表面增强拉曼光谱法定性定量检测保健食品中非法添加物西地那非

建立表面增强拉曼光谱法定性定量检测保健食品中非法添加药物西地那非。利用拉曼光谱的特异性进行西地那非定性检测;用三聚氰胺作为内标物进行西地那非定量检测,以西地那非特征峰1 234 cm~(-1)与三聚氰胺特征峰707 cm~(-1)的峰高比为纵坐标,西地那非质量浓度为横坐标进行线性回归拟合,结果表明西地那非质量浓度在0～50 mg/L范围呈现良好的线性关系,R~2=0.996 9。定性检测限为0.05 mg/L,定量检测限为5 mg/L。其固体、液体样品加标回收率分别为85.6%～92.6%、98.7%～105.2%,相对标准偏差分别为4.14%、3.25%。采用本方法对实际样品进行检测,其结果与高效液相色谱-质谱法测量结果基本一致。本方法准确、灵敏、快速,可用于保健品中非法添加西地那非类药物的定性定量检测。     

HPLC/MS定性定量检测清脑保心康胶囊中非法添加的氢氯噻嗪

目的:建立HPLC/MS方法,快速定性定量检测保健食品中非法添加的氢氯噻嗪。方法:采用高效液相色谱串联三重四级杆质谱仪进行检测。色谱条件:Agilent Eclipse Plus C18(4.6 mm×100 mm,3.5μm)色谱柱。柱温:30℃。流动相:甲醇:0.1%甲酸水溶液=80:20。流速为0.5 m L/min。质谱条件:ESI源,多反应监测(MRM)。以保留时间及定性离子对之间的相对丰度定性,以定量离子对峰面积定量。结果:氢氯噻嗪在10.29~1 028.79 ng/m L范围内呈现良好的线性关系(r0.999 7)。平均加样回收率:99.12%。定量限为10.29 ng。结论:本方法定性可靠、定量准确,且快速简便。可用于保健食品中非法添加氢氯噻嗪的检验。     

高效液相色谱对保健食品中五种违禁的性功能药物成分的同时检测

目的:建立一种同时测定保健食品中5种常见违禁添加的增强性功能药物成分的反相液相色谱方法"用二极管阵列检测器检测,利用色谱峰保留时间和物质的紫外吸收光谱综合定性,外标法定量.方法:采用反相高效液相色谱法实现5种成分完全分离.Agilent C18 4.6×250mm色谱柱,柱温30%,流动相为乙腈/0.02%磷酸,梯度洗脱,流速1.0 mL/min,检测波长为225.0nm.结果:5种违禁性功能药物成分在0.4～20.0 μg/mL浓度范围内工作曲线线性关系良好.相关系数r2≥0.9989,相对标准偏差≤0.47%(n=6),平均回收率＞91.0%,最低检出限≤56ng/mL(信噪比≥3).结论:该方法简便快速,结果准确可靠,可用于保健食品中常见添加的性功能药物成分的测定.     

高效液相色谱法检测保健食品中的违禁药物

目的建立高效液相色谱法同时测定保健食品中的3大类共10种违禁添加药物的方法。方法样品经色谱分离后采用二极管阵列(PDA)检测器检测,以紫外特征吸收谱图作为定性的依据。结果苯乙双胍、芬氟拉明、格列齐特、格列甲嗪、西力士、格列苯脲、西地那非、格列美脲、伐地那非、盐酸西布曲明的最低检出限分别为8·26、38·34、13·37、22·44、6·90、16·08、25·32、40·43、40·94、11·95ng/ml;线性关系在0·5~50μg/ml浓度范围,相关系数(r)均大于0·9998,样品加标回收率大于85%,RSD小于5·20%。结论该方法样品处理简便,结果准确可靠,重复性好,灵敏度高。     

高效液相色谱/质谱联用测定保健食品中的伐地那非

为测定保健食品中伐地那非，建立了高效液相色谱质谱联用（HPLC—ES／MS）的测定方法。该方法以紫外定量，质谱定性。色谱条件为RP—C18色谱柱，流动相：0．2％甲酸水溶液＋乙腈=65＋35（体积分数）；流速1ml／min；进样量5μl；检测波长254nm。质谱采用电喷雾正离子模式（ESI＋），扫描范围为：200-700（m/z）。伐地那非在3．6—450μg/ml范围内，浓度与峰面积呈良好的线性关系，定量检出限为18ng，最低检出限为0．9ng（S/N=6）。本方法流动相简单，分析时间短且试样预处理简单，灵敏度高，借助质谱的定性能力，可大大提高方法可靠性和抗干扰能力，能准确快速地测定保健食品中的伐地那非。     

