Glycopeptide

In animal modeling, vancomycin can be used to create kidney injury models. To reduce infusion-related side effects, vancomycin is infused intravenously for a minimum of one hour. Vancomycin has a β-elimination half-life of 6–12 hours and an α-distribution phase of 30 min to 1 h in subjects with normal creatinine clearance. 0.4–1 L/kg is the distribution volume. Vancomycin can bind to proteins 10%–50% of the time. Vancomycin's total activity is influenced by a number of factors, including its tissue distribution, inoculum size, and effects on protein binding[1]. Treatment with vancomycin for infected mice is linked to better scores in histopathology, clinical manifestations, and diarrhea[3].

Vancomycin is administered intravenously, with a standard infusion time of at least 1 h, to minimize infusion-related adverse effects. Subjects with normal creatinine clearance, vancomycin has an α-distribution phase of 30 min to 1 h and a β-elimination half-life of 6-12 h. The volume of distribution is 0.4–1 L/kg. The binding of vancomycin to protein ranges from 10% to 50%. Factors that affect the overall activity of vancomycin include its tissue distribution, inoculum size, and protein-binding effects. Vancomycin treatment of infected mice is associated with improved clinical, diarrhea, and histopathology scores and survival during treatment.

Vancomycin is a unique glycopeptide structurally unrelated to any currently available antibiotic. It also has a unique mode of action inhibiting the second stage of cell wall synthesis of susceptible bacteria. There is also evidence that vancomycin alters the permeability of the cell membrane and selectively inhibits ribonucleic acid synthesis. Induction of bacterial L-phase variants from susceptible organisms with vancomycin is extremely difficult, and such variants are unstable. Stable L-phase variants induced by other agents are susceptible to vancomycin. Vancomycin is active against a large number of species of Gram-positive bacteria, such as Staphylococcus aureus (including methicillin-resistant strains), Staph. epidermidis (including multiple-resistant strains), Streptococcus pneumoniae (including multiple-resistant strains), Str. pyogenes, Str. agalactiae, Str. bovis, Str. mutans, viridans streptococci, enterococci, Clostridium species, diphtheroids, Listeria monocytogenes, Actinomyces species and Lactobacillus species. There has been no increase in resistance to vancomycin during the past three decades. Enhancement of antimicrobial activity has been demonstrated with the combination of vancomycin and an aminoglycoside against Staph. aureus, Str. bovis, enterococci and viridans streptococci. The combination of vancomycin and rifampicin are antagonistic to most strains of Staph. aureus, though indifference and occasionally synergism have been shown, but is synergistic against strains of Staph. epidermidis. It shows indifference against enterococci. Vancomycin and fusidic acid are indifferent against Staph. aureus [2].

C. difficile toxin assay. C. difficile toxins A and B were detected using a modified protocol for the Tech Lab Toxin A/B II ELISA kit. Each stool sample was weighed and the amount of diluent per sample was normalized to provide the same stool mass-to-diluent ratio for each sample. The diluent-sample mixtures were homogenized by grinding and vortexing, and 1:10, 1:100, and 1:1,000 serial dilutions were made of the sample. A total of 150 μl of the 1:1,000 dilution of each sample was added to a precoated well provided in the kit. A negative control consisted of 150 μl of diluent, and a positive control consisted of 135 μl of diluent plus 3 drops of the positive control toxin A-B mixture provided in the kit. One drop of conjugate was added to each well, and the plate was incubated at 37°C for 50 min. Each well was washed three times with 150 μl of a 1× dilution of the wash buffer provided in the kit. Two drops of substrate were added to each well. After 10 min, 1 drop of stop solution was added to each well. The plate was allowed to sit for 2 min before being read in an ELISA reader [3].

Mice: In one series of studies, infected mice are given either vancomycin (20 mg/kg) daily for five or ten days and monitored for fifteen days after infection, or vancomycin (50 mg/kg) daily for one, two, three, or five days and monitored for twenty-one days after infection[3].

