Ghostsec420 Huge Returns

GhostSec420 Huge Returns

Hint i gave someone that has been bugging me about my method i use this to get a huge return.
AS FOLLOWS:
Buffers and stock solutions
Cytoskeletal bound proteins extract buffer
10 mM Tris, pH 7.4
100 mM NaCl
1 mM EDTA
1 mM EGTA
1 mM NaF
20 mM Na4P2O7
2 mM Na3VO4
1% Triton X-100
10% glycerol
0.1% SDS
0.5% deoxycholate

Soluble protein buffer
20 mM Tris-HCl, pH 7.5
1 mM EGTA (Ca2+ chelator)
RIPA buffer (RadioImmunoPrecipitation Assay) buffer
RIPA buffer contains the ionic detergent sodium deoxycholate as an active constituent and is particularly used for nuclear
membrane disruption for nuclear extracts. A RIPA buffer gives low background but can denature kinases. It can also disrupt
protein-protein interactions.
50mM Tris HCl pH 8
150 mM NaCl
1% NP-40
0.5% sodium deoxycholate
0.1% SDS
The 10% sodium deoxycholate stock solution (5 g into 50 ml) must be protected from light.
The 100 mM EDTA stock solution is made with 1.86 g into 40 ml H2O and then add NaOH to dissolve and adjust pH to 7.4.
Finally, adjust the total volume to 50 ml). Store the buffer at 4°C.
Nonidet-P40 (NP-40) buffer
20 mM Tris HCl pH 8
137 mM NaCl
10% glycerol
1% nonidet P-40
2 mM EDTA
Sodium orthovanadate preparation
This needs to be done under the fume hood
1. Prepare a 100 mM solution in double distilled water.
2. Set pH to 9.0 with HCl.
3. Boil until colorless.
4. Cool to room temperature.
5. Set pH to 9.0 again.
6. Boil again until colorless.
7. Repeat this cycle until the solution remains at pH 9.0 after boiling and cooling.
8. Bring up to the initial volume with water.
9. Store in aliquots at -20°C.
Note: do not permit great changes in volume during boiling; put a loose lid on the container to protect from evaporation.
Discard if the samples turn yellow.
TBS 10x (concentrated TBS)
24.23 g Trizma HCl
80.06 g NaCl
Mix in 800 ml ultra pure water.
pH to 7.6 with pure HCl.
Top up to 1 L.
TBST
For 1 L: 100 ml of TBS 10x + 900 ml ultra pure water + 1ml Tween20
Medium stripping buffer
Make fresh stripping buffer:
15 g glycine
1 g SDS
10 ml Tween20
Set the pH to 2.2
Make up to 1 L with ultrapure water
Harsh stripping buffer
To be done under the fumehood

