高效液相色谱-蒸发光散射法测定保健食品中磷脂酰丝氨酸的方法优化
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摘要
目的优化高效液相色谱-蒸发光散射法测定保健食品中磷脂酰丝氨酸含量的分析方法。方法以氯仿-甲醇溶液(9:1, V:V)为提取剂,样品经旋涡混合辅助超声提取后高效液相色谱仪测定。以硅胶柱(4.6 mm×250 mm,5μm)作为分离柱,流动相为异丙醇:水:乙酸:三乙胺(84:16:1.5:0.08, V:V:V:V),流速1.0 mL/min,等度洗脱,柱温40℃。蒸发光散射检测器选择不分流模式,漂移管温度100℃,载气(氮气)流速为2.0 L/min,增益值为200。结果磷脂酰丝氨酸在20～320mg/L浓度范围内现呈良好线性关系,相关系数为0.9999,加标回收率为95.7%～100.7%,检出限为4.0 mg/100 g,定量限为11 mg/100 g。采用该方法对14种保健食品中磷脂酰丝氨酸的含量进行测定,结果符合产品质量标准。结论该方法适用范围广,有机溶剂用量少,数据准确可靠和重现性好,可满足实验要求。
Objective To optimize the method for determination of phosphatidylserine in health foods by high performance liquid chromatography-evaporative light scattering detection(ELSD). Methods The samples were extracted with chloroform-methanol mixture(9:1, V:V) by vortex mix-assisted ultrasound and determined by high performance liquid chromatography. The separation was conducted on a silica gel column(4.6 mm×250 mm, 5 μm)with a mobile phase of isopropanol:water:acetic acid:triethylamine(84:16:1.5:0.08, V:V:V:V). The flow rate was 1.0 mL/min with a isocratic elution system and the column temperature was set at 40 ℃. The ELSD parameters were non-distributary mode, drift tube temperature of 100 ℃, flow rate of nebulizing gas(N_2) of 2.0 L/min, and gain of200, respectively. Results Phosphatidylserine had good linearity in the concentration range of 20-320 mg/L with the correlation coefficient of 0.9999. The recoveries were between 95.7% and 100.7%. The limit of detection was 4.0 mg/100 g and the limit of quantitation was 11 mg/100 g. The method was used to determine the content of PS in 14 kinds of health foods and the results were consistent with the product quality standard. Conclusion This method has the advantages of wide applicability, less solvent consumption, good accuracy and reproducibility, which can meet the requirements of experiment.
Objective To optimize the method for determination of phosphatidylserine in health foods by high performance liquid chromatography-evaporative light scattering detection(ELSD). Methods The samples were extracted with chloroform-methanol mixture(9:1, V:V) by vortex mix-assisted ultrasound and determined by high performance liquid chromatography. The separation was conducted on a silica gel column(4.6 mm×250 mm, 5 μm)with a mobile phase of isopropanol:water:acetic acid:triethylamine(84:16:1.5:0.08, V:V:V:V). The flow rate was 1.0 mL/min with a isocratic elution system and the column temperature was set at 40 ℃. The ELSD parameters were non-distributary mode, drift tube temperature of 100 ℃, flow rate of nebulizing gas(N_2) of 2.0 L/min, and gain of200, respectively. Results Phosphatidylserine had good linearity in the concentration range of 20-320 mg/L with the correlation coefficient of 0.9999. The recoveries were between 95.7% and 100.7%. The limit of detection was 4.0 mg/100 g and the limit of quantitation was 11 mg/100 g. The method was used to determine the content of PS in 14 kinds of health foods and the results were consistent with the product quality standard. Conclusion This method has the advantages of wide applicability, less solvent consumption, good accuracy and reproducibility, which can meet the requirements of experiment.
引文
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[6]师维,李煜惠,锁显,等.离子液体选择性萃取分离磷脂酰丝氨酸和磷脂酰胆碱[J].化工学报,2017,68(9):3442-3450.Shi W,Li YH,Suo X,et al.Selective separation of phosphatidylserine and phosphatidylcholine with ionic liquids as extractants[J].J Chem Ind Eng,2017,68(9):3442-3450.
