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UPLC
Chromatography was carried out with a Ultra-high performance liquid chromatography. Samples were measured by a C18 carotenoid column (100 mm × 2.1 i.d., 1.7 μm). The eluent was methyl tertiary butyl ether and methanol (V/V = 30:70) with the type of isocratic elution. And each eluent contained 0.01% BHT and 0.05% TEA (triethylamine) as modifiers in order to prevent the degradation of carotenoids on the column. The following rate was 0.5 mL/min, column temperature was 25 ℃, and the injection volume was 5 μL, analyzing with UV at 445 nm.
References：
Shen YH, Yang FY, Lu BG, Zhao WW, Jiang T, Feng L, Chen XJ, Ming R. Exploring the differential mechanisms of carotenoid biosynthesis in the yellow peel and red flesh of papaya. BMC Genomics. 2019 Jan 16;20(1):49. doi: 10.1186/s12864-018-5388-0. PMID: 30651061; PMCID: PMC6335806.
HPLC
HPLC analysis of chlorophyll and carotenoids were carried out using an Agilent 1260 instrument with a VWD detector . Samples (10 μL) were separated at 30℃ on a C18 column (150 mm × 3.9 mm i.d.; 4 μm particle size) using isopropanol and 80% acetonitrile–water at a flow rate of 0.5ml/min.
References：
Di H, Zhang Y, Ma J, Wei J, Wang Y, Li Z, Cui C, Fang P, Ma W, Li H, Sun B, Zhang F. Sucrose treatment delays senescence and maintains the postharvest quality of baby mustard (Brassica juncea var. gemmifera). Food Chem X. 2022 Mar 1;14:100272. doi: 10.1016/j.fochx.2022.100272. PMID: 35257095; PMCID: PMC8897633.
HPLC
The specific elution included phase A: acetonitrile: methanol = 3:1 (containing 0.01% butylated hydroxytoluene (BHT)); phase B: 100% methyl-tert-butyl ether (MTBE) (containing 0.01% BHT). The elution conditions were as follows: 0 min: A-B (95: 5); 0–10 min: A-B (95: 5); 10–19 min: A-B (86: 14); 19–29 min: A-B (75: 25); 29–54 min: A-B (50:50); 54–66 min: A-B (26:74); and 67 min: A-B (95:5). The mobile phase flow rate was 1 mL/min, and the detection wavelength was 450 nm; the column temperature was 35 ℃.
References：
Zhang R, Zhang J, Li C, Pan Q, Haq SU, Mosa WFA, Fang F, Zhang L, Li B. The Accumulation of Health-Promoting Nutrients from Representative Organs across Multiple Developmental Stages in Orange Chinese Cabbage. Plants (Basel). 2023 May 26;12(11):2120. doi: 10.3390/plants12112120. PMID: 37299099; PMCID: PMC10255399.
HPLC
HPLC analysis of carotenoids was carried out using an equipped with a variable wavelength detector (VWD). Samples (10 μL) were separated at 30℃ on a C18 column (150 × 3.9 mm id; 3 μm particle size) using isopropanol and 80% acetonitrile-water at a flow rate of 0.5 mL min?1 ; the absorbance was measured at 448 and 428 nm.
References：
Jian Y, Zhang C, Wang Y, Li Z, Chen J, Zhou W, Huang W, Jiang M, Zheng H, Li M, Miao H, Zhang F, Li H, Wang Q, Sun B. Characterization of the Role of the Neoxanthin Synthase Gene BoaNXS in Carotenoid Biosynthesis in Chinese Kale. Genes (Basel). 2021 Jul 24;12(8):1122. doi: 10.3390/genes12081122. PMID: 34440295; PMCID: PMC8393425.