For 100 ml:
20 ml SDS 10%
12.5 ml Tris HCl pH 6.8 0.5M
67.5 ml ultra pure water
Add 0.8ml ß-mercaptoethanol under the fumehood.
Nuclear fractionation protocol reagents
Buffer A – 10 mM HEPES, 1.5 mM MgCl2, 10 mM KCl, 0.5 mM DTT, 0.05% NP40 (or 0.05% Igepal or Tergitol) pH 7.9
To prepare 250 ml stock of buffer A –
HEPES: 1M = 238.3 g/L, therefore 10 mM = 0.59 g/250 ml
MgCl2: 1M = 203.3 g/L, therefore 1.5 mM = 0.076 g/250 ml
KCl: 1M = 74.5 g/L, therefore 10 mM = 0.187 g/250 ml
DTT: 1M = 154.2 g/L, therefore 0.5 mM = 0.019 g/250 ml
NP40 = 0.05%
Buffer B – 5 mM HEPES, 1.5 mM MgCl2, 0.2 mM EDTA, 0.5 mM DTT, 26% glycerol (v/v), pH 7.9
(2)
Extraction without chemical change.
Some solutes such as noble gases can be extracted from one phase to another without the need for a chemical reaction (see absorption). This is the simplest type of solvent extraction. When a solvent is extracted, two immiscible liquids are shaken together. The more polar solutes dissolve preferentially in the more polar solvent, and the less polar solutes in the less polar solvent. Some solutes that do not at first sight appear to undergo a reaction during the extraction process do not have distribution ratio that is independent of concentration. A classic example is the extraction of carboxylic acids (HA) into nonpolar media such as benzene. Here, it is often the case that the carboxylic acid will form a dimer in the organic layer so the distribution ratio will change as a function of the acid concentration (measured in either phase).
Chrystian Iezid MAIA-ALMEIDA FERES · Universidade Federal de Tocantins (UFT)
Hello, the others work colleague presented the methods and solvents that can really be hexane and methanol extraction solvent, hence you will have to make the partition lighter and injected in the GCMS and the heavier phenols such as HPLC or LCMS, through the balance polarities.
in addition to sending information below.
For essential oil extraction is recommended you do first distillation using water to drag the oil that can be micro-Clevenger, with a capacity of 1 g of dried vegetable matter. Collecting the oil in vials and have specific weight of the empty bottle determined scale of 4 digits and by means of the difference output. Store this oil in bottles with spigot and lid. Coat the flasks with aluminum foil and store refrigerated. For analysis by GC / MS
You can use the methods below:
The analysis: Carried Out in the GC / MS - Shimadzu/QP-5000, equipped with DB-5 column (30 m X 0.25 mm X 0.25 mM) containing the Helium carrier gas at 1.0 ml / min flow, injector at 240 ˚ C , and detector at 250 ˚ C. The Following program was used: 50 ˚ C (5 ') - until 160 ˚ C, 3 ˚ C per minute, and 160-220 ˚ C 10 ˚ C per minute, 1/35 split. The 2mg-oil sample was diluted in 1 ml ethyl acetate, from Which the-1μL aliquot was injected. To identify the Substances, Nist. Library 62 Were Compared with Those results in literature (Mclafferty et al., 1989). Kovats index retention was determined by Comparing with the date Those found in the literature (Adams, 1995). Analyses Were Performed in triplicate.
Determine the yield compared with the weight of plant material
Then, if you want to get phenolics from the same sample you must dry the samples up to 45 º C, remember to keep the content of the liquid hidrodestilador along with the plant material to dry. To maintain the integrity of some compounds.
And after drying it is possible to fine grinding of the material, add methanol and put under stirring for 2-4 hours, tests are required for adjustment.
The ratio solvent: plant material may be 1 g of dry matter vegetable to 25 ml of methanol. Consider the use of shaker table may make the most efficient extraction.
After the extraction period you can filter the solution on filter paper, an analytical standard. Collect the filtrate in 25 ml amber glass, with stopper and cap pre identified and weighed on an analytical balance to four places after the comma beyond identification with treatments etc ...
Take the rotary evaporator with adjustable pressure and temperature of the bath. For methanol the temperature is around 40 º C. Varying with applied pressure. Adjust the rotation. Note the process and avoid refluxing the glass sample into the coil rotary evaporator.
You also need a cooler of water with anti freeze solution for use in capacitors, both the extraction of oil and solvent recovery.
After removal of the solvent extract you will get almost dry. To dry it put them in the drying chamber with silica gel activated
for exemple:
