超高效液相色谱-串联质谱法检测螺旋藻保健品中7种微囊藻毒素

采用固相萃取净化结合超高效液相色谱-串联质谱法(UPLC-MS/MS)测定了螺旋藻保健品中7种微囊藻毒素(MCs)。螺旋藻保健品经70%(体积分数)的甲醇超声提取,冷冻离心沉淀杂质后,经HLB柱净化。在Waters ACQUITY UPLCBEH C18色谱柱上以乙腈和0.2 mmol/L乙酸铵水溶液为流动相进行梯度洗脱分离,采用电喷雾离子源正离子模式进行多反应离子监测,外标法定量。7种MC在线性范围内线性关系良好(相关系数(r)不小于0.995);检出限为6.7～33.3 μg/kg,定量限为20.0～100.0 μg/kg。各分析物在阴性螺旋藻保健品中的加标回收率在87.5%～97.9%之间,相对标准偏差(RSD)在1.6%～6.9%之间。该方法准确可靠,灵敏度高,可用于螺旋藻保健品中MC污染的确证定性、定量检测。     

气相色谱-串联质谱法测定土壤中的邻苯二甲酸酯

建立了土壤中6种邻苯二甲酸酯的气相色谱-串联质谱(GC-MS/MS)分析方法。土壤样品用超声提取,以二氯甲烷-丙酮(1:1, v/v)混合溶液为提取溶剂,提取液经Florisil小柱净化后,经HP-5MS色谱柱(30 m×0.25 mm×0.25 μm)分离,利用MS/MS的多反应监测(MRM)模式进行定性和定量。结果表明本方法可对样品中的邻苯二甲酸酯进行分析,在10～1000 μg/L质量浓度范围内,线性关系良好,相关系数为0.9973～0.9976;6种邻苯二甲酸酯的相对标准偏差(RSD)不大于14.3%, 2 μg/kg和10 μg/kg两个加标水平的回收率为72.9%～106.2%; 6种目标化合物的检出限为0.1～0.5 μg/kg。该方法快速准确、背景干扰较少、分析灵敏度较高,适用于土壤样品中邻苯二甲酸酯的分析。     

超高效液相色谱-串联质谱法同时测定化妆品中35种性激素

采用超高效液相色谱-串联质谱(UPLC-MS/MS)建立了同时测定化妆品中35种性激素的方法。样品经50%乙腈水溶液超声提取,采用CORTECS C_(18)(2.1 mm×150 mm,2.7μm)色谱柱分离,以乙腈-水为流动相进行梯度洗脱,多反应监测(MRM)模式测定。结果表明,35种性激素在各自的质量浓度范围内线性关系良好(相关系数r0.99),检出限(LOD)和定量下限(LOQ)分别为0.028～0.117μg/g和0.093～0.352μg/g。在0.25、0.50、1.00μg/g 3个加标水平下,水状、乳状和霜状3种基质样品的回收率为84.0%～117%,相对标准偏差(RSD,n=6)为1.5%～14%。该方法前处理简单、灵敏度高、准确性好,适用于化妆品中性激素的测定。     

高效液相色谱-串联质谱法同时测定夏天无中的4种生物碱

建立了高效液相色谱-电喷雾串联四极杆质谱同时测定夏天无中4种生物碱的分析方法。夏天无样品用甲醇超声提取,提取溶液过滤并用甲醇稀释后分析。色谱分离采用C18反相色谱柱(150 mm×2.1 mm, 3.5 μm),流动相为0.2%乙酸水溶液和乙腈,梯度洗脱。电喷雾串联质谱在多反应监测(MRM)模式下检测目标分析物,以保留时间和特征离子对(母离子和两个碎片离子)信息比较进行定性分析和定量分析。4种生物碱的检出限(LOD)为0.02～0.2 μg/L,定量限(LOQ)为0.07～0.66 μg/L,加标回收率为93.6%～103.5%,相对标准偏差小于3.8%。该方法简便、准确、灵敏,可用于夏天无中药材的质量控制。     