[7]林宏琳,陈华峰,林国斌.酰胺基亲水作用色谱-高效液相色谱-紫外法测定DHA颗粒中的磷脂酰丝氨酸[J].药物分析杂志,2017,37(11):1962-1966.Lin HL,Chen HF,Lin GB.Quantification of phosphatidylserine in DHAgranules by amide-HPLC-UV[J].Chin J Pharm Anal,2017,37(11):1962-1966.
[8]饶雪娥,林伟铃.磷脂酰丝氨酸的应用研究进展[J].海峡药学,2015,27(11):9-11.Rao XE,Lin WL.Application research advance of phosphatidylserine[J].Strait Pharm J,2015,27(11):9-11.
[9]苏永恒,张伟,马青青.高效液相色谱-蒸发光散射检测法测定保健食品中的磷脂酰丝氨酸[J].中国卫生检验杂志,2017,27(6):792-794.Su YH,Zhang W,Ma QQ.Determination of phosphatidylserine in health food by high performance liquid chromatograph with evaporative light scattering detector[J].Chin J Health Lab Technol,2017,27(6):792-794.
[10]廖惠云,刘献军,庄亚东,等.涡旋混合辅助超声提取-气相色谱-质谱联用内标法分析食品添加剂中的咪唑[J].食品科学,2013,34(24):238-242.Liao HY,Liu XJ,Zhuang YD,et al.Determination of imidazole in food additives by using vortex mixing coupled with ultrasonic extraction and GC-MS[J].Food Sci,2013,34(24):238-242.
[11]王国祥,戴琛,张春华.高效液相色谱蒸发光散射检测器测定乳制品中磷脂类化合物[J].化学工程与装备,2018,(1):243-245.Wang GX,Dai C,Zhang CH.Determination of phospholipids in dairy products by high performance liquid chromatograph with evaporative light scattering detector[J].Chem Eng Equip,2018,(1):243-245.
[12]刘彬丽,唐亚利,金绍明,等.高效液相色谱法测定大豆磷脂类保健食品中磷脂酰胆碱含量[J].食品安全质量检测学报,2019,10(1):90-94.Liu BL,Tang YL,Jin SM,et al.Determination of phosphatidylcholine from soybean phospholipid health products by high performance liquid chromatography[J].J Food Saf Qual,2019,10(1):90-94.
[13]游景水,莫毅梅,林丽,等.高效液相色谱-蒸发光散射检测法测定保健食品中磷脂酰丝氨酸[J].食品安全质量检测学报,2018,9(10):2537-2540.You JS,Mo YM,Lin L,et al.Determination of phosphatidylserine in health food by high performance liquid chromatography with evaporative light scattering detector[J].J Food Saf Qual,2018,9(10):2537-2540.
[14]石丹,王晓彦,魏显纲,等.高效液相色谱检测奶粉中磷脂酰丝氨酸分析方法的建立[J].营养学报,2015,37(1):88-92.Shi D,Wang XY,Wei XG,et al.Establishment of analytical method for detection of phosphatidylserine in milk powder by high performance liquid chromatography[J].Acta Nutr Sin,2015,37(1):88-92.
[15]潘城,郑小严,黄红霞,等.高效液相色谱/蒸发光散射检测法同时测定饲料中12种甜味剂[J].分析测试学报,2015,34(10):1107-1112.Pan C,Zheng XY,Huang HX,et al.Simultaneous determination of twelve sweeteners in feeds by high performance liquid chromatography/evaporative light scattering detection[J].J Instrum Anal,2015,34(10):1107-1112.
[2]Kato-Kataoka A,Sakai M,Ebina R,et al.Soybean-derived phosphatidylserine improves memory function of the elderly japanese subjects with memory complaints[J].J Clin Biochem Nutr,2010,47:246-255.
[3]Mcdaniel MA,Maier SF,Einstein GO.“Brain-Specific”nutrients:Amemory cure?[J].Nutrients,2003,19(11/12):957-975.
[4]Jorisse BL.Safety of soy-derived phosphatidyl-serine in elderly people[J].Nutr Neurosci,2002,5(5):337-343.
