Determination of Z/E Isomer Content in Lycopene Micelle by HPLC

doi:10.6048/j.issn.1001-4330.2022.09.015

Abstract

Abstract:

【Objective】 To determine the content of Z/E isomers in lycopene micelles. 【Method】 HPLC method， The column was YMC Carotenoid S-5（4.6 mm×250 mm）； Mobile phase A：acetonitrile B：methyl tert-butyl ether； Linear gradient elution； Flow rate：1 mL/min； Detection wavelength：472 nm； Column temperature：30℃， Complete the separation and detection of isomers within 45 minutes， the isomers in the lycopene micelles are analyzed and identified through the characteristic peaks and absorption intensity on the ultraviolet-visible absorption spectrum.【Result】 The linear ranges of Lycopene was 0.22~6.57 μg/mL （r=0.999）， Whose average recoveries were 100.06%（RSD=1.99%）. It shows that the method has strong specificity， good linear relationship， fast， accurate and simple， and is suitable for the determination of isomers in lycopene micelles.【Conclusion】 Determined that there are all-trans forms in the lycopene micelles：All-E， and three cis isomers：13Z， 9Z， 5Z.with an average relative percentage of 76.06%，10.83%，4.55%，8.56%， respectively.

Key words: lycopene; micelles; isomers; content determination

摘要：

【目的】 测定番茄红素胶束中番茄红素Z/E异构体含量。【方法】 采用HPLC法，色谱柱为YMC Carotenoid S-5（4.6 mm×250 mm）；流动相 A：乙腈B：甲基叔丁基醚；线性梯度洗脱；流速：1 mL/min；检测波长：472 nm；柱温：30 ℃，在45 min内完成异构体的分离检测，通过紫外-可见吸收光谱图上的特征峰及吸收强度,分析和鉴定番茄红素胶束中存在的异构体。【结果】 番茄红素在0.22~6.57 μg/mL 线性关系良好（r=0.999）；加样回收率为100.06%（RSD=1.99%），该方法专属性强，线性关系良好，快速、准确、简便，适用于番茄红素胶束中异构体的含量测定。【结论】 番茄红素胶束中存在全反式：All-E，以及三个顺式异构体：13 Z，9 Z，5 Z，其平均相对百分含量分别为76.06%、10.83%、4.55%、8.56%。

关键词: 番茄红素, 胶束, 异构体, 含量测定

CLC Number:

S641.2

MA Xuehong, LIU Ting, LI Chuantian, ZHU Jinfang. Determination of Z/E Isomer Content in Lycopene Micelle by HPLC[J]. Xinjiang Agricultural Sciences, 2022, 59(9): 2209-2216.

马雪红, 刘婷, 李传天, 朱金芳. HPLC法测定番茄红素胶束中番茄红素Z/E异构体含量[J]. 新疆农业科学, 2022, 59(9): 2209-2216.