液相色谱-串联质谱法测定谷物类饲料中的41种激素

建立了谷物类饲料中10种雄性激素、11种孕激素、10种糖皮质激素、5种雌性激素以及5种二羟基苯甲酸内酯类药物的液相色谱-串联质谱(LC-MS/MS)检测方法。均质样品经乙酸乙酯提取,以改良的QuEChERS吸附剂分散于提取液中实现快速净化。采用C18色谱柱(150 mm×2.1 mm, 3.0 μm)分离,分别在正、负电喷雾串联质谱多反应监测模式下检测。在优化条件下,41种化合物在5.0～200.0 μg/kg含量范围内有良好的线性相关性,相关系数均不低于0.99,定量限(S/N≥10)为0.123～2.72 μg/kg。在5、40、200 μg/kg或10、40、200 μg/kg 3个添加水平下,豆粕类饲料中各化合物的回收率为70.3%～118.1%,相对标准偏差(RSD)为0.82%～19.5%;玉米类饲料中各化合物的回收率为76.1%～122.8%, RSD为1.3%～15.0%。该方法简便、快速、准确,可用于谷物类饲料中雄性激素、孕激素、糖皮质激素、雌性激素以及二羟基苯甲酸内酯类激素的筛查和测定。     

液相色谱-串联质谱法同时检测防晒化妆品中的11种紫外吸收剂

建立了防晒化妆品中11种紫外吸收剂的液相色谱-串联质谱检测方法。样品经甲醇-乙醇-水(80:10:10, v/v/v)提取,利用Eclipse XDB-C₁₈色谱柱(150 mm×4.6 mm, 5 μm)分离,以乙腈和0.1%甲酸水溶液为流动相,梯度洗脱。采用多反应监测(MRM)模式进行质谱测定,根据保留时间及质谱图上特征离子的相对丰度比进行定性,外标法定量。结果表明:该方法11种紫外吸收剂的定量限范围为200～500 mg/kg,回收率为77.17%～98.32%,相对标准偏差为2.40%～11.11%。该方法对于膏状和乳状防晒化妆品中的11种紫外吸收剂具有较好的适用性。     

固相萃取净化-超高效液相色谱-串联质谱法测定蜂蜜中雷公藤次碱

建立了一种固相萃取净化-超高效液相色谱-串联质谱检测蜂蜜中雷公藤次碱的分析方法.样品用水溶解后经固相萃取净化,采用Hypersil GOLD C18柱(50 mm×2.1 mm, 1.9 μm)分离,以水和甲醇(含0.15%甲酸)为流动相,在流速0.25 mL/min下梯度洗脱,电喷雾离子源正离子多反应监测(MRM)模式串联质谱进行测定.考察了试样溶解溶剂、色谱和质谱测定条件等.结果表明:雷公藤次碱在0.01~2 μg/L范围内具有较好的线性关系,相关系数大于0.998.该方法定量限(S/N>10)为0.01 μg/kg,在0.01、0.05和0.5 μg/kg添加水平的回收率为76.1%~96.2%,相对标准偏差(RSD, n=6)小于10%.该方法快速、灵敏、准确,适用于蜂蜜中雷公藤相关物质残留的定性、定量分析.     

顶空气相色谱-质谱联用法测定胶粘剂中的残余单体

建立了一种测定胶黏剂中6种丙烯酸酯和甲基丙烯酸酯类残余单体的分析方法。样品经100 ℃、30 min顶空加热后,通过DB-WAX色谱柱(30 m×0.25 mm×0.25 μm)分离和质谱选择离子模式检测,内标法定量。结果表明,6种残余单体能达到良好的分离,检出限(信噪比为3)为0.069～0.096 mg/kg,定量限(信噪比为10)为0.23～0.32 mg/kg,平均回收率为96.0%～104.6%,相对标准偏差(RSD)小于7.2%。该方法操作简便、准确、重复性好、灵敏度高,可用于胶黏剂中丙烯酸酯和甲基丙烯酸酯类残余单体的快速测定。     

