Food Science and Technology Research
Online ISSN : 1881-3984
Print ISSN : 1344-6606
ISSN-L : 1344-6606
Technical papers
Measurement of glucosinolates in Brassicaceae vegetables grown in Japan
Yuta Yamanouchi, Akira Ando, Yuko Takano-Ishikawa, Hiroshi Ueda
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Keywords: Glucosinolate, Brassicaceae vegetables, Brassica oleracea, Brassica rapa, Brassica napus, Brassica juncea, Raphanus sativus
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2023 Volume 29 Issue 6 Pages 541-551

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Abstract

Brassicaceae vegetables contain glucosinolate (GSL), which is well known for its role in the plant defense system, bitter substances, and functional components. This study reported the GSL profiles of 18 GSLs in 14 Brassicaceae vegetables (kale, cabbage, kohlrabi, broccoli, cauliflower, komatsuna, pak choi, turnip, choy sum, Chinese cabbage, rutabaga, leaf mustard, leaf radish, and radish root) consisting of 64 cultivars grown in Japan. The GSL compositions showed specific trends among each species, and furthermore, there was considerable variation in the total and individual GSL contents among the cultivars and tissues of the same vegetable. Because each vegetable was grown under the same conditions, this variation was expected to primarily be due to the genetic background. It is desirable to collect additional data from commonly distributed cultivars to conduct more detailed studies on GSLs in Brassicaceae vegetables, such as investigating a negative correlation between GSL consumption and cancer risk.

Introduction

Glucosinolates (GSLs) are the major secondary metabolites in many Brassicaceae vegetables, and more than 200 GSLs have been identified (Clarke, 2010). They can be grouped into aliphatic, indole, and aromatic GSLs according to their amino acid precursor methionine, tryptophan, and phenylalanine, respectively. Intact GSLs are stored in the vacuoles of plant cells and hydrolyzed by myrosinase (EC 3.2.1.147) in the cytoplasm upon plant tissue disruption into several bioactive compounds (Bones and Rossiter, 2006). Isothiocyanates (ISNs), the major hydrolyzed products, are responsible for the protection of plants from diseases caused by bacterial or fungal and insect infections (Hopkins et al., 2009). However, ISNs have bitter and pungent flavors which are characteristic of Brassicaceae vegetables. For example, some ISNs derived from aliphatic and indole GSLs in cauliflower and broccoli affect their bitterness (Schonhof et al., 2004). In cabbage, ISNs derived from sinigrin (SIN) and progoitrin (PRO) are the characteristic pungent and bitter compounds (Drewnowski et al., 2000).

In the last decades, the health-promoting attributes of GSLs have received as much attention as their sensorial attributes. Several studies have reported that GSLs and their breakdown products affect many stages of cancer development by modulating the induction of detoxification enzymes (phase II enzymes) and inhibition of activation enzymes (phase I enzymes; Zhang et al., 1992). Sulforaphane, the hydrolysis product of glucoraphanin (GRA), is the most studied ISN, but some studies have reported that others may contribute to its anticarcinogenic effect (Staack et al., 1998). In addition, several epidemiological studies have reported a negative correlation between the consumption of Brassicaceae vegetables and the risk of cancer (Verhoeveb et al., 1996; Higdon et al., 2007). In contrast, it was previously attempted to reduce the amount of PRO in animal feed as it caused goiters, and toxicity due to ISNs has been reported in some animals (Stoewsand, 1995). Although no negative effects of the regular consumption of Brassicaceae vegetables in humans have been reported, investigating the detailed profiles of a wide range of GSLs is important to understand the amount of GSLs humans consume daily. Although several studies have been published on the GSL profiles of various Brassicaceae vegetables (Ciska et al., 2000; Verkerk et al., 2009; Chen, 2008; Wu and Pehrsson, 2022; Hill and Williams, 1987; Lee et al., 2021), it is known that they are influenced by cultivar genetic background and environmental factors, such as temperature, rainfall, nutrient supply, and growing season (Kushad et al., 1998; Ciska et al., 2000; Falk et al., 2007; Padilla et al., 2007; Velasco et al., 2007). Because there is limited information regarding the comparison between the GSLs among Brassicaceae vegetables grown in Japan, further analysis is required to determine the amount of GSLs consumed by the Japanese. Therefore, this study aimed to compare and evaluate the GSL profiles of several Brassicaceae vegetables grown in Japan.

Materials and Methods

Chemicals and reagents  SIN, sulfatase (type H-1 from Helix pomatia), and DEAE-Sephadex® A-25 were purchased from Sigma-Aldrich (St. Louis, MO, USA). GRA was purchased from Cayman Chemical Co. (Ann Arbor, MI, USA). Glucoraphenin (GRE) was purchased from LKT Laboratories (Minneapolis, MN, USA). Other GSL standards [PRO, gluconapin (GNA), glucobrassicanapin (GBN), glucoerucin (GER), glucobrassicin (GBS), 4-methoxyglucobrassicin (4-OMGBS), and gluconasturtiin (GST)] were purchased from Extrasynthese (Lyon, France). Methanol and acetonitrile [liquid chromatography-mass spectrometry (LC-MS) grade] were purchased from Kanto Chemical Co., Inc. (Tokyo, Japan).

Samples  All vegetable seeds (5 kale, 5 cabbage, 5 kohlrabi, 2 broccoli, 5 cauliflower, 5 komatsuna, 5 pak choi, 2 turnip, 5 choy sum, 5 Chinese cabbage, 5 rutabaga, 5 leaf mustard, 5 leaf radish, and 5 radish root) used in this study were obtained from the Research Center of Genetic Resources (Ibaraki, Japan) and conventionally cultivated at the Institute of Vegetable and Floriculture Science, NARO (Mie, Japan). The inedible parts were discarded, and 50–300 g of edible parts were obtained through appropriate sample reduction. For example, we cut cabbages radially and sampled those on the diagonal. Each vegetable was rapidly frozen with liquid nitrogen. The frozen samples were lyophilized, milled with a grinder mill (GM-200; Verder Scientific Co., Ltd.), and stored at −30 °C until extraction.

Sample preparation  Desulfo-GSL extraction from the lyophilized samples was conducted as a previously published HPLC protocol (Grosser and Dam, 2017) with minor modifications. Briefly, the samples (20 mg) with two small metal balls were suspended in 1 mL of 70 % methanol and heated at 70 °C for 10 min to inactivate the myrosinase. After sonication for 15 min and centrifugation at 9 500 × g for 10 min, the supernatant was transferred to Pasteur pipette columns packed with DEAE-Sephadex® A-25. After repeating this procedure thrice, the samples were washed with methanol, distilled water, and 20 mM sodium acetate buffer (pH 5.5) and desulfurized with sulfatase (8.0 units) at 37 °C for 15 h. Desulfo-GSLs were eluted with 3 × 0.5 mL distilled water and lyophilized. After lyophilization, the samples were dissolved in 1.0 mL distilled water by a whole pipette and filtered through a 0.22 µm ETFE filter. The SIN standards for the calibration curves were prepared in the same manner. The spike and recovery tests were conducted by comparing analytical data of the SIN standard and the lyophilized powder of rutabaga, adding the same concentration of SIN, which was heated, extracted, and purified in the same manner as the other samples.

HPLC analysis  Desulfo-GSLs were separated using an Agilent 1260 Infinity II LC system (Agilent, Palo Alto, CA, USA) with a Unison UK-C18 column (Impact Co., Kyoto, Japan) at 20 °C and detected by UV detection at 229 nm. Solvent A (distilled water) and solvent B (acetonitrile) were applied at a flow rate of 0.75 mL/min as follows: 0 min 98 % A and 2 % B, 15 min 86 % A and 14 % B, and 40 min 73 % A and 27 % B. GSLs, with standards available, were identified by comparing their retention times (RTs) and mass spectra from an Agilent 6120B Single Quadrupole Walkup System. The MS scans were performed in the positive-ion mode on desulfo-GSLs [M + Na]+ ions, and spectra were detected from 250–450 m/z (capillary voltage, 3000 V; nebulizer, 60 psig; dry gas temperature, 350 °C; and dry gas flow, 12 L/min). The others were estimated from RT in previous studies (Grosser and Dam, 2017; Osada, 2016) and the obtained desulfo-GSL mass spectra. The obtained analytical data were confirmed to be within the calibration curve created with SIN standards, each GSL content was calculated according to the previously their reported response factor (ISO9167, 2019). The lower limits of detection and quantification in this study were calculated based on the standard deviation of the lowest SIN standard.

Statistical analysis  The statistical analysis in this study was performed using R software (version 4.2.2). Tukey's Honestly Significant Difference (HSD) test and t-test were conducted to compare the GSL profiles among the cultivars in each vegetable. Principal component analysis (PCA) was conducted on the GSL profiles of Brassica vegetables using the prcomp function in R and visualized with the ggplot2 package.

Results

Identification of GSLs  The identification of the GSL peaks by HPLC was determined based on their RT and mass spectra. A typical GSL chromatogram of the standard mixture and sample (cabbage, “Best Of All” and rutabaga, “Green Top”) is shown in Fig. 1. The analysis of the 64 Brassicaceae vegetables identified 18 GSLs: 13 aliphatic, 4 indole, and 1 aromatic (Table 1). The GSL profiles of each Brassicaceae vegetable are shown in Tables 2–6. (Detailed information such as the standard deviations and statistical analysis results are described in Supplemental Table 1). Although the peaks for SIN and GRE overlapped, it was not considered problematic because there was no Brassicaceae vegetable that contained both SIN and GRE by MS analysis in this study. The peak at RT = 14.2 that was detected in some cultivars of pak choi, turnip leaves, choy sum, and rutabaga could not be identified (Fig. 1C). In addition, the spike and recovery tests were conducted by comparing the SIN standard and lyophilized powder of rutabaga (without SIN and GRE), adding the same concentration of SIN, which resulted in 101.6 % recovery (data not shown). The result indicated that there was minimal loss of GSLs during the extraction process.

