DETERMINATION OF BENZOIC AND SORBIC ACID IN TURKISH FOOD USING HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY

Benzoic and sorbic acid levels in different brands of jam, candied chestnut, carbonated drink, pickle, black table olive, green table olive, wholemeal/brown bread and white bread, which are available on the Turkish market, were determined by high-performance liquid chromatography with diode array detector (235–254 nm). Chromatographic separation was achieved with a C18 column and acetate buffer (pH 4.74)–methanol mixture (70:30) as a mobile phase. The levels of benzoic and sorbic acid in the analyzed samples were in the range of not detected to 662 mg/kg or L, and not detected to 432 mg/kg or L, respectively. Only one sample (jam) presented a preservative although not permitted by the legislation enforced in Turkey. The levels of preservative in the other samples were determined in legal limitations. According to the results, the utilization of benzoic and sorbic acid is lower than the certain legal limitations permitted by legislation to be used.

PRACTICAL APPLICATIONS

The amount of the sorbic and benzoic acid used as preservatives during food processing is important for consumer health. Determination of the amounts of the preservatives mentioned above will be possible in a short period of time (11 min) with the methods used in this study (high-performance liquid chromatography). According to the results obtained from study and data about food consumption, daily intake amount of these preservatives (benzoic and sorbic acid) will be able to be defined in Turkey, separately.     

超高效液相色谱测定蜂蜜及其制品中的苯甲酸、山梨酸

研究蜂蜜及其制品中的山梨酸与苯甲酸两种防腐剂的提取净化及其超高效液相色谱检测方法。样品用水溶解,超声波辅助提取样品中的山梨酸与苯甲酸。样液经高速离心,滤膜过滤后,采用超高效液相色谱梯度洗脱与二极管阵列检测器测定样品中的山梨酸与苯甲酸的含量。以BEH Shield RP18(2.1mm × 100mm,1.7μm)为色谱柱,以0.02mol/L 乙酸铵溶液和乙醇为流动相梯度洗脱,两种防腐剂在4min 中内实现较好的分离。方法检出限：苯甲酸为2.5mg/kg,山梨酸为2.0mg/kg。以加标样品做添加回收率测定,苯甲酸与山梨酸的回收率分别为94.1%～102.3% 和96.2%～99.8%,相对标准偏差均小于5%。方法简单、快速准确、灵敏度高是一种较好的定性和定量方法。     

流动相pH对苯甲酸等5种食品添加剂反相高效 液相色谱行为影响及分析方法的建立

目的研究反相高效液相色谱中流动相pH值对苯甲酸、山梨酸、脱氢乙酸、安赛蜜和糖精钠等5种食品添加剂色谱行为的影响。方法调节流动相甲醇-乙酸铵pH值,观察在不同品牌色谱柱中苯甲酸等5种化合物保留时间、分离度。结果在流动相pH=6.20±0.05条件下梯度洗脱,10个品牌的C_(18)色谱柱验证结果表明,苯甲酸等5种化合物均实现了基线分离。5种食品添加剂在0.10～100.0mg/L范围内线性关系良好(r~20.999),检出限和定量限分别为0.004～0.008 mg/L、0.01～0.04 mg/L。食品添加剂在调味品、肉制品类等6种基质中回收率范围为90.6%～109.0%,相对标准偏差为0.2%～7.2%。结论该方法具有稳定性强、准确、易操作且不需要刻意选择亲水性C_(18)色谱柱,适用于调味品、肉制品类等复杂基质的样品添加剂的检测。     

Sorbate and benzoate in Turkish retail foodstuffs

From 2008 to 2011, surveys were conducted to determine the levels of benzoic and sorbic acids and their respective salts in 983 retail food samples which included sauces, vegetable and fruit preparations, flavoured syrups, food supplements, cereals, bakery products, jelly, synthetic cream, sprays, mustards, jam and preserves, molasses, chewing gum, confectionery, non-alcoholic beverages, tea, wine, vinegar, brine and beers. The analysis involved methanol extraction of the foodstuff and direct determination by HPLC with UV detection. Quality assurance was employed with each batch of samples. Accuracy was ensured through regular participation in proficiency tests. Over this four-year period, a total of 23 samples (2.3%), some syrups, tomato sauces and fruit contained individual or combined levels of sorbic and benzoic acids above regulatory limits. Unauthorised use of benzoic acid was also detected in a syrup sample, bakery products and fruit preserves.     

