Genetic variation in leaf and stem glucosinolates in resynthesized lines of winter rapeseed (<Emphasis Type="Italic">Brassica napus</Emphasis> L.)

Glucosinolates are secondary components characteristic for the Brassicaceae with complex biological functions. Glucosinolates in the seeds are anti-nutritive when feeding animals and their inheritance have been extensively investigated. Much less is known about the genetics of glucosinolates in leaves and stems, which may attract some insects, while repelling others. They may also inhibit bacterial processes of importance when using green biomass for the production of biogas. The objective of this study was to analyse the genetic variation of total and individual glucosinolates in the green material of rapeseed. For this 28 resynthesized winter rapeseed lines were tested at two locations. There was a large variation in leaf glucosinolate content between 0.10 and 4.75 μmol/g dry matter. The predominant leaf glucosinolates are the alkenyle glucosinolates progoitrin, gluconapin and glucobrassicanapin. The line R53 is exceptional, while combining a relative high content of the indole glucosinolate glucobrassicin with low alkenyle glucosinolates in the leaves. The total glucosinolate concentration in the stems and leaves is not correlated with the seed glucosinolate concentrations. Heritabilities are above h2 = 0.60 for progoitrin, h2 = 0.65 for gluconapin, h2 = 0.30 for glucobrassicanapin and h2 = 0.52 for total glucosinolate content in the leaves. In conclusion, resynthesized rapeseed is an important genetic resource to modify the leaf glucosinolate content and composition of rapeseed.     

Carotene, tocopherol, and ascorbate contents in subspecies of Brassica oleracea

Cruciferous vegetables contain high levels of vitamins that can act as antioxidants, compounds that may protect against several degenerative diseases. The edible portions of 50 broccoli and 13 cabbage, kale, cauliflower, and Brussels sprouts accessions were assayed to determine variation in alpha-carotene, beta-carotene, alpha-tocopherol, gamma-tocopherol, and ascorbate contents within and between subspecies of Brassica oleracea. Ascorbate content was estimated in fresh samples using HPLC. Tissues for carotene and tocopherol analysis were lyophilized prior to extraction. Carotene and tocopherol concentrations were simultaneously measured using a reverse phase HPLC system. Results indicate that there is substantial variation both within and between subspecies. Kale had the highest levels of vitamins, followed by broccoli and Brussels sprouts with intermediate levels and then by cabbage and cauliflower, with comparatively low concentrations. Variability in vitamin content among the broccoli accessions suggests that potential health benefits that accrue with consumption are genotype dependent.     

Glucoraphanin and 4-hydroxyglucobrassicin contents in seeds of 59 cultivars of broccoli, raab, kohlrabi, radish, cauliflower, brussels sprouts, kale, and cabbage

The importance of dietary sulforaphane in helping maintain good health continues to gain support within the health-care community and awareness among U.S. consumers. In addition to the traditional avenue for obtaining sulforaphane, namely, the consumption of appropriate cruciferous vegetables, other consumer products containing added glucoraphanin, the natural precursor to sulforaphane, are now appearing in the United States. Crucifer seeds are a likely source for obtaining glucoraphanin, owing to a higher concentration of glucoraphanin and the relative ease of processing seeds as compared to vegetative parts. Seeds of several commonly consumed crucifers were analyzed not only for glucoraphanin but also for components that might have negative health implications, such as certain indole-containing glucosinolates and erucic acid-containing lipids. Glucoraphanin, 4-hydroxyglucobrassicin, other glucosinolates, and lipid erucic acid were quantified in seeds of 33 commercially available cultivars of broccoli, 4 cultivars each of kohlrabi, radish, cauliflower, Brussels sprouts, kale, and cabbage, and 2 cultivars of raab.     

