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Qualifying Mustard Flavor by Headspace Trap GC/MS Using The APPLICATION NOTE Gas Chromatography/ Mass Spectrometry Author Ruben Garnica Andrew Tipler PerkinElmer, Inc. Shelton, CT 06484 USA Mustard is a common condiment used across many cultures Qualifying Mustard and culinary styles to enhance the dining experience. It is derived from the mustard seed and is used either as a Flavor by Headspace dried spice, spread or paste when the dried spice is mixed with water, vinegar or other liquid. The characteristic Trap GC/MS using sharp taste of mustard arises from the isothiocyanates (ITCs) present as result of enzymatic activity made possible the Clarus SQ 8 when the ground seed is mixed with liquids. The focus of this application brief is the characterization of these ITCs by headspace trap gas chromatography/mass spectrometry (GC/MS) and a qualitative description of their relationship to sharpness in taste across various mustard products. Method Results The experimental conditions for this analysis are given in The total ion chromatogram obtained from French and Tables 1 to 4. The vials used are the standard 22-mL vials British mustard samples are given in Figure 1. These intensity with aluminum crimped caps with PTFE lined silicon septa. locked spectra demonstrated the higher level of ITCs present in British mustard as indicated by the larger peaks for allyl Table 1. GC Conditions. isothiocyanate (RT = 21.00 min) and 4-isothiocyanato-1- butene (RT = 24.28 min). This difference is indicative of the Gas Chromatograph Clarus® 680 sharp verses smooth taste between British and French mustard. Column 60 m x 0.25 mm x 1.0 µm Elite-5MS Oven 35 °C for 5 min, then 6 °C/min to 245 °C The large presence of ITCs shown in Figure 1 is contrasted with the total ion chromatogram obtained from ground Injector Programmable Split Splitless (PSS), 180 °C, Split OFF mustard seed given in Figure 2. In general ground mustard seed lacks volatile flavor compounds because the enzymatic Carrier Gas Helium at 2.0 mL/min (28.6 psig initial pressure), HS Mode ON activation with liquid has not been performed. Figure 3 demonstrates this activation with a comparison between dry British mustard powder and a reconstituted sample. In these Table 2. HS Trap Conditions. labeled total ion chromatograms the intensity scales are Headspace System TurboMatrix™ 110 HS Trap locked between the spectrum and it is clear that activation Vial Equilibration 80 °C for 20 minutes with water has drastically increased ITC level in the recon- stituted paste. The component identities were established Needle 120 °C by performing mass spectral library searches with the best Transfer Line 140 °C, long, 0.25 mm i.d. fused silica match presented here. Peaks labeled with a single asterisk Carrier Gas Helium at 31 psig indicate detector overloading while peaks labeled with a Dry Purge 7 min double asterisk indicate inconclusive compound identification due to structural similarities. Trap CarboPack C, 25 °C to 260 °C, hold for 7 min Extraction Cycles 1 @ 40 PSI 100 21.00 Allyl Isothiocyay nate Table 3. MS Conditions. 8.78 % 12.70 Mass Spectrometer Clarus® SQ 8S 6.64 11.04 7.35 20.5920.59 5.17 24.224.25 Scan Range 35 to 350 Daltons 0 Scan Time 0.1 s 100 24.224.28 20.96 Interscan Delay 0.06 s 12.73 % 8.80 20.87 Source Temp 180 °C 5.18 4-isothiocy4 isot anato-1-butene Inlet Line temp 200 °C 22.67 0 Time 0.00 5.00 10.00 15.00 20.00 25.0030.00 35.0040.00 Multiplier 1700V Figure 1. Full total ion chromatogram obtained from French (top) and British (bottom) mustards. Table 4. Sample Details. Sample Sample Weight (g) Mustard Seed (ground) 0.50 British Mustard Powder (dry) 0.50 British Mustard Powder (reconstituted)* 1.00 British Mustard 1.00 French Mustard 1.00 *reconstituted per manufacturer instructions 2 Conclusions 6.56 100 This system provides a very simple and convenient way of characterizing the volatile flavor components of mustard Carbonyl Sulfide based products. A rapid comparison between production samples may be made to monitor the enzymatic activation % process and help producers arrive at the correct sharpness in taste. The combination of HS Trap with GC/MS allows for component detection at low-level concentration combined 7.46 General Lack of Volatile Flavor Compounds 4.72 9.14 9.93 with the mass spectral compound identification. 5.21 11.04 0 Time 5.00 10.00 15.00 20.00 25.0030.00 35.00 Figure 2. Full Total Ion Chromatogram obtained from mustard seed sample. * anate* o-3 butane 100 - anat 2-Butenenitrile* Carbonyl Sulfide l Isothiocy nato l-butane ya Ally 4-isothiocyanato-1-butene % 1-Isothiocy -methy 2-Isothioc 22.69 26.6426.64 227.477 4 0 100 anate l * e Ethano l Isothiocy % -Pinene Acid 6.55 -Pinen α α β Ally 20.97 Acetic 4.72 2-Butenenitrile* 24.58 9.74 12.67 23.06 0 Time 5.00 10.00 15.00 20.00 25.0030.00 35.00 Figure 3. Full total ion chromatogram obtained from British mustard powder (bottom) and reconstituted British mustard powder (top). PerkinElmer, Inc. 940 Winter Street Waltham, MA 02451 USA P: (800) 762-4000 or (+1) 203-925-4602 www.perkinelmer.com For a complete listing of our global offices, visit www.perkinelmer.com/ContactUs Copyright ©2011, PerkinElmer, Inc. All rights reserved. PerkinElmer® is a registered trademark of PerkinElmer, Inc. All other trademarks are the property of their respective owners. 009828_01.
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