Optimization of Extraction and Determination of Carotenoids from Banana Pulp

doi:10.3969/j.issn.1000-2561.2022.09.023

Abstract

Abstract:

Large-scale evaluation of banana carotenoids germplasm resources, exploration of varieties with high carotenoid resources and improvement of carotenoid contents in main varieties are of great importance to enhance the competitiveness of banana industry. It is necessary to establish an efficient extraction. Meanwhile, it is as well important to grope a determination system for carotenoids in banana fruit pulp. Therefore, this paper primarily aims to establish an optimized protocol for extraction and determination of carotenoids from banana pulp. In this paper, Ultra Performance Liquid Chromatography (UPLC, Agilent1290) was used to determine carotenoids in banana pulp. By performing on a YMC C30 column at the time when column temperature maintaining at 20℃ and the Ultraviolet detection wavelength at 450 nm, we compared the effects of different ratios of solid-liquid (g:mL=1:6, 1:9, 1:12, 1:15), different constant volume solution (MTBE:methanol, V:V=1:0, 1:1, 1:2), different mobile phase (methanol:acetonitrile, V:V=4:0, 3:1, 2:2, 1:3, 0:4,), and different flow rates (0.6 mL/min, 0.8 mL/min, 1 mL/min, 1.2 mL/min) on the identification and content determination of carotene in banana fruit pulp. The results showed that when the extraction solution of hexane/acetone/anhydrous ethanol was 2:1:1 (V:V:V), the solid-liquid ratio of pulp to extraction reagent was 1:9 (g/mL), ultrasonic crushing was performed for 30 minutes, repeated 3 times and the ratio for constant volume solution methanol/MTBE was 1:1 (V:V), the separation effect of carotenoids component in banana fruit pulp was the best. Moreover, the main composition of carotenoids were efficiently separated and identified when we used the binary mobile phase consisting solvent A (methanol:acetonitrile=1:1) and solvent B (MTBE, 100%), under a linear gradient elution procedure, the injection volume of 10 μL and at the flow rate of 1 mL/min. The optimized system was used to detect the carotenoid contents of different kinds of banana fruit pulp. It was found that the main carotenoid compositions in banana fruit pulp were lutein, α-carotene, β-carotene, and so on. ‘French Sombre’ banana variety with high carotenoid contents contained about 9.79 μg/g, while ‘Zhong Jiao 8’ banana variety with low carotenoid contents was only about 0.201 μg/g. This also indicated that the carotenoid contents in different banana varieties was relatively large different. The method on extraction and determination of carotenoids from banana pulp which was established in this paper, is relatively simple and accurate,, providing an optimal protocol for researchers to study the function of carotenoids in bananas.

Key words: banana (Musa spp.), carotenoids, identification and determination of compositions, ultra performance liquid chromatography, system optimization

CLC Number:

S668.1

FAN Yingkang, LI Li, ZHAN Ni, KUANG Mengyu, DENG Guiming, LYU Peitao, SHENG Ou. Optimization of Extraction and Determination of Carotenoids from Banana Pulp[J]. Chinese Journal of Tropical Crops, 2022, 43(9): 1945-1952.