Correlation of glucosinolate content to myrosinase activity in horseradish (Armoracia rusticana)

Fully developed horseradish (Armoracia rusticana Gaertn., Mey., & Scherb.) roots from 27 accessions and leaves from a subset of 9 accessions were evaluated for glucosinolates and myrosinase enzyme activity. Eight different glucosinolates were detected (based on HPLC retention times as desulfoglucosinolates) in both root and leaf tissues. The sum of these glucosinolates, referred to as total, ranged from 2 to 296 micromol g(-1) of dry weight (DW) in both tissues. Four glucosinolates (sinigrin, glucobrassicin, neoglucobrassicin, and gluconasturtiin) were detected in major quantities. In fully developed roots, sinigrin concentration represented approximately 83%, gluconasturtiin approximately 11%, and glucobrassicin approximately 1% of the total glucosinolates. Approximately the same proportions of individual glucosinolates appeared in fully developed leaves, except that glucobrassicin was substituted by neoglucobrassicin and gluconasturtiin concentration was significantly lower (<1%). At least four other glucosinolates were detected in very small quantities (<1%) in both roots and leaves. Myrosinase (beta-thioglucoside glucohydrolase, EC 3.2.3.1) is the enzyme responsible for the hydrolysis of the parent glucosinolates into biologically active products. Very little is known about myrosinase activity and the correlation of its activity to total and individual glucosinolates in plant tissues. Significant differences in myrosinase activity were detected between the roots and leaves, ranging from 1.2 to 57.1 units g(-1) of DW. Data showed no correlation between myrosinase activity and total and/or individual glucosinolates in the roots. However, in the leaves, significant correlations were found between myrosinase activity and total glucosinolates (0.78 at P = 0.01) and between myrosinase activity and sinigrin (0.80 at P = 0.01). Glucosinolates content and myrosinase activity were also correlated in young and fully developed roots and leaves and during tissue crushing. Glucobrassicin concentration in the roots and neoglucobrassicin concentration in the leaves were significantly higher in young than in fully developed tissue. Crushing of the tissue resulted in rapid hydrolysis of sinigrin and glucobrassicin, as expected, from the presence of myrosinase. Likewise, myrosinase activity declined rapidly after crushing, perhaps due to inactivation by the reaction products and/or the depletion of its substrates.     

Plant-derived biomolecules in fermented cabbage

The formation of plant-derived biomolecules during sauerkraut fermentation was studied. Cabbage was fermented with a starter culture, and the results were compared to the results of spontaneous fermentation. The concentration of flavonoids and glucosinolates was analyzed by HPLC, and that of the glucosinolate breakdown products, by GC-MS. Of the 20 different flavonoids tested, only kaempferol was found (0.9 mg/ kg FW, fresh weight). The content of kaempferol remained constant in the cabbage fiber matrix over the fermentation process. The nitrite concentration was below the detection limit in both fermentations. The total glucosinolate content in the raw material was 3.71 micro mol/g DW, dry weight. Glucosinolates were totally decomposed in both fermentations during two weeks, and different types of breakdown products were formed. Isothiocyanates, indole-3-carbinol, goitrin, allyl cyanide, and nitriles were determined in the fermented cabbage. Isothiocyanates and allyl cyanide were the predominant breakdown products in both fermentations. Sulforaphane nitrile and goitrin were found only in small quantities in the end products.     

LC-MS/MS quantification of sulforaphane and indole-3-carbinol metabolites in human plasma and urine after dietary intake of selenium-fortified broccoli

This study aimed at developing a sensitive LC-MS/MS method for the quantification of sulforaphane (SFN) and indole-3-carbinol metabolites in plasma and urine after dietary intake of regular and selenium-fertilized broccoli using stable isotope dilution analysis. In a three-armed, placebo-controlled, randomized human intervention study with 76 healthy volunteers, 200 g of regular (485 μg of total glucosinolates and <0.01 μg of selenium per gram fresh weight) or selenium-fertilized broccoli (589 μg of total glucosinolates and 0.25 μg of selenium per gram fresh weight) was administered daily for 4 weeks. Glucoraphanin and glucobrassicin metabolites quantified in plasma and urine were SFN-glutathione, SFN-cysteine, SFN-cysteinylglycine, SFN-acetylcysteine, and indole-3-carboxaldehyde, indole-3-carboxylic acid, and ascorbigen, respectively. Dietary intake of selenium-fertilized broccoli increased serum selenium concentration analyzed by means of atomic absorption spectroscopy by up to 25% (p < 0.001), but affected neither glucosinolate concentrations in broccoli nor their metabolite concentrations in plasma and urine compared to regular broccoli.     

