Sodium Citrate Nanoparticles Induce Dual-Path Pyroptosis for Enhanced Antitumor Immunotherapy through Synergistic Ion Overload and Metabolic Disturbance.

Metabolic reprogramming, as one of the characteristics of cancer, is associated with tumorigenesis, growth, or migration, and the modulation of metabolic pathways has emerged as a novel approach for cancer therapy. However, the conventional metabolism-mediated apoptosis process in tumor cells exhibits limited immunogenicity and inadequate activation of antitumor immunity. Herein, phospholipid-coated sodium citrate nanoparticles (PSCT NPs) are successfully prepared, which dissolve in tumor cells and then release significant amounts of citrate ions and Na+ ions. Massive quantities of ions lead to increased intracellular osmotic pressure, which activates the caspase-1/gasdermin D (GSDMD) mediated pyroptosis pathway. Simultaneously, citrate induces activation of the caspase-8/gasdermin C (GSDMC) pathway. The combined action of these two pathways synergistically causes intense pyroptosis, exhibiting remarkable antitumor immune responses and tumor growth inhibition. This discovery provides new insight into the potential of nanomaterials in modulating metabolism and altering cell death patterns to enhance antitumor immunotherapy.

Acute effects of sodium citrate supplementation on competitive performance and lactate level of elite fitness challenge athletes: A crossover, placebo-controlled, double-blind study.

Purpose: The performance of sodium citrate has been investigated in high-intensity exercises, but fewer studies have addressed the role of citrate in weight-bearing exercises. Methods: Twenty fitness challenge athletes, aged 24-32 years, volunteered to participate in this crossover, placebo-controlled, double-blind study. Initially, ten athletes were given a placebo and asked to complete a fitness challenge (i.e., chin-ups, squat jumps, dips, walking lunges, sit-ups, and burpees-devil press). Another ten athletes were supplemented with sodium citrate 0.5 g/kg body mass supplements 3 h prior to performing the fitness challenges. The same procedures were completed two days later with the supplement and placebo dextrose groups switched in a cross-over design. Athletes and assessors were blinded for the experimental condition (placebo vs. verum). Lactate levels were measured 5 min after exercise. The athletes' performance on each item of the fitness challenge as well as their lactate levels, were compared. Differences between the means of the measured variables were contrasted using a dependent t-test. Results: Supplementing sodium citrate substantially improved athletes' performance in all six fitness challenge items (p < 0.05, 0.69

Sodium citrate increases the aggregation capacity of calcium ions during microbial mineralization to accelerate the formation of calcium carbonate.

The microbially induced carbonate precipitation (MICP) technique is widely used in soil heavy metal pollution control. Microbial mineralization involves extended mineralization times and slow crystallization rates. Thus, it is important to discover a method to accelerate mineralization. In this study, we selected six nucleating agents to screen and investigated the mineralization mechanism using polarized light microscopy, scanning electron microscopy, X-ray diffraction and Fourier-transform infrared spectroscopy. The results showed that sodium citrate removed 90.1% Pb better than traditional MICP and generated the highest amount of precipitation. Interestingly, due to the addition of sodium citrate (NaCit), the rate of crystallization increased and vaterite was stabilized. Moreover, we constructed a possible model to explain that NaCit increases the aggregation capacity of calcium ions during microbial mineralization to accelerate the formation of calcium carbonate (CaCO3). Thus, sodium citrate can increase the rate of MICP bioremediation, which is important for improving MICP efficiency.

Sodium citrate effectively used in shed mediastinal blood autotransfusion after cardiac surgery.