Fig. 1.

Chromatogram of the desulfo-GSL standard mix solution (A) and sample extraction (B: cabbage, “Best Of All”; C: rutabaga, “Green Top”) with a UV detector (229 nm). GIB, Glucoiberin; PRO, Progoitrin; GRA, Glucoraphanin; GRE, Glucoraphenin; SIN, Sinigrin; GAL, Glucoalyssin; GNL, Gluconapoleiferin; GNA, Gluconapin; GIV, Glucoibervirin; GBN, Glucobrassicanapin; GER, Glucoerucin; GBE, Glucoberteroin; 4HGBS, 4-Hydroxyglucobrassicin; GBS, Glucobrassicin; 4MGBS, 4-Methoxyglucobrassicin; NGBS, Neoglucobrassicin; GST, Gluconasturtiin.

Table 1.List of GSLs detected in Brassicaceae vegetables.

Glucosinolate Side chain structure Abbr. RT (min) m/z of desulfo-GSLs [M + Na]+
Aliphatic Glucoiberin b 3-methylsulfinylpropyl GIB 5.4 366.1
Progoitrin a 2-hydroxy-3-butenyl PRO 6.3 332.1
Glucoraphanin a 4-methylsulfinylbutyl GRA 7.0 380.1
Glucoraphenin a 4-methylsulfinyl-3-butenyl GRE 7.6 378.1
Sinigrin a 2-propenyl SIN 7.6 302.1
Glucoalyssin b 5-methylsulfinylpentyl GAL 9.3 394.1
Gluconapoleiferin b 2-hydroxy-4-pentenyl GNL 9.6 346.1
Gluconapin a 3-butenyl GNA 11.9 316.1
Glucoibervirin b 3-methylthiopropyl GIV 14.0 350.1
Glucobrassicanapin a 4-pentenyl GBN 17.2 330.1
Glucoerucin a 4-methylthiobutyl GER 18.2 364.1
Glucoraphasatin b 4-methylthio-3-butenyl GPH 19.6 362.1
Glucoberteroin b 5-methylthiopentyl GBE 23.9 378.1
Indole 4-Hydroxyglucobrassicin 4-hydroxy-3-indolylmethyl 4HGBS 15.1 407.1
Glucobrassicin a 3-indolylmethyl GBS 21.5 391.1
4-Methoxyglucobrassicin a 4-methoxy-3-indolylmethyl 4MGBS 25.0 421.1
Neoglucobrassicin b 1-methoxy-3-indolylmethyl NGBS 32.2 421.1
Aromatic Gluconasturtiin a 2-phenylethyl GST 24.6 366.1
a  GSLs identified by comparison of RTs and mass spectra with commercial standards.

b  GSLs estimated from previous studies and mass spectra obtained in this paper.

Table 2.GSLs profile in B.oleracea a

B.oleracea JP No. b Origin Cultivation period c GIB PRO GRA GRE SLN GAL GNL GNA GIV GBN GER GPH GBE 4HGBS GBS 4MGBS NGBS GST Total
Kale
 Shashamane Local 1 JP26040 Ethiopia Aug. 31, 2021 - Oct. 18, 2021 ND d LLOQe ND ND 25.6 ND ND ND ND ND ND ND ND 0.6 2.8 2.2 0.6 ND 31.9
 Knol Khol JP26050 India Aug. 31, 2021 - Oct. 18, 2021 6.8 LLOQ 21.0 ND ND ND ND ND ND ND ND ND ND 0.8 14.2 1.0 1.1 ND 45.0
 Marrow Stem JP26055 UK Aug. 31, 2021 - Oct. 18, 2021 6.5 LLOQ LLOQ ND 4.0 ND ND ND ND ND ND ND ND 0.6 47.4 1.6 2.3 ND 62.4
 B Oleracea O 179 JP26060 Turkey Aug. 31, 2021 - Oct. 18, 2021 2.1 1.6 6.3 ND LLOQ ND ND ND ND ND ND ND ND 0.9 48.0 1.5 1.5 ND 61.8
 Col/Spain/530 JP45942 Spain Aug. 31, 2021 - Oct. 18, 2021 4.1 LLOQ ND ND 5.2 ND ND ND ND ND ND ND ND 0.4 10.0 0.6 1.7 ND 22.0
Cabbage
 Red Eekaa JP25989 unkown Aug. 31, 2021 - Feb. 2, 2022 1.8 20.2 27.2 ND 1.5 ND ND 10.6 ND ND ND ND ND 1.7 8.1 1.3 LLOQ ND 72.4
 Oogata Kopenhaagen Marketto JP25994 Japan Aug. 31, 2021 - Nov. 24, 2021 6.3 11.9 6.8 ND 7.3 ND ND 8.5 LLOQ ND ND ND ND LLOQ 6.2 1.1 0.1 ND 48.2
 Kibakei Sakusesshon JP26020 Japan Aug. 31, 2021 - Dec. 7, 2021 10.9 LLOQ ND ND 3.0 ND ND ND ND ND ND ND ND 0.2 12.4 1.2 0.2 ND 27.9
 Youshin JP26027 Japan Aug. 31, 2021 - Nov. 24, 2021 18.0 3.2 2.7 ND 16.0 ND ND 0.9 LLOQ ND ND ND ND 0.2 8.5 1.4 0.2 ND 51.0
 Best Of All JP26031 UK Aug. 31, 2021 - Feb. 2, 2022 48.5 5.2 2.2 ND 47.0 ND ND 1.2 2.4 ND ND ND ND 3.2 23.0 1.8 0.4 ND 134.9
Kohlrabi
 Yue Nan Pie Lan 1 JP80075 China Aug. 2, 2022 - Nov. 14, 2022 1.8 10.4 35.8 ND 1.2 ND ND 4.6 ND ND 17.3 ND ND 1.5 10.3 LLOQ 2.0 7.3 92.2
 Yuan Pie Lan JP80076 China Aug. 2, 2022 - Nov. 7, 2022 4.9 5.5 18.9 ND 2.0 ND ND LLOQ LLOQ ND 7.3 ND ND 0.8 5.2 LLOQ 5.5 LLOQ 50.2
 Fumei JP127521 China Aug. 2, 2022 - Nov. 7, 2022 4.9 10.9 14.8 ND 5.4 ND ND 4.8 LLOQ ND 7.2 ND ND 1.5 3.1 LLOQ 4.3 2.0 58.8
 86-703 JP127522 Nepal Aug. 2, 2022 - Nov. 1, 2022 5.5 ND 9.1 ND ND ND ND ND 4.1 ND 12.0 ND ND 1.1 1.4 LLOQ 4.3 ND 37.5
 Wener Witte JP127603 Netherlands Aug. 2, 2022 - Nov. 1, 2022 18.1 ND 10.6 ND ND ND ND ND 4.6 ND 3.8 ND ND 1.0 2.7 LLOQ 3.0 ND 43.8
Broccoli
 Nakazato Broccoli JP26686 Japan Aug. 2, 2022 - Nov. 29, 2022 LLOQ ND 34.3 ND ND ND ND ND ND ND ND ND ND 2.5 22.5 3.1 6.9 ND 69.3
 5R JP88079 Poland Aug. 2, 2022 - Nov. 29, 2022 3.8 ND 13.4 ND ND ND ND ND ND ND ND ND ND 1.9 23.7 2.7 53.4 ND 99.0
Cauliflower
 Boushuu Nakate JP26673 Japan Aug. 2, 2022 - Feb. 6, 2023 16.8 LLOQ LLOQ ND 23.0 ND ND ND 3.7 ND ND ND ND 1.1 17.9 0.8 2.1 ND 65.5
 Pak-10417 JP86202 Pakistan Aug. 2, 2022 - Nov. 14, 2022 6.1 ND ND ND 7.5 ND ND ND 4.2 ND ND ND ND 0.6 8.0 LLOQ LLOQ ND 26.4
 Minaret JP182453 unkown Aug. 2, 2022 - Jan. 10, 2023 3.4 ND 5.7 ND ND ND ND ND ND ND ND ND ND 8.5 8.7 1.2 3.0 ND 30.5
 Boushuu Wase JP26674 Japan Aug. 2, 2022 - Dec. 15, 2023 18.3 ND LLOQ ND 4.1 ND ND ND 2.3 ND ND ND ND 2.1 12.2 0.7 1.1 ND 40.7
 Boushuu Chuusei JP37405 Japan Aug. 2, 2022 - Feb. 22, 2023 21.5 ND LLOQ ND 18.3 ND ND ND 6.3 ND ND ND ND 2.6 13.3 1.5 0.8 ND 64.3
a  mg/100 g Fw, values are means (n = 3).

b  ID used in the Research Center of Genetic Resources (i).

c  Cultivation period was defined as the period from the sowing date to the harvest date.

d  Not detectable.

e  Lower limit of quantitation.

GIB: Glucoiberin; PRO: Progoitrin; GRA: Glucoraphanin; GRE: Glucoraphenin; SIN: Sinigrin; GAL: Glucoalyssin; GNL: Gluconapoleiferin; GNA: Gluconapin; GIV: Glucoibervirin GBN: Glucobrassicanapin; GER: Glucoerucin; GPH: Glucoraphasatin; GBE: Glucoberteroin; 4-OHGBS: 4-Hydroxyglucobrassicin; GBS: Glucobrassicin; 4-OMGBS: 4-Methoxyglucobrassicin; NGBS: Neoglucobrassicin; GST: Gluconasturtiin.