Figures/Tables 7

References 23

[1]	陶俊, 张上隆, 张良诚, 安新民, 刘春荣. 柑橘果皮颜色的形成与类胡萝卜素组分变化的关系[J]. 植物生理与分子生物学学报, 2003, 29(2): 121-126.
	TAO J, ZHANG S L, ZHANG L C, AN X M, LIU C R. Relationship between color formation and change in composition of carotenoids in peel of citrus[J]. Journal of Plant Physiology and Molecular Biology, 2003, 29(2): 121-126. (in Chinese)
[2]	BARTLEY G E, SCOLNIK P A. Plant carotenoids: pigments for photoprotection, visual attraction, and human health[J]. The Plant Cell, 1995, 7(7): 1027-1038.
[3]	DAVEY M W, MELLIDOU I, KEULEMANS W. Considerations to prevent the breakdown and loss of fruit carotenoids during extraction and analysis in Musa[J]. Journal of Chromatography A, 2009, 1216: 5759-5762. DOI URL
[4]	魏爱兰. 畜禽营养中维生素A的特性与作用[J]. 甘肃农业, 2012(15): 63-64.
	WEI A L. Characteristics and effect of vitamin A in animal nutrition[J]. Gansu Agriculture, 2012(15): 63-64. (in Chinese)
[5]	陈敏氡, 朱海生, 温庆放, 马宏棋, 林义章. UPLC测定草莓果实中类胡萝卜素含量[J]. 果树学报, 2013, 30(4): 706-711.
	CHEN M D, ZHU H S, WEN Q F, MA H Q, LIN Y Z. Determination of carotenoids in strawberry by UPLC[J]. Journal of Fruit Science, 2013, 30(4): 706-711. (in Chinese)
[6]	Robinson J C, SAUCO V G. Bananas and plantains[M]. Wallingford: CAB International, 1996, 2: 7-16.
[7]	衡周. 香蕉果实类胡萝卜素代谢分子机理的研究[D]. 长沙: 湖南农业大学, 2017.
	HENG Z. Study on molecular mechanism of carotenoid metabolism in banana fruit[D]. Changsha: Hunan Agricultural University, 2017. (in Chinese)
[8]	DAVEY M W, SAEYS W, HOF E, RAMON H, SWENNEN R L, KEULEMANS J. Application of visible and near-infrared reflectance spectroscopy (Vis/NIRS) to determine carotenoid contents in banana (Musa spp.) fruit pulp[J]. Journal of Agricultural and Food Chemistry, 2009, 57: 1742-1751. DOI URL
[9]	金思, 马空军. 超声波辅助提取类胡萝卜素研究进展[J]. 食品研究与开发, 2017, 38(9): 192-197.
	JIN S, MA K J. Research progress in ultrasound-assisted extraction of carotenoids[J]. Food Research and Development, 2017, 38(9): 192-197. (in Chinese)
[10]	毕可海, 孙玉奉, 张伶俐, 聂小伟, 张玉清. 海带中类胡萝卜素提取工艺条件[J]. 食品工业, 2020, 41(1): 30-34.
	BI K H, SUN Y F, ZHANG L L, NIE X W, ZHANG Y Q. Extraction process of carotenoids from kelp[J]. The Food Industry, 2020, 41(1): 30-34. (in Chinese)
[11]	徐佳佳, 崔亚娟, 姜菲菲, 李全霞, 李东, 王竹, 王国栋, 门建华. 几种常见的类胡萝卜素提取技术[J]. 食品研究与开发, 2014, 35(19): 141-144.
	XU J J, CUI Y J, JIANG F F, LI Q X, LI D, WANG Z, WANG G D, MEN J H. Study on the extraction of carotenoids[J]. Food Research and Development, 2014, 35(19): 141-144. (in Chinese)
[12]	王思元, 王惜童, 孙露川阳, 吴哲, 王帅, 任丹丹, 汪秋宽, 吴文忠. 类胡萝卜素稳态化技术的研究进展[J]. 大连海洋大学学报, 2019, 34(1): 145-152.
	WANG S Y, WANG X T, SUN-LU C Y, WU Z, WANG S, REN D D, WANG Q K, WU W Z. Research progress on carotenoid steady state technology: a review[J]. Journal of Dalian Ocean University, 2019, 34(1): 145-152. (in Chinese)
[13]	郝智勇, 邱广伟, 关丽辉, 王海艳. 马铃薯类胡萝卜素含量测定方法优化[C]// 马铃薯产业与精准扶贫2017. 哈尔滨: 哈尔滨地图出版社, 2017: 419-423.
	HAO Z Y, QIU G W, GUAN L H, WANG H Y. Optimization of method for determination of carotenoids from potato[C]// Potato industry and precision of poverty 2017. Haerbin:Haerbin Map Press House, 2017: 419-423. (in Chinese)
[14]	康迎春, 尹跃, 赵建华, 刘兰英. HPLC法测定枸杞鲜果中主要类胡萝卜素组成[J]. 食品工业, 2014, 35(12): 270-272.
	KANG Y C, YIN Y, ZHAO J H, LIU L Y. Determianation of main carotenoids in fruits of lycium barbarum by HPLC[J]. The Food Industry, 2014, 35(12): 270-272. (in Chinese)
[15]	莫玉楠, 张鲁刚, 王国芳. 橙色大白菜类胡萝卜素提取与测定方法研究[J]. 西北农林科技大学学报(自然科学版), 2014, 42(3): 206-214.
	MO Y N, ZHANG L G, WANG G F. Extraction and determination of total carotenoids in orange heading Chinese cabbages[J]. Journal of Northwest A & F University (Natural Science Edition), 2014, 42(3): 206-214. (in Chinese)
[16]	王岁楼, 谢雯丽. 红辣椒中类胡萝卜素提取方法及工艺条件优化研究[J]. 农产品加工, 2020(11): 39-48.
	WANG S L, XIE W L. Study on the optimization of extraction method and technological conditions of carotenoid in red pepper[J]. Farm Products Processing, 2020(11): 39-48. (in Chinese)
[17]	AMAH D, ALAMU E, ADESOKAN M, BILJON A V, MAZIYA-DIXON B, SWENNEN R, LABUSCHAGNE M. Variability of carotenoids in a Musa germplasm collection and implications for provitamin A biofortification[J]. Food Chemistry: X, 2019, 2: 100024. DOI URL
[18]	DAVEY M W, KEULEMANS J, SWENNEN R. Methods for the efficient quantification of fruit provitamin A contents[J]. Journal of Chromatography A, 2006, 1136: 176-184. PMID
[19]	严娟, 蔡志翔, 沈志军, 张斌斌, 马瑞娟, 俞明亮. 黄肉桃果实中类胡萝卜素提取和测定方法研究[J]. 果树学报, 2015, 32(6): 1267-1274.
	YAN J, CAI Z X, SHEN Z J, ZHANG B B, MA R J, YU M L. Extraction and analytical methods of carotenoids in fruit of yellow flesh peach[J]. Journal of Fruit Science, 2015, 32(6): 1267-1274. (in Chinese)
[20]	吴青莹, 宋莹莹, 邢思熙, 韩旭, 魏子璇, 王岁楼. 香蕉皮类胡萝卜素提取工艺条件的研究[J]. 中国食物与营养, 2016, 22(4): 32-34.
	WU Q Y, SONG Y Y, XING S X, HAN X, WEI Z X, WANG S L. Research on carotenoids extraction process from banana peel[J]. Food and Nutrition in China, 2016, 22(4): 32-34. (in Chinese)
[21]	胡丽松, 吴刚, 郝朝运, 范睿, 伍宝朵, 谭乐和. 菠萝蜜类胡萝卜素检测方法研究及呈色物质分析[J]. 热带作物学报, 2017, 38(5): 950-956.
	HU L S, WU G, HAO C Y, FAN R, WU B D, TAN L H. Method of carotenoids analysis and pigments identification in jackfruit[J]. Chinese Journal of Tropical Crops, 2017, 38(5): 950-956. (in Chinese)
[22]	FU X M, CHENG S H, LIAO Y Y, HUANG B Z, DU B, ZENG W, JIANG Y M, DUAN X W, YANG Z Y. Comparative analysis of pigments in red and yellow banana fruit[J]. Food Chemistry, 2018, 239: 1009-1018. DOI PMID
[23]	杜奋澄, 许杰纯, 杨雨欣, 吴智明. 辣椒果实类胡萝卜素的提取与测定方法研究[J]. 中国农学通报, 2019, 35(5): 41-48.
	DU F C, XU J C, YANG Y X, WU Z M. Extraction and measurement methods of carotenoids from pepper fruit[J]. Chinese Agricultural Science Bulletin, 2019, 35(5): 41-48. (in Chinese)

