residual moisture raises a handful of sensible questions. This page answers them in order, starting with the fundamentals and moving to applications.
Reviewed 2025-10-30. Anything still debated is marked as such rather than presented as settled.
The physical chemistry of freezing influences whether a formulation forms an amorphous glass or a crystalline solid. Amorphous systems can collapse if product temperature rises above the glass transition temperature of the freeze concentrate. Crystalline systems may show eutectic melting, where ice and solute melt together at a fixed temperature. Formulators add bulking agents, lyoprotectants, and buffers to preserve structure and biological activity. The optimum cycle keeps product temperature below critical thresholds during primary drying while allowing efficient sublimation.
Lyophilization is a dehydration process that removes water from a frozen material by sublimation under low pressure. The material is first frozen so that water becomes ice; then the surrounding pressure is lowered below the vapor pressure of ice. Heat is applied gently so ice changes directly to vapor without passing through a bulk liquid phase. The vapor is collected on a cold condenser, leaving a dry porous matrix. This process differs from simple evaporation because the material remains frozen during the main drying stage.
The process usually has three stages: freezing, primary drying, and secondary drying. Freezing sets the ice crystal structure and can determine pore size in the final cake. Primary drying removes free ice by sublimation at low pressure and low shelf temperature. Secondary drying removes bound water by raising the shelf temperature, although some water may remain as part of the solid. Cycle parameters depend on formulation, fill volume, vial type, and equipment performance.
Quality control for freeze-dried forms includes visual inspection, water content measurement, and reconstitution time. A satisfactory cake is typically uniform, porous, and intact, although minor shrinkage or cracking may be acceptable if specifications allow. Karl Fischer titration, thermal gravimetric analysis, and near-infrared spectroscopy are used to measure water content. Reconstitution is assessed by adding a specified diluent and recording the time and ease of dissolution. Microbiological and particulate tests are added when the product is sterile or intended for injection.
Stability of a lyophilized solid depends on water content, temperature, and the physical state of the formulation. Amorphous products may slowly absorb moisture and drop below their glass transition temperature, causing collapse or crystallization. Some proteins and peptides can aggregate even in a dry state, especially when exposed to heat or moisture. Accelerated stability studies at elevated temperature and humidity help estimate shelf life, but real-time data remain the basis for expiration dating.
After drying, a lyophilized product is usually sealed under vacuum or an inert gas to limit moisture and oxygen exposure. The container closure system matters because stoppers and seals can allow moisture ingress over time. Storage conditions are selected from stability studies that track potency, cake appearance, and reconstitution behavior. Many freeze-dried materials are kept at controlled room temperature, while some require refrigeration or protection from light.
| Property | Value | Notes |
|---|---|---|
| Common synonyms | Freeze-drying, lyophilisation | Lyophilisation is the British spelling; the process is not simple evaporation. |
| Primary drying pressure | 0.05–0.3 mbar | Pressure must remain below the vapor pressure of ice at the product temperature. |
| Sublimation temperature | Below 0 °C | Ice changes directly to vapor while the product remains frozen. |
| Typical shelf temperature | −40 to −10 °C | Exact setting depends on formulation critical temperature and equipment. |
| Cycle duration | 12–72 hours | Time varies with fill volume, formulation, and dryer performance. |
A formulation often contains excipients that protect the active ingredient during freezing and drying. Bulking agents provide structure, while lyoprotectants stabilize sensitive molecules. The freezing step can produce ice crystals whose size and distribution affect the drying rate, and cycle design includes freezing, annealing, and drying phases. If the product temperature rises above a critical value, the cake may collapse or lose its porous structure. Successful lyophilization therefore depends on the interaction between formulation, equipment, and cycle design.
Lyophilization is a dehydration process that removes water from a frozen material by sublimation under low pressure. The material is first frozen to convert liquid water into ice. Next, the pressure is reduced below the triple point of water so that ice changes directly into vapor without passing through a liquid phase. This step is called primary drying. The result is a porous solid or cake that retains the original shape of the frozen solution.
