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【Objective】 To investigate the compatibility of human albumin and its internal packaging materials of Sinopharm Lanzhou Biopharmaceutical Co., Ltd. 【Methods】 One batch of inner packaging materials (medium borosilicate glass-molded injection bottle and halogenated butyl rubber plug for injection) was extracted with 4 extraction solvents to conduct the toxicological evaluation of potential inner packaging extracts. Through the simulated acceleration test, the trend analysis of the elements in the sample and the inner surface of the glass bottle were observed, and the routine drug inspection items during the long-term stability test process were determined. 【Results】 The detection results of the leaching elements of the internal packaging materials did not exceed the limit of 50%, and the organic matter safety threshold (margin of safety, MOS) was greater than 1.0, indicating that both the leaching elements and the organic matter had no safety risk to the user under the current exposure. The results of the simulated acceleration test show that the drug will not have the risk of peeling tablets after the long-term stability condition was placed for a period of time, and the routine inspection items of the long-term stability test drugs all meet the requirements of the pharmacopoeia. 【Conclusion】 The inner packaging material has no significant impact on the quality of drugs and has good overall compatibility, making it suitable for packaging human albumin.
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【Objective】 To evaluate the determination uncertainty of aluminum residues in human albumin by inductively coupled plasma mass spectrometry (ICP-MS). 【Methods】 The aluminum residues in human albumin was determined by ICP-MS, and the top-down method was applied to assess the reasons of uncertainty and calculate the uncertainty. 【Results】 The relative standard uncertainty of the aluminum content in human albumin at the three quality control levels was 0.54 ng/mL, 1.68 ng/mL and 4.54 ng/mL, respectively, which met the requirements of the guidelines for bioanalytical methods in the Chinese Pharmacopoeia(2020). 【Conclusion】 The top-down method is simple and quick to assess the uncertainty of aluminum residues in human albumin, and is suitable for the uncertainty assessment of analytical methods in biological laboratories.
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【Objective】 To investigate the outcomes of intravenous injection of human albumin (HA) in patients with both liver cirrhosis and nephrotic syndrome. 【Methods】 We retrospectively studied 101 liver cirrhosis patients with ascites and nephrotic syndrome treated in our hospital from January 2018 to November 2021. All the patients received oral diuretic and intravenous albumin therapy. Their baseline characteristics were collected and the changes in serum albumin and creatinine levels before and after treatment were evaluated. The patients with elevated albumin levels after treatment greater than the median value (1.8 g/L) were defined as response group. The rest of the patients were classified as the non-response group. And Logistic regression analysis was used to evaluate the relevant influencing factors for treatment response. 【Results】 All the patients’ symptoms of abdominal distension related to moderate to great ascites were clinically lessened at the end of treatment, and no case of acute kidney injury occurred during the treatment. Of them, 32 patients had repeated hospitalizations within six months after discharge. The serum albumin level was significantly increased after treatment [(26.5±5.9) g/L vs. (29.9±4.9) g/L, P<0.001] and there was no significant difference in serum creatinine before and after treatment [(111.9±118.4)μmol/L vs. (108.5±87.9)μmol/L, P=0.816]. Fifty-three patients were defined as treatment response group. The differences in characteristics including age, sex, etiology of cirrhosis, and proteinuria were not statistically significant. However, the serum creatinine level was significantly lower in the response group than in the non-response group [(84.1±51.2)μmol/L vs. (142.7±158.4)μmol/L, P=0.017\]. A similar trend of difference was observed with respect to urea nitrogen level \[(8.7±5.1)mmol/L vs. (11.8±9.1)mmol/L, P=0.039\]. Multivariate analyses demonstrated that the serum creatinine level was a risk factor for non-response to treatment (hazard ratio=1.025, 95% CI: 1.010-1.049, P=0.037). In addition, the correlation analysis showed that the baseline albumin levels were negatively correlated with hospital stay time (r=-0.340, P=0.001), daily HA usage (r=-0.546, P<0.001), and baseline proteinuria levels (r=-0.654, P<0.001), respectively. 【Conclusion】 Intravenous injection of HA in cirrhotic patients with nephrotic syndrome was safe and effective for the treatment of ascites. Kidney function affects serum albumin levels and response to treatment.
