Determination Of The Drug Release Of Glutamine In Its Intestines Dissolve Capsules By HPLC

Objective:To establish HPLC method for the determination of the Drug Release of L-glutamine in its intestines dissolve capsule.Method:1. The Time Limit of Collapse inspection1.1 Artificial gastric juices and artificial intestinal juices configuration Artificial gastric juices: Takes diluted hydrochloric acid 16.4mL, pepsin 10g, thins up 1000mL, namely results in.Artificial intestinal juices: Takes potassium dihydrogen phosphate 6.8g, adds water 500mL to cause the dissolution, and adjusts pH is 6.8; Takes pancreas enzyme 10g in addition, the watering dissolution; After two fluid mixes, adds water to 1000mL, namely results in.1.2 Inspection methodsHanging the hanging basket through the upper extreme stainless steel roller on the through metal, plunges in the 1000mL beaker, and adjusts the hanging basket position causes when its drop the strainer to be apart from beaker base 25mm, in the beaker has the temperature 37℃±1℃water abundantly, the adjustment water level highly causes the hanging basket rises when the strainer under the water surface 25mm place. Takes the capsule 6 grains, inspects first in the hydrochloric acid solution(9→1000) for 2 hours, each grain of pouch shell must not have the crack or the disintegration phenomenon .Continues takes up the hanging basket, each tube joins back plate at the same place respectively, again separately sets at in the hanging basket glass tube, the start disintegration meter carries on the inspection in the artificial intestinal juices, after 1h breaks up completely, then conforms to the stipulation, otherwise, rework.2. Release determination2.1 Chromatograph conditionChromatographic column: Phenomenex NUCLEOSIL 5 NH2100A (250 mm x 4. 6 mm); Flowing: acetonitrile- 0.05moL? L-1 potassium dihydrogen phosphate buffer solution (takes potassium dihydrogen phosphate 6.8 g, watering dissolves to 1000 mL, adjusts pH with phosphoric acid to 4.0) (70:30); Examination wave length 215nm;flow rate: 1mL?min-1;Specimen handling quantity: 20μL.2.2 System operational test2.2.1 Specificities test examineObtain"2.2.2"item of 0.1mg?mL-1 L-glutamine comparison solution, the specimen handling quantity 20μl, results in the L-glutamine comparison solution the highly effective liquid chromatography chart.Takes the compound prescription L-glutamine capsule contents fine dust right amount (to be equal L-glutamine 120 mg), sets at the 250mL measuring flask, the watering dissolves and dilutes to the scale division, swings uniform, filtration; Takes continues filtrates 2mL, sets at the 10mL measuring flask, adds the flowing to dilute to the scale division, the specimen handling quantity 20μl, results in the compound prescription L-glutamine solution the highly effective liquid chromatography chart.2.2.2Standard curve preparationConfiguration density is 0.025,0.05,0.10,0.15,0.20 mg?mL-1 L-glutamine comparison solution.With HPLC measured the peak area and draws up the standard curve.2.2.3 The within-day precision InspectionTake"2.2.2"item of 0.05,0.10,0.15 mg? mL-1 standard curve preparation fluid, separated the specimen handling quantity within a day (in 8h) 5 times, examines the low, middle and high 3 densities the determination accuracy.2.2.4 Accuracy inspections (recovery)The precise name takes compound prescription L-glutamine capsules contents fine dust 0.20g (to be equal to L-glutamine 120 mg) , sets at the 250mL measuring flask separately; Again distinguishes the precise name to take L- glutamine comparison 50mg and 125mg, joins in the abovemeasuring flask, thins up to the scale division, swings uniform, after 0.22μm pore filter diaphragm filtration; Takes separately continues filtrates 2mL, sets at 10 mL measuring flasks, adds the flowing to dilute to the scale division, determines and calculates recovery with the HPLC law.2.3 Stable inspectionLike under the"2.2.2"item of 0.1 mg? mL-1 L- L-glutamine comparison solution, in the 0,1,2,4,8h distinction specision handling, inspects the L- glutamine solution the laying aside stability.2.4 Release determinationTakes the compound prescription L-glutamine capsule sample, according to release measuring method operation.2.4.1 Release in AcidsQuantity to take 0.1 mg?mL-1 hydrochloric acid solution 900mL, pours into each vessel, the maintenance temperature in 37℃±0.5℃, adjustment rotational speed 100r﹒ min-1.。Takes 6 capsules to invest separately transfers in the basket, starts the instrument to revolve 2h, was stipulating sampling point absorption solution 5mL, and has filtered after 0.22μm the pore filter diaphragm, determines with the HPLC law.According to standard equation of a curve computation density, with each grain of capsule release percentage.2.4.2 Release in Buffer solutionsTo abandon in the above various vessels the acid liquor, joins phosphate buffer solution (pH6.8) 900mL immediately, continues to revolve 85min, was stipulating sampling point absorption solution 5mL, passes through 0.22μm the pore to filter immediately, takes the filtrate, determines the compound prescription L-glutamine capsule release fluid chromatograph with the HPLC law.According to standard equation of a curve computation density, with each grain of capsule release percentage.Result:1. The Time Limit of Collapse resultCompound prescription L-glutamine intestines dissolve capsule in 2 hours the intestines to dissolve the clothes level in the artificial gastric juice to be complete, also breaks up in the artificial intestinal juices in 45 minutes, namely the compound prescription L-glutamine intestines dissolvecapsule the disintegration time limit to be qualified.2. The determination release2.1 System operational test result2.1.1 Specificity examination resultResult showed the L-glutamine peak the retention time is 12.3min, the theoretical plate number according to the L-glutamine peak area is 4600.2.1.2 The release in Buffer solutions result Standard equation of a curve is: A=16.884C-0.0438 (R=0.9999) linear scope 0.5~4μg.2.1.3 The within-day precision inspection result Result Indicated the L-glutamine in high, center, low three thickly gets by the within-day precision is RSD1.72%, 0.62%, 0.97%. Conforms to the pharmacopoeia stipulation.2.1.4 Accuracy inspection result Result indicated the L-glutamine, the low 2 densities add the recovery respectively are 98.0%, 99.0%, RSD are 2.13%,1.60%.Conforms to the pharmacopoeia stipulation.2.1.5 Stable inspection result Result indicated the L-glutamin solution is stable in the 8h intensive quantity.2.2 The determination of the Drug Release result2.2.1 release in AcidsThe quantity result to indicate L-glutamin 2h release is zero, conforms to the pharmacopoeia stipulation.2.2.2 Release in Buffer solutions result Result to indicate the L-glutamin in the Buffer solutions 45min release is 40.77%, has not achieved the pharmacopoeia stipulation standard. When 65min releases atalent to achieve 71.05%, 85min reaches 78.98%.Conclusion:The HPLC method can be used in the determination of the Drug Release of L-glutamine in its intestines dissolve capsule as this article. And the Time Limit of Collapse result of the compound prescription glutamine intestines dissolve capsule conforms to the stipulation; Its release has not achieved the stipulation standard.