高效液相色谱法测定酱油及食醋中的苯甲酸和山梨酸

建立了一种用高效液相色谱法快速测定酱油及食醋中苯甲酸和山梨酸的方法.使用的色谱柱为Agillent C18柱,流动相为乙酸胺:甲醇(93:7,v/v),检测波长230nm.苯甲酸和山梨酸标准溶液在2.5mg/L～50mg/L内线性良好,其线性相关系数分别0.9991和0.9984.加标回收率分别为91.7％和98.7％,测量结果的相对标准偏差(RSD)为2.77％和3.55％,苯甲酸和山梨酸的检测限分别为0.90mg/kg和0.85mg/kg.该法简单,快速,具有良好的准确度和灵敏度.     

Optimized Dispersive Liquid–Liquid Microextraction and Determination of Sorbic Acid and Benzoic Acid in Beverage Samples by Gas Chromatography

A new rapid method for direct determination of trace levels of sorbic and benzoic acids was developed by dispersive liquid–liquid microextraction and gas chromatography with flame ionization detection. In the proposed approach, the separation procedure of sorbic and benzoic acids was performed on a general chromatographic column without any prior derivatization processes. Some effective parameters on the microextraction recovery were studied and optimized utilizing multilevel factorial and central composite experimental designs. The best concurrent extraction efficiency acquired using ethanol and chloroform as dispersive and extraction solvents. Central composite design (CCD) resulted in the optimized values of microextraction parameters as follows: 1.0 mL of dispersive and 0.1 mL of extraction solvents, ionic salt concentration of 50 g?L^?1 at pH 4. Under optimum conditions, the calibration curve was linear over the range 0.5–20 mg L^?1. Relative standard deviation was 11% and 13% for five repeated determinations for sorbic and benzoic acids, respectively. Limits of detection were acquired as 0.2 mg L^?1 for sorbic acid and 0.5 mg L^?1 for benzoic acid. The average recoveries were 31% and 39% for sorbic and benzoic acids, respectively. The method was successfully applied to the determination of sorbic and benzoic acids as preservatives in beverage samples.     

RP-HPLC法同时测定糕点中安赛蜜、苯甲酸、山梨酸、糖精钠和脱氢乙酸

建立反相高效液相色谱法(RP-HPLC同时测定糕点中安赛蜜、苯甲酸、山梨酸、糖精钠和脱氢乙酸。该方法糕点样品经处理后,采用SunFire C185μ4.6 mmmm反相色谱柱,流动相为甲醇:磷酸盐缓冲液(pH6.7=793等度洗脱,流速为1.0 mL/min检测波长230 nm柱温为30℃进样量为20μ。线性范围0.2 mg/L120 mg/L检出限0.4 mg/kg1.0 mg/kg加标回收率为87.7%105.3%相对标准偏差小于5%。结果表明,该方法操作简便、结果可靠,可用于同时检测糕点中安赛蜜、苯甲酸、山梨酸、糖精钠和脱氢乙酸。     

Sorbic,benzoic and propionic acids in fishery products: a survey of the South Korean market