Table 3.GSLs profile in B.rapa a

B.rapa JP No. b Origin Cultivation period c GIB PRO GRA GRE SIN GAL GNL GNA GIV GBN GER GPH GBE 4HGBS GBS 4MGBS NGBS GST Total
Komatsuna
 Junkei Nozawana JP26903 Japan Jun. 28, 2021 - Aug. 16, 2021 ND d 4.1 1.5 ND ND 1.6 ND 56.1 ND 7.2 3.5 ND 1.0 ND 1.0 1.1 2.5 ND 79.8
 Shin Kuro Mizuna JP26907 Japan Jun. 28, 2021 - Aug. 16, 2021 ND 18.2 0.8 ND ND 3.2 LLOQe 55.2 ND 13.4 ND ND ND ND 0.3 1.6 2.4 1.3 96.4
 Maruba Komatsuna JP37635 Japan Jun. 28, 2021 - Aug. 16, 2021 ND 2.7 ND ND ND 0.7 ND 65.5 ND 12.1 ND ND ND ND 0.4 0.9 1.2 0.6 84.2
 Uguisuna JP43250 Japan Jun. 28, 2021 - Aug. 16, 2021 ND 5.4 ND ND ND 3.5 ND 65.3 ND 14.2 LLOQ ND 0.7 ND 0.7 1.3 4.6 LLOQ 95.5
 Toukyou Aona JP37637 Japan Jun. 28, 2021 - Aug. 16, 2021 ND 3.4 ND ND ND 0.7 ND 87.2 ND 8.1 ND ND ND LLOQ 0.8 2.0 1.3 LLOQ 103.5
Pak choi
 Tokinashi Taisai JP26061 Japan Aug. 25, 2021 - Oct. 12, 2021 ND 4.7 ND ND ND 1.9 ND 17.6 ND 9.1 ND ND ND ND 0.7 1.4 3.3 1.0 39.7
 Kuroba JP26076 Japan Aug. 25, 2021 - Oct. 18, 2021 ND 5.8 ND ND ND 4.4 1.0 7.7 ND 10.7 ND ND ND ND 2.1 1.6 7.0 LLOQ 40.3
 Yukijiro Taisai JP37343 Japan Aug. 25, 2021 - Oct. 12, 2021 ND 3.4 ND ND ND 1.9 LLOQ 14.0 ND 12.5 ND ND ND ND 1.0 1.2 4.9 0.9 39.7
 Choy Sum Ex China 3 JP37372 Malaysia Aug. 25, 2021 - Oct. 5, 2021 ND 5.9 ND ND ND 4.9 LLOQ 38.9 ND 17.9 ND ND ND ND 1.4 0.9 8.0 1.0 78.8
 Fengshan Pai Tsai JP37369 pak choi Aug. 25, 2021 - Oct. 12, 2021 ND 5.7 ND ND ND 1.3 0.9 2.6 ND 3.0 ND ND ND ND 0.5 0.9 1.2 LLOQ 16.0
Turnip leave
 Kanamachi Kokabu JP26852 Japan Aug. 25, 2021 - Oct. 12, 2021 ND 0.8 ND ND ND 0.9 ND 49.4 ND 14.2 ND ND ND LLOQ 1.2 1.0 8.4 0.7 76.6
 Sugukina JP26869 Japan Aug. 25, 2021 - Oct. 12, 2021 ND 2.0 0.8 ND ND 1.0 ND 9.1 ND 2.6 2.4 ND 1.1 ND 0.6 0.6 3.2 ND 23.3
Turnip root
 Kanamachi Kokabu JP26852 Japan Aug. 25, 2021 - Oct. 12, 2021 ND 1.3 LLOQ ND ND 0.9 ND 27.4 ND 6.2 2.7 ND 2.3 2.2 1.9 1.9 9.4 8.9 65.1
 Sugukina JP26869 Japan Aug. 25, 2021 - Oct. 12, 2021 ND 5.8 LLOQ ND ND 0.8 ND 2.7 ND 1.1 9.1 ND 2.9 1.5 1.7 2.4 24.6 6.1 58.9
Choy sum
 Zen Nen Shin JP26935 China Aug. 18, 2022 - Oct. 19, 2022 ND 2.7 ND ND ND 3.1 ND 5.3 ND 4.5 ND ND ND ND 0.6 ND LLOQ LLOQ 16.2
 Za Tiao Da You Cai JP76720 China Aug. 18, 2022 - Oct. 19, 2022 ND 3.4 ND ND ND 3.3 ND 5.1 ND 3.5 ND ND ND LLOQ 1.5 LLOQ 0.9 LLOQ 17.8
 40 Days Choy Sum JP37409 Taiwan Aug. 18, 2022 - Oct. 19, 2022 ND 2.1 ND ND ND 2.2 ND 2.4 ND 1.4 ND ND ND LLOQ 1.9 LLOQ 1.1 LLOQ 11.1
 Choy Sum Ex China 1 JP37411 Malaysia Aug. 18, 2022 - Oct. 19, 2022 ND 4.3 ND ND ND 2.2 ND 2.4 ND 2.0 ND ND ND ND 0.8 LLOQ 0.8 LLOQ 12.5
 Dai Koushi JP43248 China Aug. 18, 2022 - Oct. 19, 2022 ND 1.5 ND ND ND 1.9 ND 2.9 ND 3.0 ND ND ND ND 0.5 LLOQ 0.5 ND 10.2
Chinese cabbage
 Kaga JP26708 Japan Sep. 22, 2022 - Dec. 8, 2022 ND 2.1 ND ND ND 2.0 ND 2.5 ND 3.0 ND ND LLOQ LLOQ 4.9 3.9 0.8 LLOQ 19.3
 Mikawa JP26716 Japan Sep. 22, 2022 - Dec. 8, 2022 ND 1.3 ND ND ND LLOQ ND 0.8 ND 2.1 ND ND ND ND 1.8 2.2 0.4 ND 8.7
 Shouseikou JP26751 China Sep. 22, 2022 - Dec. 2, 2022 ND 1.3 ND ND ND 2.1 ND LLOQ ND 2.5 ND ND 1.4 1.1 0.7 1.6 0.3 1.2 12.2
 Yamagata JP37454 Japan Sep. 22, 2022 - Dec. 8, 2022 ND 3.1 ND ND ND 3.4 ND 1.2 ND 2.9 ND ND 1.6 LLOQ 2.3 2.8 1.1 1.5 19.7
 Tieh Chia 2 JP37532 Taiwan Sep. 22, 2022 - Dec. 2, 2022 ND 1.5 ND ND ND 5.0 ND 1.8 ND 4.4 ND ND 1.3 ND 1.8 2.3 ND 1.9 20.1
a  mg/100 g Fw, values were means (n = 3).

b  ID used in the Research Center of Genetic Resources (i).

c  Cultivation period was defined as the period from the sowing date to the harvest date.

d  Not detectable.

e  Lower limit of quantitation.

GIB: Glucoiberin; PRO: Progoitrin; GRA: Glucoraphanin; GRE: Glucoraphenin; SIN: Sinigrin; GAL: Glucoalyssin; GNL: Gluconapoleiferin; GNA: Gluconapin; GIV: Glucoibervirin GBN: Glucobrassicanapin; GER: Glucoerucin; GPH: Glucoraphasatin; GBE: Glucoberteroin; 4-OHGBS: 4-Hydroxyglucobrassicin; GBS: Glucobrassicin; 4-OMGBS: 4-Methoxyglucobrassicin; NGBS: Neoglucobrassicin; GST: Gluconasturtiin.

Table 4.GSLs profile in B. napus a

B.napus JP No. b Origin Cultivation period c GIB PRO GRA GRE SIN GAL GNL GNA GIV GBN GER GPH GBE 4HGBS GBS 4MGBS NGBS GST Total
Rutabaga
 Green Top JP41103 unkown Aug. 25, 2021 - Oct. 16, 2021 ND d 47.2 2.1 ND ND 7.7 4.1 1.1 ND 4.1 18.0 ND 21.6 1.5 3.8 0.7 7.2 54.5 173.9
 Wilhelmsburger JP76714 USA Aug. 25, 2021 - Nov. 16, 2021 ND 19.0 2.4 ND ND 11.4 ND 3.1 ND LLOQe 6.7 ND 9.2 3.2 6.9 3.5 6.8 11.2 83.5
 Krasnoselskaya JP128131 unkown Aug. 25, 2021 - Nov. 24, 2021 ND 41.2 2.8 ND ND LLOQ ND 1.7 ND ND 6.7 ND ND 1.8 9.4 1.7 17.7 12.6 95.6
 Koukei 10-1 JP26139 Japan Aug. 25, 2021 - Nov. 16, 2021 ND 132.4 2.9 ND ND ND ND 8.0 ND ND 30.9 ND 1.2 1.8 11.9 2.4 15.2 28.0 234.8
 Koukei 11 JP26140 Japan Aug. 25, 2021 - Nov. 16, 2021 ND 78.1 2.3 ND ND ND ND 12.7 ND ND 37.5 ND 1.8 2.7 4.2 2.1 5.5 37.7 184.8
a  mg/100 g Fw, values were means (n = 3).

b  ID used in the Research Center of Genetic Resources (i).

c  Cultivation period was defined as the period from the sowing date to the harvest date.

d  Not detectable.

e  Lower limit of quantitation.

GIB: Glucoiberin; PRO: Progoitrin; GRA: Glucoraphanin; GRE: Glucoraphenin; SIN: Sinigrin; GAL: Glucoalyssin; GNL: Gluconapoleiferin; GNA: Gluconapin; GIV: Glucoibervirin GBN: Glucobrassicanapin; GER: Glucoerucin; GPH: Glucoraphasatin; GBE: Glucoberteroin; 4-OHGBS: 4-Hydroxyglucobrassicin; GBS: Glucobrassicin; 4-OMGBS: 4-Methoxyglucobrassicin; NGBS: Neoglucobrassicin; GST: Gluconasturtiin.