For the quantification of phyllanthin(lignana) was employed for high efficiency liquid chromatography (Tecnhologies Agilent 1100 series), consisting of quaternary pump, autosampler and UV absorption detector / visible column chromatography and Inertsil ODS-3 (250nm ID x 4.6, GL Sciences Inc. Tokyo, Japan). The chromatographic conditions were: acetonitrile mobile phase, water (55:45 v / v), isocratic elution, flow 1mL/min at room temperature (25øC) and a wavelength of 230 nm in pre-defined tests and supported by Results & Chan Murugaiyah (2007). The sample injection volume was 20 ìl. The mean retention time of 12.9 minutes was phyllanthin.
(3)
Pretty much any partially polar or nonpolar solvent can be used to extract hash oil, some are better than others.
So here goes:
Butane
Pros: It makes wonderful oil and it's very easy to remove every last trace of it, especially if you have a way to purge it with a vacuum (which I recommend for any solvent extract). Ready to smoke the fastest of any method listed here.
Cons: It's the most flammable thing on this list, considering ether is also on this list that's saying A LOT! Because of that it needs the most ventilation. As in outside and away from people unless you happen to have a fume hood in your house. Some of us don't have that kind of privacy. It also requires more specialized equipment, specifically a stainless steel or glass tube that you can inject butane into, which also limits the batch size. Do not use a PVC or other plastic for the tube. You can pull plasticizers out of in and into your oil!
Ether
Pros: Makes wonderful oil, also evaporates quickly (it boils at body temp!). Is just a generally extremely useful solvent.
Cons: needs to be stabilized or distilled fresh or it forms explosive peroxides with the oxygen in the air in the bottle. Requires glass or steel. It loves to catch fire. It will autoignite (no spark needed) at 160C. Hope your oven isn't on. It's almost as flammable as butane, and the only liquid on this list that gets a flammability rating of 4 on the safety diamond. Skip it if you don't have extensive lab experience.
Isopropanol
Pros: The most readily available and cheapest on this list. If a very fast wash is done it makes excellent oil, but ultimately the yield is limited by the fact that it starts pulling out chlorophyll and sugars if it stays in contact with the weed for any real period of time.
Cons: Pulls out things besides the resin, is pretty flammable despite the small amount of water (don't even bother with the 70%, it must be 91%+) due to it being a branched hydrocarbon and having an oxygen atom. Low yield because you can really only do a quick rinse or you get poor taste if any chlorophyll or sugars gets into the oil from soaking too long.
Ethanol
Pros: Less flammable than IPA. YOU CAN DRINK IT! Really I wouldn't do this to make oil but green dragon is awesome. I decarb bud before extracting to make a more potent product.
Cons: if you are making oil it's a waste of ethanol which could cause ancestral haunting if you're Irish like me smile emoticonProbably other hard drinking groups like Russian or Polish as well.
Acetone
Pros: I can't really think of any
Cons: Doesn't fully evaporate!! Doesn't fully evaporate!! Also pulls out chlorophyll and sugars like IPA. Also extremely flammable. Skip it!
Pentane
Pros: Pulls great oil of equivalent quality to butane. Evaporates very fast (it also boils at body temp, hold the flask in your hands when evaporating and watch it start to boil!). Can be used for a large batch if you have a way to minimize evaporation.
Cons: needs really good storage or you risk opening an empty bottle. Requires a glass or steel vessel to extract and evaporate in.
Hexanes
refers to n-hexane, isohexanes, or a mixture of isomers
Pros: basically the same as the other hydrocarbons as far as quality (I can't tell the difference), a little better for bulk extractions as it evaporates slower than pentane, I still wouldn't leave it that long. An hour at most.
Cons: needs to be done in glass or steel (like everything but the alcohols)
n-Heptane
Pros: that great alkane oil! Can sit for a few hours to really soak everything out of the weed. I'd give it a period stir to check on it and maximize extraction. Because n-heptane is produced by a species of pine tree it is available in very high purity for pretty cheap.
Cons: Glass or steel required, flammable. Really about as minimal as you can get for an organic solvent.
Naphtha
Pros: Rick Simpson seems to like it. Gives an ok extract.
Cons: Steel/Glass required (I wouldn't suggest a rice cooker, most have plastic parts that will come in contact with the solvent and probably end up in your oil), is a mix of light alkanes from crude oil so it's inconsistent batch to batch. Sometimes has traces of heavy material from production (bad distillation) or just added in because it drives production costs down and still retains the correct properties required to be a naphtha cut.
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Cannabinoid Boiling Points:

