ABSTRACT
OBJECTIVE:To establish the method for the simultaneous determination of 4 flavonoids in rat plasma sample, such as quercetin-3-O-sophoroside,isoquercitrin,hyperin,rutin. METHODS:8 SD rats were selected and given Poacynum hender-sonii leaf extract suspension intragastrically 0.1 g/kg(calculated by extract). The blood sample 1 mL was collected from orbit 1 h after medication. The contents of 4 flavonoids were determined by microemulsion liquid chromatography(MELC)after centrifuga-tion. The separation was performed on Zorbax SB-C18 column with mobile phase consisted of microemulsion [polyoxyethylene lau-ryl ether-n-butanol-ethyl acetate-triethylamine-water (volume ratio was 2.5:3.0:1.7:0.3:92.5,pH=5.0)] at the flow rate of 0.8 mL/min. The column temperature was set at 30 ℃,and detection wavelength was 360 nm. Sample size was 10 μL,and internal standard was bergenin. It was compared with HPLC method using organic solvent as mobile phase(mobile phase consisted of 0.2%phosphoric acid solution-acetonitrile,gradient elution,other condition same as MELC method). RESULTS:In rats plasma,the lin-ear range of quercetin-3-O-sophoroside,isoquercitrin,hyperin,rutin were 10-1000 ng/mL(r≥0.9980). The limits of quantitation was 10 ng/mL(S/N=10). RSDs of precision,stability and reproducibility tests were all below 7.5%(n=5). Average sample recov-eries were between 97.4%-98.5% and RSDs were less than 5.3%(n=5). Extraction recoveries were between 88.7%-100.3%(RSD≤6.14%,n=5). The methodological evaluation results of MELC and HPLC method were all in line with the regulations of pharmacopeia,and the results of blood concentration betweenthem were similar. Compared with HPLC method, MELCmethod could shorten detection time(10 min vs. 40 min)andreduce the amount of organic solvent (0.38 mL vs. 28 mL).CONCLUSIONS:Established MELC method is rapid,simpleand green,and can be used for 4 flavonoids from P. henderso-nii leaf in rat plasma sample.
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Objective A new hollow fiber liquid phase microextraction combined with high-performance liquid chromatography was developed to determine the concentration of berberine in rat plasma. Methods Parameters that affect the hollow fiber liquid phase microextraction processes were investigated and optimized. The optimized conditions were pH of donor and acceptor phases 12 and 2.0, respectively; extraction time 20 min; stirring speed 800 rpm; and addition of 10 % (w/v) salt. Under the optimized conditions, the preconcentration factor for berberine was 347. Moreover, a rapid and sensitive method was developed to determine berberine in rat plasma by HPLC. Results The calibration curve for berberine was linear in the range of 10-1000 ng/ml (r2≥0.9992). The limit of quantitation for the analyte was 10 ng/ml (S/N=10). The limit of detection for the analyte was 3.3 ng/ml (S/N=3). The intra-day and inter-day precision, and stability (RSD) were less than 6.3%. The average recovery was 96.7% ± 3.82%, and RSD was 4.82%. Conclusions The method is efficient, green, accurate and repeatable. It can be applied in determination of berberine in rat plasma.
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83 patients with severe pancreatitis, cancer of esophagus, gastrointestipal surgery and intestinal perforation were treated with TPN, clinical level of total protein, albumin, blood sugar and fat, body weight and nitrogen were measured. The results showed patients with TPN acquired active effects, such as fully enhancement of the utilization and adaptability of body, maintenance of positive nitrogen balance effectively, complement of the lost of albumin and Hb, improvement of metabolic condition, inhibition of glyconegenesis, reduction of the lost of fat and nitrogen and the decomposition of muscle protein. These improve the nitrogen condition, maitain the body in the positive. nitrogen balance, reduce thecllurence of complication and pro ide the better condition for the health.