Module 2: Biochemistry and Pharmacognosy PDF

REVIEW ON PROFESSIONAL SUBJECTS 1 MODULE 2: BIOCHEMISTRY & PHARMACOGNOSY Ribosome – actual site of protein synthesis BIOCHEMISTRY (70s = bacteria; 80s = human) Biochemistry...

REVIEW ON PROFESSIONAL SUBJECTS 1 MODULE 2: BIOCHEMISTRY & PHARMACOGNOSY Ribosome – actual site of protein synthesis BIOCHEMISTRY (70s = bacteria; 80s = human) Biochemistry Endoplasmic reticulum ▪ Science that deals with the chemical Rough ER – other site of protein synthesis basis of life Smooth ER – site of lipid synthesis ▪ Biomolecular study Lysosomes – suicide sac; has hydrolytic Monomer/Building Blocks enzymes that digest foreign substances. Carbohydrates Monosaccharide Peroxisomes – act as detoxifier of reactive Lipids Fatty acids oxygen species or free radicals; responsible for Proteins Amino acids oxidation of uric acid; breaks Very long chain Nucleic acids Nucleotides fatty acids. The Cell and its parts Golgi apparatus – modifies protein and lipids ▪ Structural and functional unit of all living Organelles (‘little organs’) things Organelle Function ▪ Basic unit of life Ribosomes Actual site of CHON Cell Membrane synthesis (contains phospholipid; hydrophilic polar head, Endoplasmic reticulum CHON synthesis (rough) hydrophobic non-polar tail – therefore they are Lipid synthesis (smooth) amphiphilic) Golgi apparatus Modifies CHON & lipids ▪ Fluid Mosaic Model Lysosomes Digest foreign substances Properties Peroxisomes Detoxifier of free radicals 1. Semi-permeable (not all can pass Mitochondria Energy production through) Centrosome Microtubule organizing 2. Amphiphilic (amphiprotic) center (MTOC) of animal (cell cycle regulation) ▪ Board Exam cell, cell cycle regulation Integral protein – embedded Nucleus within the cell membrane; this could serve as gateway where ions can go ▪ Control center of the cell through this type of protein ▪ Where DNA synthesis is located Peripheral protein – embedded Nucleolus – site of ribosome assembly outside the cell membrane. Act as CARBOHYDRATES receptor 1. Aka: Polyhydroxyaldehyde and Polyhydroxyketone 2. Building block: monosaccharides 3. Functional groups: a) Aldehyde b) Ketone c) Alcohol Reactions: 1. Synthesis: Dehydration (remove Cytoplasm water) thus form a glycosidic ▪ Cell contents outside the nucleus bond/linkage similar to ether linkage ▪ Cytosol = Fluid part of the cell 2. Degradation: Hydrolysis ▪ Organelles = small organs that have its Classification according to the number own function of monomer units Unit Type/Class Mitochondrion – powerhouse of the cell; in charge of energy production Monosaccharides Always 1 Always 1 Disaccharides Always 2 Up to 2 PORLANTE, Jefferson “Tino” Aguado REVIEW ON PROFESSIONAL SUBJECTS 1 MODULE 2: BIOCHEMISTRY & PHARMACOGNOSY Oligosaccharides 3 – 10 1 or more Polysaccharides > 10 1 or more Glu + Glu + Glu = 3 units/ 1 class Glu + Fru + Gal = 3 units/ 3 class Forms of Isomerism ▪ Enantiomers (mirror images and non- superimposable) D and L isomerism ▪ Anomers Forms = α (OH is at lower position), β (OH is at higher position) Reducing Sugars ▪ Rule: ✓ Monosaccharides = reducing sugars ✓ Disaccharides = reducing Structures = sugars except sucrose and Mutarotation = inversion of OH trehalose group at the anomeric Carbon ✓ Polysaccharides = non- ▪ Epimers = same all except in 1 Carbon reducing sugars (not C1) ▪ Tests: ▪ Tautomers 1. Fehling’s test Aldose-ketose isomerism 2. Benedict’s test 3. Barfoed’s test (can be used in Pyranose Furanose differentiation