[5]冯雷刚,周红梅,胡光祥,等.磷脂酰丝氨酸的应用研究[J].食品研究与开发,2006,27(9):191-193.Feng LG,Zhou HM,Hu GX,et al.Application research on phosphatidylserine[J].Food Res Dev,2006,27(9):191-193.
[6]师维,李煜惠,锁显,等.离子液体选择性萃取分离磷脂酰丝氨酸和磷脂酰胆碱[J].化工学报,2017,68(9):3442-3450.Shi W,Li YH,Suo X,et al.Selective separation of phosphatidylserine and phosphatidylcholine with ionic liquids as extractants[J].J Chem Ind Eng,2017,68(9):3442-3450.
[7]林宏琳,陈华峰,林国斌.酰胺基亲水作用色谱-高效液相色谱-紫外法测定DHA颗粒中的磷脂酰丝氨酸[J].药物分析杂志,2017,37(11):1962-1966.Lin HL,Chen HF,Lin GB.Quantification of phosphatidylserine in DHAgranules by amide-HPLC-UV[J].Chin J Pharm Anal,2017,37(11):1962-1966.
[8]饶雪娥,林伟铃.磷脂酰丝氨酸的应用研究进展[J].海峡药学,2015,27(11):9-11.Rao XE,Lin WL.Application research advance of phosphatidylserine[J].Strait Pharm J,2015,27(11):9-11.
[9]苏永恒,张伟,马青青.高效液相色谱-蒸发光散射检测法测定保健食品中的磷脂酰丝氨酸[J].中国卫生检验杂志,2017,27(6):792-794.Su YH,Zhang W,Ma QQ.Determination of phosphatidylserine in health food by high performance liquid chromatograph with evaporative light scattering detector[J].Chin J Health Lab Technol,2017,27(6):792-794.
[10]廖惠云,刘献军,庄亚东,等.涡旋混合辅助超声提取-气相色谱-质谱联用内标法分析食品添加剂中的咪唑[J].食品科学,2013,34(24):238-242.Liao HY,Liu XJ,Zhuang YD,et al.Determination of imidazole in food additives by using vortex mixing coupled with ultrasonic extraction and GC-MS[J].Food Sci,2013,34(24):238-242.
[11]王国祥,戴琛,张春华.高效液相色谱蒸发光散射检测器测定乳制品中磷脂类化合物[J].化学工程与装备,2018,(1):243-245.Wang GX,Dai C,Zhang CH.Determination of phospholipids in dairy products by high performance liquid chromatograph with evaporative light scattering detector[J].Chem Eng Equip,2018,(1):243-245.
[12]刘彬丽,唐亚利,金绍明,等.高效液相色谱法测定大豆磷脂类保健食品中磷脂酰胆碱含量[J].食品安全质量检测学报,2019,10(1):90-94.Liu BL,Tang YL,Jin SM,et al.Determination of phosphatidylcholine from soybean phospholipid health products by high performance liquid chromatography[J].J Food Saf Qual,2019,10(1):90-94.
[13]游景水,莫毅梅,林丽,等.高效液相色谱-蒸发光散射检测法测定保健食品中磷脂酰丝氨酸[J].食品安全质量检测学报,2018,9(10):2537-2540.You JS,Mo YM,Lin L,et al.Determination of phosphatidylserine in health food by high performance liquid chromatography with evaporative light scattering detector[J].J Food Saf Qual,2018,9(10):2537-2540.
[14]石丹,王晓彦,魏显纲,等.高效液相色谱检测奶粉中磷脂酰丝氨酸分析方法的建立[J].营养学报,2015,37(1):88-92.Shi D,Wang XY,Wei XG,et al.Establishment of analytical method for detection of phosphatidylserine in milk powder by high performance liquid chromatography[J].Acta Nutr Sin,2015,37(1):88-92.
[15]潘城,郑小严,黄红霞,等.高效液相色谱/蒸发光散射检测法同时测定饲料中12种甜味剂[J].分析测试学报,2015,34(10):1107-1112.Pan C,Zheng XY,Huang HX,et al.Simultaneous determination of twelve sweeteners in feeds by high performance liquid chromatography/evaporative light scattering detection[J].J Instrum Anal,2015,34(10):1107-1112.