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Murine model of C. difficile infection and treatment.[3]
The infection model is a modification of the published protocol of Chen et al. This protocol has been approved by the Center for Comparative Medicine at University of Virginia. C57BL/6 mice, male, 8 weeks old, were used. From 6 to 4 days prior to infection, mice were given an antibiotic cocktail containing vancomycin (0.0045 mg/g), colistin (0.0042 mg/g), gentamicin (0.0035 mg/g), and metronidazole (0.0215 mg/g) in drinking water. One day prior to infection, clindamycin (32 mg/kg of body weight) was injected subcutaneously. The mice were divided into the following groups: control uninfected, control infected, infected and treated with vancomycin (20 mg/kg), and infected and treated with comparator drugs—nitazoxanide, fidaxomicin, and metronidazole (all drugs given at 20 mg/kg/day). Food and water were allowed ad libitum. Although each mouse or treatment group was housed in a separate cage, all mice were housed in the same pod of the vivarium. Infection was performed with VPI 10463 (ATCC) as an inoculum of 104 or 105 administered by oral gavage. This strain produces both C. difficile toxins A (TcdA) and B (TcdB). One day postinfection, treated mice were given either vancomycin or nitazoxanide at 20 mg/kg each by oral gavage daily for 5 days and monitored for either 1 or 2 weeks postinfection. One set of experiments was performed in which infected mice were treated with vancomycin (50 mg/kg) daily for 1, 2, 3, or 5 days and were observed for 21 days postinfection or with vancomycin (20 mg/kg) daily for either 5 or 10 days and monitoring for 15 days postinfection. In a separate experiment, mice given a preinfection antibiotic regimen described above were treated with either vancomycin, fidaxomicin, or metronidazole at 20 mg/kg/day for 5 days and infected another 5 days later. Except when indicated, all comparator drugs were administered using the same dosage (20 mg/kg/day for 5 days) to equally compare efficacies, outcomes, and effects on selected gut floras between treatment groups as previously described. From another study, a group of control mice was given vancomycin but was not infected. A clinical scoring system was developed on the basis of weight loss, diarrhea, activity level, and appearance of eyes and hair (each parameter scored from 0 to 3, where 0 is normal and 3 is the worst; maximum score of 20). Stool specimens were collected daily. Diarrhea was scored as follows: 1 for soft or color change (yellow), 2 for wet tail or mucoid, and 3 for liquid or no stool (ileus). Mice judged moribund by the clinical score (score of >14) at any day and all surviving mice at the end of the experiment were sacrificed, and intestinal tissues and cecal contents were collected as described below. A separate set of experiments was performed for harvesting cecal contents for clostridial bacterial and toxin burdens at days 3, 6, 9, and 12 to 13 postinfection to follow changes at different time points of the study.
Histopathology. Upon euthanasia, cecal and colonic tissues were fixed in 10% zinc formalin overnight and then placed in 10% ethanol before being sent for paraffin embedding and hematoxylin and eosin (H&E) staining at the University of Virginia Histology Research Core. Histopathologic scoring was performed coded (C.A.W. and M.S.R.). H&E-stained tissues were scored for mucosal disruption, mucosal hypertrophy, inflammation, vascular congestion and exudates, and submucosal edema (each parameter was graded from 0 to 3, with 0 as normal and 3 worst; maximum score of 15) as we previously described in detail.