Effects of Brussels sprout juice on the cell cycle and adhesion of human colorectal carcinoma cells (HT29) in vitro

Consumption of Brassica vegetables is associated with a reduced risk of cancer of the alimentary tract in animal models and human populations. We used raw juice extracted from Brussels sprouts rich in the glucosinolate sinigrin to explore the effect of naturally occurring glucosinolate breakdown products on cell cycle progression and apoptosis in human colorectal carcinoma cells (HT29). Juice was prepared from sprout tissue immediately before use, and the glucosinolate breakdown products were determined by gas chromatography mass spectrometry and liquid chromatography mass spectrometry. The cell cycle was analyzed by flow cytometry on detached and adherent cells, and apoptosis was measured in the detached population by annexin V staining. Twenty-four hours after challenge with juice (10 microL/mL), 7-13% of adherent cells had detached from the substratum but the majority (82%) of these cells had not entered apoptosis, whereas only 33% of detached control cells were not apoptotic (p < 0.05). The main glucosinolate breakdown products were as follows: the sinigrin breakdown product, 1-cyano-2,3-epithiopropane (ca. 38 mM); the gluconapin hydrolysis product, 3-butenyl isothiocyanate (ca. 2.2.mM); the glucobrassicin metabolite, ascorbigen (ca. 8 mM); and low concentrations of other indole glucosinolate-derived hydrolysis products such as neoascorbigen and 3,3'-diindolylmethane. A variety of biologically active glucosinolate breakdown products are released by mechanical disruption of raw Brussels sprout tissue, but contrary to previous assumptions, allyl isothiocyanate is not the main compound responsible for the inhibition of cell proliferation.     

花椰菜主要生物活性物质及其抗氧化能力分析

分析了花椰菜不同器官(花球、茎、叶和根)中芥子油苷等主要生物活性物质及其抗氧化能力,发现烯丙基芥子油苷是花椰菜中主要的芥子油苷,不同器官中芥子油苷的组分与含量差异显著,根系中的总芥子油苷、总脂肪类、总芳香类以及烯丙基芥子油苷和4-甲氧基-吲哚-3-甲基芥子油苷的含量均显著高于其他器官,叶片中吲哚类芥子油苷的含量最高,花球中总芥子油苷、烯丙基芥子油苷和吲哚-3-甲基芥子油苷含量次之,茎中的总芥子油苷含量最低;叶片中的叶绿素、类胡萝卜素、维生素C和多酚含量及抗氧化能力显著高于其他器官。这些结果表明花椰菜的叶片和根系中生物活性物质含量丰富,具有进一步开发和综合利用潜力。     

Content of glucosinolates in cruciferous vegetables grown at the same site for two years under different climatic conditions

Individual glucosinolates (GLS) were determined in vegetables of three Cruciferae species: Brassica oleracea L. (white cabbage, red cabbage, Savoy cabbage, Brussels sprouts, cauliflower, kale, kohlrabi), Brassica rapa L. (turnip), and Raphanus sativus L. (red radish, black radish, and white radish) produced in two years. The cultivars were compared for the contents of total-, indole-, and aliphatic GLS. In both years, the total content of GLS was highest in black radish, and all examined R. sativus vegetables contained the greatest amount of aliphatic GLS. Neither the level nor the identity of GLS differentiated among the vegetables of the other cultivars grown in the same year. Comparison of the GLS contents of the same cultivar in two production years, which differed in temperature and rainfall rate, showed that low average 10-day rainfall and high average temperature during the vegetation period significantly increased the GLS content of vegetables. This suggests that the year x cultivar interaction modified the GLS content of vegetables.     

氨基酸对甘蓝硫代葡萄糖苷的影响

以中甘21(ZG21)、06-115两个甘蓝品种为试验材料,研究叶面喷施6种氨基酸对甘蓝硫代葡萄糖苷组分及其含量的影响。结果表明,不同种类氨基酸处理甘蓝硫代葡萄糖苷组分相同,均含有9种硫苷。氨基酸对甘蓝硫苷含量有一定的影响,半胱氨酸和甲硫氨酸显著提高06-115硫苷的总含量、脂肪族硫苷的含量和4-甲基硫氧丁基硫苷含量,其中半胱氨酸处理含量最高,与对照相比分别提高63.0％、69.6％和69.1％;半胱氨酸处理提高了3-吲哚基甲基硫苷含量,达到725.6 μmol/kg, FW;半胱氨酸处理也显著提高了中甘21硫苷的总含量、脂肪族硫苷的含量、吲哚族硫苷含量和4-甲基硫氧丁基硫苷含量,与对照相比分别提高了27.2％、39.6％、15.3％和26.2％。半胱氨酸、苯丙氨酸、甘氨酸和谷氨酸处理都提高了3-吲哚基甲基硫苷的含量,其中半胱氨酸处理最高,达到292.7 μmol/kg, FW,其次是谷氨酸和甘氨酸处理。     

Large scale determination of glucosinolates in brussels sprouts samples after degradation of endogenous glucose.

