HPLC-DAD法测定草鱼肉质中的维生素B_(1)和维生素B_(12)的含量

本文建立了高效液相色谱-二极管阵列检测法(HPLC-DAD)同时分离检测草鱼肉质中维生素B_(1)和维生素B_(12)的方法。草鱼样品经过酸消化和酶提取后释放出样品中蛋白结合和磷酸化的维生素B_(1)和维生素B_(12),再经固相萃取净化,结合HPLC-DAD对两种目标物进行检测。考察了高效液相色谱条件中流动相组成对分离效果的影响及固相萃取条件中样品溶液pH和洗脱剂对样品回收率的影响。采用HLB固相萃取柱对草鱼样品进行净化,甲醇/水(v/v=1/1)混合液洗脱维生素B_(1)和维生素B_(12)。色谱流动相分别为85%甲醇和50 mmol/L磷酸二氢钠(pH 3.00,含5 mmol/L己烷磺酸),进行梯度洗脱,柱温为40℃,检测波长为255 nm,进样体积为20µL,维生素B_(1)和B_(12)可在15 min内分离。在最佳分离检测条件下,维生素B_(1)和B_(12)的检出限(S/N=3)为0.21~0.36µg/mL,加标回收率为83.9%~95.1%。本文建立的方法可对草鱼肉质中维生素B_(1)和维生素B_(12)含量进行准确定量。

维生素C片含量测定的HPLC法及溶出度方法的研究

目的建立测定维生素C片含量的高效液相色谱(high performance liquid chromatography,HPLC)法,并通过新建的维生素C片溶出度的测定方法提高其质量控制标准。方法HPLC测定法:色谱柱为菲罗门C_(18)柱(250 mm×4.6 mm,5μm),流动相为0.1 mol·L^(-1)磷酸二氢钾溶液(用磷酸调节pH值至2.5)-甲醇(92∶8),流速为0.8 mL·min^(−1),检测波长为243 nm,柱温为30℃,进样量为10μL;溶出度测定法:按照溶出度与释放度测定法(《中华人民共和国药典》2020年版四部附录0931第一法),以0.1 mol·L^(-1)盐酸溶液1000 mL为溶剂,转速为50 r·min^(−1),20 min时取样。结果维生素C在20~140μg·m L^(-1)范围内与其峰面积线性关系良好,y=37487.239x−6860.622(r=0.99996),平均回收率为100.16%,RSD值为0.48%(n=9);其在水、0.1 mol·L^(-1)盐酸溶液、pH 4.0醋酸盐缓冲液、pH 6.8磷酸盐缓冲液各1000 mL的溶出介质中的溶出曲线差异明显,选择0.1 mol·L^(-1)盐酸溶液为溶出介质对5个企业的样品进行测定,均在10 min全部溶出并进入平台。结论所建立的方法使检测的准确性更高、专属性更强、重复性更好,具有可信、精准的优势,能更有效地对维生素C片进行质量控制。

HPLC-ELSD法测定硫酸庆大霉素碳酸铋胶囊中庆大霉素C组分及有关物质含量

目的为硫酸庆大霉素碳酸铋胶囊中庆大霉素C组分及有关物质建立质控方法。方法采用HPLC-ELSD方法,色谱柱:GRACEApollo C18柱(250 mm×4.6 mm,5μm);流动相:0.2 mol/L三氟乙酸溶液-甲醇(96:4,V/V),流速0.6 mL/min;柱温30℃;进样量20μL;Waters低温分流型蒸发光散射检测器,载气流量40 psi,漂移管温度55℃。结果所建方法通过方法学验证,庆大霉素C组分与各杂质间分离完全,庆大霉素、西索米星和小诺霉素分别在0.5~6 mg/mL、25~100μg/mL和25~100μg/mL的浓度范围内,各浓度的对数值与对应色谱峰面积的对数值呈现出良好的线性。采用所建方法对市售16家生产企业43批次硫酸庆大霉素碳酸铋胶囊样品进行测定,获得该制剂产品中各组分及有关物质的数据。结论所建方法是在中国药典方法基础上优化并验证适用于硫酸庆大霉素碳酸铋胶囊C组分及有关物质的含量测定,方法简便准确,专属性强,稳定性好。由获得的实验数据可知,该复方制剂中C组分及有关物质的含量在各厂家间或同厂家不同批次间差异显著,可能存在影响用药安全的潜在隐患,建议在该剂型质量标准中增加相关质控项。