编号No.	类胡萝卜素 Carotenoids	R_t	λ_max	线性方程 Linear equations	线性范围 Linear range/(μg·mL^?1)	相关系数 Correlation index
1	新黄质neoxanthin	11.512	468 416	y = 0.0078x + 0.1042	3.125~50	R² = 0.9999
2	叶黄素lutein	19.192	450 478	y = 0.0048x + 0.0935	3.125~50	R² = 1
3	β-阿朴-8°-胡萝卜素醛 β -apo-8°-carotenal	23.801	466	y = 0.0048x + 0.0529	3.125~50	R²= 1
4	玉米黄质zeinoxanthin	27.359	450 478	y = 0.0051x + 0.0170	3.125~50	R² = 1
5	ε-胡萝卜素ε-carotence	31.662	420 444 474	y = 0.0047x + 0.0591	3.125~50	R² = 0.9999
6	13-顺式-β-胡萝卜素13-cis-β-carotence	33.826	450 474	y = 0.0006x ? 0.0358	3.125~50	R² = 1
7	全反式-α-胡萝卜素all-trans-α-carotence	35.501	450 478	y = 0.0090x + 0.0806	3.125~50	R² = 1
8	全反式-β-胡萝卜素all-trans-β-carotence	39.213	456 484	y = 0.0041x + 0.0070	3.125~50	R² = 1
9	9-顺式-β-胡萝卜素9-cis-β-carotence	40.291	452 478	y = 0.0048x + 0.0529	3.125~50	R² = 1