LC-Q-TOF-MS高分辨质谱法测定保健食品中24种抗炎、镇静安神类违禁药物

建立保健食品中抗炎、镇静安神类违禁药物的液相色谱联用四极杆飞行时间高分辨质谱(LC-Q-TOFMS)的检测方法。对比分析溶剂提取法与QuEChERS前处理法提取违禁药物,高分辨质谱数据依存获取的采集模式(IDA-MS),获得目标物的一级母离子的精确质量数、同位素丰度比、二级碎片离子的精确质量数等信息,进行定性、定量分析24种抗炎镇痛、镇静安神类违禁药物。结果显示,QuEChERS前处理技术与IDA-MS高分辨质谱分析方法检测保健食品中抗炎、镇静安神类违禁药物选择性好,检测限和定量限低,灵敏度高,线性范围良好(r20.997),回收率在81.8%～103%之间。     

超高效液相色谱-三重四极杆/复合线性离子阱质谱法快速筛查和鉴定保健食品中非法添加的新型西地那非类衍生物

目的建立一种超高效液相色谱-三重四极杆/复合线性离子阱质谱(ultra performance liquid chromatography-triple quadrupole linear/ion trap of mass spectrometry,UPLC-Qtrap-MS/MS)检测方法,用于保健食品中西地那非类衍生物的快速筛查和鉴定。方法样品经甲醇提取,经甲醇-水(50:50,V:V)适当稀释,采用0.1%甲酸水(A)和0.1%甲酸乙腈(B)作为流动相进行洗脱,质谱(ESI+)采用母离子扫描模式(precursor ion)对西地那非类似物进行检测,利用一级、二级、三级质谱碎片进行结构解析。结果通过该方法成功发现了一种新型的西地那非类衍生物。结论该方法快速、准确、灵敏,可用于保健食品中西地那非类衍生物的筛查。     

高效液相色谱-电喷雾串联三重四级杆质谱法测定水中微囊藻毒素

目的研究利用固相提取净化、高效液相色谱-电喷雾串联三重四级杆质谱检测分析水中微囊藻毒素MC-LR和MC—RR。方法水样品经玻璃纤维滤纸过滤后,经C18固相提取柱富集净化,氮吹后定容。流动相中添加0．1％甲酸,采用梯度洗脱模式,经C18反相色谱柱分离,以电喷雾串联三重四级杆质谱定性、定量测定。结果线性定量范围0．1～5．0μg／kg,检测低限为MC-LR 0．01μg／kg,MC—RR 0．02μg／kg。结论此方法操作简便、定性定量能力较强,可作为检测水质量和评估食品安全风险的可靠方法。     

超高效液相色谱-串联质谱法测定保健食品中78种非法添加化学药物

建立了超高效液相色谱-串联质谱法快速筛查减肥、抗疲劳和增强免疫力类保健食品中非法添加78种化学药物的方法。保健食品样品经甲醇溶剂提取、QuEChERS净化，试样以C18柱分离，多反应监测（MRM）模式进行定量与定性分析，采用外标法定量。采用ACQUITY UPLC HSS T3柱（2.1×100 mm，1.7 μm）进行分离，以乙腈和乙酸铵溶液（含0.1%甲酸）作为流动相，进行梯度洗脱,质谱采用正离子和负离子同时扫描分析。在优化的色谱、质谱条件下，78种化学药物能够得到较好分离，在线性范围内线性关系良好，相关系数均大于0.9991，检出限（LOQ）为0.5 μg/kg~420 μg/kg。各组分在三个不同加标水平下平均回收率为59.9%~120.7%，相对标准偏差（RSD）为1.3%~16.2%。运用本方法对57批次保健食品进行快速筛查，检出41批次样品含有一种或多种非法添加药物,被检出的药物为西布曲明、苄基西布曲明、氯代西布曲明、酚酞、伪麻黄碱、麻黄碱、二甲双胍、咖啡因、茶碱、番泻苷A、番泻苷B、呋塞米、大黄素和西地那非。本方法样品处理过程简单，分析时间短，准确可靠，灵敏度高，适用于保健食品中非法添加化学药物的定性与定量检测及快速筛查。     