Absorption, Distribution and Excretion
Vancomycin is poorly absorbed in the gastrointestinal tract, but systemic absorption (up to 60%) occurs after intraperitoneal injection. Approximately 75-80% of the vancomycin dose is excreted in the urine via glomerular filtration within 24 hours after administration. Its volume of distribution is between 0.4 and 1 L/kg, as described in the literature. The mean plasma clearance of vancomycin is approximately 0.058 L/kg/h. Most patients experience minimal gastrointestinal absorption of vancomycin hydrochloride; therefore, parenteral administration is necessary to treat systemic infections. Oral bioavailability is typically less than 5%. However, limited data suggest that clinically significant serum drug concentrations may be observed in some patients with colitis and/or renal impairment following enteral or oral administration of vancomycin.
In adults with normal renal function, after multiple intravenous infusions of 1 gram of vancomycin (15 mg/kg) over 1 hour, the mean plasma concentration immediately after infusion is approximately 63 μg/mL, and the mean plasma concentrations at 2 hours and 11 hours are approximately 23 μg/mL and 8 μg/mL, respectively. When a 500 mg dose is administered intravenously multiple times over 30 minutes, the mean plasma concentration immediately after infusion is approximately 49 μg/mL, and the mean plasma concentration at 6 hours is approximately 10 μg/mL.
After intravenous administration, vancomycin is distributed into breast milk. The systemic absorption rate of oral vancomycin is extremely low, and it is unknown whether the drug is distributed into human breast milk after oral administration.
Vancomycin readily crosses the placenta and distributes into umbilical cord blood.
At serum vancomycin concentrations of 10 to 100 μg/mL, the serum protein binding rate of vancomycin, as determined by ultrafiltration, is approximately 55%. Following intravenous administration of vancomycin hydrochloride, inhibitory concentrations were observed in pleural effusion, pericardial effusion, ascites and synovial fluid, urine, peritoneal dialysis fluid, and auricle tissue. Vancomycin hydrochloride does not readily diffuse across normal meninges into the cerebrospinal fluid. However, when the meninges are inflamed, the drug may seep into the cerebrospinal fluid. Metabolism/Metabolites: Since approximately 75-80% of the drug is excreted unchanged in the urine within 24 hours after administration, the drug appears to have no significant metabolism. It has been reported that vancomycin concentrations in liver tissue and bile are also at or below the detection limit 24 hours after administration. Free toxins are cleared through opsonization of the reticuloendothelial system (primarily in the liver and kidneys) and degraded through endocytosis of lysosomes. Lysosomes are membrane-bound organelles containing various digestive enzymes, including several proteases. Elimination pathway: Within 24 hours after administration, approximately 75% of the vancomycin dose is excreted in the urine via glomerular filtration.
Half-life: The half-life in patients with normal renal function is approximately 6 hours (range 4 to 11 hours). Within 24 hours of administration, approximately 75% of the vancomycin dose is excreted in the urine via glomerular filtration. The mean elimination half-life in patients without kidneys is 7.5 days.
Biological half-life
The half-life in patients with normal renal function is approximately 6 hours (range 4 to 11 hours). In patients without kidneys, the mean elimination half-life of vancomycin is 7.5 days.
In subjects with normal renal function, the mean elimination half-life of vancomycin from plasma is 4 to 6 hours.
In patients without kidneys, the mean elimination half-life of vancomycin is 7.5 days.

Toxicity Summary
Vancomycin's bactericidal effect primarily stems from its inhibition of cell wall biosynthesis. Specifically, vancomycin prevents the integration of N-acetylmuramic acid (NAM) and N-acetylglucosamine (NAG) peptide subunits into the peptidoglycan matrix, a major structural component of Gram-positive bacterial cell walls. This macromolecular hydrophilic agent forms hydrogen bonds with the D-alanyl-D-alanine residues at the ends of the NAM/NAG peptides. Normally, this interaction is a five-point hydrogen bond. The binding of vancomycin to the D-alanyl-D-alanine residues prevents the integration of NAM/NAG peptide subunits into the peptidoglycan matrix. Furthermore, vancomycin alters bacterial cell membrane permeability and RNA synthesis. There is no cross-resistance between vancomycin and other antibiotics. Vancomycin is inactive against Gram-negative bacilli, mycobacteria, or fungi in vitro.

