应用高效液相色谱-质谱/质谱联用技术检测贝达喹啉血药浓度

目的釆用高效液相色谱-质谱/质谱联用(HPLC-MS/MS )技术检测人血浆中贝达喹啉浓度,并对其应用效果进行评价。方法人血浆样品经乙睛沉淀蛋口等处理后检测.液相色谱采用Agilent ZORBAX SB-C18色谱柱(2.1 mm ×100 mm. 3.5μm )为分析柱,以甲醇-5mmol/L甲酸钱水溶液(含0.1%甲酸)(85:15, V/V)为流动相,质谱使用电喷雾离子源(ESI),以正离子多反应监测(MRM)方式进行检测,分析时间为3min结果本测定方法不受血浆中内源性物质干扰,贝达喹啉母离子和定量子离子的质荷比(m/z)为555.2→58.3,标准曲线线性范围为0.1~5.0μg/ml.方法的日内、日间精密度(相对标准偏差.RSD)均小于10%,准确率为97.8%~ 107%。用建立的方法检测了 12例服用贝达喹啉的结核病患者.服药剂量为每日400 mg时,贝达喹啉峰浓度为(3.17 ± 1.14)μg/ml.谷浓度为(1.12 ±0.64) gg/ml;服药剂量为200 mg/次.每周3次时.贝达喹啉峰浓度为(2.01 ±0.87) jig/ml,谷浓度为(0.65 ± 0.30 )μg/ml,48份样本血药浓度均在检测范围内(0.1 ~ 5.0μg/ml)结论本研究建立了抗结核新药贝达喹啉血浆中药物浓度的HPLC-MS/MS联用方法,该方法适用于临床结核病患者血浆样品中贝达喹啉浓度的快速分析.

Pharmacokinetics and Absolute Bioavailability of the Sublingual Naloxone Hydrochloride Tablet in Dogs

The pharmacokinetics and absolute bioavailability of the sublingual naloxone tablet were studied with HPLC-electrochemical detection. Eight male dogs received single 5 mg dose of naloxone intravenously, the plasma concentration-time curves could be fitted to two-compartment open model, with 12.0 min of t1/2( , 143.4 min of t1/2( and 7.92 mg(min/L of AUC. The same eight dogs received 5 mg dose of the sublingual naloxone tablet after an interval of a week. The main pharmacokinetic parameters were: t1/2ka = 11.0 min, t1/2( = 15.4 min, t1/2( = 164.1 min, Tmax = 27.7 min, Cmax = 34.2 ng / ml, and AUC = 6.79 mg(min / L, respectively. The plasma concentration-time curves were fitted to the first order absorption two-compartment open model also. The mean absolute bioavailability of the sublingual naloxone tablet was 86.8 ( 10.9%. No statistically significant differences were found with t1/2(, t1/2(, ( and ( between the two routes of administration. These results indicated that the course of disposition for naloxone in dogs was similar for the two routes of administration, and the absolute bioavailability of the sublingual naloxone tablet was high. Thus satisfactory clinical effects could be expected.

The Patterns of Secretory Structure and Their Relation to Hypericin Content in Hypericum

利用整体透明、石蜡制片和半薄切片法 ,对金丝桃属 (HypericumL .) 8组 2 0种 1变种植物的分泌结构进行了比较解剖研究 ,结果表明 :该属植物的分泌结构可分为分泌细胞团和分泌囊两种类型。但在不同植物种和不同器官内 ,分泌结构的类型和分布密度存在差异。对上述植物的提取物进行薄层层析和高效液相层析检测 ,结果表明 :具有分泌细胞团的植物器官含有金丝桃素 ;而无分泌细胞团的植物器官 ,则不含金丝桃素 ,从而证明金丝桃素由分泌细胞团合成和贮藏。在前者中 ,其金丝桃素的含量与其分泌细胞团密度成正相关。