HPLC-Q-TOF/MS测定保健食品中19种抗抑郁药物

目的：建立保健食品中非法添加19种抗抑郁药物的高效液相色谱—四极杆—飞行时间质谱快速筛查定性与定量分析方法。方法：样品采用甲醇溶液进行超声提取,提取液经C₁₈色谱柱分离,以0.1%甲酸水溶液—乙腈为流动相,梯度洗脱,在电喷雾正离子模式下进行检测,检测结果以试验方法建立的19种抗抑郁药物一级精确质量数据库和二级碎片质谱库进行筛查匹配。结果：在5～100 μg/L质量浓度范围内,19种目标化合物线性关系良好,相关系数为0.988 7~0.999 9,在2,5,10 μg/kg 3个加标水平下样品的回收率为81.2%～98.6%,相对标准偏差为0.9%～6.7%。结论：所建立的方法适用于保健食品中非法添加抗抑郁药物的快速筛查定性与定量分析。     

电喷雾-离子迁移谱法快速筛查保健酒饮料中3种壮阳类药物

目的建立电喷雾-离子迁移谱(ESI-IMS)快速筛查保健酒饮料中违法添加西地那非、硫代艾地那非和伐地那非3种壮阳类违禁药物的方法。方法样品经乙腈超声提取,稀释定容后直接测定。以ESI作为电离源,空气作为迁移气体,在正离子模式进行检测,基质外标法定量。结果西地那非、伐地那非、硫代艾地那非分别在1.25~12.50、2.00~20.00、1.00~10.00 mg/L范围内线性关系良好,相关系数(r)均大于0.99,3种违禁药物的检测限分别为0.50、0.80、0.40 mg/L。平均回收率为93.33%~121.17%,相对标准偏差(RSD)为2.65%~10.00%,实际样品检测与气相色谱-质谱联用法确证结果一致。结论该方法快速简便,结果准确可靠,适用于保健酒饮料中非法添加西地那非、硫代艾地那非和伐地那非3种壮阳类违禁药物的快速筛查。     

表面增强拉曼光谱法快速测定保健酒中西地那非

目的 建立金核银壳结构纳米颗粒(Au@Ag NPs)的表面增强拉曼光谱法(surface-enhanced Raman scattering, SERS)快速检测保健酒中非法添加物西地那非的分析方法。方法 制备3种纳米粒子Ag、Au、Au@Ag NPs为SERS基底, 比较3种基底的增强效果。样品前处理基于溶剂萃取法, 利用二氯甲烷对保健酒中的西地那非进行简单提取, 通过调节体系pH值, 得到最佳提取率和SERS增强效果。结果 Au@Ag NPs的SERS增强效应优于Au NPs和Ag NPs; 用0.1 mol/L氢氧化钾调节溶液pH值可有效提高二氯甲烷的提取效果, 再用0.1 mol/L稀盐酸溶解挥发后残留物, 使得西地那非在pH调节后更有利于吸附在Au@Ag NPs表面, 获得更强的SERS信号。西地那非的检出限为0.5 mg/L, 在0.5~10 mg/L浓度范围内线性关系较好, 相关系数(r2)为0.9472, 回收率为86.0%~95.8%之间, 相对标准偏差(relative standard deviation, RSD)为3.6%~5.9%。 结论 SERS技术灵敏度高、特异性强, 可用于快速检测保健酒中的西地那非, 为快速筛查大量样品提供新方法。     

降血糖类保健食品中非法添加的8 种药物检测及实例分析

目的：建立检测降血糖类保健食品中非法添加8 种药物的方法并用于实际检测。方法：采用高效液相色谱-串联质谱法,以ZORBAX Eclipse Plus C18（2.1 mm×50 mm,1.8 μm）色谱柱,以0.1%甲酸溶液为流动相A,甲醇为流动相B,梯度洗脱。通过分子离子峰、二级碎片、色谱保留时间等信息,对宣称辅助降血糖类保健食品非法添加8 种化学药物进行鉴别和定量测定。结果：8 种药物实现了同时分离与专属鉴定,检出限为5.5～76 ng/g,精密度为0.22%～1.19%,回收率为74.0%～111.9%;本法适用于硬胶囊样品;利用本法,从15 批样品中检测出9 批含非法添加化学药物。结论：本方法专属性强、灵敏度高,结果准确可靠,可作为宣称辅助降血糖类保健食品中检测非法添加化学药物的方法参考。     