柱后氨化-超高效液相色谱-串联质谱法测定牛奶和奶粉中的三聚氰酸

建立了一种柱后氨化-超高效液相色谱-串联质谱法(UPLC-MS/MS)测定牛奶和奶粉中三聚氰酸的分析方法。样品用5%三氯乙酸溶液提取,无需净化和浓缩,用Acquity UPLC HSS T3色谱柱(50 mm×2.1 mm, 1.8 μm)进行分离,以乙腈和水作为流动相进行梯度洗脱,柱后氨化,电喷雾负离子(ESI~)模式电离,多反应监测(MRM)模式进行检测。结果表明: 三聚氰酸在10.0～160.0 μg/L范围内线性关系良好(r≥0.99)。牛奶在0.05、0.10、0.20 mg/kg的添加浓度的回收率为60%～118.5%,相对标准偏差(RSD, n=6)为8.2%～12.6%;奶粉在0.25、0.50、1.00 mg/kg的添加浓度的回收率为88%～108%, RSD(n=6)为2.5%～5.7%。牛奶中方法的定量限(信噪比为10)为0.05 mg/kg,奶粉中为0.25 mg/kg。该方法快速、灵敏、准确,适合于牛奶、奶粉等乳及乳制品中三聚氰酸的快速、高灵敏度的分析检测。     

固相萃取-气相色谱串固相萃取-气相色谱-串联质谱法测定水果中11种三唑类杀菌剂

建立了固相萃取-气相色谱-串联质谱同时测定水果中11种三唑类杀菌剂残留的方法。采用乙腈匀浆提取样品中的待测组分,经固相萃取法(SPE)净化,采用气相色谱-串联质谱在多反应监测(MRM)模式下进行测定,外标法定量。分别对水果样品进行4个水平(10、50、100、250 μg/kg)的加标回收试验,回收率为82.6%～117.1%,相对标准偏差小于10%,定量限(LOQ,以信噪比(S/N)为10计)为0.8～3.4 μg/kg。结果显示该方法的背景干扰低,灵敏度高,定量限低于国家标准及有关文献报道值,适合橘子等水果中三唑类杀菌剂的同时测定。     

Determination of Sex Hormones in Antler Velvet by High Performance Liquid Chromatography Tandem Mass Spectrometry

Eighteen sex hormones in antler velvet were determined by high performance liquid chromatography tandem mass spectrometry.The solid phase extraction was applied to eliminating the matrix effect.The exp...     

不同加工方式对鹿茸中水溶性多糖含量及单糖组成的影响

采用水提醇沉法提纯鹿茸中的水溶性多糖，苯酚-硫酸法测定其含量；多糖经水解、衍生后通过UPLC分析不同加工方式及不同部位鹿茸中单糖组成及含量的差异。结果表明，煮炸茸蜡片、粉片、纱片、骨片中水溶性多糖含量分别为1.74、1.67、1.03和1.13 g/kg，冻干茸为2.77、3.07、1.22和3.20 g/kg；排血茸4个部位水溶性多糖含量依次为1.55、1.78、0.96和0.77 g/kg，带血茸为1.69、1.64、1.01和1.31 g/kg。不同加工方式鹿茸中水溶性多糖单糖组成均检出甘露糖、氨基葡萄糖、核糖、葡萄糖醛酸、半乳糖醛酸、氨基半乳糖、葡萄糖和半乳糖8个组分，对于同一部位，煮炸茸单糖含量低于冻干茸，排血茸单糖（氨基葡萄糖和氨基半乳糖除外）含量低于带血茸；对于同一加工方式不同部位，表现出蜡片、粉片高于纱片、骨片。研究加工方式对鹿茸中水溶性多糖含量及单糖组成的影响，以期为鹿茸加工及其产品开发提供理论参考。     