Table 5.GSLs profile in B. juncea a

B. napus JP No. b Origin Cultivation period c GIB PRO GRA GRE SIN GAL GNL GNA GIV GBN GER GPH GBE 4HGBS GBS 4MGBS NGBS GST Total
Leaf mustard
 Karashina JP26131 Japan Aug. 18, 2022 - Oct. 18, 2022 ND d 2.0 ND ND 25.2 ND ND 26.9 ND ND ND ND ND LLOQe LLOQ LLOQ ND LLOQ 54.1
 Kairyou Katsuona JP26135 Japan Aug. 18, 2022 - Oct. 18, 2022 ND ND ND ND 49.7 ND ND 2.4 ND ND ND ND ND LLOQ 0.7 0.7 ND ND 53.5
 Kaichoy (Wainaw Sfrain) JP37394 USA Aug. 18, 2022 - Oct. 18, 2022 ND ND ND ND 50.9 ND ND LLOQ ND ND ND ND ND 0.4 LLOQ 0.5 0.4 ND 52.1
 Pak-85833 JP88051 Pakistan Aug. 18, 2022 - Oct. 18, 2022 ND LLOQ ND ND 19.0 ND ND 73.3 ND ND ND ND ND ND LLOQ ND ND ND 92.2
 Monehyin JP256550 Myanmar Aug. 18, 2022 - Oct. 18, 2022 ND ND ND ND 72.2 ND ND 3.4 ND ND ND ND ND ND 0.6 ND ND ND 76.2
a  mg/100 g Fw, values were means (n = 3).

b  ID used in the Research Center of Genetic Resources (i).

c  Cultivation period was defined as the period from the sowing date to the harvest date.

d  Not detectable.

e  Lower limit of quantitation.

GIB: Glucoiberin; PRO: Progoitrin; GRA: Glucoraphanin; GRE: Glucoraphenin; SIN: Sinigrin; GAL: Glucoalyssin; GNL: Gluconapoleiferin; GNA: Gluconapin; GIV: Glucoibervirin GBN: Glucobrassicanapin; GER: Glucoerucin; GPH: Glucoraphasatin; GBE: Glucoberteroin; 4-OHGBS: 4-Hydroxyglucobrassicin; GBS: Glucobrassicin; 4-OMGBS: 4-Methoxyglucobrassicin; NGBS: Neoglucobrassicin; GST: Gluconasturtiin.

Table 6.GSLs profile in R. sativus a

R.sativus JP No. b Origin Cultivation period c GIB PRO GRA GRE SIN GAL GNL GNA GIV GBN GER GPH GBE 4HGBS GBS 4MGBS NGBS GST Total
Leaf radish
 Kosena Daikon JP76693 Japan Jun. 28, 2021 - Jul. 26, 2021 ND d ND 1.1 3.6 ND ND ND ND ND ND 2.8 158.8 ND 0.3 2.0 0.4 ND ND 168.9
 Bisai JP27118 Japan Jun. 28, 2021 - Jul. 26, 2021 ND ND 1.5 4.8 ND ND ND ND ND ND 3.9 199.2 ND ND 1.0 0.4 ND ND 210.8
 Tousui JP26966 Japan Jun. 28, 2021 - Jul. 26, 2021 ND ND 2.0 3.6 ND ND ND ND ND ND 3.0 172.0 ND 0.3 0.9 0.4 ND ND 182.2
 Viole Tde Hiver De Gournay JP27016 France Jun. 28, 2021 - Jul. 26, 2021 ND ND 3.9 10.7 ND ND ND ND ND ND 1.4 48.1 ND 0.4 1.2 0.4 ND ND 66.1
 Shusen Akadaikon JP133694 China Jun. 28, 2021 - Jul. 26, 2021 ND ND 2.3 3.9 ND ND ND ND ND ND 3.1 227.6 ND 0.4 4.7 1.2 ND ND 243.2
Radish root
 Red Radish Round JP80940 USSR Aug. 25, 2021 - Sep. 27, 2021 ND ND ND 3.6 ND ND ND ND ND ND 3.3 203.7 ND 3.0 1.5 1.5 ND ND 216.6
 Kinmon Akamaru JP27262 China Aug. 25, 2021 - Oct. 5, 2021 ND ND ND 9.4 ND ND ND ND ND ND 1.8 184.8 ND 3.9 1.7 3.9 ND ND 205.5
 Red Prince JP27294 USA Aug. 25, 2021 - Oct. 5, 2021 ND ND ND 10.6 ND ND ND ND ND ND 2.1 238.8 ND 5.6 1.8 3.7 ND ND 262.6
 White Prince JP27295 USA Aug. 25, 2021 - Oct. 5, 2021 ND ND ND 1.6 ND ND ND ND ND ND 0.9 77.7 ND 1.7 0.3 0.9 ND ND 83.1
 Crimson Giant JP27288 USA Aug. 25, 2021 - Oct. 5, 2021 ND ND ND 1.1 ND ND ND ND ND ND 1.9 142.0 ND 2.8 1.2 1.6 ND ND 150.5
a  mg/100 g Fw, values were means (n = 3).

b  ID used in the Research Center of Genetic Resources (i).

c  Cultivation period was defined as the period from the sowing date to the harvest date.

d  Not detectable.

GIB: Glucoiberin; PRO: Progoitrin; GRA: Glucoraphanin; GRE: Glucoraphenin; SIN: Sinigrin; GAL: Glucoalyssin; GNL: Gluconapoleiferin; GNA: Gluconapin; GIV: Glucoibervirin GBN: Glucobrassicanapin; GER: Glucoerucin; GPH: Glucoraphasatin; GBE: Glucoberteroin; 4-OHGBS: 4-Hydroxyglucobrassicin; GBS: Glucobrassicin; 4-OMGBS: 4-Methoxyglucobrassicin; NGBS: Neoglucobrassicin; GST: Gluconasturtiin.

Variations in the GSL profiles in Brassica oleracea  In B. oleracea, 12 GSLs were detected (Table 2). In kale, the predominant GSLs were glucoiberin (GIB), PRO, GRA, SIN, and GBS. Although a high content of GBS was commonly observed in all cultivars, “Marrow Stem” had 17-fold higher GBS than “Shashamane Local 1.” In some cases, the most abundant GSL in one cultivar, such as GRA and SIN, was not detected in another. “Marrow Stem” had about 3-fold higher total GSLs than “Col/Spain/530.” In cabbage, GIB, GRA, SIN, and GBS were abundant, but the most dominant GSLs varied between each cultivar. “Best Of All” had 4-fold higher total GSLs than “Kibakei Sakusesshon.” In kohlrabi, GIB, GRA, and GBS were abundant, and “Yue Nan Pie Lan 1” had the highest GRA content of all Brassicaceae vegetables analyzed in this study. In addition, differences in the GSL composition by origin were also observed: PRO, SIN, and GNA were detected in three cultivars of Chinese origin, whereas they were not detectable in the other cultivars from other origins. For broccoli, two cultivars were measured. It is well known as a vegetable that is high in GRA, which was confirmed in this study, but there were variable differences in the content. In cauliflower, GIB and GBS were commonly found in high amounts, and SIN and glucoibervirin (GIV) also tended to be abundant in the four cultivars except for “Minaret.” “Boushuu Nakate” had about 3-fold higher total GSLs than “Pak-10417.”

Variations in the GSL profiles in Brassica rapa  In B. rapa, 13 GSLs were detected (Table 3). Komatsuna contained higher levels of PRO, GNA, and GBN regardless of the cultivar. There was no significant difference in the GSL profiles among the five cultivars measured in this study. In pak choi, there was a trend toward high levels of PRO, GNA, and GBN, as well as in komatsuna. The GSL content in “Choy Sum Ex China 3” was 4-fold higher than the total GSLs in “Fengshan Pai Tsai.” In turnips, two cultivars were analyzed for their leaves and roots. In the leaves, PRO, GNA, and GBN were abundant; in the roots, in addition to GNA, neoglucobrassicin (NGBS) and GST were abundant. For choy sum and Chinese cabbage, although the GSL contents varied about twice depending on the cultivars, the total GSL content was relatively low compared to the other vegetables. Focusing on the individual GSL, there was a trend toward relatively high levels of PRO, GNA, and GBN, as well as in komatsuna, pak choi, and turnip leaves.

Variations in the GSL profiles in Brassica napus  In B. napus (rutabaga), 13 GSLs were detected the same number as that in B. rapa (Table 4). A high amount of PRO and GST was commonly observed, but there was significant variability among the cultivars for the other GSL profiles. A high GST content was confirmed in the rutabaga, as well as the turnip root. “Koukei 10-1” had about 3-fold higher total GSLs than that of “Wilhelmsburger.”

Variations in the GSL profiles in Brassica juncea  In B. juncea (leaf mustard), 8 GSLs were detected (Table 5). SIN and GNA tended to be abundant, but there were significant differences in content among the cultivars.

Variations in the GSL profiles in Raphanus sativus  In R. sativus, 7 GSLs were detected (Table 6). GPH was the most abundant GSL, followed by the second highest amount of GRE in almost cultivars. Depending on the cultivar, these compounds accounted for 89–97 % of the total GSLs. Focusing on the other minor GSL profiles, there were no significant differences, except for GRA which was only detected in the leaf radish. There were a more than 3-fold difference in the total GSLs between the highest (“Shusen Akadaikon” and “Red Prince”) and lowest (“Violet D'hiver De Gournay” and “White Prince”) GSL cultivars in leaves and roots, respectively.