Causal relationship between circulating vitamin C and 25-hydroxyvitamin D concentrations and common mental disorders-a Mendelian randomization study

Mental disorders seriously affect people’s health and social stability.This Mendelian randomization(MR)study was designed to investigate the causal relationship between circulating vitamin C(VC)or 25-hydroxyvitamin D(25(OH)D)levels and mental disorders.The data used for the MR analysis were derived from the summary genome-wide association studies(GWAS)database for VC and 25(OH)D and from the Finn Gen consortium for fourteen mental disorders.Based on the inverse variance weighted(IVW)method,we found a potential causal association between circulating VC and anxiety disorders(IVW:OR=1.139,95%CI:1.023-1.269,P=0.018).However,no causal association was found between VC or 25(OH)D and other mental disorders(P>0.05).In the reverse MR analysis,individuals with Alzheimer’s disease was causally associated with higher concentrations of circulating VC(P=0.012),while individuals with anxiety disorders had a negative association between the concentrations of 25(OH)D(P=0.012).However,the current evidence does not support a causal relationship between VC or 25(OH)D and other mental disorders.In addition,there was no causal association between circulating VC and 25(OH)D(P>0.05).Future studies are needed to confirm these findings and to elucidate the mechanisms of potential causality.

Hemolysis attributed to high dose vitamin C:Two case reports

BACKGROUND High-dose vitamin C treatment(HVCT)can reduce the adverse effect of chemotherapy and enhance the effect of antitumor therapy,which has been considered one of the safest alternative treatments.However,the severity of its adverse effects may have been underestimated.The most serious adverse effect is hemolysis,which may result in acute kidney injury or death.Although glucose-6-phosphate dehydrogenase(G6PD)deficiency is considered to be the main cause,the probability and pathological mechanism are not completely understood,leading to a lack of effective and standardized treatment methods.CASE SUMMARY Two patients with colorectal cancer developed hemolytic anemia after using 1 g/kg HVCT.In contrast to previous cases,the lowest hemoglobin level in the two cases was<50 g/L,which was lower than previously reported.This may be because Case 1 had chronic hepatitis B for many years,which caused abnormal liver reserve function,and Case 2 had grade II bone marrow suppression.Both patients improved and were discharged after blood replacement therapy.Our cases had the most severe degree of hemolysis but the best prognosis,suggesting that our treatment may be helpful for rescue of drug-induced hemolysis.This is the first review of the literature on hemolysis caused by HVCT,and we found that all patients with G6PD deficiency developed hemolysis after HVCT.CONCLUSION G6PD deficiency should be considered as a contraindication to HVCT,and it is not recommended for patients with bone marrow suppression,moderate-tosevere anemia,hematopoietic abnormalities,or abnormal liver and kidney function.Early blood purification and steroid therapy may avoid acute kidney injury or death caused by HVCT-related hemolytic anemia.

Estimation of Vitamins B-Complex (B₂, B₃, B₅and B₆) of Some Leafy Vegetables Indigenous to Bangladesh by HPLC Method

The current experiment was conducted for the simultaneous determination of several water-soluble vitamins like ribo- flavin (vitamin B2), niacin (vitamin B3), pantothenic acid (vitamin B5) and pyridoxine (vitamin B6) in five highly con- sumed local leafy vegetables named as bottle gourd leaves (Lagenaria vulgaris) (local name Lau shak), green amaranth leaves (Amaranthus viridis) (local name Data shak), red amaranth leaves (Amaranthus gangeticuss) (local name Lal shak), Indian spinach (Basella alba) (local name Pui shak) and bitter gourd leaves (Momordica charantia) (local name Korola shak). The analyses were performed by HPLC using an analytical reversed phase C-18 (ODS column, 250 × 4.6 mm, 5 μm, Phenomenex, Inc.) column with the mobile phase consisting of a mixture of buffer (hexane sulphonic acid sodium, potassium dihydrogen phosphate and triethylamine, pH 3.0) and methanol in the ratio of 96:4 (v/v) at a flow rate of 1 mL/min with UV detection at 210 nm. The retention times for the vitamins were obtained as 3.61 min, 6.37 min, 9.51 min and 11.51 min for Vitamins B2, B3, B5 and B6, respectively. These obtained values of the vitamins were compared with the values available in published literatures of Deshio Khaddar Pustiman (DKPM), Indian food value (IFV) and United States Department of Agriculture (USDA).