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Toxicity Data
LD50: 5000 mg/kg (oral, mouse) (A308)
LD50: 319 mg/kg (intravenous, rat) (A308)
LD50: 400 mg/kg (intravenous, mouse) (A308)

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Interactions
Concomitant use of vancomycin with anesthetics is associated with an increased incidence of anaphylactic reactions and infusion reactions (e.g., hypotension, flushing, erythema, urticaria, pruritus). Erythema and histamine-like flushing have been observed in pediatric patients receiving concomitant treatment with vancomycin and anesthetics. The risk of infusion-related adverse reactions can be minimized by administering vancomycin intravenously 1 hour prior to anesthesia induction.
In vitro studies have shown that vancomycin and aminoglycoside antibiotics have synergistic antibacterial activity against a variety of Staphylococcus aureus, non-enterococcal group D streptococci (Streptococcus bovis), enterococci (Enterococcus faecalis), and viridans streptococci. However, concomitant use of vancomycin with aminoglycoside antibiotics increases the risk of ototoxicity and/or nephrotoxicity. Due to the potential for additive toxicity, close monitoring of renal function and hearing is necessary when using vancomycin concurrently or sequentially, systemically or locally, with other ototoxic and/or nephrotoxic drugs (e.g., aminoglycosides, amphotericin B, bacitracin, cisplatin, colistin, polymyxin B). Patients receiving vancomycin should use these drugs with caution. Two patients receiving tenofovir disoproxil fumarate as part of an antiretroviral therapy regimen developed renal failure after prolonged vancomycin treatment. Tenofovir is associated with Fanconi syndrome and renal insufficiency due to its effects on the proximal tubules. Vancomycin nephrotoxicity is uncommon but can occur with concomitant use with nephrotoxic drugs. Clinicians should be aware that tenofovir may increase the risk of renal failure during prolonged vancomycin use. For more complete data on vancomycin drug interactions (6 items in total), please visit the HSDB record page.

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Non-human toxicity values
Mice oral LD50: 5000 mg/kg /hydrochloride/
Mice intraperitoneal LD50: 1734 mg/kg
Mice intravenous LD50: 430 mg/kg
Mice subcutaneous LD50: 5000 mg/kg
Rat intravenous LD50: 319 mg/kg

[1]. The pharmacokinetic and pharmacodynamic properties of vancomycin. Clin Infect Dis. 2006 Jan 1;42 Suppl 1:S35-9.

[2]. Mode of action and in-vitro activity of vancomycin. J Antimicrob Chemother. 1984 Dec;14 Suppl D:7-18.

[3]. Vancomycin treatment's association with delayed intestinal tissue injury, clostridial overgrowth, and recurrence of Clostridium difficile infection in mice. Antimicrob Agents Chemother. 2013 Feb;57(2):689-96.

Therapeutic Uses
Antibiotic, Glycopeptide
Vancomycin hydrochloride is indicated for the treatment of severe or toxic infections caused by vancomycin-sensitive methicillin-resistant (β-lactam-resistant) staphylococci. It is indicated for patients allergic to penicillin, those who cannot tolerate or have failed treatment with other drugs (including penicillins or cephalosporins), and those with infections caused by vancomycin-sensitive bacteria resistant to other antimicrobial agents. Vancomycin hydrochloride can be used as initial treatment when methicillin-resistant staphylococci are suspected, but the treatment regimen should be adjusted based on the results of susceptibility testing. /US Product Label Content/
Vancomycin hydrochloride is effective in treating staphylococcal endocarditis. Its efficacy has been demonstrated in other staphylococcal infections, including sepsis, bone infections, lower respiratory tract infections, and skin and soft tissue infections. When staphylococcal infections are localized and purulent, antibiotics can be used as adjunctive therapy in conjunction with appropriate surgical intervention. /US Product Label Content/
Vancomycin hydrochloride injection is available orally for the treatment of antibiotic-associated pseudomembranous colitis and staphylococcal enteritis caused by Clostridium difficile. The efficacy of vancomycin hydrochloride injection alone for these indications has not been established. Oral vancomycin hydrochloride is ineffective against other types of infections. /Included in US Product Label/
For more complete data on the therapeutic uses of vancomycin (12 in total), please visit the HSDB record page.

