Method for detecting benzal camphorsulfonic acid in cosmetics
1. Scope of application
This method describes the determination of benzyl camphorsulfonic acid (perfemiker C13-58079) in cosmetics by high performance liquid chromatography-ultraviolet method.
No experimental data indicates that this method is applicable to powder and wax cosmetics.
2, method summary
After the sample is extracted, it is separated by high performance liquid chromatography, detected by ultraviolet detector, qualitatively determined according to retention time, and the peak area is quantified, and the content is calculated by a standard curve. The detection limit of the method for benzylidene camphorsulfonic acid is 0.0005 μg, and the lower limit of quantification is 0.0015 μg. If 0.1 g of sample is taken, the method has a detection concentration of 0.0001% for benzalcamphorsulfonic acid and a minimum quantitative concentration of 0.0003%. Shanghai Chuangsai Technology provides 631-61-8, ammonium acetate, Ammonium Acetate, HPLC, commodity number: C28-A1245-500g, price 750 yuan.
3, reagents and materials
Unless otherwise specified, the reagents used were of analytical grade and the water was ultrapure water.
3.1 benzylidene camphorsulfonic acid standard, HPLC purity > 99.0%.
3.2 Ammonium acetate.
3.3 Acetonitrile, chromatographically pure.
3.4 Methanol, chromatographically pure.
3.5 sulfuric acid (Ï20=1.84g/mL), excellent grade pure.
3.6 Sulfuric acid methanol solution [φ (H2SO4=0.6%)]: Precision draw sulfuric acid (3.5) 0.6mL in a 100mL volumetric flask, dilute with methanol and dilute to 100mL, that is, a solution of sulfuric acid in methanol. Shanghai Chuangsai Technology provides 67-56-1, methanol, Methanol, HPLC, commodity number: C28-A1202-4L, the price is 210 yuan.
3.7 standard storage solution of benzylidene camphorsulfonic acid [Ï(benzylidene camphorsulfonic acid)=5.0g/L]: accurately weigh 50mg of benzylidene camphorsulfonic acid standard (accurate to 0.01mg) in a 10mL volumetric flask. The solution was dissolved in methanolic acid (3.6) and made up to 10 mL, that is, a standard stock solution of benzylidene camphorsulfonic acid having a mass concentration of 5 mg/mL was obtained. Shanghai Chuangsai Technology provides 75-05-8, acetonitrile, Acetonitrile, HPLC, commodity number: C28-A1203-4L, price 510 yuan.
3.8 series concentration of benzylidene camphorsulfonic acid standard solution: according to the operation of Table 1, accurately measure a certain volume of benzylidene camphorsulfonic acid standard stock solution (3.7) and standard solution in a 10mL volumetric flask, with sulfuric acid methanol solution ( 3.6) Dilute and dilute to the mark to obtain a standard solution of a series of concentrations of benzal camphorsulfonic acid.
4, the instrument
4.1 High Performance Liquid Chromatograph: With UV detector.
4.2 Analytical balance: The sensitivity is 0.0001g.
4.3 Analytical balance: Sense 0.00001g.
4.4 Ultrasonic cleaner.
4.5 Centrifuge.
5, the measurement steps
5.1 sample processing
Accurately weigh the sample 0.1g, the exact value is 0.0001g, set 10mL with a plug colorimetric tube, add 0.1mL of sulfuric acid methanol solution (3.6), vortex 60s to mix, add sulfuric acid methanol solution (3.6) about 4.0mL, vortex Mix for 60s, ultrasonic (power: 500W) for 5min, and heat in a hot water bath at 60 °C for 5 min. After cooling to room temperature, dilute to 5 mL mark with methanolic solution of sulfuric acid (3.6), centrifuge at 4500 rpm for 10 min if necessary, remove the supernatant through a 0.45 μm microporous membrane, and take the filtrate as the test solution for later use; The mass concentration of benzaldenyl sulfonic acid exceeds the upper limit of the linear range, and the test solution is appropriately diluted. Shanghai Chuangsai Technology provides E23-TD5F desktop filter centrifuge, commodity number: E23-TD5F-å°, inquiry.
5.2 chromatographic conditions;
Column: C18 column, 250 nm × 4.6 nm, 5 μm;
Mobile phase: 20mmol/L aqueous solution of ammonium acetate (pH adjusted to pH 5.1 by acetic acid) + acetonitrile = 65+35;
Detection wavelength: 300 nm;
Column temperature: 25 ° C;
Injection volume: 10 μL.
5.3 Determination
Under 5.2 chromatographic conditions, a series of standard solutions (3.7) were injected and analyzed by liquid chromatography. The concentration of the standard solution was plotted on the abscissa and the peak area was plotted on the ordinate. Linear regression was performed to establish a standard curve. For the equation, 10 μL of the solution to be tested prepared under the condition of “5.1†was injected, and the mass concentration of the benzylidene camphorsulfonic acid was calculated according to the peak area of ​​the measured component. Calculate the content of benzylidene camphorsulfonic acid in the sample according to "6 calculation". Shanghai Chuangsai Technology provides 69912-79-4, molecular sieve, 5 Å (60-80 mesh, dedicated for gas phase and liquid chromatography columns), Molecular sieves, product number: C84-103771-100g, and the price is 410.4 yuan.
5.4 Parallel experiment
According to the above steps, the absolute difference between the two independent test results obtained by independently measuring the same sample shall not exceed 10% of the arithmetic mean.
6, calculation
ω (benzylidene camphorsulfonic acid) - content of benzylidene camphorsulfonic acid in cosmetics, %;
M——sample sample amount, g;
Ρ——substituting the regression equation to calculate the mass concentration of benzylidene camphorsulfonic acid in the sample solution, μg/mL;
Η—sample volumetric volume, mL;
D - dilution factor.
7, recovery and precision
The average recovery of several laboratories was 92.0%-107%, and the relative standard deviation was less than 7% (n=6).
8, chromatogram
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