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【Objective】 To compare the effects of human albumin products from different domestic manufacturers on human stem cell culture. 【Methods】 Human CD34+ cells were cultured by supplementing human albumin from different manufacturers to serum-free medium, and the expansion ratios of cells within 15 days were counted. Post translational modifications of human albumin products were studied by LC-MS. 【Results】 Supplementing plasma-derived human albumin(pd-alb) to serum-free medium, the expansion ratios of cells could reach up to 20.6±5.7, while the recombinant human albumin(rhAlb) resulted in no significant expansion within 15 days. LC-MS showed significant differences in post-translational modifications from different human albumin products. 【Conclusion】 Different human albumin products showed significantly different effects in the expansion of stem cells due to different sources, processes and stabilizers. Pd-alb products were better for stem cell culture than rhAlb products. There are significant differences between pd-albs and rhAlbs in post-translational modification, but whether these differences are related to stem cell expansion remains to be studied.
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【Objective】 To study the quality control of human albumin intermediates content in production process using High Performance Liquid chromatography (HPLC) method. 【Methods】 In accordance with the general principles of The Chinese Pharmacopoeia (2015 edition), 3121 human albumin polymer was used to determine the component V precipitation solution, human albumin stock solution, human albumin semi-finished products and human albumin products in the production process of human albumin. The chromatographic retention time and the chromatographic peak area percentage corresponding to the retention time were analyzed. 【Results】 The polymer content of component V precipitation solution, human albumin stock solution, human albumin semi-finished products, and human albumin products were was 0.2% vs 0.0% vs 1.9% vs 1.7%, 0.2% vs 0.0% vs1.8% vs1.5% and 0.1% vs 0.0% vs 1.6% vs 1.5%, respectively. 【Conclusion】 HPLC can be used as a monitoring method for human albumin production.
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AIM: To provide reference for rational application of human albumin in pediatrics. METHODS: On the basis of the human albumin instructions, refer to the relevant guidelines and literature on the use of human albumin in pediatrics, draft the first draft of the Drug Use Evaluation standard and list it into an expert consultation questionnaire, which is finalized through two rounds of expert discussions. This standard was used to retrospectively evaluate the medical records of hospitalized children who used human albumin in the Pediatric Department of our hospital from June 2019 to January 2020. RESULTS: The standards established in this study included three primary indicators of medication indications, medication process, medication results and 6 secondary indicators of indications, contraindications, treatment process monitoring, drug application, efficacy, and adverse reactions. A total of 269 medical records of hospitalized children were included in this evaluation, of which newborns accounted for the highest proportion (56.88%). 229 cases met the indications, accounting for 85.13%; 251 cases were tested for serum albumin concentration before medication, accounting for 93.31%; after medication, the relevant indicators reached the standard, and the symptoms improved significantly in 226 cases, accounting for 84.01%. CONCLUSION: The Drug Use Evaluation criteria for pediatric human albumin established by our hospital has strong practicability, which is conducive to discovering problems or deficiencies in clinical medication and promoting rational clinical use of medication.
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OBJECTIVE: To establish a method for detecting the impurity components in human serum albumin by electrospray ionization mass spectrometry (ESI-Q-TOF MS) and investigate the impurity components in 28 batches of post-marketing albumin products from 27 companies. METHODS: According to Chinese Pharmacopoeia′s General Principles 4121, gel filtration column (Hiprep16/60 SepharylS-200HR) was used to separate the protein components. The dimer and multimer components were collected, then desalted, concentrated and digested by trypsin. The peptide was eluted by BEH C18(2.1 mm×100 mm,130) column with mobile phase consisting of water (0.1% formic acid)-acetonitrile in gradient elution mode. Chromatography-mass spectrometry was used to determine the secondary sequences of the peptides and the searching software (PLGS3.0) was used to identify the impurity protein components in albumin. RESULTS: A total of 23 impurity proteins were detected, among which apolipoprotein A-II, haptoglobin, and hempoexin, α-1B-glycoprotein and α-2-HS-glycoprotein were the impurity proteins common to all enterprise products; α-albumin (VE binding protein), N-acetyl-L-alanine amidase, haptoglobin-related proteins, hemoglobin (β subunit, α subunit), transthyretin and other proteins were found in the products of most companies. The number of heterologous proteins in each enterprise was between 8-17, and the numbers of impurity proteins in 27 companies were quite different. CONCLUSION: This method can be used for the investigation of unknown components in human serum albumin, which provides a guidance for the optimization of human albumin production process.