Handling and storage practices aim to keep the cake intact and dry. Vials are typically stored upright at controlled temperatures, often between 2 °C and 8 °C or at -20 °C for longer-term use. Reconstitution involves adding a suitable diluent and gently mixing until the solid dissolves. Shaking or rapid injection of diluent can create foam or damage sensitive molecules. Once reconstituted, the product may require refrigeration and use within a defined period.
Residual moisture is a key quality attribute for lyophilized products. Water that remains after secondary drying can affect chemical stability, cake structure, and shelf life. Karl Fischer titration is a common method for measuring water content in the dried solid. The target range varies by product, but many biologics are dried to between 0.5% and 3% water by weight. Acceptable limits are set during development and confirmed by stability studies.
A typical cycle begins with freezing, which fixes the material into a solid and determines ice crystal size. Primary drying then raises heat under vacuum so ice sublimes, often near or below the collapse temperature of the formulation. Secondary drying removes bound water that remains after ice is gone, usually by gently warming the product. Each stage balances heat input against pressure to avoid melting or structural damage. Temperature probes and pressure sensors guide the transition between stages.
In practice, lyophilization is slower and more energy intensive than simple drying. Cycle times can range from hours to several days depending on load, container, and formulation. Amorphous materials may require excipients that help preserve structure during freezing and drying. The method is widely used for biological materials, pharmaceuticals, and foods where heat drying would cause unacceptable change. Open questions remain about scaling cycles between laboratory and production equipment, and this gap affects technology transfer.
Lyophilization removes water by freezing a material and then lowering pressure so ice changes directly to vapor. The process relies on sublimation, the phase transition from solid to gas without an intermediate liquid state. Because the material remains frozen during primary drying, the structure often stays porous. This porous matrix can rehydrate quickly when water is added back. The low pressure also allows vapor to leave the solid matrix without boiling.
After lyophilization, a product's quality depends on residual moisture, cake appearance, and reconstitution time. Residual moisture is often measured by Karl Fischer titration or thermogravimetric analysis. A low moisture content can slow chemical degradation, but overly dry cakes may be brittle or slow to dissolve. Stability studies track these attributes over months under defined temperature and humidity conditions. Batch records link these measurements to specific process runs and help identify trends before a product fails specification.
Storage conditions for dried products usually aim to exclude moisture and oxygen. Vials are sealed under vacuum or with an inert gas, and stoppers must maintain a barrier during transport. Temperature recommendations vary; some materials remain stable at room temperature, while others need refrigeration or frozen storage. Humidity control is critical because dried cakes can absorb water rapidly once a container is opened. Desiccant packs and moisture-barrier bags add further protection during shipping.
Upon learning of Milea's death, Ceaușescu appointed Victor Stănculescu minister of defence. He accepted after a brief hesitation. Stănculescu, however, ordered the troops back to their quarters without Ceaușescu's knowledge, and also persuaded Ceaușescu to leave by helicopter, thus making the dictator a fugitive. At that same moment angry protesters began storming the Communist Party headquarters; Stănculescu and the soldiers under his command did not oppose them. By refusing to carry out Ceaușescu's orders (he was still technically commander-in-chief of the army), Stănculescu played a central role in the overthrow of the dictatorship. "I had the prospect of two execution squads: Ceaușescu's and the revolutionary one!" confessed Stănculescu later. In the afternoon, Stănculescu "chose" Ion Iliescu's political group from among others that were striving for power in the aftermath of the recent events.