Study on Relative Bioavailability of Famotidine Sustained-release Tablets in Healthy Volunteers

The relative bioavailability of famotidine sustained release (SR) tablets was studied in 16 healthy male volunteers. Famotidine plasma concentration was determined by HPLC method, and the plasma concentration time data were processed with the method provided by USP XXIII. For single dose administration the peak plasma concentration occurring at 8 13±0 34 h was 69 52±3 00 ng/ml and the relative bioavailability was 112 4±8 6%. For multiple dose administration the peak plasma concentration of SR tablet was 86 14±2 95 ng/ml and the degree of fluctuation (DF) was 140 6±13 5% at steady state. Two one sided tests were performed in bioequivalence assessment. The results showed that the sustained release tablets were basically bioequivalent to the immediate release (IR) tablets on sale.

Pharmacokinetics and Bioavailability of 2-Amino-6-Cyclopropylamino-9-(2,3 -Dideoxy-β-D-glyceropent-2-enofuranosyl) Purine (Cyclo-D4G) in Rats

AimTo characterize the pharmacokinetics of 2 -amino-6-cyclopropylamino-9-(2,3-dideoxy-β-D-glyceropent-2-enofuran osyl) purine (Cyclo-D4G) following intravenous administration and oral administ ration to rats. MethodsThe concentrations of Cyclo-D4G in rat (Sprague-Dawley male rats) plasma and urine were analyzed by high performance liquid chromatography (HPLC). ResultsFollowing intravenous adm inistration to rats, concentrations of Cyclo-D4G in plasma declined with a term inal phase half-life of 0 78±0 14 h (±s). Total clearance was 0 90±0 21 L·h -1 ·kg -1 . Renal excretion of unchanged Cyclo-D4G accounted for approximately 20% of total clearance. Steady state volume of distr ibution was 0 91±0 07 L·kg -1 . After oral administration to rats, conce ntrations of Cyclo-D4G in plasma declined with a terminal phase half-life of 0 83±0 13 h (±s). Total clearance was 3 81±2 03 L·h -1 ·kg -1 . Renal excretion of unchanged Cyclo-D4G accounted for approximat ely 9% of total clearance. Oral bioavailability of Cyclo-D4G in rat was 26 9%. ConclusionThe favorable pharmacokinetic profiles and lower to xicity provide support for further development of Cyclo-D4G clinical trials.

Economic and Qualitative Traits of Italian Alps Saffron

Saffron, obtained from the flower stigmas of Crocus sativus L., is one of the most expensive food spices. The introduction of saffron in alpine areas could help to broaden and diversify the activities of mountain multifunctional farms, with a positive impact on economy and land management. According to ISO 3632(2010/2011), saffron can be classified into three categories of quality(I, II, III) depending on the concentration of the three main metabolites responsible for its characteristic colour, flavor and aroma: Crocin, Picrocrocin and Safranal. This study represents the first investigation of the quality of saffron produced in the Italian Alps evaluated with spectrophotometry, HPLC, solid-phase microextraction(SPME), and gas chromatographic analysis combined with mass spectrometry(GC/MS). The experiments used Crocus sativus stigmas produced in 2012-2013 in different areas of the Central Italian Alps were located at an altitude between 720 and 1200 m a.s.l.. Results obtained were compared to commercial saffron. The analyses confirmed that all samples can be classified in the first quality category according to the ISO classification. This high quality is also confirmed by HPLC analysis. Moreover, the SPME-GC/MS analysis identified some differences in the aromatic profile of saffron samples, in particular regarding safranal concentration. A preliminary assessment of the economic viability of high quality saffron production for local markets was also performed. Our study provides valid information regarding the quality and economic sustainability of saffron production in the alpine area confirming this crop as a good candidate for a new source of income for multifunctional farms in mountain areas.

