Scope of Application and Technical Background
This standard specifies the high performance liquid chromatography (HPLC) method for the determination of salicylic acid (C7H6O3), dehydroacetic acid (C8H8O4), and 4-chlorobenzoic acid (C7H5ClO) in exported wines. The limits of detection are 2 mg/L, 3 mg/L, and 3 mg/L, respectively, and the limits of quantification are 7 mg/L, 10 mg/L, and 10 mg/L, respectively. This method was developed with reference to the relevant standards of the International Organization of Vine and Wine (OIV) and is applicable to the quality and safety supervision of exported wines.
Core detection process
| Steps | Key parameters | Technical requirements |
| Sample pretreatment | Sampling volume, pH adjustment | Take 5.0mL of wine sample and dilute to 50mL with 0.02mol/L acetic acid solution, and filter through a 0.45μm filter membrane |
| Chromatographic conditions | Chromatographic column, mobile phase | C18 column (250mm×4.6mm, 5μm), methanol-acetic acid buffer (pH5.0) (30:70), flow rate 1.0mL/min |
Mixed working solution: Mix the three standard stock solutions in proportion to prepare a 100 mg/L mixed standard solution Standard curve: Gradient dilution to a series of concentrations of 20, 10, 5.0, 2.0, 1.0, and 0.5 mg/L
Method Validation Data
Standard validation results showed that within the spiked concentration range of 7-70 mg/L, the recoveries of the three substances were 97.3%-101.6%, with relative standard deviations (RSDs) ≤8%.
- The recovery rate of salicylic acid at a spike level of 10 mg/L is 98.8%-100.0%
- The recovery rate of dehydroacetic acid at a spike level of 20 mg/L is 98.9%-100.4%
- The recovery rate of 4-chlorobenzoic acid at a spike level of 100 mg/L is 97.6%-99.9%
Implementation Tips
Precautions
1. The pH of the acetate buffer solution must be accurately adjusted to 5.0, as deviation will affect the retention time.
2. Organic filter membranes must be used for sample filtration to avoid adsorption loss.
3. The retention time drift of the chromatographic system should be checked regularly with standard solutions.
Technological Evolution
Compared with the early spectrophotometric method, this HPLC method has the following advantages:
• Simultaneous detection of three additives
• Stronger anti-interference ability
• Sensitivity increased by more than 10 times
Sample only — not a preview of SN/T 4675.15-2016
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