Discussion

Comparison of identified GSL and GSL profiles with previous studies  We identified the GSL profiles of 14 Brassicaceae vegetables from 64 cultivars grown in Japan. The 18 detected GSLs were those that had already reported in previous studies (Ciska et al., 2000; Verkerk et al., 2009; Chen, 2008; Wu and Pehrsson, 2022; Hill and Williams, 1987; Lee et al., 2021), and the GSL profiles and contents of each vegetable obtained in this study were similar to those obtained in previous studies, but some differences were observed. For example, Ciska et al. (2000) reported the presence of GRE in kohlrabi and cauliflower. However, they were not detected in our data (Table 2) and not for the mass spectrum of desulfo-GRE (m/z = 378.1). Also, glucoberteroin (GBE) was detected in the turnip root in our analysis but was not mentioned in the report by Chen (2007). These discrepancies may be due to differences in the cultivars used in each study, but differences in the analysis methods may also cause them. Some studies have identified GSLs by only comparing RT with commercial standards or another research data. However, doing so may overlook certain GSLs due to overlapping peaks or misidentify them as other GSLs with similar RTs. In fact, when considering the analytical conditions in this study, the separation of GBE, GST, and 4MGBS was found to vary significantly depending on the column temperature, with these peaks overlapping at 40 °C (Fig. 2C). Therefore, when considering the analytical conditions for GSLs in Brassicaceae vegetables, it may be desirable to combine mass spectra with the RT data, as was done in this study.

Fig. 2.

Effect of column temperatures on the separation of GBE, GST, and 4MGBS. Turnip root (“Sugukina”) was analyzed under the same conditions, changing only the column temperatures (A: 40 °C; B: 30 °C; C: 20 °C). GBE, Glucoberteroin; GBS, Glucobrassicin; 4MGBS, 4-Methoxyglucobrassicin; GBN, Glucobrassicanapin; GER, Glucoerucin; GST, Gluconasturtiin.

The GSL detected at RT = 14.2 could not be identified by our analytical method (Fig. 1C). Its mass spectrum showed a peak at m/z = 318.1 (i.e., its molecular weight as GSL is predicated to be 375.1), and detection was between GIV and GBN, suggesting that it may be one of the GSLs reported in previous studies with a butyl, isobutyl, or 1-methylpropyl side chain structure (Osada, 2016). Therefore, its identification requires using NMR to accurately identify these structural isomers, as was done in a previous study (Carl et al., 2016).

Differences in GSL profiles among each Brassicaceae vegetable  Brassicaceae vegetables are known to contain genus specific GSLs such as glucosativin and dimeric 4-mercaptobutyl GSLs in the genus Eruca and sinalbin in the genus Sinapis (Bell et al., 2015; Agerbink et al., 2008). The GRE and GPH detected in the genus Raphanus analyzed in this study were not detected in other vegetables of the genus Brassica, indicating that they are likely to be genus specific GSLs. On the other hand, comparing the GSL profiles within the Brassica genus, GIB, GRA, SIN, and GBS were abundant in B. oleracea (Table 2); PRO, GNA, and GBN in B. rapa (Table 3); PRO and GST in B. napus (Table 4); and SIN and GNA in B. juncea (Table 5). To confirm these trends, PCA was performed on the profile of 16 GSLs detected in the 4 Brassica species, which revealed that principal components (PC) 1 and PC2 explained 24.3 and 18.7 % of the total variance, respectively (Fig. 3A). Focusing on the loading plots, the abundant GSLs in each species, such as GIB and GBS in B. oleracea and GNA and GBN in B.rapa, highly contributed to either or both PC1 and PC2 (Fig. 3B). In addition, some GSLs were located close together, such as GIV, GIB, and SIN with side chains of 3 carbons, and GNA and GBN with alkene side chains, share a common chemical feature. Therefore, these features may characterize the GSL profile of each Brassica species.

Fig. 3.

Scores (A) and loading plots (B) of PC 1 and 2 of the principal component analysis results obtained from GSL profiles on 4 Brassica vegetables. GIB, Glucoiberin; PRO, Progoitrin; GRA, Glucoraphanin; GRE, Glucoraphenin; SIN, Sinigrin; GAL, Glucoalyssin; GNL, Gluconapoleiferin; GNA, Gluconapin; GIV, Glucoibervirin; GBN, Glucobrassicanapin; GER, Glucoerucin; GBE, Glucoberteroin; 4HGBS, 4-Hydroxyglucobrassicin; GBS, Glucobrassicin; 4MGBS, 4-Methoxyglucobrassicin; NGBS, Neoglucobrassicin; GST, Gluconasturtiin.

The highest GRA content was 35.8 mg/100 g FW in “Yue Nan Pie Lan 1.” SIN, GBS, and GST had the highest content of 72.2 mg/100 g FW (“Monehyin”), 48.0 mg/100 g FW (“B Oleracea O 179”), and 54.5 mg/100 g FW (“Green Top”), respectively, as the other GSLs with reported functionality (Staack et al., 1998). Some epidemiological studies have indicated that the consumption of Brassicaceae vegetables effectively reduces the risk of cancer (Verhoeveb et al., 1996; Higdon et al., 2007). In addition, a study on Japanese participants reported a decrease in the rate of lung cancer due to the consumption of cabbage (Gao et al., 1993). Therefore, vegetables with higher GSL content, such as “Yue Nan Pie Lan 1” and “Monehyin” may have health-promoting effects at lower levels of intakes. In contrast, cabbage with its high levels of SIN and PRO was reported to have an increased bitterness and more pungency (Drewnowski et al., 2000). Because these characteristics are undesirable to consumers basically, it would be better to focus not only on the functionality but also taste when breeding with a focus on GSLs.

Differences in GSL profiles among cultivars and between tissues of the same Brassicaceae vegetables  Focusing on the total GSL content among the cultivars of the same Brassicaceae vegetable, a 3- to 4-fold difference was frequently observed between the cultivars with the highest and lowest GSL content. Furthermore, individual GSL content also differed among the cultivars, with a stronger trend in B. oleracea, as mentioned in previous reviews (Ishida et al., 2014; Cartea and Velasco, 2008). Because each Brassicaceae vegetable used in this study, except for cabbage and cauliflower, was grown under the same conditions during the same period, these differences may be attributed to their genetic background. The biosynthesis pathway of GSLs, including the related genes, was clarified by studies using Arabidopsis thaliana (Grubb and Abel, 2006; Sønderby et al., 2010). Therefore, to investigate the cause of the differences among the cultivars, it is necessary to study the details of the gene expression involved in the GSL biosynthesis pathway. This variation in GSL composition among the cultivars may be challenging to accurately determine the GSL intake for epidemiological studies. To conduct a more accurate survey, it is necessary to create a more extensive dataset of the commonly distributed cultivars of the Brassicaceae vegetables of interest, as in previous studies (McNaughton and Marks, 2003; Wu and Pehrsson, 2022).

The results for the radish and turnip showed that the GSL profiles varied depending on the tissues, even within the same cultivar, which is consistent with previous reports (Ko et al., 2017; Bhandari et al., 2015). However, these studies reported that higher GLS levels were observed in the roots than the leaves among some vegetables, but this was not the same as our results. Concerning radishes, many studies have focused on the root (Ishida et al., 2012; Yi et al., 2016), which is the central edible part, and a few have analyzed the GSLs in the leaves in detail (Lee et al., 2021). Meanwhile, it was clarified that the leaf radish measured in this study had as many GSLs as the roots. It is probably related to the reason that of the leaf radish analyzed in this study, “Kosena Daikon” and “Bisai” were cultivars whose leaves, and not the roots, are edible, whereas others were harvested to use their leaves but before the roots had matured. Furthermore, in the GSL biosynthesis pathway, GER is a common precursor of GRA and GPH.

Because GER and GRA are increased in radish leaves which lack 2-oxoglutarate-dependent dioxygenase (Kakizaki et al., 2017), the enzyme that converts GER into GPH, and thus enhancing GRA in leaf radish may be a potential breeding target.

Acknowledgements  We thank Drs. Nobuko Fukino and Mitsuyo Kawasaki (Institute of Vegetable and Floriculture Science, NARO) for offering the Brassicaceae vegetables used in this study.

Conflict of interest  There are no conflicts of interest to declare.

Supplemental Table 1
Detailed information of GSL profiles in Brassicaceae vegetables a