BACKGROUND: We used sodium citrate as an alternative anticoagulation agent to heparin in the procedure of autologous blood transfusion with patients with postoperative haemorrhage after CPB. The aim of study was to evaluate the efficacy and safety of sodium citrate used in shed mediastinal blood autotransfusion after cardiac surgery. METHODS: Ninety-three patients were divided into two groups in this study. In the control group, 52 patients' shed mediastinal blood was discarded. The reinfusion group consisted of 41 patients receiving a reinfusion of washed autologous red cells from shed mediastinal blood. Each 400 mL shed blood sample was anticoagulated by 140 mL of 1.6% diluted sodium citrate in the washing procedure using a blood recovery machine. Hemoglobin (Hb), hematocrit (Hct), and electrolyte concentrations in both the patients and shed mediastinal blood were measured before and after this procedure. RESULTS: The mean volume of autotransfused shed blood was 239.5 ± 54.6 mL.The Hct of the washed red cells was 56.8 ± 6.1%. Significantly, fewer units of allogeneic blood were required per patient in the reinfusion group at 24 h postoperatively (2.91 ± 1.34 vs 4.03 ± 0.19 U, p = 0.002). At 24 h postoperatively, Hb and Hct levels were higher in the reinfusion group than in the control group. The calcium ion concentration was very low in the shed mediastinal blood, 0.25 ± 0.08 mmol/L, and was lower after washing, 0.15 ± 0.04 mmol/L. CONCLUSIONS: Sodium citrate, as an alternative anticoagulant agent, can be used in autologous shed mediastinal blood transfusion after CPB cardiac surgery. This procedure can effectively reduce the amount of allogeneic blood for patients with haemorrhage.

Influence of blood sample storage and different types of anticoagulants on results of NK cytotoxicity tests.

Introduction: Natural killer (NK) cells are important players in the human immune response. Impaired NK function may lead to serious, life-threatening conditions. Defects may be consequences of genetic mutations or results of secondary factors such as infections, malignancies and autoimmune diseases. The cytotoxicity test is very useful, but its accessibility is limited to special immunological laboratories. Blood samples are often transported to remote centers, which takes time and requires special conditions.The aim of this study was to compare cytotoxicity assay results between samples preserved with three different anticoagulants to standardize the diagnostic procedure. Material and methods: Peripheral blood from healthy donors was taken with three anticoagulants: heparin, K2EDTA and citrate. Peripheral blood mononuclear cells (PBMC) were isolated and tested directly after blood drawing and after 24-hour storage. Cytotoxic abilities of NK cells were tested in 4 h co-culture with K562. NK cytotoxicity was measured by flow cytometry. Results: In most cases of analyzed healthy donors, cytotoxicity results were similar regardless of type of anticoagulant. However, the highest mean values were obtained in samples with citrate. There was a significant decrease in cytotoxicity after 24 hours of storage of the whole blood at ambient temperature. The mean drop in cytotoxicity results was substantial for all anticoagulants: 76% for heparin, 67% for citrate and 70% for EDTA. Conclusions: Results of spontaneous NK cytotoxicity seem to be affected by the anticoagulants used for blood protection. Commercial instant cytotoxicity testing and delayed analysis after blood storage gave the highest results in blood with sodium citrate.

Zuragen™ antiseptic catheter lock: A bridge too far?

When sodium citrate is used as an anticoagulant catheter lock the best concentration is 7%, since this provides a density approximately the same as blood. Our laboratory found that the addition of methylene blue and parabens greatly augmented antibacterial properties. Ash Access sponsored a randomized clinical trial in 400 dialysis patients with tunneled CVC, and this showed a significant decrease in incidence of catheter-related bloodstream infection (CRBSI) defined by stringent criteria. The FDA decided that the study missed its primary endpoint, and that the product was mis-classified, so they did not give approval to market. The licensee decided not to appeal to the decision to the physician panel and ended support of the project. Rights to market the catheter lock eventually returned to Ash Access.

Stability of benzylpenicillin for continuous intravenous infusions: An isotonic formulation for therapeutic use and a low-dose formulation for clinical trial.

