Chem 133L Lab Finals Discussion EXP 10 - 12 PDF
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This document provides a detailed description of laboratory experiments related to digestive processes. It outlines procedures and tests, with a focus on salivary and intestinal digestion. The document includes diagrams and chemical formulas.
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INALS DISCUSSION F XP 10 SALIVARY DIGESTION —-------------------------------------------------- E — ALIMENTARY DIGESTION Ingested food materials are madeassimilable - Transported through intestinal mucosa and absorbed into portal venous...
INALS DISCUSSION F XP 10 SALIVARY DIGESTION —-------------------------------------------------- E — ALIMENTARY DIGESTION Ingested food materials are madeassimilable - Transported through intestinal mucosa and absorbed into portal venous system Series ofenzymatic hydrolysis reactions - Convert Colloidal food molecules to simple, non colloidal solutes — HYDROLYTIC ENZYMES Involved indigestion Naturally occurring foodstuff → assimilable forms Catalyzehydrolysisof - Proteins → amino acids - Starches → monosaccharides - Fats → acyl glycerols — SALIVA Waterysecretion Produced by - 3 pairs of large salivary glands - Many minute glands (buccal glands) _ mucosal lining in the mouth Speeds up chemical changes incarbohydrates No chemical effects on proteins and fats OMPOSITION OF SALIVA C WATER ~99% of saliva ALIVARY AMYLASE S Digestive enzymewhich hydrolyses starch to maltose MINERAL SALTS E.g sodium hydrogen carbonate Maintain pH ~ 6.5 to 7.5 - Optimal for salivary amylase UCIN M Sticky material (glycoprotein) Bindingof food particles Lubricationto assist swallowing H OF SALIVA p pH (pH paper) ~ 8.0 Actual pH = 6.5 to 7.5 - Salivary amylase functions best - Slightlyalkaline, but basically neutral due tosodiumbicarbonate NZYME ACTION E MOUTH AND ELSEWHERE Enzyme action dependent on pH of surroundings E.g pHoptimum, salivary amylase : 6.5 - 7.5 1 CHEM 133L | AY 2024 - 2025 FINALS DISCUSSION TOMACH S Very acidic - pH 1.0 - 3.5 Salivary amylase inactivated - No carb digestion in stomach PROCEDURE 1 - Determine approximate pH of saliva with pH paper 2 - Test for mucin Positive result (+) : precipitate UCIN M Sticky Highlyglycosylated, extended and hydrated protein Extensive insalivary secretions Increasesviscosityof fluids in saliva Bind andlubricatefood particles (assist swallowing) 3 - Test for Protein (Biuret test) Positive result (+) : violet solution Coordinate complex Intensity of color is dependent on number ofpeptidebonds 4 - Inorganic Matter EST FOR CHLORIDES T Positive result (+) :white precipitate Ag+ + Cl– → AgCl (white ppt) EST FOR PHOSPHATES T Positive result (+) :yellow ppt(ammonium molybdate) 2 CHEM 133L | AY 2024 - 2025 FINALS DISCUSSION TEST FOR SULFATES Positive result (+) :white precipitate TEST FOR CALCIUM Positive result (+) :white precipitate I MPORTANCE OF Ca IN SALIVA Maintenance orstabilizationof compact conformationof ∝ - amylase Maintenance of enzymaticallyactive conformation 5 - Test for Thiocyanates (ferric chloride test) Positive result (+):pale yellow with red ppt Confirmation with Mercuric Chloride (HgCl2) NS C Metaboliteof CN– End product ofdetoxification of CN– containing compounds - Enzyme: rhodanese NS IN HUMAN SALIVA C Research: present in saliva sources: - Smoking - Food that containscyanide 6 - Digestion of Starch Paste Starch formsviolet complex with I2 - Causehelical formof amylose and amylopectin Starch –(digestion by saliva) → (smaller saccharide units) Salivary digestion completes if violet colored product is not formed Longer duration means lightening of color of violet-colored complex - ↑ duration ↑ lightening of complex 3 CHEM 133L | AY 2024 - 2025 FINALS DISCUSSION CHROMATIC POINT A Reaction mixture - No more color changewith iodine solution - Complete breakdownof starch - Starch to dextrin (disaccharides, monosaccharides) Starch → dextrins → maltose → glucose ALIVARY AMYLASE S