B.oleracea JP No. b Origin Cultivation period c GIB PRO GRA GRE SIN GAL GNL GNA GIV GBN GER GPH GBE 4-OHGBS GBS 4-OMGBS NGBS GST Total GSL
 Kale
   Shashamane Local 1 JP26040 Ethiopia Aug. 31, 2021 – Oct. 18, 2021 ND d LLOQ e ND ND 25.6 ± 3.5 a ND ND ND ND ND ND ND ND 0.6 ± 0.1 ab 2.8 ± 0.3 b 2.2 ± 0.1 a 0.6 ± 0.2 b ND 31.9 ± 4.1 b
   Knol Khol JP26050 India Aug. 31, 2021 – Oct. 18, 2021 6.8 ± 0.5 a LLOQ 21.0 ± 1.1 * ND ND ND ND ND ND ND ND ND ND 0.8 ± 0.0 ab 14.2 ± 2.1 b 1.0 ± 0.0 c 1.1 ± 0.3 b ND 45.0 ± 3.4 ab
   Marrow Stem JP26055 UK Aug. 31, 2021 – Oct. 18, 2021 6.5 ± 1.4 a LLOQ LLOQ ND 4.0 ± 0.6 b ND ND ND ND ND ND ND ND 0.6 ± 0.1 ab 47.4 ± 2.6 a 1.6 ± 0.1 b 2.3 ± 0.5 a ND 62.4 ± 2.3 a
   B Oleracea O 179 JP26060 Turkey Aug. 31, 2021 – Oct. 18, 2021 2.1 ± 1.4 b 1.6 ± 0.2 6.3 ± 1.8 ND LLOQ ND ND ND ND ND ND ND ND 0.9 ± 0.2 a 48.0 ± 11.9 a 1.5 ± 0.0 b 1.5 ± 0.2 ab ND 61.8 ± 14.5 a
   Col/Spain/530 JP45942 Spain Aug. 31, 2021 – Oct. 18, 2021 4.1 ± 0.9 ab LLOQ ND ND ND 5.2 ± 0.9 b ND ND ND ND ND ND ND ND 0.4 ± 0.1 b 10.0 ± 1.0 b 0.6 ± 0.0 a 1.7 ± 0.5 ab ND 22.0 ± 3.3 b
 Cabbage
   Red Eekaa JP25989 unkown Aug. 31, 2021 – Feb. 2, 2022 1.8 ± 0.4 d 20.2 ± 5.8 a 27.2 ± 4.1 a ND 1.5 ± 0.2 c ND ND 10.6 ± 2.2 a ND ND ND ND ND 1.7 ± 0.3 b 8.1 ± 1.1 bc 1.3 ± 0.2 b LLOQ ND 72.4 ± 8.6 b
   Oogata Kopenhaagen Marketto JP25994 Japan Aug. 31, 2021 – Nov. 24, 2021 6.3 ± 3.0 cd 11.9 ± 6.2 ab 6.8 ± 2.2 b ND 7.3 ± 1.8 bc ND ND 8.5 ± 4.0 ab LLOQ ND ND ND ND LLOQ 6.2 ± 1.6 c 1.1 ± 0.0 b 0.1 ± 0.0 b ND 48.2 ± 13.3 bc
   Kibakei Sakusesshon JP26020 Japan Aug 31, 2021 – Dec. 7, 2021 10.9 ± 2.9 bc LLOQ ND ND 3.0 ± 0.7 c ND ND ND ND ND ND ND ND 0.2 ± 0.1 c 12.4 ± 2.7 b 1.2 ± 0.1 b 0.2 ± 0.0 b ND 27.9 ± 6.4 c
   Youshin JP26027 Japan Aug. 31, 2021 – Nov. 24, 2021 18.0 ± 1.7 b 3.2 ± 1.3 b 2.7 ± 0.5 b ND 16.0 ± 6.8 b ND ND 0.9 ± 0.6 b LLOQ ND ND ND ND 0.2 ± 0.0 c 8.5 ± 1.3 bc 1.4 ± 0.1 ab 0.2 ± 0.0 b ND 51.0 ± 8.0 bc
   Best Of All JP26031 UK Aug. 31, 2021 – Feb. 2, 2022 48.5 ± 4.1 a 5.2 ± 0.8 b 2.2 ± 0.1 b ND 47.0 ± 3.8 a ND ND 1.2 ± 0.2 b 2.4 ± 0.4 ND ND ND ND 3.2 ± 0.2 a 23.0 ± 1.9 a 1.8 ± 0.1 a 0.4 ± 0.0 a ND 134.9 ± 5.0 a
 Kohlrabi
   Yue Nan Pie Lan 1 JP80075 China Aug 2, 2022 – Nov. 14, 2022 1.8 ± 1.3 b 10.4 ± 3.5 a 35.8 ± 4.4 a ND 1.2 ± 0.9 b ND ND 4.6 ± 3.4 ND ND 17.3 ± 8.0 a ND ND 1.5 ± 0.3 a 10.3 ± 8.7 a LLOQ 2.0 ± 0.1 a 7.3 ± 7.5 92.2 ± 8.9 a
   Yuan Pie Lan JP80076 China Aug. 2, 2022 – Nov. 7, 2022 4.9 ± 0.7 b 5.5 ± 1.8 a 18.9 ± 1.0 b ND 2.0 ± 0.5 b ND ND LLOQ LLOQ ND 7.3 ± 1.5 a ND ND 0.8 ± 0.2 a 5.2 ± 1.3 a LLOQ 5.5 ± 1.0 a LLOQ 50.2 ± 2.1 b
   Fumei JP127521 China Aug. 2, 2022 – Nov. 7, 2022 4.9 ± 3.8 b 10.9 ± 3.3 a 14.8 ± 3.6 b ND 5.4 ± 3.9 a ND ND 4.8 ± 1.3 LLOQ ND 7.2 ± 2.2 a ND ND 1.5 ± 0.4 a 3.1 ± 1.0 a LLOQ 4.3 ± 2.6 a 2.0 ± 1.5 58.8 ± 9.4 b
   86-703 JP127522 Nepal Aug. 2, 2022 – Nov. 1, 2022 5.5 ± 2.2 b ND 9.1 ± 2.6 b ND ND ND ND ND 4.1 ± 2.0 ND 12.0 ± 3.5 a ND ND 1.1 ± 0.1 a 1.4 ± 0.4 a LLOQ 4.3 ± 1.1 a ND 37.5 ± 11.0 b
   Wener Witte JP127603 Netherlands Aug 2, 2022 – Nov. 1, 2022 18.1 ± 2.8 a ND 10.6 ± 6.8 b ND ND ND ND ND 4.6 ± 1.8 ND 3.8 ± 2.0 a ND ND 1.0 ± 0.3 a 2.7 ± 0.6 a LLOQ 3.0 ± 0.4 a ND 43.8 ± 5.8 b
 Broccoli
   Nakazato Broccoli JP26686 Japan Aug. 2, 2022 – Nov. 29, 2022 LLOQ ND 34.3 ±8.1 * ND ND ND ND ND ND ND ND ND ND 2.5 ± 0.9 * 22.5 ± 7.1 3.1 ± 0.1 6.9 ± 1.7 * ND 69.3 ± 4.2 *
   5R JP88079 Poland Aug. 2, 2022 – Nov. 29, 2022 3.8 ± 0.2 ND 13.4 ± 1.1 ND ND ND ND ND ND ND ND ND ND 1.9 ± 0.2 23.7 ± 5.2 2.7 ± 0.3 53.4 ± 8.3 ND 99.0 ± 13.2
 Cauliflower
   Boushuu Nakate JP26673 Japan Aug. 2, 2022 – Feb. 6, 2023 16.8 ± 1.3 ab LLOQ LLOQ ND 23.0 ± 3.8 a ND ND ND 3.7 ± 0.8 ab ND ND ND ND 1.1 ± 0.5 bc 17.9 ± 3.9 a 0.8 ± 0.3 a 2.1 ± 1.1 ab ND 65.5 ± 6.6 a
   Pak-10417 JP86202 Pakistan Aug. 2, 2022 – Nov. 14, 2022 6.1 ± 1.4 bc ND ND ND 7.5 ± 2.9 b ND ND ND 4.2 ± 0.7 ab ND ND ND ND 0.6 ± 0.1 c 8.0 ± 1.4 b LLOQ LLOQ ND 26.4 ± 6.1 b
   Minaret JP182453 unkown Aug. 2, 2022 – Jan. 10, 2023 3.4 ± 2.7 c ND 5.7 ± 2.0 ND ND ND ND ND ND ND ND ND ND 8.5 ± 0.4 a 8.7 ± 2.7 b 1.2 ± 0.4 a 3.0 ± 0.6 a ND 30.5 ± 3.3 b
   Boushuu Wase JP26674 Japan Aug 2, 2022 – Dec. 15, 2023 18.3 ± 2.9 ab ND LLOQ ND 4.1 ± 2.4 b ND ND ND 2.3 ± 0.6 b ND ND ND ND 2.1 ± 0.3 bc 12.2 ± 1.9 ab 0.7 ± 0.1 a 1.1 ± 0.3 ab ND 40.7 ± 4.7 b
   Boushuu Chuusei JP37405 Japan Aug. 2, 2022 – Feb. 22, 2023 21.5 ± 7.7 a ND LLOQ ND 18.3 ± 3.8 a ND ND ND 6.3 ± 1.7 a ND ND ND ND 2.6 ± 0.8 b 13.3 ± 0.8 ab 1.5 ± 0.3 a 0.8 ± 0.2 b ND 64.3 ± 10.3 a
B.rapa JP No. Origin Cultivation period GIB PRO GRA GRE SIN GAL GNL GNA GIV GBN GER GPH GBE 4-OHGBS GBS 4-OMGBS NGBS GST Total GSL
 Komatsuna
   Junkei Nozawana JP26903 Japan Jun. 28, 2021 – Aug. 16, 2021 ND 4.1 ± 1.3 b 1.5 ± 0.6 ND ND 1.6 ± 0.4 ab ND 56.1 ± 5.2 b ND 7.2 ± 2.5 a 3.5 ± 2.3 ND 1.0 ± 0.4 ND 1.0 ± 0.3 a 1.1 ± 0.3 c 2.5 ± 0.6 ab ND 79.8 ± 4.9 a