INTRODUCTION: The objectives of this study were to develop a stability-indicating high performance liquid chromatography (HPLC) assay for benzylpenicillin (BPC) in pharmaceutical fluids, and to investigate the stability of (i) isotonic citrate-buffered BPC solutions at the clinically relevant concentration of 30 mg/mL, and (ii) low concentration citrate-buffered BPC intravenous infusions (5-30 µg/mL). METHODS: The stability of isotonic BPC solutions containing 3.4 or 7.2 mg/mL sodium citrate was compared against contemporary hypertonic solutions. The HPLC assay was shown to be stability-indicating following acidic, alkali, oxidative and elevated temperature stress testing. RESULTS: After 7 d storage at 4 °C and 24 h at 35 °C, the concentrations of isotonic BPC 30 mg/mL solutions containing 3.4 and 7.2 mg/mL sodium citrate were 96% and 95% respectively, compared to day 0. After 3 d at 4 °C and 24 h at room temperature (22 °C), the concentrations of isotonic BPC solutions with 3.4 and 7.2 mg/mL sodium citrate were 99% and 96% respectively, compared to day 0. These data were comparable to the hypertonic solutions and meet pharmacopeial stability requirements. Low concentration BPC infusions showed 0.5% and 2.5% degradation after 24 h storage at 22 °C and 35 °C, respectively. CONCLUSIONS: The isotonic BPC 30 mg/mL formulation is simple to prepare and may offer clinical benefits in settings where hypertonic solutions are problematic. This study provides assurance that high- and low-dose isotonic BPC infusions are stable at room temperature and our findings may be applicable to in vitro studies of BPC.

Effects of Various Anticoagulants on Biochemistry Analytes From Domestic Pigeons (Columba livia domestica).

The objective of this study was to compare the effects of anticoagulant and no anticoagulant on routine biochemical analytes in domestic pigeons (Columba livia domestica). Blood samples were obtained from 8 clinically healthy pigeons. The sample obtained from each bird was divided into 4 blood collection tubes containing either ethylenediaminetetraacetic acid (EDTA), lithium heparin, sodium citrate, or no anticoagulant. The concentrations of creatinine, uric acid, triglyceride, total cholesterol, glucose, phosphorus, calcium, magnesium, total protein, albumin, and iron, and the activities of alkaline phosphatase (ALP), alanine aminotransferase (ALT), and aspartate aminotransferase (AST), were measured in blood from each of the blood collection tubes. The values of the measured parameters, with the exception of iron, were significantly lower in the citrated plasma samples compared with the serum samples, even after correcting for dilution. In the lithium heparin plasma samples, significant decreases in albumin, triglyceride, calcium, total cholesterol, and ALP, and a significant increase in iron, were observed compared with the values in the serum samples. The concentrations of total protein, creatinine, glucose, total cholesterol, triglyceride, magnesium, calcium, and phosphorus, as well as the activities for AST and ALP, were significantly lower in the EDTA plasma samples compared with the serum samples. In conclusion, the anticoagulants had significant effects on most of the measured parameters compared with serum. The findings of the present study suggest that a lithium heparin sample is the most appropriate plasma sample for the measurement of blood biochemical parameters in the domestic pigeon.

Sodium citrate and biochar synergistic improvement of nanoscale zero-valent iron composite for the removal of chromium (â ¥) in aqueous solutions.

Sodium citrate (SC) is a widely-used food and industrial additive with the properties of complexation and microbial degradation. In the present study, nano-zero-valent iron reaction system (SC-nZVI@BC) was successfully established by modifying nanoscale zero-valent iron (nZVI) with SC and biochar (BC), and was employed to remove Cr(Ⅵ) from aqueous solutions. The nZVI, SC-nZVI and SC-nZVI@BC were characterized and compared using X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), thermogravimetric analyses (TGA), vibrating sample magnetometer (VSM), scanning electron microscope (SEM), X-ray diffraction (XRD) and X-ray photoelectron spectroscopy (XPS). The results showed that nZVI was successfully loaded on the biochar, and both the agglomeration and surface passivation problems of nanoparticles were well resolved. The dosage of SC, C:Fe, initial pH and Cr(Ⅵ) concentration demonstrated direct effects on the removal efficiency. The maximum Cr(Ⅵ) removal rate and the removal capacity within 60 min were 99.7% and 199.46 mg/g, respectively (C:Fe was 1:1, SC dosage was 1.12 mol.%, temperature was 25°C, pH = 7, and the original concentration of Cr(Ⅵ) was 20 mg/L). The reaction confirmed to follow the pseudo-second-order reaction kinetics, and the order of the reaction rate constant k was as follows: SC-nZVI@BC > nZVI@BC > SC-nZVI > nZVI. In addition, the mechanism of Cr(Ⅵ) removal by SC-nZVI@BC mainly involved adsorption, reduction and co-precipitation, and the reduction of Cr(Ⅵ) to Cr(Ⅲ) by nano Fe0 played a vital role. Findings from the present study demonstrated that the SC-nZVI@BC exhibited excellent removal efficiency toward Cr(Ⅵ) with an improved synergistic characteristic by SC and BC.