HPLC法测定赖氨酸维B_(12)颗粒中维生素B_(12)的含量及含量均匀度

目的建立HPLC方法测定赖氨酸维B_(12)颗粒中维生素B_(12)含量及含量均匀度。方法采用高效液相色谱法,紫外检测器,以C_(18)色谱柱为分离柱,以甲醇-0.028 mol·L^(-1)磷酸氢二钠溶液(磷酸调节pH值至3.5)(20∶80)为流动相,在检测波长361 nm处测定维生素B_(12)含量。流速为1.0 mL·min^(-1),进样体积为100μL。结果维生素B_(12)浓度在0.2~5.0μg·mL-1范围内与峰面积呈良好的线性关系,平均回收率为105.80%(n=9)。按此方法分析了3个厂家5个批次样品,样品中维生素B_(12)的测定值为标示量的43.8%~93.3%,同时进行了含量均匀度评价试验。结果显示,某些批次样品中的维生素B_(12)存在含量偏低、含量均匀度结果不符合规定的现象,表明厂家的生产工艺可能存在一定的问题。结论本法结果准确可靠,重现性好,可作为赖氨酸维B_(12)颗粒中维生素B_(12)含量及含量均匀度测定方法。

Quantification and Comparison of Sugars, Carboxylic Acids and Vitamin C Components of Various Citrus Species by HPLC Techniques

Six orange varieties （Washington Navel, Navelina, Navelate, Lane Late, Cara Cara, Valencia）, four mandarin varieties （Okitsu Wase, Dobashi Beni, Ortanique, Satsuma Owari） and two grapefruit varieties （Star Ruby and Rio Red） were evaluated for their fruit quality characteristics such as fructose, glucose, sucrose, total sugars, malic acid, citric acid and Vitamin C contents. Sugars, carboxylic acids, and vitamin C contents of citrus juices were identified and quantified by HPLC （High Performance Liquid Chromatography） technique. The predominating individual sugars were detected as sucrose in all mandarin and orange varieties except grapefruit varieties. Whereas fructose and glucose were found to be higher in grapefruit varieties, citric acid was considered as a major carboxylic acid in all citrus species and detected in oranges higher concentration than mandarin and grapefruit varieties. Furthermore, vitamin C content of grapefruit juices were detected higher concentration than the oranges （except cv. Valencia） and mandarins. Our results showed that itrus species differed in not only species but also varieties based on the sugar and acid characteristics.

HPLC测定丙肝新药索非布韦维帕他韦片的含量

建立高效液相色谱法(HPLC)测定丙肝治疗新药索非布韦维帕他韦片中索非布韦和维帕他韦含量的方法。采用高效液相色谱法,使用Agilent Eclipse plus C18(4.6×150 mm,5μm)色谱柱,以乙腈-0.1%磷酸溶液(30∶70)为流动相,流速为1.0 mL/min,检测波长为210 nm,柱温30℃,进样体积10μL。在该色谱条件下,索非布韦和维帕他韦线性关系均良好,精密度、重复性、稳定性等均符合规定,该方法可以作为索非布韦维帕他韦片中索非布韦和维帕他韦含量测定的方法。