   Shin Kuro Mizuna JP26907 Japan Jun. 28, 2021 – Aug. 16, 2021 ND 18.2 ± 5.7 a 0.8 ± 0.4 ND ND 3.2 ± 1.3 a LLOQ 55.2 ± 4.1 b ND 13.4 ± 1.9 a ND ND ND ND 0.3 ± 0.0 b 1.6 ± 0.1 ab 2.4 ± 0.7 ab 1.3 ± 0.2 * 96.4 ± 6.7 a
   Maruba Komatsuna JP37635 Japan Jun. 28, 2021 – Aug. 16, 2021 ND 2.7 ± 1.3 b ND ND ND 0.7 ± 0.3 b ND 65.5 ± 6.4 b ND 12.1 ± 5.3 a ND ND ND ND 0.4 ± 0.1 b 0.9 ± 0.1 c 1.2 ± 0.1 b 0.6 ± 0.2 84.2 ± 7.9 a
   Uguisuna JP43250 Japan Jun. 28, 2021 – Aug. 16, 2021 ND 5.4 ± 0.5 b LLOQ ND ND 3.5 ± 0.5 a ND 65.3 ± 4.4 b ND 14.2 ± 0.5 a LLOQ ND 0.7 ± 0.2 ND 0.7 ± 0.1 ab 1.3 ± 0.1 bc 4.6 ± 1.1 a LLOQ 95.5 ± 7.0 a
   Toukyou Aona JP37637 Japan Jun. 28, 2021 – Aug. 16, 2021 ND 3.4 ± 1.2 b ND ND ND 0.7 ± 0.1 b ND 87.2 ± 9.3 a ND 8.1 ± 1.2 a ND ND ND LLOQ 0.8 ± 0.1 ab 2.0 ± 0.1 a 1.3 ± 0.9 b LLOQ 103.5 ± 9.6 a
 Pak choi
   Tokinashi Taisai JP26061 Japan Aug. 25, 2021 – Oct. 12, 2021 ND 4.7 ± 0.3 a ND ND ND 1.9 ± 0.4 a ND 17.6 ± 1.8 b ND 9.1 ± 1.3 b ND ND ND ND 0.7 ± 0.1 c 1.4 ± 0.2 ab 3.3 ± 0.4 cd 1.0 ± 0.2 a 39.7 ± 2.1 b
   Kuroba JP26076 Japan Aug. 25, 2021 – Oct. 18, 2021 ND 5.8 ± 0.9 a ND ND ND 4.4 ± 2.5 a 1.0 ± 0.0 7.7 ± 1.2 bc ND 10.7 ± 1.0 b ND ND ND ND 2.1 ± 0.1 a 1.6 ± 0.1 a 7.0 ± 0.9 ab LLOQ 40.3 ± 4.6 b
   Yukijiro Taisai JP37343 Japan Aug. 25, 2021 – Oct. 12, 2021 ND 3.4 ± 0.5 a ND ND ND 1.9 ± 0.7 a LLOQ 14.0 ± 2.3 bc ND 12.5 ± 1.7 b ND ND ND ND 1.0 ± 0.3 bc 1.2 ± 0.3 ab 4.9 ± 0.1 bc 0.9 ± 0.0 a 39.7 ± 4.1 b
   Choy Sum Ex China 3 JP37372 Malaysia Aug. 25, 2021 – Oct. 5, 2021 ND 5.9 ± 0.6 a ND ND ND 4.9 ± 1.1 a LLOQ 38.9 ± 8.8 a ND 17.9 ± 1.9 a ND ND ND ND 1.4 ± 0.2 b 0.9 ± 0.1 b 8.0 ± 1.2 a 1.0 ± 0.2 a 78.8 ± 8.9 a
   Fengshan Pai Tsai JP37369 Taiwan Aug. 25, 2021 – Oct. 12, 2021 ND 5.7 ± 1.1 a ND ND ND 1.3 ± 0.4 a 0.9 ± 0.3 2.6 ± 0.5 c ND 3.0 ± 0.1 c ND ND ND ND 0.5 ± 0.1 c 0.9 ± 0.0 b 1.2 ± 0.2 d LLOQ 16.0 ± 1.8 c
 Turnip leave
   Kanamachi Kokabu JP26852 Japan Aug. 25, 2021 – Oct. 12, 2021 ND 0.8 ±0.1 * ND ND ND 0.9 ± 0.2 ND 49.4 ± 1.6 * ND 14.2 ± 3.1 * ND ND ND LLOQ 1.2 ± 0.1 * 1.0 ± 0.1 * 8.4 ± 1.0 * 0.7 ± 0.1 76.6 ± 5.0 *
   Sugukina JP26869 Japan Aug. 25, 2021 – Oct. 12, 2021 ND 2.0 ± 0.5 0.8 ± 0.6 ND ND 1.0 ± 0.3 ND 9.1 ± 2.4 ND 2.6 ± 0.6 2.4 ± 1.7 ND 1.1 ± 1.0 ND 0.6 ± 0.2 0.6 ± 0.1 3.2 ± 0.3 ND 23.3 ± 2.9
 Turnip root
   Kanamachi Kokabu JP26852 Japan Aug. 25, 2021 – Oct. 12, 2021 ND 1.3 ± 0.3 * LLOQ ND ND 0.9 ± 0.4 ND 27.4 ± 0.6 * ND 6.2 ± 2.0 * 2.7 ± 0.3 ND 2.3 ± 0.6 2.2 ± 0.2 1.9 ± 0.6 1.9 ± 0.2 9.4 ± 1.0 * 8.9 ± 1.8 65.1 ± 5.7
   Sugukina JP26869 Japan Aug. 25, 2021 – Oct. 12, 2021 ND 5.8 ± 1.3 LLOQ ND ND 0.8 ± 0.7 ND 2.7 ± 0.9 ND 1.1 ± 0.4 9.1 ± 4.9 ND 2.9 ± 1.0 1.5 ± 0.5 1.7 ± 0.3 2.4 ± 0.2 24.6 ± 3.3 6.1 ± 1.6 58.9 ± 6.0
 Choy sum
   Zen Nen Shin JP26935 China Aug. 18, 2022 – Oct. 19, 2022 ND 2.7 ± 0.3 a ND ND ND 3.1 ± 0.1 a ND 5.3 ± 1.0 a ND 4.5 ± 0.4 a ND ND ND ND 0.6 ± 0.0 a ND LLOQ LLOQ 16.2 ± 1.0 a
   Za Tiao Da You Cai JP76720 China Aug. 18, 2022 – Oct. 19, 2022 ND 3.4 ± 0.6 a ND ND ND 3.3 ± 0.6 a ND 5.1 ± 2.2 a ND 3.5 ± 1.9 a ND ND ND LLOQ 1.5 ± 1.2 a LLOQ 0.9 ± 0.9 a LLOQ 17.8 ± 3.2 a
   40 Days Choy Sum JP37409 Taiwan Aug. 18, 2022 – Oct. 19, 2022 ND 2.1 ± 1.1 a ND ND ND 2.2 ± 0.7 a ND 2.4 ± 1.3 a ND 1.4 ± 1.9 a ND ND ND LLOQ 1.9 ± 1.0 a LLOQ 1.1 ± 0.8 a LLOQ 11.1 ± 2.8 a
   Choy Sum Ex China 1 JP37411 MALAYSIA Aug. 18, 2022 – Oct. 19, 2022 ND 4.3 ± 2.2 a ND ND ND 2.2 ± 1.0 a ND 2.4 ± 2.1 a ND 2.0 ± 2.8 a ND ND ND ND 0.8 ± 0.3 a LLOQ 0.8 ± 0.5 a LLOQ 12.5 ± 6.0 a
   Dai Koushi JP43248 China Aug. 18, 2022 – Oct. 19, 2022 ND 1.5 ± 0.8 a ND ND ND 1.9 ± 0.7 a ND 2.9 ± 0.3 a ND 3.0 ± 0.5 a ND ND ND ND 0.5 ± 0.0 a LLOQ 0.5 ± 0.1 a ND 10.2 ± 2.2 a
 Chinese cabbage
   Kaga JP26708 Japan Sep. 22, 2022 – Dec. 8, 2022 ND 2.1 ± 1.0 a ND ND ND 2.0 ± 0.7 a ND 2.5 ± 1.8 a ND 3.0 ± 0.7 a ND ND LLOQ LLOQ 4.9 ± 0.4 a 3.9 ± 0.2 a 0.8 ± 0.3 ab LLOQ 19.3 ± 2.0 a
   Mikawa JP26716 Japan Sep. 22, 2022 – Dec. 8, 2022 ND 1.3 ± 0.9 a ND ND ND LLOQ ND 0.8 ± 0.4 a ND 2.1 ± 0.8 a ND ND ND ND 1.8 ± 0.5 b 2.2 ± 0.2 bc 0.4 ± 0.1 ab ND 8.7 ± 2.3 b
   Shouseikou JP26751 China Sep. 22, 2022 – Dec. 2, 2022 ND 1.3 ± 0.8 a ND ND ND 2.1 ± 0.5 a ND LLOQ ND 2.5 ± 0.7 a ND ND 1.4 ± 0.3 a 1.1 ± 0.2 0.7 ± 0.3 b 1.6 ± 0.3 c 0.3 ± 0.2 b 1.2 ± 0.4 a 12.2 ± 2.6 ab
   Yamagata JP37454 Japan Sep. 22, 2022 – Dec. 8, 2022 ND 3.1 ± 0.3 a ND ND ND 3.4 ± 1.4 a ND 1.2 ± 0.6 a ND 2.9 ± 0.2 a ND ND 1.6 ± 0.5 a LLOQ 2.3 ± 0.5 b 2.8 ± 0.2 b 1.1 ± 0.2 a 1.5 ± 0.2 a 19.7 ± 1.1 a
   Tieh Chia 2 JP37532 Taiwan Sep. 22, 2022 – Dec. 2, 2022 ND 1.5 ± 0.9 a ND ND ND 5.0 ± 2.6 a ND 1.8 ± 0.3 a ND 4.4 ± 1.2 a ND ND 1.3 ± 0.8 a ND 1.8 ± 0.9 b 2.3 ± 0.4 bc ND 1.9 ± 0.6 a 20.1 ± 5.5 a
B.napus JP No. Origin Cultivation period GIB PRO GRA GRE SIN GAL GNL GNA GIV GBN GER GPH GBE 4-OHGBS GBS 4-OMGBS NGBS GST Total GSL
 Rutabaga
   Green Top JP41103 unkown Aug. 25, 2021 – Oct. 16, 2021 ND 47.2 ± 14.0 c 2.1 ± 0.1 a ND ND 7.7 ± 1.2 * 4.1 ± 0.8 1.1 ± 0.6 c ND 4.1 ± 1.4 18.0 ± 8.1 bc ND 21.6 ± 7.3 a 1.5 ± 0.2 c 3.8 ± 1.3 b 0.7 ± 0.1 c 7.2 ± 1.7 bc 54.5 ± 22.7 a 173.9 ± 52.6 ab