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OBJECTIVE: To understand the quality status and problems of human albumin. METHODS: A total of 153 batchs human albumin samples from 33 manufacturers were analyzed according to the legal quality standard. Three exploratory researches on human parvovirus B19 screening, protein impurity and influencing factors of alum inumion content were conducted. RESULTS: The qualified rate of the 153 batches of samples was 100.0%. The positive rate of B19 DNA in 60 batches from 33 manufacturers was zero. Twenty-three types of other plasma protein were identified in 28 batches of samples from 27 manufacturers. The increase in aluminum contents was related with glass bottle and citrate ion content.CONCLUSION: The overall quality of human albumin is good. However, the content of aluminium ion increases significantly during storage. It is suggested to conduct further study on the influencing factors of aluminium ion content.
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OBJECTIVE: To develop and verify a magnetic beads-based extraction combined with quantitative PCR(q-PCR)method for determination of residual host cell DNA in recombinant human albumin products expressed in Pichia pastoris. METHODS: The residual Pichia pastoris host cell DNA in samples were extracted by magnetic beads-based extraction method and then determined by Taqman probe-based q-PCR. The residual DNA content was calculated according to the standard curve. The developed method was verified for accuracy and precision with different derivation albumin matrixes and concentrations, and the residual DNA of 3 batches of recombinant human albumin products expressed in Pichia pastoris were detected. RESULTS: The minimum detection limit of Pichia pastoris residual DNA by the developed method was 3 fgμL-1, the linear range was 3 fgμL-1-300 pgμL-1, and the correlation coefficient(r2) was 0.998 3. The recovery rates of spiked samples in rHA matrix were 93.58%(RSD 19.6%, n=4)at 100 fgμL-1 and 215.56%(RSD 42.9%, n=4) at 10 fgμL-1, respectively. The recovery rates of spiked samples in HSA matrix were 67.09%(RSD 6.9%,n=3)at 100 fgμL-1 and 113.40%(RSD 11.1%, n=3) at 10 fgμL-1, respectively. The residual Pichia pastoris DNA contents in 3 batches of recombinant human albumin products expressed in Pichia pastoris determined by the developed method were 5.98, 4.16, 4.49 fgμL-1(n=7) respectively and not more than 1 ng per 10 g protein. CONCLUSION: Magnetic beads extraction method combined with fluorescence quantitative PCR method solves the technical problem of quantitative determination of trace DNA in recombinant human albumin products with ultra-high concentration protein. The method is accurate and reproducible, and can be used for quantitative determination of DNA residue in recombinant human albumin expressed by Pichia pastoris.
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OBJECTIVE: To evaluate the applicability of UPLC/MS method for the identification test of human serum albumin (HSA) products including plasma derived and recombinant HSA samples. METHODS: ACQUITY UPLC with Vion IMS QT of LC/MS system was used combined with on-line HSA sample desalting with ACQUITY UPLC BEH C18 column. The acquired multiplycharged mass spectrum was processed with MaxEnt1 automatic protein deconvolution software in UNIFI, which can transfer the raw mass spectrometry data to zero charge molecular mass or mass distribution of the intact protein. RESULTS: Intact protein mass analysis not only provided the accurate mass of HSA, but also provided an overall view of the heterogeneity of HSA and the relative amounts of various forms. From this study, a very specific mass signal [(66 437 ± 1), which is the theoretical average MW of human serum albumin ]was obtained from all the six HSA samples. And the characteristic spectra of different samples were also got. CONCLUSION: UPLC/MS method has very good specificity and high sensitivity and can distinguish HSA products made by different manufacturers and processes. The total analytical time is 10 min, which is ideal for the QC identification test of HSA products.