A method was developed for the determination of the glucosinolate content in glucose-rich samples of Brassica vegetables such as Brussels sprouts. Glucose in the samples was enzymatically degraded by the enzyme glucose oxidase (GOD). The resulting hydrogen peroxide and the enzyme GOD were thereafter respectively dissociated and inactivated by a heat treatment at 100 degrees C. After the degradation of endogenous glucose the glucosinolates were converted into glucose and related metabolites with the enzyme thioglucosidase originating from Brussels sprouts seeds. Glucose released was determined enzymatically with a glucose oxidase/peroxidase assay as a measure for the glucosinolate content of samples. The method was used to study the influence of harvest time, crop production location, and the choice of parental lines on the glucosinolate content of Brussels sprouts F1-hybrids. The sum of sinigrin and progoitrin of F1-hybrids was found to be significantly correlated to the glucosinolate content.     

Glucosinolates in mixed-packaged mini broccoli and mini cauliflower under modified atmosphere

Contents of total and individual glucosinolates of mini broccoli cv. Milady and mini cauliflower cv. Clarke were assessed to determine the effect of modified atmosphere packaging on postharvest glucosinolate dynamics of mixed mini Brassica vegetables. Therefore, mixed-packaged mini broccoli and mini cauliflower stored in food trays sealed with two different microperforated biaxial-oriented polypropylene films for up to 7 days at 8 degrees C were analyzed. The results indicate that modified atmosphere at 8% O2 + 14% CO2 was a suitable gaseous combination to maintain aliphatic and indole glucosinolates in mini broccoli for 7 days after an initial decrease at 4 days. In contrast, modified atmosphere at 1% O2 + 21% CO2 resulted in the best retention of indole glucosinolates of mini cauliflower for 7 days and also of aliphatic glucosinolates after an initial decrease at 4 days. Thus, to maintain glucosinolates and external appearance and to prevent off-odor, mini broccoli and mini cauliflower should be packed separately in suitable altered gas composition.     

喷施蔗糖和葡萄糖对小白菜硫代葡萄糖苷含量的影响

以小白菜"上海青"为试验材料,研究了喷施蔗糖和葡萄糖对小白菜生长和硫代葡萄糖苷含量的影响。结果显示,蔗糖和葡萄糖喷施对小白菜地上部鲜重和干重均没有显著影响,喷施蔗糖均能够显著增加小白菜总脂肪族硫代葡萄糖苷、总吲哚族硫代葡萄糖苷和总硫代葡萄糖苷含量,其中,0.1mol/L蔗糖喷施显著提高了3-丁烯基硫代葡萄糖苷和吲哚-3-甲基硫代葡萄糖苷含量,0.2mol/L蔗糖喷施显著增加了吲哚-3-甲基硫代葡萄糖苷和4-甲氧基吲哚-3-甲基硫代葡萄糖苷含量。0.2mol/L葡萄糖喷施显著增加总硫代葡萄糖苷及单个硫代葡萄糖苷含量。蔗糖和葡萄糖喷施均有增加总吲哚族硫代葡萄糖苷相对百分含量的趋势。研究表明外源喷施糖对小白菜硫代葡萄糖苷含量和组成具有显著的影响。     

Variation of glucosinolates in wild radish (Raphanus raphanistrum) accessions

Glucosinolate composition was determined in wild radish accessions from eight states in the northeastern and southern United States to determine the variability of production among accessions. Glucosinolates were evaluated from roots, leaves, flowers, primary, and secondary branches. Seventeen glucosinolates were identified, with glucoerucin, glucoraphenin, glucobrassicin, and gluconasturtiin contributing 90% to 100% of the total glucosinolates. Flowers contained the highest glucosinolate concentrations, 12.07 to 55.36 μmol/g, but flowers contributed only 5.3 to 21.3% to the total glucosinolates. Of the eight accessions, the Mississippi accession produced significantly higher levels of total glucosinolates and glucosinolates which can be degraded to isothiocyanates per plant, totals of 618.97 and 563.53 μmol/plant, respectively. Total plant biomass did not differ between accessions indicating a difference in the ability of the Mississippi accession to produce glucosinolates. Further studies are needed to determine if this accession would consistently produce higher glucosinolate levels under different environmental conditions.     