of (6 membered ring) (5 membered ring) monosaccharide and disaccharide) Beta Polysaccharides ▪ Contain > 10 monomer units Alpha ▪ “Glycans” ▪ 2 types: Homopolysaccharides (one type of sugar is produced after hydrolysis) Glucosan = after hydrolysis, the Glycosidic bond product is pure glucose Epimers Ex. Starch, glycogen, cellulose Fructosan = the product is pure fructose Ex. Inulin Heteropolysaccharides Gums, mucilages Glycosaminoglycans (GAGs) Homopolysaccharides ▪ Same type of monomer units ▪ Types Glucosan Fructosan PORLANTE, Jefferson “Tino” Aguado REVIEW ON PROFESSIONAL SUBJECTS 1 MODULE 2: BIOCHEMISTRY & PHARMACOGNOSY Heteropolysaccharides Glycogenolysis vs Glycogenesis ▪ Contains 2 or more types of monomer Parameter Glycogenolysis Glycogenesis units Description Glycogen → Glucose → ▪ Examples of GAGs Glucose Glycogen 1. Heparin Type of reaction Catabolic Anabolic 2. Heparan reaction reaction 3. Chondroitin Hormone Glucagon Insulin ▪ Examples of Gums regulator (Epinephrine, (Amylin, ✓ Acacia gum Cortisol, growth Incretin) ✓ Xanthan gum hormone) (can be used in ✓ Dextran management in Glycosaminoglycans Diabetes 1. Chondroitin SO4 – most common and mellitus) abundant; found in human body – Glucose level Hyperglycemia Hypoglycemia cartilage, ligaments, aorta, tendons) 2. Keratan SO4 – most heterogenous 3. Dermatan SO4 – present in skin, heart Bioenergetics valve, blood vessel Parameter Substrate Level Oxidative 4. Hyaluronic acid – unsulfated and not Phosphorylation Phosphorylation covalently bonded; used as lubricant, Oxygen No Yes shock absorber requirement 5. Heparan SO4 – only extracellular GAG; ATP Yes (1 ATP) Yes (2 – 3 ATP) generation located in cell surface 6. Heparin SO4 (anticoagulant) Examples PEP → Pyruvate Pyruvate → Acetyl CoA Biochemical Metabolism Through the enzyme, pyruvate Through the ▪ Metabolism kinase enzyme, pyruvate ✓ Totality of chemical reaction There’s formation of dehydrogenase that occur in an organism ATP Intermediate ▪ Biochemical Metabolism process 1. Catabolism NADH will be produced 2. Anabolism Enzyme Kinase Dehydrogenase 3. Amphibolism involvement ✓ FADH2 = 2 ATP Metabolism ✓ NADH = 3 ATP Parameter Catabolism Anabolism Keywords “Break down” “build-up” (Complex to GLYCOLYSIS simpler) Exerts (releases) Gain (absorbs) Old name of glucose: Glycose energy energy Important precursor: Glucose Aerobic: Glucose → Pyruvate “Exergonic” “Endergonic” Release ▪ Embden-Mayerhoff pathway Examples “-lysis” “-genesis” ▪ End product: Glycolysis, Glycogenesis, Glycogenolysis Gluconeogenesis, Aerobic: Pyruvate Proteolysis Lipogenesis Anaerobic: Lactate ▪ Cellular location: Cytosol Enzyme Involvement: Glycolysis *Amphibolism = intermediate reaction of catabolism and anabolism; Krebs cycle/TCA ▪ Kinase: ± PO4-3 (addition or removal of (tricarboxylic acid) cycle phosphate) ▪ Isomerase: Aldose ↔ Ketose Applying principles of Thermodynamics in Aldose Ketose exergonic and endergonic: Glucose Fructose G < 0 (exergonic) Glyceraldehyde (3C) Dihydroxyacetone (3C) G > 0 (endergonic) PORLANTE, Jefferson “Tino” Aguado REVIEW ON PROFESSIONAL SUBJECTS 1 MODULE 2: BIOCHEMISTRY & PHARMACOGNOSY ▪ Aldolase: number of Carbon is divided by 1. Glycerol 2 2. Amino acid Ex. Fructose (6 Carbon) divided by 2 = 3 Carbon 3. Pyruvate (Glyceraldehyde and Dihydroxyacetone (DHA)) 4. Oxaloacetate ▪ Dehydrogenase: produces ATP; NADH 5. Sugar alcohol producer; NADH will be transported using G3P shuttle, malate aspartate shuttle Almost reversible of glycolysis ▪ Mutase: change the location of the Glycolysis Gluconeogenesis phosphate group Pyruvate kinase reaction (a) Pyruvate carboxylase Ex. from position 3 to position 2 Pyruvate → ▪ Enolase: water removal; process of Oxaloacetate (b) PEP carboxykinase dehydration OAA → PEP PFK reaction Fructose bisphosphatase reaction Hexokinase reaction Glucose-6-phosphatase In Glycolysis 11 steps (refer to as anaerobic), 8 reversible, 3 irreversible (if there’s kinase, however there are 4 steps that have kinases, 1 is reversible which is Step 7 Phosphoglycerate kinase) G3P shuttle: 2 ATPs Malate-Aspartate shuttle: 3 ATPs 6 – 8 ATP was produced Anaerobic Glycolysis Oxygen is not required ▪ ATP production: 2 ATP ▪ Pyruvate becomes Lactate ▪ Oxygen requiring? NO Gluconeogenesis Glucose new formation ▪ Definition: formation of new glucose ▪ Confined in the liver, intestinal epithelium, kidney ▪ Sources: PORLANTE, Jefferson “Tino” Aguado REVIEW ON PROFESSIONAL SUBJECTS 1 MODULE 2: BIOCHEMISTRY & PHARMACOGNOSY Kreb’s Cycle Electron Transport Chain ▪ Other names ▪ Location: Inner mitochondrial membrane 1. Tricarboxylic acid cycle (TCA) ▪ Final electron acceptor: Oxygen 2. Citric acid cycle ▪ Partial reduction: Free radicals (O2-, H2O2) ▪ Occurrence: Mitochondria until it reduces to H2O ▪ Net reaction: ▪ Complete reduction: H2O Acetyl CoA + 3NAD + FAD + GDP → 2CO2 + FADH2 + 3NADH + GTP (12 ATP or 10 ATP per Complexes in ETC 1 Acetyl CoA) Responsible for transport of electrons If dehydrogenase is not encountered, oxidoreductase is used 1 FADH2 = 2 ATP (1.5 ATP) 1 NADH = 3 ATP (2.5 ATP) ▪ Complex I: NADH dehydrogenase Complex 1 GTP = 1 ATP ▪ Complex II: Succinate dehydrogenase Complex ▪ Complex III: Cytochrome b-c1 complex ▪ Complex IV: Cytochrome oxidase enzyme ▪ Complex V: ATP synthase (synthase means to produce) *Connecting links: ✓ Ubiquinone (Q) – mobile carrier of electron ✓ Cytochrome C These three can produce NADH or FADH2 o Glycolysis has NADH on step 6 o Krebs cycle has NADH on steps 3,4,8 o B-oxidation of fatty acid Oxidation ✓ Addition of Oxygen ✓ Removal of hydrogen ✓ Valence increase ✓ Loss of electron ✓ Substance that oxidized: Reducing agent *Thiokinase (Succinyl CoA synthetase) Reduction Step/s with NADH: 3,4,8 = 9 ATP/7.5 ATP ✓ Addition of Hydrogen Step/s with FADH2: 6 = 2 ATP/1.5 ATP ✓ Removal of oxygen Step/s with GTP: 5 = 1 ATP ✓ Valence decrease Total ATP: 12 ATP/10 ATP ✓ Gain of electron 1 mole of glucose, how many ATPs? ✓ Substance that reduced: Oxidizing agent In every complex, there are 4H+ that can be pumped. In Complex IV, 4H should be pumped, PORLANTE, Jefferson “Tino” Aguado REVIEW ON PROFESSIONAL SUBJECTS 1 MODULE 2: BIOCHEMISTRY & PHARMACOGNOSY however 2H was utilized in reducing Oxygen which results to H2O as final product. Reduced GSH → Oxidized GSH Which complex is not responsible for pumping (protective) (non-protective) of proton/hydrogen? Complex II ATP synthase: Oxidized GSH - it doesn’t provide In 4H+ = 1 ATP protection against hemolysis due to free radicals. For NADH Complex I = 4H+ = 1 ATP Thus, NADPH is needed to maintain Complex III = 4H+ = 1 ATP reduced GSH form, which is produced Complex IV = 2H+ = ½ ATP via PPP. Once produced, the H adds to Therefore, 1 NADH = 2.5 ATP the oxidized GSH, it will be in reduced form. For FADH2, found in Complex II Complex II = FADH2 → FAD+ + 2e Favism (G6PD deficiency) – patient lacks Glucose-6-phopshate 2e will be transferred to Q, Cytochrome iii, dehydrogenase, which is helpful in PPP. Cytochrome