review on health benefits and biological action of honey, propolis and royal jelly. J Med Plant. 2021; 9(5):01-13.
  • [13] Ajitha Nath KG, Adithya Krishna SB, Smrithi S, Jayakumaran Nair A, Sugunan VS. Biochemical characterization and biological evaluation of royal jelly from Apis cerana. J Food Tech Food Chem. 2019; 2:101.
  • [14] Hu FL, Bíliková K, Casabianca H, Daniele G, Espindola FS, Feng M, Guan C, Han B, Kraková TK, Li JK, Li L, Li XA, Simuth J, Wu LM, Wu YQ, Xue XF, Xue YB, Yamaguchi K, Zeng ZJ, Zheng HQ, Zhou JH. Standard methods for Apis mellifera royal jelly research. J Apicult Res. 2019; 58(2):1-68. https://doi.org/10.1080/00218839.2017.1286003.
  • [15] Balkanska R. Correlations of physicochemical parameters, antioxidant activity and total polyphenol content of fresh royal jelly samples. Int J Curr Microbiol Appl Sci. 2018; 7(4):3744-3750. https://doi.org/10.20546/ijcmas.2018.704.421.
  • [16] Nagai T, Inoue R, Suzuki N, Nagashima T. Antioxidant properties of enzymatic hydrolysates from royal jelly. J Med Food. 2006; 9(3):363-367. https://doi.org/10.1089/jmf.2006.9.363.
  • [17] Rajasekaran A, Sivagnanam G, Xavier R. Nutraceuticals as therapeutic agents: a review. Res J Pharm Tech. 2008; 1(4): 328–340.
  • [18] Kocot J, Kiełczykowska M, Luchowska-Kocot D, Kurzepa J, Musik I. Antioxidant potential of propolis, bee pollen, and royal jelly: Possible medical application. Oxid Med Cell Longev. 2018; 7074209. https://doi.org/10.1155%2F2018%2F7074209.
  • [19] Kanelis D, Tananaki C, Liolios V, Dimou M, Goras G, Rodopoulou MA, Karazafiris E, Thrasyvoulou A. A suggestion for royal jelly specifications. Arh Hig Rada Toksikol. 2015; 66(4): 275–284. https://doi.org/10.1515/aiht-2015-66-2651.
  • [20] Kolayli S, Sahin H, Can Z, Yildiz O, Malkoc M, Asadov A. A member of complementary medicinal food: anatolian royal jellies, their chemical compositions, and antioxidant properties. J Evid Based Complementary Altern Med. 2016; 21(4):NP43–48. https://doi.org/10.1177/2156587215618832.
  • [21] Oršolić N. Royal jelly: component efficiency, analysis, and standardisation. Arh Hig Rada Toksikol. 2013; 64(3): 445–461. https://doi.org/10.2478/10004-1254-64-2013-2332.
  • [22] Wytrychowski M, Daniele G, Casabianca H. Combination of sugar analysis and stable isotope ratio mass spectrometry to detect the use of artificial sugars in royal jelly production. Anal Bioanal Chem. 2012; 403(5): 1451–1456. https://doi.org/10.1007/s00216-012-5934-6.
  • [23] Liu JR, Yang YC, Shi LS, Peng CC. Antioxidant properties of royal jelly associated with larval age and time of harvest. J Agric Food Chem. 2008; 56(23): 11447–11452. https://doi.org/10.1021/jf802494e.
  • [24] El-Guendouz S, Machado AM, Aazza S, Lyoussi B, Miguel MG, Mateus MC, Figueiredo AC. Chemical characterization and biological properties of royal jelly samples from the Mediterranean area. Nat Prod Commun. 2020; 15(2): 1–13. https://doi.org/10.1177/1934578X20908080.
  • [25] Park MJ, Kim BY, Deng Y, Park HG, Choi YS, Lee KS, Jin BR. Antioxidant capacity of major royal jelly proteins of honeybee (Apis mellifera) royal jelly. J Asia-Pac Entomol 2020; 23(2):445-448. https://doi.org/10.1016/j.aspen.2020.03.007.
  • [26] Hindi NKK, Al-Charrakh AH, Naher HS, Abbas AS. Study of chemical analysis of Iraqi propolis and active component of propolis. J Sci. 2015; 5(11): 1095–1103.
  • [27] Pavel CI, Mărghitaş LA, Dezmirean DS, Tomoş LI, Bonta V, Şapcaliu A, Buttstedt A. Comparison between local and commercial royal jelly—use of antioxidant activity and 10-hydroxy-2-decenoic acid as quality parameter. J Apicult Res. 2014; 53(1):116-123. https://doi.org/10.3896/IBRA.1.53.1.12.
  • [28] Liming W, Jinhui Z, Xiaofeng X, Yi L, Jing Z. Fast determination of 26 amino acids and their content changes in royal jelly during storage using ultra-performance liquid chromatography. J Food Comp Anal. 2009; 22(3):242- 249. https://doi.org/10.1016/j.jfca.2008.10.022.
  • [29] Kokotou MG, Mantzourani C, Babaiti R, Kokotos G. Study of the royal jelly free fatty acids by liquid chromatography-high resolution mass spectrometry (LC-HRMS). Metabolites. 2020; 10(1):40. https://doi.org/10.3390/metabo10010040.
  • [30] Duong HA, Vu MT, Nguyen TD, Nguyen MH, Mai TD. Determination of 10-hydroxy-2-decenoic acid and free amino acids in royal jelly supplements with purpose-made capillary electrophoresis coupled with contactless conductivity detection. J Food Comp Anal. 2020; 87:103422. https://doi.org/10.1016/j.jfca.2020.103422.
  • [31] Matsui M. Analysis of 10-hydroxydecenoic acid in commercial royal jelly products by gas chromatography. Food Hyg Safe Sci. 1986; 27(3):212-217. https://doi.org/10.3358/shokueishi.27.212.
  • [32] Qi D, Ma C, Wang W, Zhang L, Hao J, Li J. Gas chromatography-mass spectrometry analysis reveals the differences in volatile components of royal jelly from different honeybee stocks. LWT. 2020; 124:109143. https://doi.org/10.1016/j.lwt.2020.109143.
  • [33] Sesta G. Determination of sugars in royal jelly by HPLC. Apidologie. 2006; 37(1):84-90. https://doi.org/10.1051/apido:2005061.
  • [34] Xue XF, Zhou JH, Wu LM, Fu LH, Zhao J. HPLC determination of adenosine in royal jelly. Food Chem. 2009; 115(2):715-719. https://doi.org/10.1016/j.foodchem.2008.12.003.
  • [35] Ciulu M, Floris I, Nurchi VM, Panzanelli A, Pilo MI, Spano N, Sanna G. HPLC determination of pantothenic acid in royal jelly. Anal Method. 2013; 5(23):6682-6685. https://doi.org/10.1039/C3AY41284A.
  • [36] Guo H, Ekusa A, Iwai K, Yonekura M, Takahata Y, Morimatsu F. Royal jelly peptides inhibit lipid peroxidation in vitro and in vivo. J Nutr Sci Vitaminol. 2008; 54(3):191-195. https://doi.org/10.3177/jnsv.54.191.
  • [37] Jamnik P, Goranovič D, Raspor P. Antioxidative action of royal jelly in the yeast cell. Exp Gerontol. 2007; 42(7):594-600. https://doi.org/10.1016/j.exger.2007.02.002. This is an open access article which is publicly available on our journal’s website under Institutional
Toplam 37 adet kaynakça vardır.