HPLC法测定赖氨酸维B_(12)颗粒中香草醛的含量

目的:建立HPLC法测定赖氨酸维B_(12)颗粒中香草醛含量的方法。方法:取赖氨酸维B_(12)颗粒1.0 g,置10 mL刻度离心管中,加水1 mL溶解后,涡旋1 min,加甲醇至刻度后,继续涡旋1 min进行提取。采用Agilent C 18(250 mm×4.6 mm,5μm)色谱柱,流动相为甲醇-水(35:65),检测波长为277 nm。结果:本方法香草醛检出限为0.2μg·g^(-1),浓度在1.962~49.04μg·mL^(-1)的范围内呈线性关系(r=0.9999),低、中、高三水平回收率在91.86%~101.61%之间,RSD为2.9%(n=9)。测定样品6批,A企业4批,香草醛含量在55.18~79.55μg·g^(-1),B企业和C企业均未检出。结论:该方法操作简便,结果可靠,可用于赖氨酸维B 12颗粒中香草醛含量的测定。

Is Titration as Accurate as HPLC for Determination of Vitamin C in Supplements? <br/>—Titration versus HPLC for Vitamin C Analysis

Herein, the iodometric titration and HPLC-RP methods were compared for the determination of vitamin C in vitamin and mineral supplements. The methods were validated in terms of linearity, limits of detection (LOD), limits of quantification (LOQ), precision, and recovery by using vitamin standards and a reference material SRM 3280 (multivitamin/multimineral tablets) obtained from the National Institute of Standards and Technology (NIST). A total of 22 samples of vitamin supplements, randomly acquired in local markets of Sao Paulo (Brazil) were evaluated for content of vitamin C by these two methodologies. The precision expressed as RSD was lower than 5% for both methods. LOD was 3.6 μg/mL for HPLC and 1.0 mg for titration, while LOQ were 12.0 μg/mL and 3.0 mg for HPLC and titration, respectively. Percent recoveries (%) of spiked samples ranged from 98.7 to 100.5 for HPLC and from 98 to 104 for titration. The contents of vitamin C in SRM 3280 (Certified value = 42.2 ± 3.7 mg/g) were 40.2 and 42.1 mg/g when determined by HPLC and titration respectively. Statistically, there was no difference in the analysed vitamin C content for half of the samples, irrespective of the method applied. HPLC was more sensitive, but the titrimetric method was faster and consumed less reagent. Although both methods were accurate in determining the vitamin C content in SRM 3280, the matrix constituents of some vitamin supplements may have interfered with the analysis.

SIMULTANEOUS DETERMINATION OF VITAMIN B COMPLEX (THIAMINE, RIBOFLAVIN, NIACIN, PYRIDOXINE AND FOLIC ACID) IN FOODS BY HPLC

A high performance liquid chromatographic assay has been developed for the simultaneous determination of five water soluble vitamins:thimine, riboflavin, niacin, pyridoxine and folic acid in cereal products and fresh vegetables. Food samples were hydrolyzed in 0.4 mol/L HCl, autoclaved at 120℃ 15 psi for 20 minutes, using a μ-Bondapak column (3.9×300mm, Waters Co.), a mobile phase of methanol-water (30:70) (0.005 mol/L heptanesulfonic acid) and a flow rate of 1.0 ml/min gave the most satisfactory separation of the five water soluble vitamins. A double channel deteetion was used: four vitamins (B_5, folic acid, B_2, B_1) were detected hy UV spectrophotometry (254 nm) first, pyridoxine (B_6) was detected by fluoromelry (EX 290nm, EM 395 nm) afterwards. Detection limits were 2, 5, 5, 2and 5ng, linear ranges were 5-10ng, 5-50ng, 5-40ng, 5-50ng and 10-50ng for B_1, B_2, B_5, B_6 and folic acid respectively. Recoveries were 92-100%(B_5), 51-52% (folio acid), 103—105% (B_2), 99—100% (B_6) and 91.3—102% (B_1) respectively. In comparison with a reference method and checking with food composition tables, very satisfactory results were obtained by this method.