Investigation of Metal Wired Coloured Historical Textile Using Scanning Electron Microscopy and HPLC-DAD

SEM-EDS （scanning electron microscopy-energy dispersive spectrometry） and HPLC （high performance liquid chromatography） with DAD （diode array detection） were used to examine metal-wrapped coloured textiles. The sample was provided from Azerbaijan History Museum. Material included silk and thin wire. SEM micrographs showed that the metal strip had 250μm width and 6 9m thickness. SEM-EDS results indicated elemental composition of the wrapping： the presence of gold, silver as the components of the gilt metals. Furthermore, sulphur and chlorine were determined as contaminants. EDS analysis also revealed that both sides were gildered. Colouring compounds in the same sample were identified by HPLC-DAD. The major colouring compound found in the red fibre sample in large quantity was carminic acid and fuchsin components. The presence of fuchsin, which is a synthetic dye, dates the sample between the end of 19th and beginning of 20th century. Fisetin, which is a natural dye, was detected in the yellow fibre.

Simultaneous determination of four saponins in Shugan Quzhi Capsule by UHPLC-MS/MS

Shugan Quzhi capsule is a hospital preparation of Xinhua Hospital affiliated to School of Medicine, Shanghai Jiaotong University. It has been used for the treatment of adult patients with fatty liver caused by obesity, high cholesterol and other factors. In the present study, we established an ultra-high pressure liquid chromatography-tandem mass spectrometry （UHPLC-MS/MS） method to simultaneously determine 4 saponin ingredients including Notoginsenoside R1, Ginsenoside Rbl, Ginsenoside Re and Ginsenoside Rgl present in Shugan Quzhi capsule. The chromatographic separation was conducted on ZORBAX SB-C18 （2.1 mm×50 mm, 1.8 μm）. The mobile phase was composed of acetonitrile and aqueous solution consisted of 0.05% formic acid and 5 mM ammonium acetate. Gradient elution rate was 0.3 mL/min, the column temperature was 40 ℃ MS was conducted using electrospray ionisation （ESI） and multiple reaction monitoring （MRM） coupled with positive and negative scanning switch. With which, Ginsenoside Re and Ginsenoside Rgl were detected using negative ion mode detection while Notoginsenoside R1, Ginsenoside Rbl and internal standard （Ginsenoside F1） were detected using positive ion mode detection. The limits of quantification （LOQ） for Notoginsenoside R1, Ginsenoside Rbl, Ginsenoside Re and Ginsenoside Rgl were 6.54× 10-4, 2.57×104, 0.11 and 6.91 × 10-3 ng/mL, respectively. The limits of detection （LOD） for these compounds were 1.96×10-4, 7.70×10-5, 3.45× 10-2, and 2.07× 10-3 ng/mL, respectively. All calibration curves showed a good linearity （r2〉0.9633） within the test range. The intra- and inter-day precisions （relative standard deviation, RSD） were lower than 5% and the average recoveries were in the range of 80%-120%. With this method, four kinds of saponins were separated and detected in 6 min. This method is simple, rapid and shows high sensitivity and accuracy.

Application of UHPLCMS/ MS method in determining the content of multiingredient in hospital preparations