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OBJECTIVE: To determine glycoprotein content in recombinant human albumin from different expression systems with different methods. METHODS: Recombinant human albumin samples from Saccharomyces cerevisiae expression system, Pichia pastoris expression system, Oryza sativa expression system as well as plasma derived human albumin were investigated by phenol sulfuric acid method, HPLC peak area method and ConA combining elution HPLC method. RESULTS: For 10 batches of samples expressed in pichia pastoris expression system, the total contents of mannose were 2.7 mg•g(Pro)-1 (A manufacturer, n = 4) and 1.7 mg•g (Pro)-1 (B manufacturer, n = 6), respectively. The HPLC peak area percentages of ConA binding protein in recombinant human albumin from Pichia pastoris expression system were the highest, which showed 2.65% (A manufacturer) and 0.71% (B manufacturer) respectively, the peak area percentage of ConA binding protein in Oryza sativa expression system was 0.05% (E manufacturer, n = 3), and the ConA binding protein peak area of plasma derived human albumin was 0.01% (W manufacturer). The results of ConA binding and elution analysis with HPLC method for Quantitative determination of ConA binding protein showed that the ConA binding protein contents in the samples from pichia pastoris expression system were much higher: 27.58 mg•g(Pro)-1 (A manufacturer), 21.48 mg•g(Pro)-1 (B manufacturer), 32.02 mg•g(Pro)-1 (C manufacturer); the ConA binding protein content in the sample from Saccharomyces cerevisiae expression system was lower, 2.29 mg•g(Pro)-1 (D manufacturer); the ConA binding protein content in the samples from oryza sativa expression system was the lowest, 1.27 mg•g(Pro)-1 (E manufacturer); the plasma derived human albumin ConA binding protein content was 31.16 mg•g(Pro)-1 (S manufacturer). CONCLUSION: In terms of the results of the samples and methods involved in this study, there were glycosylated or glycol forms of protein in all recombinant human albumin samples from different expression systems; the glycosylated protein content in samples of Pichia pastoris expression system is higher than Saccharomyces cerevisiae expression system, while the glycoformed protein in samples of Oryza sativa expression system is the lowest. Plasma derived human albumin also contains glycoprotein or glycosylated protein.
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OBJECTIVE: To determine the citrate ion and aluminum contents in human albumin products and investigate the related factors of aluminum content. METHODS: Sixty-five batches of human albumin products which were very close to the end of shelf life or would be expired within not more than 2 m, including 54 batches of imported ones and 11 batches of domestic ones, were analyzed. The content of citrate ion was determined using the enzyme reaction method, and aluminum content was determined using the atomic absorption method. Factors related to human albumin production, such as ultrafiltration time, glass bottles, sample storage conditions, and the initial value of aluminum content were investigated and analyzed, especially the relationship with the aluminum content at the end of the shelf life. RESULTS: The overall mean content of citrate ion in 65 batches of human albumin was 24 μmol•L-1 (0 - 144 μmol• L-1), the mean content of citrate ion in 54 batches of domestic human albumin was 24 μmol•L-1, and that in 11 batches of imported human albumin was 23 μmol•L-1. The linear correlation coefficients between final aluminum content and citrate ion content of 65 batches of human albumin, 54 batches of domestic human albumin and 11 imported human albumin were 0.315 5, 0.331 8 and 0.746 6, respectively. The linear correlation coefficients between the final aluminum content and storage time of 65 batches of human albumin, 54 batches of domestic human albumin and 11 imported human albumin were 0.102 6, 0.059 3 and - 0.037 4, respectively. The linear correlation coefficient between the final and initial aluminum contents of 54 batches of domestic human albumin was 0.325 5. The linear correlation coefficients between final aluminum content and ultrafiltration process time, citrate ions and ultrafiltration process time of 54 batches of domestic human albumin were 0.011 8 and - 0.108 2, respectively. CONCLUSION: Citrate ion content in human albumin retention samples are lower than 150 μmol•L-1. Citrate ion content and final aluminum content are weakly correlated, however, the correlation coefficient between the two indexes of imported human albumin is much higher than that of domestic samples. The initial and final aluminum contents shows low correlation, and ultrafiltration time shows very weak correlation with storage time.