Comparison of isothiocyanate metabolite levels and histone deacetylase activity in human subjects consuming broccoli sprouts or broccoli supplement

Increased consumption of cruciferous vegetables such as broccoli may reduce the risk of various cancers. Myrosinase is required to convert dietary glucosinolates from broccoli into bioactive isothiocyanates. We evaluated isothiocyanate excretion profiles in healthy subjects who consumed broccoli sprouts or broccoli supplement (no myrosinase) with equivalent glucosinolate content. Urinary metabolites of two major isothiocyanates, sulforaphane and erucin, were measured by liquid chromatography coupled with tandem mass spectrometry. Peak excretion of sulforaphane and erucin was higher and occurred sooner in subjects who consumed broccoli sprouts as compared to subjects who consumed the supplement. A subject-dependent shift in the ratio of urinary sulforaphane to erucin metabolites was observed in both groups, indicating conversion of sulforaphane to erucin. Lower histone deacetylase activity was observed in the peripheral blood mononuclear cells only in subjects consuming sprouts. Fresh broccoli sprouts differ from broccoli supplements in regards to excretion of isothiocyanates and bioactivity in human subjects.     

Aqueous extract from Spanish black radish (Raphanus sativus L. Var. niger) induces detoxification enzymes in the HepG2 human hepatoma cell line

Spanish black radish (Raphanus sativus L. var. niger) is a member of the Cruciferae family that also contains broccoli and Brussels sprouts, well-known to contain health-promoting constituents. Spanish black radishes (SBR) contain high concentrations of a glucosinolate unique to the radish family, glucoraphasatin, which represents >65% of the total glucosinolates present in SBR. The metabolites of glucosinolates, such as isothiocyanates, are implicated in health promotion, although it is unclear whether glucosinolates themselves elicit a similar response. The crude aqueous extract from 0.3 to 3 mg of dry SBR material increased the activity of the phase II detoxification enzyme quinone reductase in the human hepatoma HepG2 cell line with a maximal effect at a concentration of 1 mg/mL. Treatment of HepG2 cells with the crude aqueous extract of 1 mg of SBR per mL also significantly induced the expression of mRNA corresponding to the phase I detoxification enzymes: cytochrome P450 (CYP) 1A1, CYP1A2, and CYP1B1 as well as the phase II detoxification enzymes: quinone reductase, heme oxygenase 1, and thioredoxin reductase 1. Previous studies have shown that the myrosinase metabolites of different glucosinolates vary in their ability to induce detoxification enzymes. Here, we show that while glucoraphasatin addition was ineffective, the isothiocyanate metabolite of glucoraphasatin, 4-methylthio-3-butenyl isothiocyanate (MIBITC), significantly induced phase II detoxification enzymes at a concentration of 10 microM. These data demonstrate that the crude aqueous extract of SBR and the isothiocyanate metabolite of glucoraphasatin, MIBITC, are potent inducers of detoxification enzymes in the HepG2 cell line.     

Influence of nitrogen and sulfur on biomass production and carotenoid and glucosinolate concentrations in watercress (Nasturtium officinale R. Br.)