Feasibility of Citrate Dialysis in Hyponatremia: A Case Series.

BACKGROUND: Correcting hyponatremia too quickly can lead to osmotic demyelination syndrome. During citrate dialysis, a significant sodium load is brought to the prefilter. We reviewed the impact of this sodium load on the evolution of sodium levels in patients undergoing continuous renal replacement therapy with citrate anticoagulation. MATERIALS AND METHODS: The medical records of 5 patients with hyponatremia who received dialysis with citrate anticoagulation, over a 10-year period, were reviewed. The sodium of the dialysate and of the reinjection fluid was adapted according to the serum sodium level recommended by the guidelines of the time. Data from the first 24 h after initiation of dialysis was evaluated. RESULTS: The difference in serum sodium levels between day 1 and day 2 was statistically significant, with a rise of 7.8 ± 3.7 mmol/L. DISCUSSION: The mean serum sodium increase in our series of patients did not exceed the increase of 10-12 mEq/L/day permitted by the guidelines. The excess sodium was absorbed by the filter. CONCLUSION: In this small series of patients, with adjustment of the sodium concentration of dialysate and reinjection fluid, the use of citrate was found to be safe.

Synergistic degradation of pyrene by Pseudomonas aeruginosa PA06 and Achromobacter sp. AC15 with sodium citrate as the co-metabolic carbon source.

Two pyrene-degrading strains, Pseudomonas aeruginosa PA06 and Achromobacter sp. AC15 were co-incubated in equal proportions as a microbiological consortium and could enhance the degradation of pyrene. The enzymatic activities of the catechol 1,2-dioxygenase (C12O) and 2,3-dioxygenase activities (C23O) were produced complementary expression by P. aeruginosa PA06 and Achromobacter sp. AC15, respectively. Meanwhile, results showed that pyrene degradation was sufficiently promoted in the presence of sodium citrate as a co-metabolic carbon source, likely a result of enhanced biomass and biosurfactant production. The optimized dosage and ideal initial pHs were 1.4 g L-1 and 5.5, respectively. We also analyzed the rate constant of pyrene degradation, cell growth, and enzyme activity. Results show that P. aeruginosa PA06 had a better effect than Achromobacter sp. AC15 in bacterial growth. However, the C23O or C12O activity produced by Achromobacter sp. AC15 continued at a similar or even faster than that of P. aeruginosa PA06. The mixed bacteria had a better effect than any single bacteria, suggesting the strains worked synergistically to enhance the degradation efficiency. In the co-metabolism system of 600 mg/L pyrene and 1.4 g/L sodium citrate, pyrene degradation reached 74.6%, was 1.57 times, 2.06 times, and 3.89 times that of the mix-culture strains, single PA06 and single AC15 without sodium citrate, respectively. Overall, these findings are valuable as a potential tool for the bioremediation of high-molecular-weight PAHs.

Intranasal sodium citrate in quantitative and qualitative olfactory dysfunction: results from a prospective, controlled trial of prolonged use in 60 patients.