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OBJECTIVE: To evaluate the quality of human albumin products, and determine the content of aluminumion in the upcoming expired human albumin samples of both domestic and imported products for batch release. METHODS: Statistics was carried out for domestic and imported human albumin products in 2017 about the approved registration, import approval number, storage conditions, aluminumion content of batch release and other information. Aluminumion contents of the samples were detected by National Institutes for Food and Drug Control which banded with 7 authorized agencies for batch release of blood products according to the atomic absorption spectrometry or verified inductively coupled plasma mass spectrometry (ICP-MS)method for the determination of aluminumion content in China Pharmacopoeia 2015. RESULTS: There were a total of 28 domestic manufacturers of blood products and 158 human albumin drug approval numbers (totally 40 for daily production, 2017), among which 21 were approved for the storage condition of "room temperature" and 7 for "refrigerate". A total of 185 batches of human albumin samples from 25 domestic manufacturers were sampled. The mean aluminumion content in the batch release reports was 61 μg·L-1(8-134 μg·L-1), while that in the samples of the upcoming expired products was 137 μg·L-1(20-487 μg·L-1). The mean aluminumion content increased by about 1 time after the storage period. There were about 17.8% (33/185)samples having aluminumion content over 200 μg·L-1. There were a total of 13 manufacturers of imported human albumin with a total of 17 import specifications in 2017. The approved storage condition was all "room temperature". A total of 78 batches of human albumin products for batch release from 11 import enterprises were sampled. The mean aluminumion content in the batch release reports was 27 μg·L-1(7-60 μg·L-1), while that in the samples of the upcoming expired products was 50 μg·L-1(1-175 μg·L-1). The overall mean increased about 1 time after the storage period, which was consistent with the domestic human albumin products. But the two parameters of the imported products were both lower than the domestic ones. None (0/78)of the samples having aluminumion content over 200 μg·L-1. CONCLUSION: The aluminumion contents of some batches of upcoming expired domestic human albumin products are over 200 μg·L-1. The overall aluminumion contents in the initial batch release and the upcoming expired products in the import albumin products are lower than the domestic ones.
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OBJECTIVE: To prepare the first batch of Chinese national standard for human albumin used for the test of batch release or other quality control of human albumin products. METHODS: The first batch of national standard for human albumin was prepared with certificated human albumin products, mixed and filled under aseptic conditions. The standards were distributed to 11 laboratories for cooperative calibration according to the unified methods published on China Pharmacopeia. Seven test items including total protein content, sodium caprylate, polymers content, pH, absorbance, sodium content, and aluminium content were detected. RESULTS: The limits of the six test items except aluminium content were established as follows after the data from 11 collaboration laboratories were received and statistically analyzed: total protein(193.30 ± 5.08)g·L-1;sodium caprylate (0.162 4 ± 0.009 2) mmol· g(Pro)-1; polymers content (2.72 ± 0.29)% (HPLC-SEC);pH(6.71 ± 0.08) [temperature (22 ± 3)℃];absorbance 0.030 ± 0.005;sodium content (134.9 ± 23.6)mmol ·L-1. The range of initially established aluminium content was (88.4 ± 30.5)μg·L-1. However, it was observed to increase obviously after 21 months according to the trend analysis, so it was deleted from the test items for the national standard for human albumin ultimately. CONCLUSION: The prepared national standard for human albumin met the relevant requirements and may be served as the first generation of national standard for human albumin products.
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Purpose: To investigate the in vitro effect of human albumin (HA) and fresh frozen plasma (FFP) added to prime solution on platelet functions and coagulation in patients undergoing cardiopulmonary bypass (CPB) surgery. Methods: Sixty consecutive patients receiving elective cardiopulmonary bypass with open heart surgery were enrolled in the study. Patients were divided into three equal groups. Group 1: with 2 units of fresh frozen plasma added to prime solution. Group 2: With 100 cc 25% human albumins added to prime solution. Group 3: (control group) with no FFP or HA added to prime solution. PFA-100 platelet function analyzer and platelet aggregation tests were investigated pre-induction, during and after CPB and on the 1st day postoperatively. Results: Postoperative drainage was significantly higher in groups 2 and 3 compared to Group 1 (p<0.01). The compromise in platelet functions in groups 1 and 2 improved, while in Group 3 preoperative values were not attained at the end of the 1st day postoperatively. There was a significant difference between groups 2 and 3 in terms of erythrocyte suspension (ES) used in intensive care (p<0.01). Greater hemorrhage occurred in the postoperative period in Group 3 and more ES was used. In addition, lengths of stay in intensive care differed significantly between groups 2 and 3 (p<0.01). Conclusion: FFP used in CPB causes reduced drainage in the postoperative period and necessitates less use of blood and blood products.