In December 2010, Chipotle hired chef Nate Appleman to develop new cuisine. Appleman has won Rising Star Chef from the James Beard Foundation, was named "Best New Chef" by the Food & Wine magazine, and competed on The Next Iron Chef. In 2010, U.S. Immigration and Customs Enforcement (ICE) audited Chipotle's Minneapolis restaurants, and found that some employees had been hired using fraudulent documents. In December, Chipotle fired 450 employees from its Minneapolis restaurants as a result of the audit, resulting in protests by local groups. In February 2011, ICE expanded the audit to include 60 restaurants in Virginia and Washington, D.C. which resulted in 40 workers being fired. In April 2011, the criminal division of the attorney general's office in Washington, D.C., joined the case, and ICE agents began interviewing employees at 20 to 25 restaurants in other locations, such as Los Angeles and Atlanta. In response to the government investigations, Chipotle hired former director of ICE Julie Myers Wood and high-profile attorneys Robert Luskin and Gregory B. Craig. In April 2014, Chipotle announced an increase in menu prices for the first time in nearly three years, due to increasing costs for steak, avocados, and cheese. The price increase was expected to be rolled out from the end of the second quarter of 2014 through the end of the third quarter. In late 2015, Chipotle expanded its mobile strategy through delivery partnerships with tech startups like Tapingo, a delivery service that targets college campuses.
== Efficacy == The COVID-19 mRNA vaccines from Moderna and Pfizer–BioNTech had short-term efficacy rates of over 90 percent against the original SARS-CoV-2 virus. Prior to mRNA, drug trials on pathogens other than COVID-19 were not effective and had to be abandoned in the early phases of trials. The reason for the efficacy of the new mRNA vaccines is not clear. Physician-scientist Margaret Liu stated that the efficacy of the new COVID-19 mRNA vaccines could be due to the "sheer volume of resources" that went into development, or that the vaccines might be "triggering a nonspecific inflammatory response to the mRNA that could be heightening its specific immune response, given that the modified nucleoside technique reduced inflammation but hasn't eliminated it completely", and that "this may also explain the intense reactions such as aches and fevers reported in some recipients of the mRNA SARS-CoV-2 vaccines". These reactions though severe were transient and another view is that they were believed to be a reaction to the lipid drug delivery molecules. In June 2021, the US Food and Drug Administration added a warning about the possibility of increased risk of myocarditis and pericarditis for some people.
=== Degradation of DNA during bisulfite treatment === A major challenge in bisulfite sequencing is the degradation of DNA that takes place concurrently with the conversion. The conditions necessary for complete conversion, such as long incubation times, elevated temperature, and high bisulfite concentration, can lead to the degradation of about 90% of the incubated DNA. Given that the starting amount of DNA is often limited, such extensive degradation can be problematic. The degradation occurs as depurinations resulting in random strand breaks. Therefore, the longer the desired PCR amplicon, the more limited the number of intact template molecules will likely be. This could lead to the failure of the PCR amplification, or the loss of quantitatively accurate information on methylation levels resulting from the limited sampling of template molecules. Thus, it is important to assess the amount of DNA degradation resulting from the reaction conditions employed, and consider how this will affect the desired amplicon. Techniques can also be used to minimize DNA degradation, such as cycling the incubation temperature. In 2020, New England Biolabs developed NEBNext Enzymatic Methyl-seq, an alternative enzymatic approach to minimize DNA damage. Instead of bisulfite, APOBEC is used to convert C into U. Distinction between C, 5mC, and 5hmC is granted by the further modifications that "protect" the modified bases from APOBEC.
Sources: en.wikipedia.org
==== Rebuilding links with the West ==== In 1999, Libya began secret talks with the British government to normalize relations. In 2001, Gaddafi publicly condemned the September 11 attacks on the US by al-Qaeda, expressing sympathy with the victims and calling for Libyan involvement in the US-led war on terror against militant Islamism. His government continued suppressing domestic Islamism, at the same time as Gaddafi called for the wider application of sharia law. Libya also cemented connections with China and North Korea, being visited by Chinese President Jiang Zemin in April 2002. However, relations with China became strained in May 2006 due to a visit to Tripoli by Taiwanese President Chen Shui-bian. Influenced by the events of the Iraq War, in December 2003, Libya renounced its possession of weapons of mass destruction, decommissioning its chemical and nuclear weapons programs. Relations with the US improved as a result. British Prime Minister Tony Blair visited Gaddafi in March 2004; the pair developed close personal ties. In 2003, Libya paid US$2.7 billion to the families of the victims of the Lockerbie bombing as it was the condition the US and UK had made for terminating the remaining UN sanctions. Libya continued to deny any role in the bombing. In 2009, Gaddafi attempted to strong-arm global energy companies operating in Libya to cover Libya's settlement with the families of the victims of Lockerbie.