This study was conducted to provide basic data as part of a project to distinguish naturally occurring organic acids from added preservatives. Accordingly, we investigated naturally occurring levels of sorbic, benzoic and propionic acids in fish and their processed commodities. The levels of sorbic, benzoic and propionic acids in 265 fish and their processed commodities were determined by high-performance liquid chromatography–photodiode detection array (HPLC-PDA) of sorbic and benzoic acids and gas chromatography-mass spectrometry (GC/MS) of propionic acid. For propionic acid, GC-MS was used because of its high sensitivity and selectivity in complicated matrix samples. Propionic acid was detected in 36.6% of fish samples and 50.4% of processed fish commodities. In contrast, benzoic acid was detected in 5.6% of fish samples, and sorbic acid was not detected in any sample. According to the Korean Food and Drug Administration (KFDA), fishery products and salted fish may only contain sorbic acid in amounts up to 2.0 g kg^?1 and 1.0 g kg^?1, respectively. The results of the monitoring in this study can be considered violations of KFDA regulations (total 124; benzoic acid 8, propionic acid 116). However, it is difficult to distinguish naturally generated organic acids and artificially added preservatives in fishery products. Therefore, further studies are needed to extend the database for distinction of naturally generated organic acids and added preservatives.     

多样品同时萃取-气相色谱法测定食品中山梨酸和苯甲酸

研究了多样品同时萃取/气相色谱法对食品中山梨酸和苯甲酸的快速测定。以醋酸乙酯+正己烷(1+1)混合溶剂对样品和标准系列进行一次性萃取,用SE-30宽口径毛细柱作分析柱对萃取相进行分析。方法条件下,样品中山梨酸和苯甲酸的检出限为1mg/kg,线性范围25-500 mg/kg,回收率93.2%、102.4%,相对标准偏差(RSD)为1.9-4.1%。方法操作简便快捷易于控制,使用试剂少成本低,用于批量样品的山梨酸和苯甲酸快速测定获得良好效果。     

毛细柱气相色谱法测定饮料中的山梨酸、苯甲酸

探讨了采用毛细柱气相色谱法测定饮料中作为防腐剂的山梨酸、苯甲酸，此方法可使各组分得到良好分离，分析时间短，准确度高，山梨酸、苯甲酸测定的检出限为0．1mg／kg，相对标准偏差小于3％，样品加标回收率为89％～99％。     

The response of Gluconobacter oxydans to sorbic and benzoic acids

The minimal inhibitory concentrations (MIC) of sorbic and benzoic acids for Gluconobacter oxydans were 1000 mg/l and 900 mg/l respectively at pH 3.8. A reduction in the pH of the test medium to 3.3 reduced the MIC of both preservatives by about 300 mg/l. When G. oxydans was grown in the presence of sublethal concentrations of sorbic or benzoic acids before the MIC was determined, the MIC of both compounds increased substantially within 1 h. Growth of G. oxydans was modified in several ways by the presence of sorbic acid in the medium. The duration of the lag phase increased and there was a substantial decrease in the viable count during the lag phase in the presence of high concentrations. The generation time increased and the viable count at the end of the logarithmic phase was reduced. At 1 degree C, G. oxydans grew in the absence of sorbic acid but was inactivated by 400 mg sorbic acid/l. At 37 degrees C the viable count of suspensions of G. oxydans decreased in both the absence and presence of sorbic acid. Sorbic acid increased the death rate. Growth of G. oxydans was prevented by eliminating air from culture vessels, combined with the addition of ascorbate to the medium containing 400 mg sorbic acid/l.     

Simultaneous determination of sorbic and benzoic acids in milk products using an optimised microextraction technique followed by gas chromatography

