6 8 Vol. 6 No. 8 2015 8 Journal of Food Safety and Quality Aug., 2015 张喜金 *, 苏昭仑, 黄康惠 (, 519040) 摘要 : 目的 方法, / (V:V=60:40), 1.2 ml/min, Waters Spherisorb C 18, 40 ; 203 nm 结果 0.1023~0.5118 mg/ml, r>0.999, 0.0025%,, 100.5%, 0.6%(n=9) WS-1001-(HD-080-18)-2002, 结论,, 关键词 : ; ; Determination of squalene in squalene soft capsule by high performance liquid chromatography ZHANG Xi-Jin *, SU Zhao-Lun, HUANG Kang-Hui (By-Health Co., Ltd., Zhuhai 519040, China) ABSTRACT: Objective To establish a method for the determination of squalene in squalene soft capsules by high performance liquid chromatography. Methods Samples were extracted with methanol, and being separated by waters spherisorb C 18 column with the mobile phase of methanol/acetonitrile (V:V=60:40), and the flow rate was 1.2 ml/min, the column temperature was 40, the content of squalene were determined by HPLC under the detection wavelength at 203 nm. Results The method showed a good linearity in the range of 0.1023~0.5118 mg/ml for squalene with the coefficient r>0.999, the detection limit was 0.0025%, the average recovery of different adding levels was 100.5%, and the relative standard deviation was 0.6%(n=9). Compared with the WS-1001-(HD-080-18)-2002 squalene gas chromatography method of National Drug Standards, there was no statistical difference between the two results. Conclusion HPLC method is more quick and easy, saving testing cost, and applicable to measure the content of squalene in health foods. KEY WORDS: squalene; high performance liquid chromatography; health food 1 引言,, 1914 squalene, 2,6,10,15,19,23- -2,6,10,14,18,22-,,, (SOD), [1-5] * 通讯作者 :, E-mail: 215865748@qq.com *Corresponding author: ZHANG Xi-Jin, By-Health Co., Ltd., Zhuhai 519040, China. E-mail: 215865748@qq.com
8, : 3109,,,,,, [6-8],,,,,,, 2 材料与方法 2.1 实验仪器 15C ( ), C 18 (5 μm, 4.6 mm 150 mm), XP-205 ( ) 2.2 标准与试剂 (SUPELCO, 99.0%), (, 2) 2.3 色谱条件 2.3.1 液相色谱条件 : Waters Sphrisorb C 18 (5 μm, 4.6 mm 150 mm); : 40 ; : 203 nm; : 1.2 ml/min; : - (V:V=60:40) 2.3.2 气相色谱条件 : Agilent 7890A, FID( ); : 280, 10:1, 1 μl; : Agilent, HP-5, 30 m 320 μm 0.25 μm, 2 ml/min; : 250, 14 min; : 280, 30 ml/min, 400 ml/min, 30 ml/min 2.4 对照品溶液的制备 10.34 mg 10 ml,, 1.0 2.0 3.0 4.0 5.0 ml 10 ml, 0.10237 0.20473 0.30710 0.40946 0.51183 mg/ml 2.5 样品溶液的制备 10.00 mg( 0.01 mg) 50 ml,,,, 0.45 μm, 3 结果与讨论 3.1 色谱条件选择 3.1.1 波长的选择 ( 1) 203 nm,, 203 nm, 203 nm 3.1.2 流动相和柱温选择,,,, /, : =60:40(V:V), 2 203 nm, 1.0 1.2 ml/min 30 40, : =60:40(V:V) 203 nm 1.2 ml/min 40, 3 4.0 min, 3.2 方法性能指标 3.2.1 工作曲线和线性范围, 5,, 0.10 mg/ml 0.50 mg/ml [9,10] Y=8 10-8 X-0.021, r 2 =0.9999, GB/T 27404-2008, GB/T 27404-2008 r 2 0.99 [11] 1 4 3.2.2 检出限实验, (S/N) 3, 0.005 μg/ml, 10 mg,
3110 6 Fig. 1 1 Ultraviolet absorption curve of squalene standard Fig. 2 2 Chromatogram of squalene standard Fig. 3 3 Chromatogram of the sample
8, : 3111 Tapble 1 表 1 对照品浓度与峰面积的关系 The relationship between the concentration of the reference and the peak area /mg ml -1 STD1 0.102366 1532512 STD2 0.204732 2836243 STD3 0.307098 4056251 STD4 0.409464 5359604 STD5 0.511830 6590887 Y=8 10-8 X-0.021 (r 2 ) 0.9999 Fig. 4 4 Standard curve of squalene 50 ml, 0.0025% 3.2.3 重复性实验 ( 20140701E), 6, 10 mg, 2.5, 2.3, 98.5%, RSD 0.2% 3.2.4 供试品含量测定 3, 2.5, 2.3, 2 3.2.5 回收率实验 10.00 mg ( 98.5 %)( 9 ), 50mL, ( 2.67795 mg/ml)2.4 ml 3.0 ml 3.6 ml,,, 0.45 μm, 100.5 %, (RSD) 0.6 %, GB/T 27404-2008( ) [11-14], 3 3.2.6 稳定性实验, 0 4 8 12 24 48 h,, RSD 0.2 % (n=6), 48 h Taple 2 表 2 不同样品中角鲨烯含量测定结果 Squalene content determination results in different samples /mg -1 / % 500 20150701E 98.5 1400 406130203 71.2 250 141101 99.2 Table 3 表 3 回收率实验 The recovery of experiment /mg /mg /mg /% /% RSD/% 8.5411 6.4271 14.9740 100.09 8.6198 6.4271 15.1515 100.62 8.5708 6.4271 15.0295 100.49 8.0196 8.0338 16.1185 100.81 8.2164 8.0338 16.2365 99.82 8.2853 8.0338 16.3535 100.42 8.6789 9.6406 18.3405 100.21 8.9642 9.6406 18.6515 100.48 8.8462 9.6406 18.5710 100.87 100.5 0.6
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