OBJECTIVES: We have previously shown that treatment with intranasal sodium citrate may be beneficial in post-infectious olfactory dysfunction. Sodium citrate reduces free intranasal calcium and is, therefore, thought to prevent calcium-mediated feedback inhibition at the level of the olfactory receptor. We aimed to determine whether treatment with a 2-week course of intranasal sodium citrate improves quantitative olfactory function in patients with post-infectious impairment. We also aimed to determine whether sodium citrate is beneficial in treating qualitative olfactory dysfunction. METHODS: We performed a prospective, controlled study. Patients applied intranasal sodium citrate solution to the right nasal cavity for 2 weeks. The left nasal cavity was untreated and, therefore, acted as an internal control. Monorhinal olfactory function was assessed using the "Sniffin' Sticks" composite 'TDI' score, before and after treatment. The presence of parosmia and phantosmia was also assessed. RESULTS: Overall, there was a significant increase in TDI after treatment (using the best of right and left sides). Treatment with sodium citrate did not significantly improve quantitative olfactory function, compared to control. The proportion of patients reporting parosmia did not change significantly after treatment. However, there was a significant reduction in the proportion of patients reporting phantosmia, at the end of the study period. CONCLUSIONS: Treatment with intranasal sodium citrate for a period of 2 weeks does not appear to improve quantitative olfactory function in patients with post-infectious impairment, compared to control. It may, however, be beneficial in treating phantosmia, which should be further addressed in future work.

Extracellular buffer choice influences acid-base responses and gastrointestinal symptoms.

To compare the bicarbonate kinetics and gastrointestinal (GI) symptom responses between an equal dose of sodium bicarbonate and sodium citrate using delayed-release capsules. Thirteen active males (age 20.5 ± 2.1 y, height 1.8 ± 0.1 m and body mass [BM] 76.5 ± 9.6 kg) consumed either 0.3 g.kg-1 BM sodium bicarbonate, sodium citrate or a placebo, using a double-blind, randomized crossover design. Blood bicarbonate ion (HCO3-) concentration, pH and GI symptoms were measured pre-consumption and every 10 min for 180 min post-consumption. Blood HCO3- concentration (P < 0.001) and pH (P = 0.040) were significantly higher in the sodium bicarbonate condition compared with sodium citrate condition up to 3 h post-consumption. Peak blood HCO3- concentration was significantly higher with the sodium bicarbonate compared with citrate (P < 0.001). Mean GI symptom scores were lower (P = 0.037) for sodium citrate (1.5 ± 1.8 AU) than bicarbonate (2.6 ± 3.1 AU), with considerable inter-individual variability. No GI symptoms were reported following consumption of the placebo. Both substances increase HCO3- values significantly, with sodium bicarbonate causing significantly higher pH and HCO3- values than the same dose of sodium citrate, but results in slightly more severe GI symptoms.

A comparison of the use of different swab materials for optimal diagnosis of amoebic gill disease (AGD) in Atlantic salmon (Salmo salar L.).

Routine gill swabbing is a non-destructive sampling method used for the downstream qPCR detection and quantitation of the pathogen Neoparamoeba perurans, a causative agent of amoebic gill disease (AGD). Three commercially available swabs were compared aiming their application for timelier AGD diagnosis (Calgiswab® (calcium alginate fibre-tipped), Isohelix® DNA buccal and cotton wool-tipped). Calcium alginate is soluble in most sodium salts, which potentially allows the total recovery of biological material, hence a better extraction of target organisms' DNA. Thus, this study consisted of (a) an in vitro assessment involving spiking of the swabs with known amounts of amoebae and additional assessment of retrieval efficiency of amoebae from agar plates; (b) in vivo testing by swabbing of gill arches (second, third and fourth) of AGD-infected fish. Both in vitro and in vivo experiments identified an enhanced amoeba retrieval with Calgiswab® and Isohelix® swabs in comparison with cotton swabs. Additionally, the third and fourth gill arches presented significantly higher amoebic loads compared to the second gill arch. Results suggest that limiting routine gill swabbing to one or two arches, instead of all, could likely lead to reduced stress-related effects incurred by handling and sampling and a timelier diagnosis of AGD.

Air-Jet Spun Corn Zein Nanofibers and Thin Films with Topical Drug for Medical Applications.