A rapid and reliable method for direct determination of sorbic and benzoic acids in milk products was developed by dispersive liquid–liquid microextraction (DLLME) and gas chromatography with flame ionisation detector (GC-FID). A response surface methodology (RSM) based on a central composite design (CCD) was applied for optimisation of the main variables, such as volume of extraction and dispersive solvents, pH and salt effect. The primary extraction of sorbic and benzoic acids were performed in 8 mL NaOH (0.1 M) in a closed-vessel system. Carrez solutions (potassium hexaferrocyanide and zinc acetate) were used for protein sedimentation. The best simultaneous extraction efficiency was identified using acetone and 1-octanal as dispersive and extraction solvents, respectively. For DLLME, central composite design resulted in the optimised values of microextraction parameters as follows: 475 µL of dispersive and 60 µL of extraction solvents, 2 g NaCl at pH 2.5. Under optimum conditions, the calibration curve was linear over the range 0.1–50 μg mL^?1 and the square of correlation coefficient (R²) was 0.9992 for sorbic acid and 0.9994 for benzoic acid. Relative standard deviation (RSD %) was 6.1% and 3.1% (n = 5) for sorbic and benzoic acids, respectively. Limits of detection were 150 ng g^?1 for sorbic acid and 140 ng g^?1 for benzoic acid and recoveries were 88% and 103.7% respectively. Good reproducibility (RSD %), short extraction time and no matrix interference were advantages of the proposed method which was successfully applied to the determination of sorbic and benzoic acids in milk products.     

气相色谱法测定糕点中山梨酸和苯甲酸的含量

目的建立了糕点中山梨酸和苯甲酸的毛细管气相色谱检测方法。方法样品加有机溶剂并用高速匀浆的方法提取后,通过改变酸碱度而改变样品在有机相和水相中分配比的方法,去除脂肪等杂质的干扰,用FFAP毛细管色谱柱分离,FID检测器进行检测。结果本法的变异系数为1.2%～3.9%,回收率为89.0%～99.2%,最低检出限为1mg/kg。结论本法具有步骤简单、准确度和灵敏度高的特点,适用于糕点中山梨酸和苯甲酸的检测。     

发酵食品中多种防腐剂检测方法的建立

该实验建立了高效液相色谱法同时测定苯乳酸、苯甲酸、山梨酸的方法。在以C18柱为分离柱,流动相为甲醇-0.020 mol/L乙酸铵（20∶80,V/V）,色谱柱温为25 ℃,流速为1.0 mL/min,检测波长为220 nm条件下,12 min内可实现苯乳酸、苯甲酸、山梨酸的分离。该方法在0.25～50.00 mg/L范围内具有良好的线性关系（相关系数R2＞0.998）,检出限（LOD）＜50.00 ng/mL,回收率为96.93%～118.42%,精密度试验结果相对标准偏差（RSD）为0.39%～6.21%。该方法操作简便、结果准确度高,可用于发酵食品中苯乳酸、苯甲酸和山梨酸的同时分析检测。     

HPLC同时测定豆制品中常见的3种食品添加剂

采用反相高效液相色谱法快速、准确测定豆制品中的苯甲酸、山梨酸和糖精钠。3种添加剂在Zorbax sb-C18色谱柱上分离,甲醇-乙酸铵溶液(5∶95,V∶V)为流动相洗脱,流速1.0mL/min,15min内实现良好分离。方法检出限为0.5mg/kg,变异系数为0.67%～1.23%,回收率在97.7%～99.7%。该方法简便准确,适合豆制品的日常检测。     

配制酒中的苯甲酸和山梨酸的快速测定

采用高效液相色谱法建立了快速分析配制酒中苯甲酸和山梨酸的方法。最佳色谱条件为：柱温25℃,流动相体系为甲醇与0.02 mol/L乙酸铵溶液体积比20＋80,流速1 mL/min。此方法快捷可靠,精密度高,适用于大量样品的快速测定。     

肉制品中苯甲酸、山梨酸测定方法的研究

采用高效液相色谱法可以同时完成苯甲酸、山梨酸的测定。色谱条件:流动相为甲醇∶乙酸铵=7∶93,流速1.0mL/min,于230nm波长处同时测定肉制品中苯甲酸、山梨酸。因肉制品成分复杂,在苯甲酸出峰处干扰峰多,非常容易引起误判,所以采用改变流动相pH值的方法来建立苯甲酸、山梨酸的确认方法。该法的回收率大于90%,采用气-质联机确认,二者结果一致。本方法也同样适用于其它复杂食品的测定。     