SIMULTANEOUS DETERMINATION OF FAT SOLUBLE VITAMINS A, D_3, E AND K_1 IN FORTIFIED MILK POWDERS BY HPLC

A high performance liquid chromatographic method for the simultaneous determination of four fat soluble vitamins: retinol (vitamin A), cholecalciferol(vitamin D), tocopherol(vitamin E) and phylloquinone(vitamin K_1) in fortified milk powders and egg yolk has been developed. The method requires enzymatic hydrolysis of lipid component of the sample as a pretreatment. Several factors which influence the enzymatic hydrolysis were studied-Separation was achieved using μ-Bondapak C-18 column(3.9×300mm), 98% methanol as mobile phase, a double channel detection was selected; vitamins D_3 E_1 K_1 were detected by UV spectrophotometry (265 nm) first, then vitamin A by fluorometry (EX 325nm, EM 480mm). The retention times of vitamin A_1 D_3, E and K_1 4.87, 9.00, 10.58 and 15.45 min respectively. Detection limit were 0.64, 0.25, 0.50 and 0.07 ng; and the recoveries were 90.5%～103.6% 90.0%～95.6%, 91.7%～98.8%, 91.5%～98.6%, respectively. The vitamins A, D_0, E, K_1 contents in foods were determined satisfactorily.

HPLC同时测定维C银翘片中绿原酸、对乙酰氨基酚、维生素C、马来酸氯苯那敏的含量

目的：建立维C银翘片中绿原酸、对乙酰氨基酚、维生素C、马来酸氯苯那敏的含量测定方法。方法：采用HPLC法,使用C18柱,流动相：甲醇-乙腈-0.02%磷酸（5∶10∶85）;流速：1.0 mL/min;柱温：35℃;检测波长：310 nm和219 nm。结果：绿原酸、对乙酰氨基酚、维生素C、马来酸氯苯那敏进样量分别在0.03～0.13μg、0.94～4.68μg、0.49～2.44μg、0.01～0.05μg范围内呈良好线性关系,平均回收率（n=5）均不低于98%。结论：本法简便、快捷,结果令人满意,可作为维C银翘片的质量控制方法之一。

复方三维亚油酸胶丸I中甲基橙皮苷和维生素C的HPLC分析

目的:建立测定复方三维亚油酸胶丸Ⅰ中甲基橙皮苷和维生素 C 的反相高效液相色谱法,并用该方法测定2种不同厂家的药品。方法:采用反相高效液相色谱法,以 Zorbax SB-C_(18)(150 mm×4.6mm,3.5μm)为色谱柱;甲醇-0.03mol·L^(-1)磷酸二氢钠水溶液(pH=2.5)(53:47)为流动相;流速为1.0 mL·min^(-1);检测波长为220 nm 和242 nm;柱温为30℃。结果:甲基橙皮苷在0.64～3.84μg范围内线性关系良好,其高、中、低3个量的平均加样回收率分别为99.40%,98.18%,97.82%;RSD 分别为0.6%,0.9%,1.0%。维生素 C 在0.50～3.00μg范围内线性关系良好,其高、中、低3个量的平均加样回收率分别为98.67%,99.22%,96.00%;RSD 分别为0.3%,0.7%,0.4%。结论:该法是一种快速、简便、准确的分析方法,可以为复方三维亚油酸胶丸Ⅰ的质量控制提供科学依据。

用高效液相色谱(HPLC)法测定血清维生素C含量

目的 : 建立更简便、快速、准确、灵敏的测定血清维生素 C(VC)含量的方法 ,并对血清 VC稳定性进行研究。方法 : 血清样品经 1 0 %偏磷酸溶液提取 ,旋涡混匀 3 0 s,离心 1 0 min(4 0 0 0 r/min,4℃ )后 ,用带二极管阵列检测 (DAD)的高效液相色谱 (HPLC)仪对 VC进行定性定量分析。流动相为 1 0 0 mmol/L KH2 PO4 ,p H值为 3 .0 ,检测波长为 2 43 nm,流速为 1 1 ml/min。结果 : 在上述色谱条件下 ,血清中 VC能得到良好分离 ,该法的日内变异系数和日间变异系数分别为 0 .42 %和 2 .1 1 % ;回收率 82 %～ 88.5 % ;VC的最低检测限为 0 .0 5 μg/ml。在对血清 VC稳定性的研究中 ,发现血清 VC在放置过程中很容易被氧化降解 ,其偏磷酸提取液相对稳定 ,但在 5 d后含量也开始明显下降 ,到第 44d后血清和提取液中 VC含量均几乎完全消失。结论 : 建立了高速、高效、高灵敏度的测定血清 VC含量的 HPLC法。血清 VC应立即测定 ,或将样品用偏磷酸提取 ,保存在 -70℃ ,5 d以内分析完毕。