=== Protists === A common protist eukaryotic expression system is the slime mold, Dictyostelium discoideum, and is unique as it has a circular plasmid, packaged similarly to chromatin. As a simple eukaryotic haploid organism, it can grow in high concentrations without the expensive conditions of mammalian cell culture, and perform post-translational modifications. The protein itself is expressed in several forms including as membrane attached, secreted, or cell associated, and can glycosylate protein product.
== Treatment == The primary treatment of PPID is pergolide, a dopamine agonist that provides suppression to the pars intermedia in place of the dysfunctional hypothalamus. Horses should be reassessed in 30 days following the start of treatment, through evaluation of clinical signs and by baseline diagnostic testing, to ensure the appropriate dose is being prescribed. Results from that test dictate changes in dose. Horses that are responding to treatment should be retested every 6 months, including a test in the autumn when a seasonal increase in ACTH occurs, to ensure their ACTH levels are appropriately suppressed during this time. Drug side effects include a transient decrease in appetite, typically seen when first introducing the medication or increasing the dose, which can be reduced by slowly increasing the dose to therapeutic levels, and by breaking up the daily dose into twice-daily administrations. Attitude, activity levels, hyperglycemia, and increased drinking and urination are usually improved within 30 days of initiating treatment. Other clinical signs, such as hirsutism, potbellied appearance, muscle wasting, laminitic episodes, and increased predisposition to infection, usually take between 30 days and a year to improve. Cyproheptadine may be added to the treatment regimen in horses that are inadequately responding to pergolide, but is usually only used in horses with advanced PPID on high doses of pergolide.
==== Cell-based scheme of coagulation ==== A newer model of coagulation mechanism explains the intricate combination of cellular and biochemical events that occur during the coagulation process in vivo. Along with the procoagulant and anticoagulant plasma proteins, normal physiologic coagulation requires the presence of two cell types for formation of coagulation complexes: cells that express tissue factor (usually extravascular) and platelets. The coagulation process occurs in two phases. First is the initiation phase, which occurs in tissue-factor-expressing cells. This is followed by the propagation phase, which occurs on activated platelets. The initiation phase, mediated by the tissue factor exposure, proceeds via the classic extrinsic pathway and contributes to about 5% of thrombin production. The amplified production of thrombin occurs via the classic intrinsic pathway in the propagation phase; about 95% of thrombin generated will be during this second phase.
== Classification == Cerebrospinal fluid leaks are classified into two distinct disorders: cranial leaks, and spinal leaks. Where there has been no preceding surgery or medical procedure that may have caused a CSF leak it is called a spontaneous CSF leak (sCSF leak).
Sources: en.wikipedia.org
== See also == Control of water pollution – Contamination of water bodies Clean Water Act – 1972 U.S. federal law regulating water pollution Peak water – Concept on the quality and availability of freshwater resources Pulsed-power water treatment – Using electro-magnetic fields on cooling water Solar water disinfection – Portable water purification powered by sunlight Raw water#Treatment – Untreated water found in a natural environment Water purification – Process of removing impurities from water Water quality – Assessment against standards for use Water softening – Removing positive ions from hard water Water supply – Provision of water by public utilities, commercial organisations or others
== Canada, United States of America, Europe == Abdul Rahman Ibrahima Sori (c. 1762 – 1829) – Son of Ibrahim Sori Mawdo of Futa-Jallon. Enslaved in Natchez, Mississippi but freed and repatriated to Liberia. Alhaji Mohammed Ayuba Suleiman Diallo (also known as Job ben Solomon) – Trader, then slave. Freed and repatriated to his homeland in Boundou, Senegal Bill Hamid Diaryatou Bah - French female activist. Moussa Diallo Elhadj - Senator, Belgian Senate. Philippe Diallo - President of French Football Federation Nafissatou Thiam - Three time heptathlon Olympic Gold Medalist, Hepthathlon World Record Holder. Satou Sabally - German-American Female Basketballer, Three time WNBA All star, WNBA Most Improved Player (2023), All-WNBA First Team (2023). Nyara Sabally - German Female Basketballer, WNBA champion 2024. Abou Diaby - former Arsenal and French National team Player. Romane Dicko - Two time single judo olympics bronze medalist, two time mixed olympics Gold medalist. Ousmane Dembélé - french national team player, Ballon D'or 2025 Elladj Baldé - Canadian figure skater. He won the 2015 Nebelhorn Trophy, an ISU Challenger Series event. He is the 2008 Canadian Junior champion. Boris Diaw - French basketball player Alexander Bah - Danish football player Djibril Sidibé (footballer, born 1992) - French World cup winner.