超高效液相色谱-大气压化学电离-三重四极杆质谱法同时测定保健食品中14种性激素类药物

建立了超高效液相色谱-大气压化学电离-三重四极杆质谱（UPLC-APCI-MS/MS）测定保健食品中14种性激素类药物的方法。样品用乙腈提取2次，再用HLB固相萃取柱净化处理。采用Hypersil Gold C18色谱柱（100 mm×2.1 mm，1.9 μm）分离，以乙腈-10 mmol/L乙酸铵溶液为流动相进行梯度洗脱，采用APCI-三重四极杆质谱检测，外标法定量。结果表明，14种性激素类药物在各自范围内线性关系良好，相关系数（r）≥0.996，检出限为0.0990~2.09 μg/kg，定量限为0.495~5.23 μg/kg；在低、中、高3个水平下的平均加标回收率为65.8%~118.8%，精密度为0.6%~8.7%（n=6）。该方法前处理简单，灵敏度高，回收率良好，适用于保健食品中性激素类非法添加物的定量测定。     

Rapid qualitative and quantitative evaluation of deer antler (Cervus elaphus) using near-infrared reflectance spectroscopy

Near-infrared (NIR) spectroscopy has been applied for both the qualitative and quantitative evaluation of the velvet deer antler. The most important parameters of determining the quality of velvet antler are the habitat (the country of origin) and ash content. Conventionally, the habitat is determined by examining the appearance of samples (by human eye), which lacks objectivity. Ash content is measured by an ignition method (measurement ash residue), however, it is too slow (4–5 h) to be used for rapid at-site measurement. Velvet antlers from three different habitats (China, New Zealand, and Russia), albeit the same species of Cervus elaphus, were evaluated in this paper. Soft independence modeling of class analogies (SIMCA) and partial least squares (PLS) were used for classification of habitat and determination of ash content. The habitat was successfully identified with over 80% accuracy, and the ash content prediction result using PLS regression showed good correlation with the reference ignition method with a standard error of prediction (SEP) of 1.264%.     

麇鹿茸样品的制备和化学成分分析

建立了麋鹿茸样品制备的工艺路线和方法,获得了易保存的干燥粉末样品,对麋鹿茸中的水分、粗蛋白、粗脂肪、膳食纤维、氨基酸、必需无机元素以及维生素的含量进行了测定,并与马鹿和梅花鹿茸进行了比较。结果表明,麋鹿茸与马鹿和梅花鹿茸的化学成分及含量相近,麋鹿茸中膳食纤维和必需无机元素含量高于其它两种鹿茸。     

Comparison of Structure and Biological Activity of Natural Polypeptide from Velvet Antlers of Cervus elaphus with Those of Synthesized Polypeptide

Biochemical techniques including ion-exchange chromatography, gel filtration chromatography and reversed-phase high-performance liquid chromatography(RP-HPLC) were used to isolate and purify the natural polypeptide from velvet antler(nVAP) of Cervus elaphus(C. elaphus), which has a molecular weight of 3215.8 and the primary structure of VLSAADKSNVKAAWGKVGGNAPAFGAEALLRM. The homology of the protein sequence in nVAP with known protein sequence is less than 50%, suggesting that nVAP appears to be a new bioactive substance. At a level of 0.4―50 μg/mL, nVAP promotes mitosis in epidermal cells, chondrocytes and NIH3T3 fibroblasts primarily cultured in a significant way. Given that a yield of high-purity nVAP isolated from C. elaphus is 0.001%, nVAP is artificially synthesized to prepare synthetic velvet antler polypeptide(sVAP) according to its primary structure. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) of sVAP shows a single band, and its HPLC spectrum displays a single peak. The matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF-MS) was used to identify sVAP to be of a molecular weight of 3200 and the consistency between primary structures of sVAP and nVAP. Bioactivity test shows that at a dose of 5―40 μg/mL, sVAP promotes the proliferation of primarily cultured epidermal cells and NIH3T3 cell line. From the traditional Chinese medicine theory, velvet antler from Cervus nippon(C. nippon) and velvet antler from C. elaphus are considered as the same medicine, but differences between biochemical base and pharmacological effect of these two velvet antlers have been observed. We compared the total polypeptide mapping of the two velvet antlers, discovering that nVAP is active polypeptide and only exists in the velvet antler of C. elaphus. sVAP is similar to nVAP in physicochemical property and biological activity. These studies extend the possible utility of sVAP to be the promising compound to prepare velvet antler polypeptide of C. elaphus.     