Δ-9-tetrahydrocannabinol (THC)
Boiling point: 157*C / 314.6 degree Fahrenheit
Properties: Euphoriant, Analgesic, Antiinflammatory, Antioxidant, Antiemetic Antibiotic Anticancer.

Cannabidiol (CBD)
Boiling point: 160-180*C / 320-356 degree Fahrenheit
Properties: Anxiolytic, Analgesic, Antipsychotic, Antiinflammatory, Antioxidant, Antispasmodic.
Cannabinol (CBN)
Boiling point: 185*C / 365 degree Fahrenheit
Properties: Oxidation, breakdown, product, Sedative, Antibiotic.
cannabichromene (CBC)
Boiling point: 220*C / 428 degree Fahrenheit
Properties: Antiinflammatory, Antibiotic, Antifungal.

cannabigerol (CBG)
Boiling point: MP52
Properties: Antiinflammatory, Antibiotic, Antifungal.

Δ-8-tetrahydrocannabinol (?-8-THC)
Boiling point: 175-178*C / 347-352.4 degree Fahrenheit
Properties: Resembles ?-9-THC, Less psychoactive, More stable Antiemetic.

tetrahydrocannabivarin (THCV)
Boiling point: < 220*C / <_428 degree="degree" fahrenheitbr="fahrenheitbr" _="_">Properties: Analgesic, Euphoriant
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Terpenoid essential oils, their boiling points, and properties:

ß-myrcene
Boiling point: 166-168*C / 330.8-334.4 degree Fahrenheit
Properties: Analgesic. Antiinflammatory, Antibiotic, Antimutagenic.

ß-caryophyllene
Boiling point: 119*C / 246.2 degree Fahrenheit
Properties: Antiinflammatory, Cytoprotective (gastric mucosa), Antimalarial.
d-limonene
Boiling point: 177*C / 350.6 degree Fahrenheit
Properties: Cannabinoid agonist?, Immune potentiator, Antidepressant, Antimutagenic.

linalool
Boiling point: 198*C / 388.4 degree Fahrenheit
Properties: Sedative, Antidepressant, Anxiolytic, Immune potentiator.

pulegone
Boiling point: 224*C / 435.2 degree Fahrenheit
Properties: Memory booster?, AChE inhibitor, Sedative, Antipyretic.

1,8-cineole (eucalyptol)
Boiling point: 176*C / 348.8 degree Fahrenheit
Properties: AChE inhibitor, Increases cerebral, blood flow, Stimulant, Antibiotic, Antiviral, Antiinflammatory, Antinociceptive.

a-pinene
Boiling point: 156*C / 312.8 degree Fahrenheit
Properties: Antiinflammatory, Bronchodilator, Stimulant, Antibiotic, Antineoplastic, AChE inhibitor.

a-terpineol
Boiling point: 217-218*C / 422.6-424.4 degree Fahrenheit
Properties: Sedative, Antibiotic, AChE inhibitor, Antioxidant, Antimalarial.

terpineol-4-ol
Boiling point: 209*C / 408.2 degree Fahrenheit
Properties: AChE inhibitor. Antibiotic.

p-cymene
Boiling point: 177*C / 350.6 degree Fahrenheit
Properties: Antibiotic, Anticandidal, AChE inhibitor.
(6)
Flavonoid and phytosterol components, their boiling points, and properties:

apigenin
Boiling point: 178*C / 352.4 degree Fahrenheit
Properties: Anxiolytic, Antiinflammatory, Estrogenic

quercetin
Boiling point: 250*C / 482 degree Fahrenheit
Properties: Antioxidant, Antimutagenic, Antiviral, Antineoplastic

cannflavin A
Boiling point: 182*C / 359.6 degree Fahrenheit
Properties: COX inhibitor, LO inhibitor

ß-sitosterol
Boiling point: 134*C / 273.2 degree Fahrenheit
Properties: Antiinflammatory, 5-a-reductase, inhibitor

Please Remember that the "lesser known" Cannabinoids as well as the "other" compounds found in Cannabis have an entourage effect that causes them to work symbiotically together.