=== Therapy assistants === Physical therapy assistant (PTA) Associate of Applied Science in Physical Therapist Assistant Occupational therapy assistant Associate of Applied Science in Occupational Therapy Assistant(AAS OTA) (COTA)
=== Bioorthogonal reactions === Successful labeling of a molecule of interest requires specific functionalization of that molecule to react chemospecifically with an optical probe. For a labeling experiment to be considered robust, that functionalization must minimally perturb the system. Unfortunately, these requirements are often hard to meet. Many of the reactions normally available to organic chemists in the laboratory are unavailable in living systems. Water- and redox- sensitive reactions would not proceed, reagents prone to nucleophilic attack would offer no chemospecificity, and any reactions with large kinetic barriers would not find enough energy in the relatively low-heat environment of a living cell. Thus, chemists have recently developed a panel of bioorthogonal chemistry that proceed chemospecifically, despite the milieu of distracting reactive materials in vivo. The coupling of a probe to a molecule of interest must occur within a reasonably short time frame; therefore, the kinetics of the coupling reaction should be highly favorable. Click chemistry is well suited to fill this niche, since click reactions are rapid, spontaneous, selective, and high-yielding. Unfortunately, the most famous "click reaction," a [3+2] cycloaddition between an azide and an acyclic alkyne, is copper-catalyzed, posing a serious problem for use in vivo due to copper's toxicity. To bypass the necessity for a catalyst, Carolyn R. Bertozzi's lab introduced inherent strain into the alkyne species by using a cyclic alkyne.
Diazepam is a long-acting "classical" benzodiazepine. Other classical benzodiazepines include chlordiazepoxide, clonazepam, lorazepam, oxazepam, nitrazepam, temazepam, flurazepam, bromazepam, and clorazepate. Diazepam has anticonvulsant properties. Benzodiazepines act via micromolar benzodiazepine binding sites as calcium channel blockers and significantly inhibit depolarization-sensitive calcium uptake in rat nerve cell preparations. Diazepam inhibits acetylcholine release in mouse hippocampal synaptosomes. This has been found by measuring sodium-dependent high-affinity choline uptake in mouse brain cells in vitro, after pretreatment of the mice with diazepam in vivo. This may play a role in explaining diazepam's anticonvulsant properties. Diazepam binds with high affinity to glial cells in animal cell cultures. Diazepam at high doses has been found to decrease histamine turnover in mouse brain via diazepam's action at the benzodiazepine-GABA receptor complex. Diazepam also decreases prolactin release in rats.
Sources: en.wikipedia.org
The main change is sublimation, in which ice becomes water vapor without melting into liquid water. This occurs when the chamber pressure is held below the vapor pressure of ice while mild heat is supplied. The result is a dry, porous solid that retains much of its original shape.
Freezing determines ice crystal size, solute distribution, and the pore network left after drying. A slow or fast freezing rate can produce different cake structures and affect reconstitution. It also sets whether the formulation follows an amorphous or crystalline drying path.
It removes most free water during primary drying and part of the bound water during secondary drying. A small residual moisture content often remains and is specified for each product. Complete removal is generally neither practical nor desirable for stability.
Sealed vials or containers should be kept at the temperature specified by stability data, often controlled room temperature or 2–8 °C. Moisture and oxygen barriers are important because both can degrade sensitive materials. Opened containers may need immediate use or protection from ambient humidity.