Diabetic patients are especially susceptible to chronic wounds of the skin, which can lead to serious complications. Sodium citrate is one potential therapeutic molecule for the topical treatment of diabetic ulcers, but its viability requires the assistance of a biomaterial matrix. In this study, nanofibers and thin films fabricated from natural corn zein protein are explored as a drug delivery vehicle for the topical drug delivery of sodium citrate. Corn zein is cheap and abundant in nature, and easily extracted with high purity, while nanofibers are frequently cited as ideal drug carriers due to their high surface area and high porosity. To further reduce costs, the 1-D nanofibers in this study were fabricated through an air jet-spinning method rather than the conventional electrospinning method. Thin films were also created as a comparative 2-D material. Corn zein composite nanofibers and thin films with different concentration of sodium citrate (1-30%) were analyzed through FTIR, DSC, TGA, and SEM. Results reveal that nanofibers are a much more effective vehicle than films, with the ability to interact with sodium citrate. Thermal analysis results show a stable material with low degradation, while FTIR reveals strong control over the protein secondary structures and hold of citrate. These tunable properties and morphologies allow the fibers to provide a sustained release of citrate and then revert to their structure prior to citrate loading. A statistical analysis via t-test confirmed a significant difference between fiber and film drug release. A biocompatibility study also confirms that cells are much more tolerant of the porous nanofiber structure than the nonporous protein films, and lower percentages of sodium citrate (1-5%) were outperformed to higher percentages (15-30%). This study demonstrated that protein-based nanofiber materials have high potential as vehicles for the delivery of topical diabetic drugs.

Response surface methodology for the extraction of wedelolactone from Eclipta alba using aqueous two-phase extraction.

Aqueous two-phase extraction of wedelolactone from Eclipta alba was studied using the polymer-salt system. The system consisted of polyethylene glycol (PEG) as a top phase (polymer) and sodium citrate as a bottom phase (salt). Process parameters such as PEG concentration, PEG molecular weight, salt concentration, and pH have been optimized using response surface methodology (RSM) with the help of central composite design (CCD). The optimized conditions for aqueous two-phase system (ATPS), in the case of one factor at a time approach, were found as PEG 6000, PEG concentration 18% (w/v), salt concentration 16% (w/v), and pH 7; with maximum extraction yield of 6.52 mg/g. While, RSM studies showed maximum extraction yield of 6.73 mg/g with the optimized parameters as PEG 6000, PEG concentration 18% (w/v), salt concentration 17.96% (w/v), and pH 7. ATPS was found to give a 1.3 fold increase in the extraction yield of wedelolactone as compared to other conventional extraction methods.

Tolerance to Sodium in Patients With CKD-Induced Metabolic Acidosis: Does the Accompanying Anion Matter?

Patients with chronic kidney disease (CKD) continue to produce endogenous acids but have a reduction in net acid excretion, resulting in a primary decrease in serum bicarbonate concentration, which is termed chronic metabolic acidosis. Recent prospective studies, along with retrospective cohort analyses, demonstrate a higher risk for CKD progression with untreated metabolic acidosis. To normalize serum bicarbonate levels, acidemic patients are often treated with sodium bicarbonate (NaHCO3) or sodium citrate, which have been shown to slow the progression of CKD. However, studies using this approach have routinely excluded patients with common sodium-sensitive comorbid conditions, such as poorly controlled hypertension, congestive heart failure, volume overload, or edema. This article examines the effect of the anion that accompanies sodium delivered with these therapies. Do the negative effects on blood pressure (BP) and sodium retention, as measured by an increase in edema, weight gain, and congestive heart failure, observed with oral administration of sodium chloride (NaCl) differ when a similar amount of sodium is given with bicarbonate or citrate in this patient population? A review of the literature suggests that NaHCO3 does not increase BP or sodium retention when administered to patients with CKD during a concurrent severe NaCl dietary restriction (∼10 mEq/d). However, this degree of NaCl restriction is feasible only under strict control in clinical research environments. In contrast, when NaHCO3 is given to patients without severe dietary NaCl restriction, there is an increase in BP and sodium retention. Thus, unless patients with CKD can tolerate a diet virtually devoid of NaCl, additional sodium, regardless of the accompanying anion, appears to increase BP and sodium retention.

Impact of sodium citrate on structural properties of gluten.