Watercress (Nasturtium officinale R. Br.) is a perennial herb rich in the secondary metabolites of glucosinolates and carotenoids. 2-phenethyl isothiocyanate, the predominate isothiocyanate hydrolysis product in watercress, can reduce carcinogen activation through inhibition of phase I enzymes and induction of phase II enzymes. Sulfur (S) and nitrogen (N) have been shown to influence concentrations of both glucosinolates and carotenoids in a variety of vegetable crops. Our research objectives were to determine how several levels of N and S fertility interact to affect watercress plant tissue biomass production, tissue C/N ratios, concentrations of plant pigments, and glucosinolate concentrations. Watercress was grown using nutrient solution culture under a three by three factorial arrangement, with three S (8, 16, and 32 mg/L) and three N (6, 56, and 106 mg/L) fertility concentrations. Watercress shoot tissue biomass, tissue %N, and tissue C/N ratios were influenced by N but were unaffected by changes in S concentrations or by the interaction of NxS. Tissue pigment concentrations of beta-carotene, lutein, 5,6-epoxylutein, neoxanthin, zeaxanthin, and the chlorophyll pigments responded to changes in N treatment concentrations but were unaffected by S concentrations or through N x S interactions. Watercress tissue concentrations of aromatic, indole, and total glucosinolate concentrations responded to changes in N treatments; whereas aliphatic, aromatic, and total glucosinolates responded to changes in S treatment concentrations. Individual glucosinolates of glucobrassicin, 4-methoxyglucobrassicin, and gluconasturriin responded to N fertility treatments, while gluconapin, glucobrassicin, and gluconasturiin responded to changes in S fertility concentrations. Increases in carotenoid and glucosinolate concentrations through fertility management would be expected to influence the nutritional value of watercress in human diets.     

Influence of temperature and ontogeny on the levels of glucosinolates in broccoli (Brassica oleracea Var. italica) sprouts and their effect on the induction of mammalian phase 2 enzymes

Broccoli inflorescences have been recognized as components of healthy diets on the basis of their high content of fiber, vitamin C, carotenoids, and glucosinolates/isothiocyanates. Broccoli sprouts have been recently shown to have high levels of glucoraphanin (4-methylsulfinylbutyl glucosinolate), the precursor of the chemoprotective isothiocyanate, sulforaphane. This study evaluated the effects of temperature and developmental stage on the glucosinolate content of broccoli sprouts. Seedlings cultivated using a 30/15 degrees C (day/night) temperature regime had significantly higher glucosinolate levels (measured at six consecutive days postemergence) than did sprouts cultivated at lower temperatures (22/15 and 18/12 degrees C; p < 0.001). Both higher (33.1 degrees C) and lower (11.3 degrees C) constant temperatures induced higher glucosinolate levels in sprouts grown to a uniform size. Glucosinolate levels were highest in cotyledons and lowest in roots of sprouts dissected both early and late in the 11 day developmental span investigated. Nongerminated seeds have the highest glucosinolate levels and concordantly greater induction of mammalian phase 2 detoxication enzymes. Levels decline as sprouts germinate and develop, with consistently higher glucosinolate content in younger developmental stages, independent of the temperature regime. Temperature stress or its associated developmental anomalies induce higher glucosinolate levels, specific elevations in glucoraphanin content, and parallel induction of phase 2 chemoprotective enzymes.     

Glucosinolate composition of young shoots and flower buds of capers (Capparis species) growing wild in Turkey

The content and glucosinolate composition of young shoots and raw flower buds of Capparis spinosa var. spinosa and Capparis ovata Desf. var. canescens at three different sizes (x 13 mm) were investigated by HPLC with UV detection. Samples were harvested in August 2001 in Turkey. Twelve different glucosinolates were identified in the young shoots and buds of both species. Total content of glucosinolates ranged from 6.55 micromol/g (large buds of C. spinosa) to 45.56 micromol/g (young shoots of C. ovata). The main glucosinolate was glucocapperin, which amounted to approximately 90% of the total glucosinolates. In both species the total glucosinolate content varied in dependence on the bud size, whereas a greater variability was given for buds from C. spinosa.     

Determination of glucosinolates using their alkaline degradation and reaction with ferricyanide.

Glucosinolates, a group of naturally occurring thioglucosides, are significant factors impairing the nutritional quality of rapeseed and postextraction rapeseed meal, restricting its use as high-quality protein animal feed. Currently, the European Community standards and Canola definition are being brought in line recommending cultivation and marketing of rapeseed with a glucosinolate content below 18 micromol of total glucosinolates per gram of seeds. Furthermore, some glucosinolates are of increasing interest in Brassica vegetables due to their proven cancer-preventing activities. A novel approach to the analysis of total glucosinolates is reported in this paper based on their alkaline degradation and subsequent reaction of released 1-thioglucose with ferricyanide. The reaction was followed spectrophotometrically using sinigrin and glucotropeaolin as model glucosinolates. The applicability of the method was demonstrated using rapeseed extracts after reducing the interfering effect of phenolics by their adsorption onto polyvinylpolypyrrolidone. Good agreement with official ISO methods was shown.