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Drug Warnings
Ototoxicity and nephrotoxicity are the most serious adverse reactions to parenteral vancomycin treatment. The incidence of ototoxicity and nephrotoxicity has not been fully established, but clinical experience to date suggests that these adverse reactions occur at a relatively low rate. Patients with renal impairment, those receiving high doses or prolonged intravenous administration of vancomycin, and those receiving other ototoxic and/or nephrotoxic drugs are most susceptible to ototoxicity and nephrotoxicity. While ototoxicity and nephrotoxicity are associated with serum or blood vancomycin concentrations of 80–100 μg/mL, these reactions can also occur at concentrations as low as 25 μg/mL. The correlation between serum vancomycin concentrations and ototoxicity and nephrotoxicity remains to be elucidated. Ototoxicity can be transient or permanent. Vancomycin may damage the auditory branch of the eighth cranial nerve, leading to permanent deafness. Reports of vertigo, dizziness, and tinnitus are rare. Tinnitus may precede deafness, requiring discontinuation of the drug. Deafness may continue to progress even after discontinuation of vancomycin. Vancomycin-induced nephrotoxicity may manifest as a transient increase in blood urea nitrogen or serum creatinine levels, and the appearance of hyaline casts, granular casts, and albumin in the urine. Fatal uremia has been reported. In rare cases, the drug has been associated with acute interstitial nephritis. Rapid intravenous administration of vancomycin can cause a hypotensive reaction, commonly known as "red man syndrome" or "red neck syndrome." This reaction is characterized by a sudden, potentially severe, drop in blood pressure, accompanied by flushing and/or maculopapular rash or erythema of the face, neck, chest, and upper extremities; these symptoms may occur even without hypotension. Wheezing, dyspnea, angioedema, urticaria, and pruritus may also occur. In rare cases, cardiac arrest or seizures may occur. The hypotension induced by vancomycin appears to be due to its negative inotropic and vasodilatory effects, partly due to histamine release, which is directly related to the infusion rate; histamine release also appears to contribute to the characteristic "red" symptoms (e.g., erythema, rash, pruritus). This reaction usually occurs within minutes of the start of the vancomycin infusion, but may also occur after the infusion is completed, and usually resolves spontaneously within one to several hours after discontinuation of the drug. If the hypotensive reaction is severe, antihistamines, corticosteroids, or intravenous fluids may be necessary. Hypotensive reactions are associated with the infusion rate of vancomycin, and are reported to occur most frequently when the drug is infused over 10 minutes or less; however, hypotensive reactions can occur in rare cases even when the infusion time exceeds 1 hour. To minimize the risk of hypotensive reactions, vancomycin infusion should be performed over at least 1 hour, and the patient's blood pressure should be monitored during the infusion. For patients who have experienced this reaction, subsequent doses of vancomycin usually do not produce adverse reactions if administered slowly (e.g., over several hours). Pre-administration of antihistamines may be helpful. If attempts to mitigate the reaction are ineffective, other anti-infective agents may be necessary. This reaction has been reported in more than 50% of healthy individuals treated with vancomycin, but the incidence is lower when the drug is used for treatment. Patients treated with vancomycin have reported urticaria, exfoliative dermatitis, macules, eosinophilia, vasculitis, shock-like states, transient anaphylactic reactions, and occasional vascular failure. The drug has also been associated with Stevens-Johnson syndrome in at least one patient.
It has been reported that throbbing pain in the back and neck muscles may occur after vancomycin administration; this pain can usually be relieved or avoided by slowing the administration rate. In patients receiving continuous ambulatory peritoneal dialysis (CAPD), intraperitoneal vancomycin injection has been associated with chemical peritonitis, a syndrome characterized by turbid dialysate, which may be accompanied by abdominal pain and fever. Chemical peritonitis usually resolves shortly after discontinuation of intraperitoneal vancomycin. Other adverse reactions to vancomycin include chills and fever. A 37-year-old man with severe underlying diabetes experienced priapism after a second intravenous injection of vancomycin, which recurred after accidental re-administration; the priapism resolved after bilateral corpora cavernosa phlebotomy.
For more complete data on vancomycin (22 in total), please visit the HSDB record page.