Figures/Tables 10

References 32

[1]	Millaydet, Bull. Processing of tomatoes and solanombin extraction[J]. Soc Naney, 1875, 2(1):21-23.
[2]	阚义, 罗星, 李华. 烟台、新疆地区的番茄中番茄红素含量的差异[J]. 山东农业大学学报(自然科学版), 2013, 44(2):176-180.
	Kan Yi, Luo Xing, Li Hua. Thediversityoftomatoes' lycopenecontentbetween Yantai and Xinjiang[J]. Journal of Shandong Agricultural University (Natural Science Edition), 2013, 44(2):176-180.
[3]	Zu K, Mucci L, Rosner B A, et al. Dietary lycopene, angiogenesis, and prostate cancer: A prospective study in the prostate-specific antigen era[J]. Journal of the National Cancer Institute, 2014, (2): 195-196.
[4]	Gajendragadkar P R, Hubsch A, Mäki-Petäjä K M, et al. Effects of oral lycopene supplementation on vascular function in patients with cardiovascular disease and healthy volunteers: a randomised controlled trial[J]. Plos One, 2014, 9(6): 1-13.
[5]	Li X N, Lin J, Xia J, et al. Lycopene mitigates atrazine-induced cardiac inflammation via blocking the NF-κB pathway and NO production[J]. Journal of Functional Foods, 2017, 29: 208-216. DOI URL
[6]	Cheng H M, Koutsidis G, Lodge J K, et al. Tomato and lycopene supplementation and cardiovascular risk factors: A systematic review and meta-analysis[J]. Atherosclerosis, 2017, 257: 100-108. DOI PMID
[7]	Bansal P, Gupta S K, Ojha S K, et al. Cardioprotective effect of lycopene in the experimental model of myocardial ischemia-reperfusion injury[J]. Molecular and Cellular Biochemistry, 2006, 289(1-2): 1-9. DOI PMID
[8]	Ardawi M S M, Badawoud M H, Hassan S M, et al. Lycopene treatment against loss of bone mass, microarchitecture and strength in relation to regulatory mechanisms in a postmenopausal osteoporosis model[J]. Bone, 2016, 83(5): 127-140. DOI URL
[9]	Pankaj Bansal, Suresh Kumar Gupta, Shreesh Kumar Ojha, et al. Cardioprotective effect of lycopene in the experimental model of myocardial ischemia-reperfusion injury[J]. Molecular and Cellular Biochemistry, 2006, 289(1-2). DOI PMID
[10]	Shi J, Le M M. Lycopene in tomatoes: chemical and physical properties affected by food processing[J]. Critical Reviews in Biotechnology, 2000, 20(4): 293-334. DOI PMID
[11]	Koe B K, Zechmeister L. In vitro conversion of phytofluene and phytoene into carotenoid pigments.[J]. Arch Biochem Biophys. 1952, 41(1): 236-243. PMID
[12]	Joseph Schierle, Werner Bretzel, Ilme Bühler, et al. Content and isomeric ratio of lycopene in food and human blood plasma[J]. Food Chemistry, 1997, 59(3):459-465. DOI URL
[13]	John Shi, Yuzhu Dai, Yukio Kakuda, et al. Effect of heating and exposure to light on the stability of lycopene in tomato purée[J]. Food Control, 2007, 19(5):514-520. DOI URL
[14]	Colle Ines J P, Lemmens Lien, Tolesa Getachew N, et al. Lycopene degradation and isomerization kinetics during thermal processing of an olive oil/tomato emulsion.[J]. Journal of agricultural and food chemistry, 2010, 58(24):12784-12792. DOI PMID
[15]	Masaki Honda, Kazuya Murakami, Yo Watanabe, et al. The E / Z isomer ratio of lycopene in foods and effect of heating with edible oils and fats on isomerization of (all-E )-lycopene[J]. European Journal of Lipid Science and Technology, 2017, 119(8).
[16]	Rao A V, Ray M R, Rao L G. Lycopene[J]. Advances in Food & Nutrition Research, 2006, 51(6): 99-164.
[17]	Shi J, Maguer M L, Bryan M, et al. Kinetics of Lycopene Degradation in Tomato Puree by Heat and Light Irradiation[J]. Journal of Food Process Engineering, 2003. 25(6): 485-498. DOI URL
[18]	Kazuya Murakami, Masaki Honda, Ryota Takemura, et al. Effect of thermal treatment and light irradiation on the stability of lycopene with high Z -isomers content.[J]. Food Chemistry, 2018, 250:253-258. DOI PMID
[19]	Honda Masaki, Takahashi Naoto, Kuwa Takahiro, et al. Spectral characterisation of Z-isomers of lycopeneformed during heat treatment and solvent effects on the E/Z isomerisation process.[J]. Food chemistry, 2015, 171:323-329. DOI PMID
[20]	Meléndez-Martínez Antonio J, Paulino Margot, Stinco Carla M, et al. Study of the time-course of cis/trans (Z/E) isomerization of lycopene, phytoene, and phytofluene from tomato.[J]. Journal of agricultural and food chemistry, 2014, 62(51):12399-12406. DOI PMID
[21]	M. T. Lee, B. H. Chen. Separation of lycopene and its cis isomers by liquid chromatography[J]. Chromatographia, 2001, 54(9-10): 613-617. DOI URL
[22]	Hanh Phan-Thi, Yves Waché. Isomerization and increase in the antioxidant properties of lycopene from Momordica cochinchinensis (gac) by moderate heat treatment with UV-Vis spectra as a marker.[J]. Food chemistry, 2014, 156:58-63. DOI PMID
[23]	C.H Lin, B.H Chen. Determination of carotenoids in tomato juice by liquid chromatography[J]. Journal of Chromatography A, 2003, 1012(1):103-109. DOI PMID
[24]	Mario G Ferruzzi, Minhthy L Nguyen, Lane C Sander, et al. Analysis of lycopene geometrical isomers in biological microsamples by liquid chromatography with coulometric array detection[J]. Journal of Chromatography B: Biomedical Sciences and Applications, 2001, 760(2):289-299.
[25]	李京, 惠伯棣. 番茄红素在体内代谢中的几何异构体组成变化[J]. 食品科学, 2008,(9):591-594.
	LI Jing, HUI Bodi. Variation in Geometrical Isomer Composition of Lycopene in Rat Body[J]. Food science, 2008,(9):591-594.
[26]	Curt Emenhiser, Lane C,. Sander, Steven J. Schwartz. Capability of a polymeric C 30 stationary phase to resolve cis-trans carotenoid isomers in reversed-phase liquid chromatography[J]. Journal of Chromatography A, 1995, 707(2):205-216. DOI URL
[27]	Lane C,. Sander, Katherine E. Sharpless,Matthias Pursch. C 30 Stationary phases for the analysis of food by liquid chromatography[J]. Journal of Chromatography A, 2000, 880(1):189-202. DOI URL
[28]	李京. 天然番茄红素的分析、制备及保健功能研究[D]. 北京, 首都师范大学, 2007.
	LI Jing. Analysis, Preparation and Health Care Function of Natural Lycopene[D]. Beijing, Capital Normal University, 2007.
[29]	惠伯棣, 李京, 裴凌鹏. 应用C30-HPLC-PDA分离与鉴定番茄红素几何异构体[J]. 食品工业科技, 2006,(7):49-51,54.
	HUI Bodi, LI Jing, PEI Lingpeng. The separation and identification of lycopene geometrical isomers by C30-HPLC-PDA[J]. Food industry technology, 2006, (7):49-51,54.
[30]	Ishida B K, Ma J, Chan B. A simple, rapid method for HPLC analysis of lycopene isomers. 2001, 12(3):194-198.
[31]	徐媛. 红葡萄柚番茄红素加工降解机制及其定量构效关系研究[D]. 武汉: 华中农业大学, 2013.
	XU Yuan. Studies on degradation mechanism of lycopene from red grapefruit during processing and quantitative structure-activity relationship [D]. Wuhan: Huazhong Agricultural University. 2013.
[32]	Jessica L,. Cooperstone, David M,. Francis, Steven J. Schwartz. Thermal processing differentially affects lycopene and other carotenoids in cis -lycopene containing, tangerine tomatoes.[J]. Food Chemistry. 2016, 210:466-472. DOI PMID

