Performance Evaluation of Automated Magnetic Beads Extraction Method for Extraction of Serum Fat-soluble Vitamins by LC-MS/MS Detection / 现代检验医学杂志

Objective To evaluate the performance of the automated magnetic beads extraction method for extraction of serum fat-soluble vitamins by liquid chromatography-tandem mass spectrometry(LC-MS/MS).Methods A total of 120 clinical residual serum samples were collected automated magnetic beads extraction method was applied to extract serum fat-soluble vitamins A,D2,D3,E and K.The linearity,limit of quantification,precision,accuracy,carryover and matrix effect of fat-soluble vitamins A,D2,D3,E and K were detected by LC-MS/MS.The consistency of the method and conventional extraction method was compared.Results The linear correlation coefficients for extracting fat-soluble vitamins A,D2,D3,E and K by automated magnetic beads extraction method were all greater than 0.99,and the quantitative limits for these five fat-soluble vitamins were 5,0.25,0.25,125 and 0.025 ng/ml.The intra-batch precision and inter-batch precision were 0.66%～4.83%and 0.15%～3.70%,respectively,and the average spike recovery rate was 87.05%～111.11%.The matrix effect was 95.43%～99.07%,and the difference between the mean value of the cyclic injection results of high-low value samples and the mean value of the cyclic injection results of low-low value samples was less than 3s of the mean value of the cyclic injection results of low-low value samples.The statistical analysis showed that the correlation between the results of the automated magnetic beads extraction method and the conventional extraction method was good(r＞0.99),and there was no significant bias in the detection results of the two methods.Conclusion The automated magnetic beads extraction method for extraction of fat-soluble vitamins had good detection performance,which may be expected to improve sample flux and analysis efficiency.

Simultaneous Detection of Fat-soluble Vitamins in Health Foods by UPLC-MS/MS and Its Application / 中国药师

Objective: To develop an ultra-performance liquid chromatography-tandem mass spectrometry ( UPLC - MS/MS ) method for the determination of nine fat-soluble vitamins in health foods. Methods:The samples were extracted by ultrasonic in organic solvents followed by the separation on a Waters ACQUITY UPLC BEH C18 column(100 mm × 2. 1 mm, 1. 7 μm) with gradient elution of 0. 1% formic acid in methanol and 0. 1% formic acid solution as the mobile phase and in a mode of atmosphere pressure chemical i-onization multiple reaction monitoring (MRM). Results:The method exhibited a good linear relationship (r≥0. 99) with the detection limit between 0. 9 and 38. 3μg/100 g for the nine fat-soluble vitamins. The recoveries were 78. 5%-114. 9% and the RSDs were 1. 57%-11. 1%(n=6). Conclusion:The analytical method is simple, rapid, accurate and sensitive, and suitable for the determina-tion of fat-soluble vitamins in nutritional supplements.

Simultaneous Detection of Fat-soluble Vitamins in Health Foods by UPLC-MS/MS and Its Application / 中国药师

Objective: To develop an ultra-performance liquid chromatography-tandem mass spectrometry ( UPLC - MS/MS ) method for the determination of nine fat-soluble vitamins in health foods. Methods:The samples were extracted by ultrasonic in organic solvents followed by the separation on a Waters ACQUITY UPLC BEH C18 column(100 mm × 2. 1 mm, 1. 7 μm) with gradient elution of 0. 1% formic acid in methanol and 0. 1% formic acid solution as the mobile phase and in a mode of atmosphere pressure chemical i-onization multiple reaction monitoring (MRM). Results:The method exhibited a good linear relationship (r≥0. 99) with the detection limit between 0. 9 and 38. 3μg/100 g for the nine fat-soluble vitamins. The recoveries were 78. 5%-114. 9% and the RSDs were 1. 57%-11. 1%(n=6). Conclusion:The analytical method is simple, rapid, accurate and sensitive, and suitable for the determina-tion of fat-soluble vitamins in nutritional supplements.