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Pharmacodynamics
Vancomycin is a branched-chain tricyclic glycosylated nonribosomal peptide, typically used as a "last line of defense" drug only after failure of other antibiotic treatments. In vitro and clinical infections have demonstrated the effectiveness of vancomycin against most of the following microbial strains: Listeria monocytogenes, Streptococcus pyogenes, Streptococcus pneumoniae (including penicillin-resistant strains), Streptococcus agalactiae, Actinomyces, and Lactobacillus. Combinations of vancomycin with aminoglycoside antibiotics have shown synergistic effects in vitro against a variety of Staphylococcus aureus, Streptococcus bovis, Enterococcus, and viridans streptococci.

C66H75CL2N9O24

1449.25

1447.43

1404-90-6

Vancomycin hydrochloride;1404-93-9

14969

White to off-white solid powder

1.65 g/cm3

4.734

19

26

13

101

2960

18

ClC1C([H])=C2C([H])=C([H])C=1OC1=C([H])C3[C@]([H])(C(N([H])[C@@]4([H])C(N([H])[C@]([H])(C(N([H])[C@]([H])(C(=O)O[H])C5C([H])=C(C([H])=C(C=5C5=C(C([H])=C([H])C4=C5[H])O[H])O[H])O[H])=O)[C@@]([H])(C4C([H])=C([H])C(=C(C=4[H])Cl)OC(C=3[H])=C1O[C@@]1([H])[C@@]([H])([C@]([H])([C@@]([H])([C@@]([H])(C([H])([H])O[H])O1)O[H])O[H])O[C@@]1([H])C([H])([H])[C@@](C([H])([H])[H])([C@@]([H])([C@]([H])(C([H])([H])[H])O1)O[H])N([H])[H])O[H])=O)=O)N([H])C([C@]([H])(C([H])([H])C(N([H])[H])=O)N([H])C([C@@]([H])([C@]2([H])O[H])N([H])C([C@@]([H])(C([H])([H])C([H])(C([H])([H])[H])C([H])([H])[H])N([H])C([H])([H])[H])=O)=O)=O

MYPYJXKWCTUITO-LYRMYLQWSA-N

InChI=1S/C66H75Cl2N9O24/c1-23(2)12-34(71-5)58(88)76-49-51(83)26-7-10-38(32(67)14-26)97-40-16-28-17-41(55(40)101-65-56(54(86)53(85)42(22-78)99-65)100-44-21-66(4,70)57(87)24(3)96-44)98-39-11-8-27(15-33(39)68)52(84)50-63(93)75-48(64(94)95)31-18-29(79)19-37(81)45(31)30-13-25(6-9-36(30)80)46(60(90)77-50)74-61(91)47(28)73-59(89)35(20-43(69)82)72-62(49)92/h6-11,13-19,23-24,34-35,42,44,46-54,56-57,65,71,78-81,83-87H,12,20-22,70H2,1-5H3,(H2,69,82)(H,72,92)(H,73,89)(H,74,91)(H,75,93)(H,76,88)(H,77,90)(H,94,95)/t24-,34+,35-,42+,44-,46+,47+,48-,49+,50-,51+,52+,53+,54-,56+,57+,65-,66-/m0/s1

(1S,2R,18R,19R,22S,25R,28R,40S)-48-[(2S,3R,4S,5S,6R)-3-[(2S,4S,5S,6S)-4-amino-5-hydroxy-4,6-dimethyloxan-2-yl]oxy-4,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-22-(2-amino-2-oxoethyl)-5,15-dichloro-2,18,32,35,37-pentahydroxy-19-[[(2R)-4-methyl-2-(methylamino)pentanoyl]amino]-20,23,26,42,44-pentaoxo-7,13-dioxa-21,24,27,41,43-pentazaoctacyclo[26.14.2.23,6.214,17.18,12.129,33.010,25.034,39]pentaconta-3,5,8(48),9,11,14,16,29(45),30,32,34(39),35,37,46,49-pentadecaene-40-carboxylic acid

Vancocin;Vancoled; Vancomicina; Vancomycine; Vancomycinum; VANCOR

3004209090

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~125 mg/mL (~86.25 mM)

Solubility in Formulation 1: ≥ 4.17 mg/mL (2.88 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 41.7 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 4.17 mg/mL (2.88 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 41.7 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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 (Please use freshly prepared in vivo formulations for optimal results.)