In the present study, we established an ultra-high pressure liquid chromatography-tandem mass spectrometry （UHPLC-MS/MS） method to simultaneously determine 11 compounds in two hospital preparations （Shegan mixture and Gandi capsules）, including ephedrine, caffeic acid, ferulic acid, rutin, scutellarin, belamcandin, baicalin, baicalein, astragaloside IV, irisflorentin and wogonin. The chromatographic separation was conducted on ZORBAX SB-C18 （2.1 mm×50 mm, 1.8 μm）. Eleven analytes and IS were detected using ESI and MRM combined with positive and negative scanning switch. Caffeic acid, ferulic acid, scutellarin and belamcandin were detected using negative ion mode detection, whereas ephedrine, rutin, baicalin, baicalein, astragaloside IV, irisflorentin and wogonin were detected using positive ion mode detection. The quantification limits of ephedrine, caffeic acid, scutellarin, belamcandin, baicalin, baicalein, irisfiorentin and wogonin were 4.90×10^-3 ng/mL, 7.80 ng/mL, 6.8 ng/mL, 5.3×10^-2 ng/mL, 4.20×10^-3 ng/mL, 4.6×10^-2 ng/mL, 1.44×10^-4 ng/mL, 4.85 ng/mL, 0.23 ng/mL, 3.18× 10^-4 ng/mL and 2.95× 10^-4 ng/mL, respectively. The detection limits of these components were 2.90× 10^-4 ng/mL, 0.77 ng/mL, 2.0 ng/mL, 0.016 ng/mL, 1.3×10^-3 ng/mL, 3.33×10^-4 ng/mL, 4.32×10^-5 ng/mL, 1.46 ng/mL, 0.07 ng/mL, 9.5×10^-5 ng/mL and 8.84× 10^-5 ng/mL, respectively. All calibration curves showed good linearity （R^2〉0.99） within the test range. The RSD values of intra-day and inter-day precision were less than 5%, and the average recovery rates of the 11 components ranged fi＇om 80% to 120%. In conclusion, our newly developed method was simple, rapid, sensitive and accurate. It could be used to determine ephedrine, caffeic acid, ferulic acid, rutin, scutellarin, belamcandin, baicalin, baicalein, astragaloside IV, irisflorentin and wogonin in Shegan mixture and Gandi capsules, which was helpful for the quality control of drugs.

Combination of reversed phase liquid chromatography and zwitterion exchange-reversed phase-hydrophilic interaction mixed-mode liquid chromatography coupled with mass spectrometry for the analysis of antibiotics and their impurities

In this study, a system involving two-dimensional, column-switching high-performance liquid chromatography （HPLC） coupled with tandem mass spectrometry （LC-MS/MS） was developed and optimized for the analysis of antibiotics and their related substances. In the first-dimensional chromatography, the analytes were separated on a zwitterion exchange-reversed phase-hydrophitic interaction （ZIC-RP-HILIC） mixed-mode column coupled with tandem mass spectrometry （LC-MS/MS）. A commonly used reversed phase LC column was employed in the second-dimensional chromatography. The mobile phase for the ZIC-RP-HILIC mixed-mode chromatography consisted of a volatile buffer that was compatible with LC-MS/MS, which enhanced the efficiency of ionization for structure elucidation. The antibiotic impurities eluted in the ion-pairing reversed phase chromatography were directly characterized by the ZIC-RP-HILIC-MS system, and the orthogonal separation of ZIC-RP-HILIC mixed-mode chromatography and reversed phase LC provided extra confidence that no impurity was missed. The efficiency of this method was demonstrated in the analysis of penicillin V potassium, oxacillin sodium, ceftriaxone sodium, and their impurities. In addition, this method is convenient for impurity profiling of antibiotics, and may be used for the analysis of other pharmaceutical ingredients.

A novel catalytic application of heteropolyacids:chemical transformation of major ginsenosides into rare ginsenosides exemplified by Rg1

High performance liquid chromatography coupled with quadruple-time-of-flight mass spectrometry(HPLC-Q-TOF-MS)method was developed for analyzing the hydrolytic mixtures of ginsenoside R_(g1) in acidic conditions(pH 3). Three catalysts, a heteropolyacid(H_4SiW_(12)O_(40), SiW_(12) for short), its complex with γ-CD(SiW_(12)/γ-CD for short) and formic acid, were used for comparison. The chemical transformation products were identified based on the accurate mass measurement and the fragment ions obtained from tandem mass spectrometry. It was concluded that the catalytic efficiency of SiW_(12)(≈SiW_(12)/γ-CD) is ca. 410 times higher than that of formic acid, thus becoming the most efficient catalyst for chemical transformations of ginsenosides.