Ayrıntılar

Birincil Dil İngilizce
Konular Farmasotik Kimya
Bölüm Articles
Yazarlar

Rahul Dubey

Lohidasan Sathiyanarayanan 0000-0001-9892-4794

Laxmi Rao

Sandeep Sankaran 0000-0002-0064-3263

Yayımlanma Tarihi 28 Haziran 2025
Yayımlandığı Sayı Yıl 2023 Cilt: 27 Sayı: 3

Kaynak Göster

APA Dubey, R., Sathiyanarayanan, L., Rao, L., Sankaran, S. (2025). Investigation of Nutraceutical Potential, in vitro Antioxidant and Free Radical Scavenging Activity of Indian Royal Jelly. Journal of Research in Pharmacy, 27(3), 1289-1300.
AMA Dubey R, Sathiyanarayanan L, Rao L, Sankaran S. Investigation of Nutraceutical Potential, in vitro Antioxidant and Free Radical Scavenging Activity of Indian Royal Jelly. J. Res. Pharm. Haziran 2025;27(3):1289-1300.
Chicago Dubey, Rahul, Lohidasan Sathiyanarayanan, Laxmi Rao, ve Sandeep Sankaran. “Investigation of Nutraceutical Potential, in Vitro Antioxidant and Free Radical Scavenging Activity of Indian Royal Jelly”. Journal of Research in Pharmacy 27, sy. 3 (Haziran 2025): 1289-1300.
EndNote Dubey R, Sathiyanarayanan L, Rao L, Sankaran S (01 Haziran 2025) Investigation of Nutraceutical Potential, in vitro Antioxidant and Free Radical Scavenging Activity of Indian Royal Jelly. Journal of Research in Pharmacy 27 3 1289–1300.
IEEE R. Dubey, L. Sathiyanarayanan, L. Rao, ve S. Sankaran, “Investigation of Nutraceutical Potential, in vitro Antioxidant and Free Radical Scavenging Activity of Indian Royal Jelly”, J. Res. Pharm., c. 27, sy. 3, ss. 1289–1300, 2025.
ISNAD Dubey, Rahul vd. “Investigation of Nutraceutical Potential, in Vitro Antioxidant and Free Radical Scavenging Activity of Indian Royal Jelly”. Journal of Research in Pharmacy 27/3 (Haziran 2025), 1289-1300.
JAMA Dubey R, Sathiyanarayanan L, Rao L, Sankaran S. Investigation of Nutraceutical Potential, in vitro Antioxidant and Free Radical Scavenging Activity of Indian Royal Jelly. J. Res. Pharm. 2025;27:1289–1300.
MLA Dubey, Rahul vd. “Investigation of Nutraceutical Potential, in Vitro Antioxidant and Free Radical Scavenging Activity of Indian Royal Jelly”. Journal of Research in Pharmacy, c. 27, sy. 3, 2025, ss. 1289-00.
Vancouver Dubey R, Sathiyanarayanan L, Rao L, Sankaran S. Investigation of Nutraceutical Potential, in vitro Antioxidant and Free Radical Scavenging Activity of Indian Royal Jelly. J. Res. Pharm. 2025;27(3):1289-300.