Validação e aplicação de métodos cromatográficos para determinação de vitaminas em suplementos / Validation and application of chromatographic methods for determination of vitamins in supplements

Foram propostas duas metodologias para realizar a determinação de vitaminas em suplementos porcromatografia líquida de alta eficiência (CLAE): uma para a determinação simultânea de vitaminas lipossolúveis (acetato de retinol, palmitato de retinol, acetato de α-tocoferol e β-caroteno) e outra para a determinação simultânea de vitaminas hidrossolúveis (B1, vitamina C, nicotinamida, ácidonicotínico, B6 e ácido pantotênico). A validação das metodologias foi realizada utilizando-se materialde referência certificado SRM 3280 do NIST e padrões de vitaminas. Os limites de detecção (LDs) e de quantificação (LQs) variaram entre 0,3 e 4,3 µg/mL e entre 0,5 e 14,0 µg/mL, respectivamente. Os percentuais de recuperação dos padrões adicionados nas matrizes variaram entre 92 % e 109 % e entre 86 % e 108 % no material de referência. A repetitividade foi calculada utilizando-se o desvio padrão relativo (RSD); e foram detectados valores entre 0,2 % e 9,6 %. Os métodos validados foramaplicados para a determinação de vitaminas A, E, B1, C, niacina, B6 e ácido pantotênico em 10 amostras de suplementos vitamínicos. Ambos os métodos são adequados para a análise de vitaminas em suplementos e suas aplicações serão imprescindíveis, visto a necessidade urgente de efetuarmonitoramento e fiscalização destes produtos.

Two methodologies were proposed to carry out the determination of vitamins in high-performance liquid chromatography (CLAE) supplements: one for the simultaneous determination of liposoluble vitamins (retinol acetate, retinol palmitate, α-tocopherol acetate and β-carotene) and another For the simultaneous determination of water soluble vitamins (B1, vitamin C, nicotinamide, acidicotinic, B6 and pantothenic acid). Validation of methodologies was performed using NIST SRM certified reference material 3280 and vitamin standards. Limits of detection (LDs) and quantification (LQs) varied between 0.3 and 4.3 μg / mL and between 0.5 and 14.0 μg / mL, respectively. The recovery percentages of the standards added in the matrices varied between 92% and 109% and between 86% and 108% in the reference material. Repeatability was calculated using the relative standard deviation (RSD); And values ​​between 0.2% and 9.6% were detected. The validated methods were applied for the determination of vitamins A, E, B1, C, niacin, B6 and pantothenic acid in 10 samples of vitamin supplements. Both methods are suitable for the analysis of vitamins in supplements and their applications will be essential, since there is an urgent need for monitoring and inspection of these products.

Validation and application of chromatographic methods for determination of vitamins in supplements / Validação e aplicação de métodos cromatográficos para determinação de vitaminas em suplementos

Two methodologies were proposed for determining vitamins in supplements by means of high performance liquid chromatography (HPLC): one for simultaneous determination of fat soluble vitamins (retinyl acetate, retinyl palmitate, -tocopheryl acetate and -carotene), and the other for the simultaneous determination of water soluble vitamins (B1, C, nicotinamide, nicotinic acid, B6 and pantothenic acid). The methodologies were validated using certified reference material SRM 3280 from NIST and vitamins standards. The limits of detection (LODs) and the limits of quantification (LOQs) ranged from 0.3 to 4.3 µg/mL and from 0.5 to 14.0 µg/mL, respectively. The recoveries of spiked vitamin standards in supplements ranged from 92 % to 109 %, and from 86 % to 108 % in the reference material. The repeatability was calculated by the relative standard deviation (RSD), with values from 0.2 % to 9.6 %. The validated methods were applied for determining vitamins A, E, B1, C, niacin, B6 and pantothenic acid in 10 multivitamin supplements samples. Both methods are suitable for determining vitamins in multivitamin supplements, and their applications will be essential, considering the urgent need for monitoring and for surveying these products.

Foram propostas duas metodologias para realizar a determinação de vitaminas em suplementos por cromatografia líquida de alta eficiência (CLAE): uma para a determinação simultânea de vitaminas lipossolúveis (acetato de retinol, palmitato de retinol, acetato de -tocoferol e -caroteno) e outra para a determinação simultânea de vitaminas hidrossolúveis (B1, vitamina C, nicotinamida, ácido nicotínico, B6 e ácido pantotênico). A validação das metodologias foi realizada utilizando-se material de referência certificado SRM 3280 do NIST e padrões de vitaminas. Os limites de detecção (LDs) e de quantificação (LQs) variaram entre 0,3 e 4,3 µg/mL e entre 0,5 e 14,0 µg/mL, respectivamente. Os percentuais de recuperação dos padrões adicionados nas matrizes variaram entre 92 % e 109 % e entre 86 % e 108 % no material de referência. A repetitividade foi calculada utilizando-se o desvio padrão relativo (RSD); e foram detectados valores entre 0,2 % e 9,6 %. Os métodos validados foram aplicados para a determinação de vitaminas A, E, B1, C, niacina, B6 e ácido pantotênico em 10 amostras de suplementos vitamínicos. Ambos os métodos são adequados para a análise de vitaminas em suplementos e suas aplicações serão imprescindíveis, visto a necessidade urgente de efetuar monitoramento e fiscalização destes produtos.