Simultaneous determination by UPLC-MS/MS of seven bioactive compounds in rat plasma after oral administration of Ginkgo biloba tablets: application to a pharmacokinetic study

A rapid, reliable, and sensitive method was developed using ultra-performance liquid chromatography- tandem mass spectrometry （UPLC-MS/MS） with an electrospray ionization （ESI） source for determination of seven bioactive compounds in rat plasma after oral administration of Ginkgo biloba tablets （GBTs）. The method simultane- ously detects bilobalide （BB）, ginkgolide A （GA）, ginkgolide B （GB）, ginkgolide C （GC）, quercetin （QCT）, kaempferol （KMF）, and isorhamnetin （ISR） for pharmacokinetic study. The analytes and internal standard （IS） were extracted from rat plasma by acetidin. An MS/MS detection was conducted using multiple reaction monitoring （MRM） and operating in the negative ionization mode. The calibration curve ranges were 5-500, 5-500, 2.5-250, 1-100, 1-100, 1-100, and 1-100 ng/ml for BB, GA, GB, GC, QCT, KMF, and ISR, respectively. The mean recovery of the analytes ranged from 68.11% to 84.42%. The intra- and inter-day precisions were in the range of 2.33%-9.86% and the accuracies were between 87.67% and 108.37%. The method was used successfully in a pharmacokinetic study of GBTs. The phar- macokinetic parameters of seven compounds were analyzed using a non-compartment model. Plasma concentrations of the seven compounds were determined up to 48 h after administration, and their pharmacokinetic parameters were in agreement with previous studies.

Protective effect of Sijunzi decoction on neuromuscular junction ultrastructure in autoimmune myasthenia gravis rats

OBJECTⅣE: To investigate the protective role of Sijunzi decoction in neuromuscular junction(NMJ)and muscle cell mitochondria ultrastructure; as well as its effects on the amount of adenosine triphosphate(ATP) and the activities of mitochondrial respiratory chain complexes I, Ⅱ, Ⅲ, and Ⅳ in autoimmune myasthenia gravis rats.METHODS: An experimental autoimmune myasthenia gravis(EAMG) rat model was established by inoculating rats with acetylcholine receptors extracted from Torpedo. Rats were divided into three groups: model, prednisone, and Sijunzi decoction, and were fed physiological saline, prednisone, or Sijunzi decoction, respectively. NMJ and muscle cell mitochondria ultrastructure were observed by transmission electron microscope. The amount of ATP was assessed by high performance liquid chromatography. The activities ofmitochondrial respiratory chain complexes I, Ⅱ, Ⅲ,and Ⅳ was determined using the Clark oxygen electrode method.RESULTS: In the model group, there were sparse muscle fibers, with decreased mitochondria, and sparse, diffluent, or absent NMJ folds. After intervention with Sijunzi decoction, the above pathology changes were improved: muscle fiber structure was clear and complete; the mitochondria count was higher; and the NMJ structure was close to normal. Gastrocnemius muscle mitochondria in the model group produced significantly less ATP than those in the prednisone group(P<0.01). Conversely, the ATP of Sijunzi decoction group was significantly higher than prednisone group(P<0.01). The activities of gastrocnemius muscle mitochondrial respiratory chain complexes I, Ⅱ, Ⅲ, and Ⅳ in both the prednisone and Sijunzi decoction groups was dramatically higher compared with the model group(P<0.05). The activities of complexes I and Ⅲ in the Sijunzi decoction group were significantly higher than those in the prednisone group(P<0.05), but there was no obvious difference in complex Ⅱ or Ⅳ activities between the two groups(P>0.05).CONCLUSION: Sijunzi decoction improved pathological changes in muscle mitochondria and NMJ,enhanced the amount of ATP in gastrocnemius muscle mitochondria, and improved the activities of respiratory chain complexes I, Ⅱ, Ⅲ, and Ⅳ(especially I and Ⅲ) of the EAMG rats.