三种水果中维生素C含量的HPLC法测定与比较

建立了西红柿,猕猴桃,冬枣中维生素C的高效液相色谱法分析方法,并对三者维生素C含量进行比较分析。采用岛津LC-20A高效液相色谱系统对样品进行定量分析,色谱柱为ODS;0.1%草酸溶液为流动相;检测波长为240nm;柱温35℃;流速1.2mL·min-1。该条件下目标组分与其他组分有效分离(R>1.5),进样量在0.144-1.44μg范围内线性关系良好,相关系数r=0.9998,平均回收率:99.1%,RSD:0.3%。该方法专属性强、分离效果好、分析速度快、灵敏度高、简便易行、结果准确可靠。分析结果可为猕猴桃、西红柿、冬枣及其它果蔬的维生素C含量测定提供参考依据。

HPLC法测定野生蔬菜中维生素C的含量

[目的]应用反相高效液相色谱法对江苏地区5种野生蔬菜中维生素C含量进行测定。[方法]反相高效液相色谱测定条件为:色谱柱Agilent Zobax C-8(5μm,4.6 mm×150.0 mm);流动相0.1%草酸-水;流速0.8 ml/min;检测波长251 nm。[结果]维生素C在浓度为50～750μg/ml范围内线性良好,R2=0.999 4,回收率为99.5%～101.8%,RSD=1.07%。该法操作简便,结果准确。对5种蔬菜的维生素C含量测定结果表明,荠菜和芦蒿中的维生素C含量较高,分别为50.9和46.5 mg/100 g;菊花脑中维生素C含量最低,为18.6 mg/100 g。[结论]该研究结果为科学开发利用野菜资源提供了一定的理论依据。

利用蒸发光散射检测器 RP-HPLC 同步分析水溶性维生素和脂溶性维生素

目的:建立用蒸发光散射检测器反相 HPLC 同时分析水溶性与脂溶性维生素的方法。方法:首先,不启动梯度,注射脂溶性维生素样品,流动相流速为1.5mL·min^(-1),保持5min;然后,注射水溶性维生素样品,并启动梯度从100%水相过渡到100%有机相;以典型的反相顺序洗脱所有的维生素。色谱柱为 Alltech Prevail-C_(18)(150mm×4.6mm,5μm);流动相 A:pH为3.2的水和甲酸的缓冲溶液,流动相 B:乙腈-甲醇(83∶17);流速:1.5mL·min^(-1)。ELSD 条件:温度:40℃,气体流量:1.5L·min^(-1);撞击器模式:开。结果:采用反相 C_(18)色谱柱和两次进样技术,优化梯度和 pH 条件,可以在30min 内实现对15种水溶性和脂溶性维生素进行同时分析,在30min 内获得了较为理想的分离与峰形。结论:建立了反相 HPLC 分析测定两类维生素的方法,避免了使用多种类型的色谱柱,较长的平衡时间和样品消耗。

HPLC法测定银州柴胡中五种皂苷的含量

首次建立HPLC法同时测定银州柴胡中柴胡皂苷a、c、d、e和f五种皂苷含量的方法。流动相为乙腈-水梯度洗脱,柱子为HibarRT RP-18(4.6 mm×250 mm,5μm),流速1.0 mL/min,柱温30℃,检测波长210 nm。柴胡皂苷a、c、d、e和f的线性范围及相关系数分别为:21.78～43.56μg(r=0.9993),3.94～9.85μg(r=0.9994),20.68～51.70μg(r=0.9998),1.67～5.57μg(r=0.9997),1.91～6.70μg(r=0.9992);柴胡皂苷a、d的加样回收率分别为101.5%和98.5%。该方法操作简便,结果准确,重现性好,为柴胡药材多指标质量分析方法的建立和银州柴胡药用提供了参考依据。