The effect of sodium citrate on gluten-starch separation and physicochemical properties of gluten was studied. The results showed that the addition of sodium citrate to the dough caused to an improvement in the aggregation of gluten and increased gluten yield in comparison to the control by augmentation pH and approaching to the isoelectric point of glutenin and gliadin. Also, sodium citrate led a shift to larger particle size distribution of glutenin macropolymer (GMP). These observations demonstrated that under the influence of sodium citrate more thiol groups were oxidized and formed disulfide bonds, which increased the storage modulus and resistance to extension. Furthermore, in this sample the GMP gel was more elastic and stiffer.

Effects of pH and concentration of sodium citrate anticoagulant on platelet aggregation measured by light transmission aggregometry induced by adenosine diphosphate.

The 2013 ISTH-SSC guidelines for the standardization of light transmission aggregometry (LTA) were largely based on expert consensus, as studies directly comparing LTA methodologies were lacking. We experimentally tested the cogency of ISTH-SSC recommendations pertaining to use of anticoagulant, in particular whether: (1) buffered citrate (BC) is preferable to unbuffered citrate (C); (2) the two recommended concentrations of sodium citrate (109 and 129 mM) are equivalent in terms of platelet aggregation (PA). Blood from 16 healthy volunteers was collected into BC and C (109 and 129 mM). PA was measured by LTA in platelet-rich plasma (PRP) stimulated by adenosine diphosphate (ADP) (2 µM) immediately after PRP preparation and up to 4 hr after blood collection; pH and platelet counts in PRP were measured in parallel. pH in PRP increased with time up to about 8 for all anticoagulants, although it was lower in BC than in C at all times. In BC, PA was lower at 45 min, but equivalent at all other times. PA was higher and more stable in sodium citrate 109 mM than in 129 mM at all times. The extent of PA did not change for up to 2 hr after blood collection, and subsequently dramatically decreased. In contrast with ISTH-SSC recommendations, (1) BC does not show advantages compared to C; (2) 109 mM citrate is preferable to 129 mM, because it better supports PA; and (3) LTA studies should be completed within 2 hr of blood collection, instead of the recommended 4 hr.

Effects of sodium citrate, citric acid and lactic acid on human blood coagulation.

INTRODUCTION: Citric acid infusion in extracorporeal blood may allow concurrent regional anticoagulation and enhancement of extracorporeal CO2 removal. Effects of citric acid on human blood thromboelastography and aggregometry have never been tested before. METHODS: In this in vitro study, citric acid, sodium citrate and lactic acid were added to venous blood from seven healthy donors, obtaining concentrations of 9 mEq/L, 12 mEq/L and 15 mEq/L. We measured gas analyses, ionized calcium (iCa++) concentration, activated clotting time (ACT), thromboelastography and multiplate aggregometry. Repeated measure analysis of variance was used to compare the acidifying and anticoagulant properties of the three compounds. RESULTS: Sodium citrate did not affect the blood gas analysis. Increasing doses of citric and lactic acid progressively reduced pH and HCO3- and increased pCO2 (p<0.001). Sodium citrate and citric acid similarly reduced iCa++, from 0.39 (0.36-0.39) and 0.35 (0.33-0.36) mmol/L, respectively, at 9 mEq/L to 0.20 (0.20-0.21) and 0.21 (0.20-0.23) mmol/L at 15 mEq/L (p<0.001). Lactic acid did not affect iCa++ (p=0.07). Sodium citrate and citric acid similarly incremented the ACT, from 234 (208-296) and 202 (178-238) sec, respectively, at 9 mEq/L, to >600 sec at 15 mEq/L (p<0.001). Lactic acid did not affect the ACT values (p=0.486). Sodium citrate and citric acid similarly incremented R-time and reduced α-angle and maximum amplitude (MA) (p<0.001), leading to flat-line thromboelastograms at 15 mEq/L. Platelet aggregometry was not altered by any of the three compounds. CONCLUSIONS: Citric acid infusions determine acidification and anticoagulation of blood similar to lactic acid and sodium citrate, respectively.