放置时间（h） Put time （h）	含量（μg/mL） Content （μg/mL）	平均含量（μg/mL） Average content （μg/mL）	RSD（%）
0	376	371	0.69
1	369
2	369
4	372
6	370
8	372
12	370
24	368

放置时间（h） Put time （h）	含量（μg/mL） Content （μg/mL）	平均含量（μg/mL） Average content （μg/mL）	RSD（%）
0	376	371	0.69
1	369
2	369
4	372
6	370
8	372
12	370
24	368

分组 Grouping	样品中量 Sample volume （μg）	加入量 Amount added （μg）	测得量 Measured amount （μg）	回收率 Recovery rate （%）	平均值 Average value （%）	RSD （%）
低 Low	185	148	328	97.30	100.06	1.99
	185	148	332	99.46
	185	148	337	102.16
中 Middle	185	185	371	100.54
	185	185	370	100.00
	185	185	372	101.08
高 High	185	222	401	96.76
	185	222	412	102.70
	185	222	408	100.54

分组 Grouping	样品中量 Sample volume （μg）	加入量 Amount added （μg）	测得量 Measured amount （μg）	回收率 Recovery rate （%）	平均值 Average value （%）	RSD （%）
低 Low	185	148	328	97.30	100.06	1.99
	185	148	332	99.46
	185	148	337	102.16
中 Middle	185	185	371	100.54
	185	185	370	100.00
	185	185	372	101.08
高 High	185	222	401	96.76
	185	222	412	102.70
	185	222	408	100.54

批号 Batch number	含量（μg/mL） Content	平均含量（μg/mL） Average content	批内 RSD（%） RSD within batch	批间 RSD（%） RSD between batches
20200801	373	371	0.41	0.54
	372
	370
20200802	372	369	0.63
	368
	368
20200803	373	372	0.56
	370
	369