   Wilhelmsburger JP76714 USA Aug. 25, 2021 – Nov. 16, 2021 ND 19.0 ± 1.0 d 2.4 ± 0.1 a ND ND 11.4 ± 0.8 ND 3.1 ± 0.3 c ND LLOQ 6.7 ± 0.2 c ND 9.2 ± 0.3 b 3.2 ± 0.3 a 6.9 ± 1.7 ab 3.5 ± 0.6 a 6.8 ± 1.2 c 11.2 ± 1.5 b 83.5 ± 6.2 c
   Krasnoselskaya JP128131 unkown Aug. 25, 2021 – Nov. 24, 2021 ND 41.2 ± 8.6 cd 2.8 ± 0.5 a ND ND LLOQ ND 1.7 ± 1.6 c ND ND 6.7 ± 1.6 c ND ND 1.8 ± 0.3 bc 9.4 ± 3.5 ab 1.7 ± 0.4 bc 17.7 ± 4.9 a 12.6 ± 0.3 b 95.6 ± 19.1 bc
   Koukei 10-1 JP26139 Japan Aug. 25, 2021 – Nov. 16, 2021 ND 132.4 ± 2.9 a 2.9 ± 0.3 a ND ND ND ND 8.0 ± 0.4 b ND ND 30.9 ± 1.9 ab ND 1.2 ± 0.0 b 1.8 ± 0.1 bc 11.9 ± 0.3 a 2.4 ± 0.0 ab 15.2 ± 0.5 ab 28.0 ± 1.0 ab 234.8 ± 5.3 a
   Koukei 11 JP26140 Japan Aug. 25, 2021 – Nov. 16, 2021 ND 78.1 ± 8.3 b 2.3 ± 0.1 a ND ND ND ND 12.7 ± 1.1 a ND ND 37.5 ± 3.7 a ND 1.8 ± 0.1 b 2.7 ± 0.5 ab 4.2 ± 1.2 b 2.1 ± 0.8 ac 5.5 ± 1.8 c 37.7 ± 6.4 ab 184.8 ± 21.1 a
B. juncea JP No. Origin Cultivation period GIB PRO GRA GRE SIN GAL GNL GNA GIV GBN GER GPH GBE 4-OHGBS GBS 4-OMGBS NGBS GST Total GSL
 Leaf mustard
   Karashina JP26131 Japan Aug. 18, 2022 – Oct. 18, 2022 ND 2.0 ± 1.0 ND ND 25.2 ± 3.2 b ND ND 26.9 ± 0.6 b ND ND ND ND ND LLOQ LLOQ LLOQ ND LLOQ 54.1 ± 4.4 c
   Kairyou Katsuona JP26135 Japan Aug. 18, 2022 – Oct. 18, 2022 ND ND ND ND 49.7 ± 4.2 ab ND ND 2.4 ± 0.2 c ND ND ND ND ND LLOQ 0.7 ± 0.1 0.7 ± 0.1 * ND ND 53.5 ± 4.3 c
   Kaichoy(Wainaw Sfrain) JP37394 USA Aug. 18, 2022 – Oct. 18, 2022 ND ND ND ND 50.9 ± 3.7 b ND ND LLOQ ND ND ND ND ND 0.4 ± 0.1 LLOQ 0.5 ± 0.0 0.4 ± 0.1 ND 52.1 ± 3.8 c
   Pak-85833 JP88051 Pakistan Aug. 18, 2022 – Oct. 18, 2022 ND LLOQ ND ND 19.0 ± 1.5 b ND ND 73.3 ± 1.9 a ND ND ND ND ND ND LLOQ ND ND ND 92.2 ± 2.9 a
   Monehyin JP256550 Myanmar Aug. 18, 2022 – Oct. 18, 2022 ND ND ND ND 72.2 ± 5.0 a ND ND 3.4 ± 0.3 c ND ND ND ND ND ND 0.6 ± 0.0 ND ND ND 76.2 ± 5.3 b
R.sativus JP No. Origin Cultivation period GIB PRO GRA GRE SIN GAL GNL GNA GIV GBN GER GPH GBE 4-OHGBS GBS 4-OMGBS NGBS GST Total GSL
 Leaf radish
   Kosena Daikon JP76693 Japan Jun. 28, 2021 – Jul. 26, 2021 ND ND 1.1 ± 0.2 c 3.6 ± 0.2 b ND ND ND ND ND ND 2.8 ± 0.2 b 158.8 ± 6.2 b ND 0.3 ± 0.0 a 2.0 ± 0.3 b 0.4 ± 0.0 b ND ND 168.9 ± 6.9 b
   Bisai JP27118 Japan Jun. 28, 2021 – Jul. 26, 2021 ND ND 1.5 ± 0.1 bc 4.8 ± 0.2 b ND ND ND ND ND ND 3.9 ± 0.2 a 199.2 ± 28.1 ab ND ND 1.0 ± 0.1 bc 0.4 ± 0.0 b ND ND 210.8 ± 28.5 ab
   Tousui JP26966 Japan Jun. 28, 2021 – Jul. 26, 2021 ND ND 2.0 ± 0.1 b 3.6 ± 1.0 b ND ND ND ND ND ND 3.0 ± 0.5 ab 172.0 ± 21.2 ab ND 0.3 ± 0.0 a 0.9 ± 0.3 c 0.4 ± 0.0 b ND ND 182.2 ± 19.6 b
   Viole Tde Hiver De Gournay JP27016 France Jun. 28, 2021 – Jul. 26, 2021 ND ND 3.9 ± 0.5 a 10.7 ± 1.5 a ND ND ND ND ND ND 1.4 ± 0.2 c 48.1 ± 6.4 c ND 0.4 ± 0.1 a 1.2 ± 0.1 bc 0.4 ± 0.1 b ND ND 66.1 ± 8.6 c
   Shusen Akadaikon JP133694 China Jun. 28, 2021 – Jul. 26, 2021 ND ND 2.3 ± 0.3 b 3.9 ± 0.5 b ND ND ND ND ND ND 3.1 ± 0.4 ab 227.6 ± 14.0 a ND 0.4 ± 0.1 a 4.7 ± 0.5 a 1.2 ± 0.1 a ND ND 243.2 ± 14.7 a
 Radish root
   Red Radish Round JP80940 USSR Aug. 25, 2021 – Sep. 27, 2021 ND ND ND 3.6 ± 0.1 b ND ND ND ND ND ND 3.3 ± 0.2 a 203.7 ± 8.0 ab ND 3.0 ± 0.4 bc 1.5 ± 0.5 a 1.5 ± 0.1 b ND ND 216.6 ± 8.2 ab
   Kinmon Akamaru JP27262 China Aug. 25, 2021 – Oct. 5, 2021 ND ND ND 9.4 ± 1.2 a ND ND ND ND ND ND 1.8 ± 0.2 b 184.8 ± 33.8 ab ND 3.9 ± 1.2 ab 1.7 ± 0.1 a 3.9 ± 0.1 a ND ND 205.5 ± 36.1 ab
   Red Prince JP27294 USA Aug 25, 2021 – Oct. 5, 2021 ND ND ND 10.6 ± 0.9 a ND ND ND ND ND ND 2.1 ± 0.1 b 238.8 ± 19.6 ab ND 5.6 ± 0.2 ab 1.8 ± 0.2 a 3.7 ± 0.3 a ND ND 262.6 ± 20.2 ab
   White Prince JP27295 USA Aug 25, 2021 – Oct. 5, 2021 ND ND ND 1.6 ± 0.4 bc ND ND ND ND ND ND 0.9 ± 0.1 c 77.7 ± 8.6 c ND 1.7 ± 0.0 c 0.3 ± 0.1 b 0.9 ± 0.1 c ND ND 83.1 ± 8.9 c
   Crimson Giant JP27288 USA Aug. 25, 2021 – Oct. 5, 2021 ND ND ND 1.1 ± 0.4 c ND ND ND ND ND ND 1.9 ± 0.2 b 142.0 ± 18.6 bc ND 2.8 ± 0.4 bc 1.2 ± 0.4 ab 1.6 ± 0.1 b ND ND 150.5 ± 19.6 bc
a  mg/100 g Fw, values are means &#00B1; SD (n = 3).

b  ID used in the Research Center of Genetic Resources (i).

c  Cultivation period was defined as the period from the sowing date to the harvest date.

d  Not detectable.

e  Lower limit of quantitation.

Different letters within each vegetable indicate a significant difference (p<0.05, Tukey's HSD test). The asterisk (*) indicate significant differences between two samples (p<0.05, t -test).

GIB: Glucoiberin; PRO: Progoitrin; GRA: Glucoraphanin; GRE: Glucoraphenin; SIN: Sinigrin; GAL: Glucoalyssin; GNL: Gluconapoleiferin; GNA: Gluconapin; GIV: Glucoibervirin; GBN: Glucobrassicanapin; GER: Glucoerucin; GPH: Glucoraphasatin; GBE: Glucoberteroin; 4-OHGBS: 4-Hydroxyglucobrassicin; GBS: Glucobrassicin; 4-OMGBS: 4-Methoxyglucobrassicin; NGBS: Neoglucobrassicin; GST: Gluconasturtiin.

References
 
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