Nutritional status of patients with biliary atresia and autoimmune hepatitis related to serum levels of vitamins A, D and E / Estado nutricional de pacientes com atresia biliar e hepatite autoimune e relação com os níveis séricos de vitaminas A, D e E

CONTEXT: Chronic liver disease may induce to malabsorption of lipids and fat-soluble vitamins, leading to injury of nutritional status. OBJECTIVES: To evaluate the nutritional status of pediatric-age patients with autoimmune hepatitis and biliary atresia related to serum levels of vitamins A, D and E and the disease severity. METHODS: This controlled transverse study, evaluated the patients with autoimmune hepatitis and biliary atresia and a reference group paired by sex and age. The patients underwent anthropometric evaluation, alimentary inquiry and determination of serum levels of vitamins A, D and E by high performance liquid chromatography. The Mann-Whitney test, Spearman correlation coefficients and variance analysis (ANOVA) were utilized for data treatment, regarding significant difference if P<0.05. RESULTS: The highest nutritional deficit was observed in patients with biliary atresia, mainly with cholestasis. The serum levels of vitamins A and E for the reference group changed as a function of age. The serum levels of vitamins A, D and E were higher in reference group than in patients with biliary atresia and autoimmune hepatitis together or separately. There were not difference in the serum levels of vitamins A, D and E between biliary atresia groups with cholestasis and without cholestasis. It was verified correlation between weight/age, triceps skinfold thickness, subscapular skinfold thickness, midarm circumference, midarm fat area values and vitamin A serum levels, as well as between all anthropometric indicators and vitamin E in patients with autoimmune hepatitis and biliary atresia. CONCLUSION: The patients with biliary atresia and cholestasis presented the highest nutritional injury. The patients with biliary atresia and autoimmune hepatitis presented lower serum levels of vitamins A, D and E that in control group. There is a directly proportional correlation between vitamin serum levels, mainly vitamin E, and all anthropometric...

CONTEXTO: As doenças hepáticas crônicas podem induzir à má absorção de lipídios e vitaminas lipossolúveis e levar ao comprometimento do estado nutricional. OBJETIVOS: Avaliar o estado nutricional e relacionar com os níveis séricos de vitaminas (A, D e E) e a gravidade da doença em pacientes com atresia biliar e hepatite autoimune na faixa etária pediátrica. MÉTODOS: O estudo foi transversal controlado e foram avaliados os pacientes com hepatite autoimune e atresia biliar e um grupo controle pareado por sexo e idade. Foi realizada avaliação antropométrica, aplicação do inquérito alimentar e determinação dos níveis séricos das vitaminas A, D e E pela técnica de cromatografia líquida de alta eficiência. Foram empregados os testes de Mann-Whitney, o coeficiente de correlação de Spearman e análise de variância (ANOVA), sendo considerada diferença significativa se P<0,05. RESULTADOS: O déficit nutricional mais grave foi observado nos pacientes com atresia biliar, principalmente com colestase. Em relação às vitaminas, no grupo controle, constatou-se que os níveis séricos das vitaminas A e E variaram com a idade. Os níveis séricos das vitaminas A, D e E foram maiores no grupo controle em relação aos pacientes com atresia biliar e hepatite autoimune em conjunto ou separadamente. Verificou-se a correlação do peso/idade, prega cutânea tricipital, prega cutânea subescapular, circunferência braquial, área adiposa braquial com a vitamina A e de todos os indicadores antropométricos com a vitamina E nos pacientes com hepatite autoimune e atresia biliar em conjunto. CONCLUSÕES: Os pacientes com atresia biliar e colestase apresentaram o maior comprometimento nutricional. Os pacientes com atresia biliar e hepatite autoimune possuíram menores níveis séricos das vitaminas A, D e E do que o grupo controle. Existe uma correlação diretamente proporcional, principalmente da vitamina E com todos as variáveis antropométricas do grupo de AB e HAI em conjunto.

Investigation of Serum Level of Fat-soluble Vitamins Among Urban Children in Zhejiang Province / 中国药房

0.05), but significantly different between the cities(P