Microwave-accelerated derivatization prior to GC-MS determination of sex hormones

A new microwave-accelerated derivatization method was developed for rapid determination of 13 natural sex hormones in feeds. Sex hormones were isolated from the sample matrix by ultrasonic extraction, followed by solid-phase extraction, derivatized under microwave irradiation, and then analyzed directly by gas chromatography-mass spectrometry (GC-MS) in selective ion monitoring (SIM) mode. The key parameters affecting derivatization efficiency, including microwave irradiation time, microwave power, and reaction solvent were studied. Under microwave power of 360 W and microwave irradiation for 3 min, 13 natural sex hormones were simultaneously derivatized using heptafluorobutyric acid anhydride (HFBA) as derivatization reagent. This method was applied to the determination of 13 natural sex hormones in different feed samples, and the obtained results were compared with those obtained by the traditional thermal derivatization. The recoveries from 58.1 to 111% were obtained at sex hormone concentrations of 10-300 μg/kg with RSDs ≤12.0%. The results showed that the proposed method was fast, simple, efficient and can be applied to the determination of 13 natural sex hormones in different feed samples.     

A Novel Polypeptide from Cervus elaphus Linnaeus

A novel polypeptide having stimulant effect on some cell proliferation was isolated from the velvet antler (Cervus elaphus Linnaeus). The velvet antler polypeptide consists of a single chain of 32 amino acid residues. Amino acid sequence of the polypeptide was identified as:VLSAADKSNVKAAWGKVGGNAPAFGAEALLRM.     

植物激素的反相高效液相色谱法分离和测定

探讨了分离和测定８种植物生长激素的反相高效液相色谱法。结果表明：以含有０．８％醋酸的甲醇和水作为流动相,采用梯度洗脱,可在１６ｍｉｎ内将８种物质完全分离并可进行定量测定,线性关系良好。方法具有快速、简便、准确的特点。     

梅花鹿茸多肽的化学结构及生物活性

在马鹿茸活性多肽结构与功能研究基础上, 从新鲜梅花鹿茸中分离纯化了活性单体多肽, 确定了其化学结构, 并与马鹿茸多肽进行结构与活性比较. 利用离子交换层析、 凝胶过滤层析及反相高效液相色谱层析等生物化学技术, 从梅花鹿茸中分离得到1个新多肽, SDS-PAGE电泳显示为一条带, HPLC图谱为单一峰, MALDI-TOF MS给出该多肽的精确分子量为3263.4, 其等电点pI=8.15. 一级结构研究表明, 该多肽是由32个氨基酸残基组成的直链多肽, 不含半胱氨酸, 富含缬氨酸、 赖氨酸、 亮氨酸和甘氨酸, 氨基酸序列为VLSATDKTNVLAAWGKVGGNAPAFGAEALERM. 生物活性检测结果表明, 该多肽可促进原代培养的表皮细胞和软骨细胞增殖, 也能刺激NIH3T3成纤维细胞株的分裂. 梅花鹿茸多肽与马鹿茸多肽在结构上均为32个氨基酸残基组成的直链多肽, 但第5, 8, 11和30位氨基酸残基不同. 2种多肽结构上的变化并未影响其促细胞增殖生物活性.