NY/T 1381-2007 Determination of Capsaicin by High Performance Liquid Chromatography NY/T1381-2007 Standard download decompression password: www.bzxz.net
This standard specifies the high performance liquid chromatography method for the determination of capsaicin. This standard is applicable to the determination of capsaicin and dihydrocapsaicin in peppers and foods produced with them as raw materials. The detection limit of this method is 0.13 mg/kg for capsaicin and 0.04 mg/kg for dihydrocapsaicin. The linear range of this method is 0.10 mg/L to 160 mg/L for capsaicin and 0.04 mg/L to 160 mg/L for dihydrocapsaicin.

ICS67.080.20
Agricultural Industry Standard of the People's Republic of China
NY/T1381—2007
Determination of Capsaicin
High Performance Liquid Chromatography
Determination of Capsaicin by High Performance Liquid Chromatography2007-06-14Issued
2007-09-01Implemented
Ministry of Agriculture of the People's Republic of China
1Scope
Determination of Capsaicin by High Performance Liquid Chromatography
This standard specifies the determination method of capsaicin by high performance liquid chromatography. NY/T1381—2007
This standard is applicable to the determination of capsaicin and dihydrocapsaicin in peppers and foods produced with them as raw materials. The detection limits of this method are 0.13mg/kg for capsaicin and 0.04mg/kg for dihydrocapsaicin. The linear range of this method is 0.10mg/L~160mg/L for capsaicin and 0.04mg/L~160mg/L for dihydrocapsaicin. 2 Normative references
The clauses in the following documents become the clauses of this standard through reference in this standard. For all references with dates, all subsequent amendments (excluding errata) or revisions are not applicable to this standard. However, parties reaching an agreement based on this standard are encouraged to study whether the latest versions of these documents can be used. For all undated references, the latest versions are applicable to this standard. GB/T6682 Specifications and test methods for water for analytical laboratories 3 Principle
The sample is extracted with methanol-tetrahydrofuran solution, passed through a microporous filter membrane, injected, separated by reverse phase chromatography, detected by a fluorescence detector, and quantified by the external standard method.
4 Reagents and materials
Unless otherwise specified, only chromatographically pure reagents are used, and the experimental water should meet the requirements of GB/T6682 Grade 1 water. 4.1 Methanol (CHOH).
4.2 Tetrahydrofuran (C4H8o).
4.3 Methanol-tetrahydrofuran solution (1+1): Take 200mL methanol and 200mL tetrahydrofuran and mix them evenly for use. 4.4 Anhydrous sodium sulfate (NaSO4), analytical grade, dry at 400℃ for 30min before use, and grind. 4.5 Capsaicin standard, purity greater than 95%. 4.6 Dihydrocapsaicin standard, purity greater than 90%. 4.7 Capsaicin standard stock solution: weigh 0.0100g capsaicin standard, dissolve in methanol, dilute to 50mL, capsaicin mass concentration is 200mg/L.
4.8 Dihydrocapsaicin standard stock solution: weigh 0.0100g dihydrocapsaicin standard, dissolve in methanol, dilute to 50mL, dihydrocapsaicin mass concentration is 200mg/L.
5 Instruments and equipment
5.1 High performance liquid chromatograph: with fluorescence detector. 5.2 Balance: sensitivity ±0.0001g, ±0.01g. 5.3 Ultrasonic extractor.
5.4 Tissue crusher.
5.5 Electric grinder.
5.6 Constant temperature water bath.
NY/T1381—2007
5.7 Filter membrane: 0.45μm, organic phase.
6 Analysis steps
6.1 Extraction
6.1.1 Chili powder, dried chili and solid chili products Grind the sample with an electric grinder, weigh 0.2g accurately to 0.0001g, add 25mL methanol-tetrahydrofuran solution to a 100mL stoppered conical flask, extract with an ultrasonic extractor for 30min under 60℃ water bath conditions, filter, and collect the filtrate. Extract the residue and filter paper with 25mL methanol-tetrahydrofuran solution through an ultrasonic extractor for 10min, and repeat once. Combine the filtrates collected by three filtrations, concentrate to about 30mL in a constant temperature water bath at 70℃~75℃, transfer to a 50mL volumetric flask, and make up to volume with methanol-tetrahydrofuran solution. Filter through a 0.45um organic phase filter membrane and perform chromatographic analysis. According to the capsaicin content in the sample, dilute if necessary to make the capsaicin content in the sample between 0.13mg/L and 160mg/L, and the dihydrocapsaicin content between 0.04mg/L and 160mg/L. 6.1.2 Fresh pepper, chili sauce and pepper products with high water content Crush the sample with a tissue crusher, weigh 10.00g of the sample in a 100mL stoppered conical flask, add 3.0g of anhydrous sodium sulfate and 25mL of methanol-tetrahydrofuran solution, shake, extract with an ultrasonic extractor for 30min, filter, and collect the filtrate. Thereafter, operate the same as 6.1.1 "Re-extract the filter residue together with the filter paper".
6.1.3 Oily products such as chili oil
Homogenize the sample with a tissue crusher, weigh 2.00g of the sample in a 100mL stoppered conical flask, add 25mL of methanol-tetrahydrofuran solution, extract with an ultrasonic extractor for 30min, filter, and collect the filtrate. Afterwards, repeat the operation with the filter paper together with the filter residue as in 6.1.1. 6.2 Chromatographic reference conditionsbZxz.net
Chromatographic column: Cl8150mm×4.6mm, particle size 5μm. Mobile phase: methanol+water=80+20, pass through 0.45um filter membrane before use, degas. Flow rate: 0.6mL/min.
Fluorescence detector: excitation wavelength 229nm, emission wavelength 320nm. Injection volume: 10μL.
6.3 Drawing of standard curve
Dilute the standard stock solutions of capsaicin and dihydrocapsaicin with methanol to a series of standard working solutions with mass concentrations of 0.2mg/L, 1mg/L, 10mg/L, 20mg/L, 40mg/L, 80mg/L and 160mg/L, respectively, and determine according to the chromatographic conditions in 6.2. With the mass concentration of capsaicin and dihydrocapsaicin as the ordinate and the corresponding peak area integral value as the abscissa, calculate the standard curve or find the linear regression equation. 6.4 Determination of test solution
Determine the prepared test solution under the chromatographic conditions of 6.2, perform single-point or multi-point calibration, and quantify by the peak area integral value. 7 Calculation and expression of results
The content of capsaicin and dihydrocapsaicin in the sample is expressed as mass fraction w, expressed in milligrams per kilogram (mg/kg), and is calculated according to formula (1):
AxpsxV
Wherein:
A is the peak area integral value of capsaicin and dihydrocapsaicin in the sample; As is the peak area integral value of capsaicin and dihydrocapsaicin in the standard working solution; Ps is the mass concentration of capsaicin and dihydrocapsaicin in the standard working solution, in milligrams per liter (rng/L); V is the final fixed volume of the sample, in milliliters (mL);
is the mass of the sample, in grams (g);
The calculation result shall be rounded to three significant figures.
8 Precision
NY/T1381—2007
The absolute difference between two independent test results obtained under repeatability conditions shall not exceed 10% of the arithmetic mean of the two measured values, provided that the absolute difference is greater than 10% of the arithmetic mean of the two measured values ​​and does not exceed 5%. 9 Spectrum
The chromatogram of the standard solution of capsaicin and dihydrocapsaicin is shown in Figure 1. 0.4
Minutes
Figure 1 Chromatogram of the standard solution of capsaicin (1.6mg/L) and dihydrocapsaicin (0.8mg/L) 14
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