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ObjectiveTheaimofthisstudywastoprovideabetterpositivecontrolforallergictestbycomparing the allergic effect of two kinds of positive materials , human albumin and ovalbumin , on active systemic anaphylaxis in guinea pig.Methods Guinea pigs were randomly divided into 14 groups, and were given human albumin , ovalbumin (2, 10, 100 mg/animal), or 0.9%sodium chloride injection as test substances , to assess the symptoms and incidence of systemic allergic responses induced by different sensitizing substances in different challenge doses and different challenge intervals.Results In the range of 2 to 100 mg/animal, the guinea pigs showed a 100%incidence rate of positive allergic reaction to human albumin and ovalbumin , the severity of anaphylactic symptoms was increasing along with the increase of sensitizing doses and challenge doses , and the allergic reaction was more strong induced by the same dose of ovalbumin than human albumin .Conclusions Our findings indicate that in the active systemic anaphylaxis test in guinea pigs , we recommend ovalbumin as the positive control in a dose of 2 mg/animal.
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Objective To evaluate the clinical application of human serum albumin for inpatients and provide evidence to its rational administration in our hospital .Methods 172 cases were randomly selected from the inpatients that treated with human serum albumin during the period from January to December 2010 by Mei-Kang clinical pharmacy workstation .To get the relating clinical ap-plication information from the choosing cases which were retrospectively analyzed .A comprehensive analysis was performed on age , sex, clinical departments, diagnosis, course of treatment, reasons of use, albumin concentration before using drugs etc.Results Most dosages of human serum albumin were used in the liver and gall surgery (26.1%), mainly for critical patients and cancer , Common individual consumption quantity ranged from 10 to 20 g.Most of patients adopted this drug for hypoproteinemia (38.9%), and most of them had a baseline serum albumin level at 10~30 g/L before using human serum albumin .Conclusion The application of human albumin in our hospital was far from perfect .The principle of effective , safe, economical and rational use of drugs should be adhere to reach the best efficacy application of human albumin .
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Objective To establish a method of a new type of liver fibrosis model in rats induced by repeated injection of rabbits' liver homogenate. Methods Female Wistar rats were randomly divided into a normal control group (8 rats), a human albumin induced liver fibrosis model group (15 rats) and a rabbits'liver homogenate induced liver fibrosis group (15 rats). The induction of liver fibrosis began with an immune sensitizing period (4 weeks) and was followed by an immune attacking period (8 weeks). After 8 weeks'attacking, all rats were sacrificed under anesthesia. Liver enzymes in serum and hydroxyproline in liver tissue were measured by standard methods and pathological scores were assessed by pathologists. Results The rats' liver weight, ratio of liver weight to body weight in the model group of liver homogenate were significantly increased compared with the normal control group. Serum globulin, tissue hydroxyproline were significantly increased, whereas serum albumin was significantly decreased in the homogenate group. There was only 20.0 percent of liver fibrosis score (2/10) exceeding a degree of 3 in the albumin group whereas 73.3 percent of that (11/15) were exceeding a degree of 3 in the homogenate group and the difference was significant (x2 = 4. 87,P = 0. 027). Conclusion In the study, we established a method of a new type of experimental liver fibrosis model in rats. The method has a significantly high success rate and this model can be used to study the mechanism of liver fibrosis and the efficacy of antifibrotic medicine.
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Background: Human albumin was produced and used in many countries. Cohn's technique had been used to precipitate albumin from human plasma. This technique was easy and cheap and the quality of the product was good. In Vietnam, human albumin had to import, but the prices was very expensive. Vietnam was having good plasma in large quantity and high quality. That\u2019s why research on production plasma albumin was essential.\r\n", u"Objectives: This study aimed at using Cohn's technique improved by Drohan and Van - Aken to produce standard albumin from human plasma. Subjects and method: Human plasma detected VIII-factor was used for present study. Plasma \ufffd?albumin was precipitated by ethanol at low temperature and pH. The collected albumins have been liophilizated and storage at 40C. The quality and quantity of Albumin was evaluated by quantitative analysis and protein \ufffd?electrophoresis. Results: The 418g of albumin powder was produced from 16 liters of plasma detected F \ufffd?VIII. The quality of this albumin come up to standard (>95%) and quantity of albumin collected from one liter of this plasma was 26g. Conclusion: In the Vietnamese condition, the technique of Cohn can be used to produce standard albumin for treatment.\r\n", u'
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OBJECTIVE:To determine the content of C 3 F 8 in human albumin microspheres filling-with C 3 F 8 .METHODS:By Tekmar3000purge and trap concentrator coupled with a Hewlett-Packard5972GC/MSD.RESULTS:C 3 F 8 contents in samples were119.1~433.5?g/ml.CONCLUSION:The method is feasible and suitable for quality control.