Cleaves alpha-1,4 linkages in starch Givesdifferent colorsin iodine - Amylodextrin:purple - Erythrodextrin:red - Achrodextrin:colorless - Maltose:colorless FEHLING’S TEST Positive result (+) :blue soln with brick red ppt Forreducing sugars BENEDICT’S TEST Positive result (+):green, red, or yellow ppt Forreducing sugars - All sugars except sucrose Aldehydes. Ketones -[O] → alkaline copper solution 7 - Separation Of Salivary Digestion Products HENYLHYDRAZINE REACTION P Salivary digestion of starch → monosaccharides Monosaccharides + phenylhydrazine reagent → Osazone crystals 4 CHEM 133L | AY 2024 - 2025 FINALS DISCUSSION 8 - Influence of Free Acid ↑ concentration of dilute HCl ↑ intensity of blue color Free acidhinders digestionof starch by salivaryamylase 9 - Influence of Alkali ↑ concentration of dilute Na2CO3 ↑ intensity of bluecolor Same trend with dilute base 5 CHEM 133L | AY 2024 - 2025 FINALS DISCUSSION EXP 11 URINALYSIS —----------------------------------------------------------------- Waste products of the differentmetabolic processesin the body are excreted in various ways NOWING THE COMPOSITION OF URINE K Helps in understanding thoroughend productsformed and excretions of urine ROCESSES OF BODY METABOLISM P Provide evidence through presence of abnormalsubstancesor substances in abnormal amounts - Aid in the diagnosis and treatment of pathological states IDNEYS K Responsible for the excretion of most soluble solid waste products Expression of excess water through urine PROCEDURE — PHYSICAL PROPERTIES OF URINE OLOR C NORMAL URINE Yellow to amber Due tourochrome(pigment) - Urobilin & peptide Will darken if set aside undisturbed - Urobilin is released BNORMAL URINE A Varies Nearly colorless to black depending on pathological condition PPEARANCE OF ABNORMAL URINE A Cloudy - Phosphates - Urates - WBCs - Bacteria - Epithelial cells - Fat DOR O NORMAL AND FRESH URINE Slightlyaromatic Ammonia(NH3) odor if set aside AUSES OF ODOR CHANGE IN URINE C Drugs Food pathological conditions PECIFIC GRAVITY S Within 24 hours 1.0.15 to 1.025 1.015 < Excessfluid intake Diabetes insipidus or chronic renal failure 6 CHEM 133L | AY 2024 - 2025 FINALS DISCUSSION 1.025 > Limitedfluid intake Fever Kidney inflammation OLUME EXCRETED PER DAY V 0.5 to 1.5L ↑ large amounts of lipids ↓ loss of water (perspiration) H p 4.5 to 8-2 ACTORS OF pH F Diet Drugs Pathological conditions — ORGANIC COMPONENTS / NITROGENOUS COMPONENTS OF URINE REA U End product of protein metabolism Protein → urea → kidneys → urine BIURET TEST (proteins) Positive result(+): deep blue / purple color Copper (Cu) ion complex with amide RIC ACID U End product ofpurine metabolism(human) High levels in gout patients Amount produced in human urine (~0.5 - 1.0 g/h) MUREXIDE TEST Positive result (+) :bluish violet color Uric acid -[O] → dialuric acid, alloxan Alloxantin reacts with ammonium hydroxide to from purpuric acid - Deposit turns bluish violet die to formation of sodium purporate 7 CHEM 133L | AY 2024 - 2025 FINALS DISCUSSION REATININE C Excreted by normal healthy adults - 1.2 - 1.7 g/h Excreted unchanged in diet For muscular development - Higher levels in men than in women and children NITROPRUSSIDE TEST (WEYL TEST) Positive result (+):red solution For free thiol (--SH), ketones, or ketoacids Light sensitive Contraindicated in patients with renal failure For suspected cystinuria - Blood in the urine - Severe pain the side or back (always on one side) - nausea and vomiting PICRIC ACID Positive result (+):deep orange / red colored solution For reducing sugars IUPAC name: 2,4,6-trinitrophenol (TNP) Yellow crystalline solid One of the most acidic phenols 8 CHEM 133L | AY 2024 - 2025 FINALS DISCUSSION I NDICAN Comes from putrefactive decomposition of tryptophan in the intestine OBERMEYER’S TEST Positive result (+): greenish blue Colorimetric method For intestinal putrefaction — PATHOLOGICAL COMPONENTS OF URINE Dysfunction of liver and kidneys Metabolic disorders LBUMIN (PROTEIN) A Proteinuria - Presence of protein in urine - Indicate kidney damage or nephritis Albuminuria - Presence of albumin in urine Globulins - Heterogeneous group of large serum proteins other than albumin - Sometimes found in urine with albumin (larger molecules) - Indicate more extensive kidney damage BOILING TEST NITRIC ACID TEST (HELLER’S) Positive result (+):white ring at junction of fluids Test for proteins (protein in solution) –HNO3→ protein ppt (ring at junction) 9 CHEM 133L | AY 2024 - 2025 FINALS DISCUSSION LUCOSE G Glycosuria - Sugar in urine Glycosuria - Glucose - Excessive carb intake - Diabetes mellitus BENEDICT’S TEST FOR GLUCOSE Positive result (+):blue to red color Cu2+ –[R]→ Cu2O (cuprous oxide) ≤ 0.1% glucose: precipitate may not appear until cooling BILE ACETONE BODIES CETONE A SODIUM NITROPRUSSIDE TEST (LEGAL’S) Positive result (+): ruby red solution Acetone and b-hydroxybutyric acid --x’ss acetic acid → more intense color ETONE BODIES K BETA - HYDROXYBUTYRIC ACID From incomplete oxidation of fats Accumulation of acetyl CoA and acetoacetyl CoA Indicates acidosis Abnormal amounts excreted (b-hydroxybut. & acetoec. acid) → depletion of Na+ and K+ 10 CHEM 133L | AY 2024 - 2025 FINALS DISCUSSION ENCE-JONES PROTEIN B Monoclonal globulin In the blood or urine Precipitates at low temperature Indicate blood plasma disorder - Multiple myeloma - Waldenstrom’s macroglobulinemia (IgM) - Chronic lymphocytic leukemia (MBCs) - Amyloidosis (protein amyloid) 11 CHEM 133L | AY 2024 - 2025 FINALS DISCUSSION EXP 12 INTESTINAL DIGESTION —------------------------------------------------ combined action ofthree secretions - pancreatic juice - Bile - intestinal juice (succous entericus) IGESTIVE ABILITY D maximal in the2nd stage of the duodenum process continues throughout the small intestine. — PANCREATIC JUICE fluid secreted by thepancreatic gland - Present in the form of inactive zymogens - Changed to active form by action of water or acids or other enzymes Extract from the gland by means of warm water may be acid in the reaction - due to the presence of lactic acid contains four ferments that aid in the digestion of food - Erepsin & Trypsin : decompose the nitrogenous constituents - Lipase: acts upon the fats - Amylopsin : converts starch into maltose Reaction of the juice isalkaline - presence of sodium carbonate (Na2C O3) ANCREATIN P Commercial mixtures of enzymes - Amylase - Lipase - protease Secreted by normal human pancreas Extracted from pigs For problems digesting fats, starches & proteins — SMALL INTESTINE Most important part is theduodenum 3 juices enter the intestine (through duodenum) - Pancreatic juice - Bile - Intestinal juice — LIPASE Family of enzymes for hydrolysis of fats Substrate - Triglycerides - Cholesterol - Phospholipids - Lipid-Soluble vitamins, etc. TO PTIMUM = 37℃ Same as trypsin (functionally disabled) - Denatured at high temperature - Rigid at lower temperatures 12 CHEM 133L | AY 2024 - 2025 FINALS DISCUSSION LIPASE VS WHIPPED CREAM ILE SALT B ↑SAlipid Emulsifying agent for lipids Improves efficiency of hydrolysis At low temperatures, it lowers the activity of lipase TEAPSIN S pancreatic lipase relatively inactive in the form in which it is secreted Activated (non specific)in presence of Bile Salt egg albumin calcium salt calcium soaps — TRYPSIN Hydrolyzes peptide linkages involving Arg and Lys Optimum pH is 7-9 secreted in theinactive form trypsinogen - found in the intestinal mucosa Activated by enteropeptidase (aka enterokinase) REPSIN E aids trypsin in its proteolytic action Hydrolyze the so-called resistant peptide linkage - not acted on by trypsin TRYPSIN VS BAPNA 13 CHEM 133L | AY 2024 - 2025 FINALS DISCUSSION FFECT OF TEMPERATURE(T) ON TRYPSIN E TOPTIMUM, TRYPSIN: 40℃ IGH TEMPERATURE H Trypsin denaturation Results to inactivation Other enzymes and proteins also denature OW TEMPERATURE L Trypsin is rigid and less active Cause decrease in mobility of atoms and molecules PROCEDURE 14 CHEM 133L | AY 2024 - 2025 FINALS DISCUSSION
