HPLC法测定保健食品中叶酸含量及稳定性研究
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摘要
目的建立测定保健食品中叶酸含量的液相色谱法,并用于考察保健食品中叶酸的稳定性研究。方法样品中的叶酸经0.5%氨水热水浴20分钟提取后,经ZORBAX SB-C18(150mm×4.6mm,5μm)分离,流动相为磷酸盐缓冲液-甲醇(70∶30,V/V),流速:1.0mL/min;紫外检测波长为280nm,根据保留时间定性,外标峰面积法定量。结果叶酸在4.685μg/mL～74.960μg/mL内线性关系良好(r=1),样品的加标回收率为94.8%～97.6%,平均回收率为96.0%,RSD为1.1%;检出限为0.18μg/mL;定量限为0.60μg/mL。样品经过3个月的加速破坏性试验后,检测其中的叶酸含量平均下降比率为2.6%。结论该方法具有操作简便快速、分离效果好、精密度和准确性高、重现性好的特点,可为叶酸类保健食品的质量控制提供方法参考。同时经稳定性试验发现样品中叶酸的含量变化不大,具有良好的稳定性。
The folic acid in the sample was extracted with 0.5%ammonia in hot water for 20 minutes,the separation was performed on a ZORBAX SB-C18 column(150 mm × 4.6 mm,5μm).The mobile phase of phosphate buffer-methanol(70∶30,V/V)was used at a flow rate of 1.0 ml/min and the UV detection wavelength was set at 280 nm.According to the retention time,the external standard method based on peak areas was used for quantification.Folic acid had a good linearity(r= 1)in the range of4.685—74.960μg/ml.The recoveries were in the range of 94.8%—97.6%;the average recovery was96.0%and the RSD was 1.1%.The detection limit was 0.18μg/ml while the limit of quantification was0.6μg/ml.The average decrease rate of folic acid was 2.6%after three months of accelerated destructive tests on the samples.The method is simple,fast,reproducible and of good separation effect,high precision and accuracy.Meanwhile,the contents of folic acid in the samples did not change significantly,which indicated that the folic acid was stable in the sample.
The folic acid in the sample was extracted with 0.5%ammonia in hot water for 20 minutes,the separation was performed on a ZORBAX SB-C18 column(150 mm × 4.6 mm,5μm).The mobile phase of phosphate buffer-methanol(70∶30,V/V)was used at a flow rate of 1.0 ml/min and the UV detection wavelength was set at 280 nm.According to the retention time,the external standard method based on peak areas was used for quantification.Folic acid had a good linearity(r= 1)in the range of4.685—74.960μg/ml.The recoveries were in the range of 94.8%—97.6%;the average recovery was96.0%and the RSD was 1.1%.The detection limit was 0.18μg/ml while the limit of quantification was0.6μg/ml.The average decrease rate of folic acid was 2.6%after three months of accelerated destructive tests on the samples.The method is simple,fast,reproducible and of good separation effect,high precision and accuracy.Meanwhile,the contents of folic acid in the samples did not change significantly,which indicated that the folic acid was stable in the sample.
引文
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[3]郭丽琼,曹秋旭,吴厚玖.果蔬中叶酸分析方法研究的进展[J].食品工业科技,2012,33(10):402-406
[4]马景宏,张旭,王为黎.微生物分析法测定保健食品中叶酸[J].中国微生态学杂志,2007,16(6):576-577.
[5]彭科怀,杨长晓,刘广辉.高效液相色谱法测定添加维生素保健食品中叶酸[J].预防医学情报杂志,2015,31(2):123-125.
[6]董娇,曹宁阳,乔玲,王烁,岑琴.高效液相色谱法测定保健食品中叶酸条件优化[J].农业科技与装备,2017,277(7):52-53.
[7]郭丽仪,黄永连,梁咏瑜,等.高效液相色谱法测定维生素软糖中叶酸的含量[J].食品安全导刊,2017,(15):134-136.
[8]陈璇,白小红.固相萃取-光化学荧光-高效液相色谱测定血浆中叶酸含量[J].中国药学杂志,2005,40(17):1346-1348.
[9]战英,刘思洁,姜楠,等.高效液相色谱法测定保健食品中的叶酸[J].中国卫生工程学,2007,6(5):289-290.
[10]宋金春,陆迅,曾俊芬,等.HPLC测定注射用13种复合维生素中维生素H、叶酸及维生素B12含量[J].中国药学杂志,2007,42(17):1345-1347.
[11]杨玉柱,王储炎,焦必宁.叶酸的研究进展[J].农产品加工学刊,2006,(5):31-32.
[12]石丹,贾云虹,包怡红,等.叶酸检测方法的研究现状及发展趋势[J].中国乳品工业,2009,37(3):42-43.
[13]陈璇,白小红.固相萃取光化学荧光高效液相色谱测定血浆中叶酸含量[J].中国药学杂志,2005,40(17):1346-1348.
[14]中华人民共和国药典委员会.中国药典一部[M].北京:中国医药科技出版社,2015.
[15]孙丽华,江月仙,张舟艺,等.保健食品5-氨基乙酰丙酸磷酸盐的离子对高效液相色谱法及稳定性试验[J].中国卫生检验杂志,2016,26(10):1377-1379.