高效液相色谱法测定月饼中3种防腐剂

利用高效液相色谱法测定5种月饼样品中的3种常见防腐剂苯甲酸、山梨酸和脱氢乙酸的含量。样品经提取、脱蛋白、膜过滤后,用高效液相色谱进行测定。结果表明,3种防腐剂的测定线性相关性均在0.99以上,检测线性范围为10μg/mL～200μg/mL。该方法应用于5种月饼中苯甲酸、山梨酸和脱氢乙酸的测定结果表明,5种样品均检出山梨酸,3种样品检出有脱氢乙酸,均未检出苯甲酸。     

Estimated intake of benzoic and sorbic acids in Denmark

The monitoring of food additives and recent dietary surveys carried out in Denmark have earlier been used to estimate the intake of sweeteners and nitrite in relation to acceptable daily intakes. The ubiquitous use of the preservatives benzoic and sorbic acids raises the question of the magnitude of the intake of these preservatives in relation to acceptable daily intakes. This area is explored in this paper. The content of benzoic and sorbic acids in all food groups, where they are allowed, was monitored in Denmark 17 times between 2001 and 2006 with a total of 1526 samples. Transgressions of maximum limits, illegal use or declaration faults were found in about 3% of samples. From repeated investigations on fat-based foods (salads and dressings), marmalade and stewed fruit, it is concluded that the amounts used in industry have been relatively stable throughout the whole period, although limited data for marmalade show some variation. Most foods in the categories soft drinks, dressings, fat-based salads, pickled herrings, and marmalade contain benzoic and sorbic acid, and sliced bread also contains in some cases sorbic acid. The median daily intake and intake distribution of benzoic and sorbic acids were calculated with data from the Danish National Survey of Dietary Habits and Physical Activity (age from 4 to 75 years) conducted in 2000–2004 with 5785 participants. The median intakes of both benzoic acid and sorbic acid are well below the acceptable daily intakes of 0–5 and 0–25 mg kg^?1 body weight (bw) day^?1 for benzoic and sorbic acid, respectively. However, the 90th percentile based on the average of the samples with a content of benzoic acid is higher than the acceptable daily intake for both men and women, with the highest value of 16 mg kg^?1 bw day^?1 for both boys and girls in the 4–6-year-old age group. Based on the average of all samples, the 95th percentile is over the acceptable daily intake for men up to 34 years and for women up to 24 years, and the 90th percentile for men up to 18 years and for women up to 10 years. Soft drinks, salads and dressings are the main contributors to benzoic acid intake. The sorbic acid intake based on the average of all samples is well below the acceptable daily intake. However, for the intake based on the average of samples with content, the 95th percentile exceeds the acceptable daily intake. This is caused by the dominating contribution to the intake of sorbic acid from sliced bread, but since only seven out of 42 samples have added sorbic acid, the calculation based on the average of samples with content will exaggerate the intake. With a built-in safety factor of 100 in the acceptable daily intakes and judging from the literature, the high intakes of benzoic acid should not cause any concern for ill-effects. However, there must be a reason to reconsider the maximum limits especially for benzoic acid in soft drinks, dressings and salads and for sorbic acid in sliced bread.     

香蕉汁中苯甲酸、柠檬黄和日落黄标准物质的研制

目的 建立香蕉汁中苯甲酸、柠檬黄和日落黄标准物质的研制与定制方法。方法 采用单因素方差分析均匀性, 线性回归趋势分析稳定性, 多家实验室合作定制, 并计算其不确定度。结果 标准值及扩展不确定度(k=2): 苯甲酸(209.96±7.52) mg/kg; 柠檬黄(302.18±4.98) mg/kg; 日落黄(41.52±0.66) mg/kg。结论 研制的标准物质均匀性和稳定性较好, 可用于食品安全监测及相关饮料的质量控制。