超高效液相色谱-二极管阵列检测器法快速测定保健食品中违法添加的14 种性功能药物

建立超高效液相色谱-二极管阵列检测器检测方法同时、快速测定抗疲劳类及增强免疫力类保健食品中违法添加的14 种性功能类化学药物（那红地那非、红地那非、伐地那非、羟基豪莫西地那非、枸橼酸西地那非、豪莫西地那非、氨基他达拉非、他达拉非、硫代艾地那非、伪伐地那非、那莫西地那非、康力龙、丙酸睾酮、苯丙酸诺龙）。采用ZORBAX SB-C18色谱柱,流动相A：乙腈-甲醇（90﹕10,V/V）,B：0.7%三乙胺溶液（pH 2.8）,梯度洗脱,流速0.35 mL/min。采集230 nm波长处的色谱图进行初筛和定量。根据14 种药物的光谱特征建立光谱库,确证检出化合物。结果表明：14 种性功能药物在15.0 min内可有效分离,在8～80 μg/mL的线性范围内相关系数均大于0.999 5,平均回收率为90.1%～106.5%,相对标准偏差为0.7%～4.9%,检测限为2～3 ng。该方法简便、快速、灵敏度高且重复性好,可作为检测抗疲劳类及增强免疫力类保健食品中违法添加化学药物的快速测定方法。     

Development and evaluation of an immunochromatographic strip for rapid screening of sildenafil-type compounds as illegal additives in functional foods

Sildenafil is a phosphodiesterase-5 inhibitor (PDE-5) for the treatment of erectile dysfunction. Undeclared sildenafil and related analogues adulterated in functional foods are a threat to public health. To screen these illegal drugs rapidly in herbal samples, an immunochromatographic (IC) assay was developed based on polyclonal antibodies specific to both sildenafil and its analogues. A group that is pharmacological necessary for sildenafil and its analogues was employed as a representative hapten for the generation antibodies against the target compounds. The desired antisera showed satisfactory specificities to sildenafil and major analogues with IC₅₀ values ranging from 19.3 to 34.6 ng ml^–1 in a referring enzyme-linked immunosorbent assay (ELISA). The optimised IC assay showed detection thresholds in the range 5.0–20 μg g^–1 for sildenafil and major analogues in herbal samples. Sixty herbal food supplements were screened and six were found to be positive using the IC strip. It was confirmed by ELISA and UPLC-PDA-MS/MS that positive samples contain target illegal additives in levels of 10–40 mg g^–1 (1–4%). In this range, sensitivity of the IC strip is adequate to screen sildenafil-type compounds in herbal commodities under a dilution ratio of 1:10³. Thus, the current IC assay is a suitable tool for screening sildenafil and its analogues as illegal additives in herbal food supplements.     

高效液相色谱法检测保健品中西地那非的含量的不确定度评定

目的 评定高效液相色谱法(high performance liquid chromatography,HPLC)检测保健品中西地那非的含量的不确定度。方法 以保健品中西地那非非法添加为例,采用高效液相色谱对样品进行定性筛查,定量分析,幵对定量结果分析不确定度来源,计算测量结果的相对扩展不确定度。结果 当保健品中西地那非含量为14.0mg/g,在95%的置信区间下,其结果的扩展不确定度为0.36mg(k=2)。结论 在影响检测结果的各分量中,标准溶液配制过程引入的不确定度对结果影响较大。     