编号No.	类胡萝卜素 Carotenoids	R_t	λ_max	线性方程 Linear equations	线性范围 Linear range/(μg·mL^?1)	相关系数 Correlation index
1	新黄质neoxanthin	11.512	468 416	y = 0.0078x + 0.1042	3.125~50	R² = 0.9999
2	叶黄素lutein	19.192	450 478	y = 0.0048x + 0.0935	3.125~50	R² = 1
3	β-阿朴-8°-胡萝卜素醛 β -apo-8°-carotenal	23.801	466	y = 0.0048x + 0.0529	3.125~50	R²= 1
4	玉米黄质zeinoxanthin	27.359	450 478	y = 0.0051x + 0.0170	3.125~50	R² = 1
5	ε-胡萝卜素ε-carotence	31.662	420 444 474	y = 0.0047x + 0.0591	3.125~50	R² = 0.9999
6	13-顺式-β-胡萝卜素13-cis-β-carotence	33.826	450 474	y = 0.0006x ? 0.0358	3.125~50	R² = 1
7	全反式-α-胡萝卜素all-trans-α-carotence	35.501	450 478	y = 0.0090x + 0.0806	3.125~50	R² = 1
8	全反式-β-胡萝卜素all-trans-β-carotence	39.213	456 484	y = 0.0041x + 0.0070	3.125~50	R² = 1
9	9-顺式-β-胡萝卜素9-cis-β-carotence	40.291	452 478	y = 0.0048x + 0.0529	3.125~50	R² = 1

品种名 Cultivar name	叶黄素 Lutein	13-顺式-β-胡萝卜素 13-cis-β-carotence	全反式-α-胡萝卜素 All-trans-α-carotence	全反式-β-胡萝卜素 All-trans-β-carotence
中蕉8号	0.131±0.039^c	—	0.053±0.007^c	0.016±0.004^c
Poingo	0.172±0.019^bc	0.010±0.001^b	2.186±0.111^b	1.147±0.057^b
French Sombre	0.242±0.040^a	0.031±0.002^a	7.172±0.578^a	2.351±0.195^a

品种名 Cultivar name	叶黄素 Lutein	13-顺式-β-胡萝卜素 13-cis-β-carotence	全反式-α-胡萝卜素 All-trans-α-carotence	全反式-β-胡萝卜素 All-trans-β-carotence
中蕉8号	0.131±0.039^c	—	0.053±0.007^c	0.016±0.004^c
Poingo	0.172±0.019^bc	0.010±0.001^b	2.186±0.111^b	1.147±0.057^b
French Sombre	0.242±0.040^a	0.031±0.002^a	7.172±0.578^a	2.351±0.195^a