TLC-UPLC-MS/MS法检测网红保健品中非法添加的8种化学药物

建立薄层色谱-超高效液相色谱-串联质谱法同时检测网红保健品中非法添加的8 种化学药物。样品采用薄层色谱法进行前处理,采用Kinetex 100 RP-18（100 mm×2.1 mm,1.6 μm）色谱柱分离,以乙腈-1 mmol/L乙酸铵（含0.1%乙酸）溶液为流动相梯度洗脱,采用多重反应监测方式检测西布曲明、洛伐他汀、辛伐他汀、麻黄碱、咖啡因、酚酞、苯乙双胍和西地那非等8 种药物。结果表明,8 种化合物在0.5～100 μg/L范围内线性关系良好,相关系数≥0.995 8,各组分的检出限在0.09～0.29 μg/kg之间,定量限在0.31～0.79 μg/kg之间。待测物在不同样品中的平均回收率介于67.3%～101.1%,变异系数≤16.5%。45 批保健食品检出23 批非法添加阳性样品。本法经方法学验证,可用于网红保健品中8 种非法添加药物的快速筛查和定量测定。     

Monitoring by LC-MS/MS of 48 compounds of sildenafil,tadalafil, vardenafil and their analogues in illicit health food products in the Korean market advertised as enhancing male sexual performance

More than 46 phosphodiesterase type 5 (PDE5) inhibitor analogues have been found to be present as illegal adulterants in various forms of health food products (powder, tablet, capsule, etc.), thereby placing the health of consumers at risk through product intake. In this study, 164 samples advertised to be effective at enhancing male sexual performance were collected over a 4-year period (2009–2012) from the Korean on-line or off-line market and screened. An LC-MS/MS method was employed to screen for the presence of 48 compounds including sildenafil, tadalafil, vardenafil and their analogues. Method validation established LOQs (0.30–10.00 ng ml^?1 or ng g^?1) and recoveries (spiked in liquid sample, 84–112%; spiked in solid sample, 83–110%). Most of the illicit products screened were adulterated with 14 of the PDE5 derivatives under examination, including considerable amounts of sildenafil and tadalafil; of the 48 compounds, tadalafil was the most frequent adulterant (42.6%), followed by sildenafil (27.9%). Specifically, tadalafil concentration ranges (mg g^?1) in the samples collected over the 4-year period were determined as follows: 2.91–52.20 (2009), 4.50–108.10 (2010), 0.37–101.40 (2011), and 0.08–138.69 mg g^?1 (2012). The concentration ranges (mg g^?1) of sildenafil were also at high levels: 4.90–117.96 (2009), 1.30–369.93 (2010), 0.03–241.77 (2011), and 18.34–297.91 mg g^?1 (2012). The results of screening for PDE5 inhibitor pharmaceuticals as adulterants in illicit health food products are of great significance with respect to the protection of public health and consumer safety.     

Simultaneous Determination of Seven Adulterants in Slimming Functional Foods by HPLC�CESI�CMS/MS

A high-performance liquid chromatography?Celectrospray ionization?Ctandem mass spectrometric method for simultaneous determination of seven adulterants including pseudoephedrine, norpseudoephedrine, caffeine, strychnine, fenfluramine, sildenafil, and amfepramone in slimming functional foods was established. For tablet formulation, the target chemicals were extracted with ammoniated methanol, while for liquid samples, plant powder, or capsules formulations, these chemicals were extracted with a mixture solution of ammoniated methanol-diethyl ether (2:1 v/v). After anhydrous sodium sulfate being added, the extracts were centrifuged and then the supernatant was evaporated to dryness under condition of a nitrogen gas flow. The residue was reconstituted with acetonitrile?Cwater (1:9 v/v) to produce a test solution. Chromatographic separation was accomplished using a RP-C18 column with a gradient elution procedure using 0.03% formic acid in acetonitrile and 0.01% formic acid solution. Seven chemicals were separated and detected in 10 min. Clenbuterol was used as an internal standard. The recoveries of seven targeted chemicals in different formulations are from 81.2% to 110.3%. Limits of detection of the method are from 4.2 to 16.7 ??g kg^?1 with relative standard deviations of 1.1?C8.4%. The linearity of the method ranges from 2.0 to 500 ng mL^?1 for all chemicals, with linear correlation coefficients varying from 0.9982 to 0.9992. The method has been used for determining the target chemicals in nine slimming functional foods, and satisfactory results are achieved. Among these tested samples, norpseudoephedrine and fenfluramine were not detected for all samples. Some other components, such as pseudoephedrine, amfepramone, strychnine, and sildenafil were detected in one or more samples, while caffeine was detected in almost all of these tested samples.