C, Cytochrome IV, accepted by the No G6PD, decrease PPP, decreased oxygen production of NADPH, glutathione can’t Complex III = 4H+ = 1 ATP be converted back to its reduced form. Complex IV = 2H+ = ½ ATP Therefore, 1 FADH2 = 1.5 ATP Primaquine, Sulfonamides = agents that will be able to hemolyze the RBC BLOCKER Patients with favism suffer from Complex I Barbiturate hemolytic anemia (Patient could die 3. Interconvert hexose and pentose from respiratory ▪ G6PD deficiency depression when he Aka Favism or she use too much Result: Hemolytic anemia amount of Glycogen Storage Disease Barbiturate) GSD Type Other name Enzyme deficient Complex II Malonate Type 0 - Glycogen synthase Complex III BAL (British Anti- Type I Von Gierke’s Glucose-6- Lewisite) disease phosphatase (Ia) Complex IV Carbon monoxide, G-6-translocase (Ib) Cyanide, Hydrogen Type II Pompe’s α-1,4-glucosidase or sulfide disease acid maltase Type III Cori’s disease Debranching enzyme Pentose Phosphate Pathway Type IIIa (α-1,6-glucosidase) ▪ Aka: Hexose monophosphate (HMP) Shunt Type IIIb ▪ Occurrence: Cytosol Type IIIc ▪ Goals: Type IIId 1. Produce ribose-5-phosphate Type IV Andersons Branching enzyme 2. Produce NADPH (needed to facilitate Disease steroid and fatty acid synthesis; Type V McArdle’s Muscle maintaining the integrity of RBC by disease phosphorylase maintaining the reduced form of GSH) Type VI Hers disease Glycogen phosphorylase H2O2 (free radical) will attempt to hemolyze the blood PORLANTE, Jefferson “Tino” Aguado REVIEW ON PROFESSIONAL SUBJECTS 1 MODULE 2: BIOCHEMISTRY & PHARMACOGNOSY Type VII Tarui’s disease Muscle Linoleic acid = 18:2; ω-6 dietary precursor of phosphofructokinase prostaglandin Type VIII - Liver phosphorylase Test for Carbohydrates Linolenic acid = 18:3; ω-3 QUALITATIVE TEST VISIBLE RESULT (Carbohydrates) General Test for Carbohydrates Arachidonic acid (Eicosanoid) = 20:4; ω-6; ✓ Molisch test ✓ Purple ring (junction) precursor of prostaglandin ✓ Anthrone test ✓ Green/blue green Test for Pentoses ✓ Tauber’s Benzidine ✓ Cherry red Simple lipids test ✓ Bial’s orcinol test ✓ Blue green ▪ Waxes ✓ Tollen’s ✓ Red color ▪ Sterols Phloroglucinol test ▪ Fats (TAGs) Test for Ketoses ▪ Fixed Oils ✓ Seliwanoff’s test ✓ Red color ✓ Tauber’s Amino ✓ Bright reddish purple Waxes High MW alcohol and fatty acids guanidine test Solid, except jojoba oil Test for Reducing Sugars Fats & Fats → solid, except cod liver oil ✓ Fehling’s A – CuSO4 ✓ Brick red precipitate oils ✓ Fehling’s B – NaK Fixed oils → liquid, except tartrate Theobroma oil ✓ Benedict’s test ✓ Brick red precipitate Sterols Nucleus: CPPP Test to differentiate monosaccharide from Cyclopentanoperhydrophenantrene disaccharide ✓ Barfoed’s test ✓ Brick red precipitate ✓ Animal: Cholesterol (wide occurring Mucic acid test sterol) ✓ Specific test for Mucic acid – the only ✓ Plant: Phytosterol Galactose insoluble disaccharide in ✓ Galactose + HNO3 = water Sitosterol – most abundant plant Galactaric acid sterol (Mucic acid) Stigmasterol ✓ Fungi: Ergosterol ✓ Feces: Coprosterol LIPIDS Complex Lipids ▪ Heterogenous group of compounds related more by their physical rather than chemical ▪ Phospholipids properties ▪ Glycolipids ▪ Important property: hydrophobic (non- ▪ Sulfolipids polar) ▪ Lipopolysaccharides (lipid w/ complex carbohydrates) When added with water, it will result to repulsion Phospholipids Important Lipids ▪ Glycerophospholipid Saturated fats = ic Glycerol + FA + phosphoric acid Unsaturated fats = eic, enic, onic ▪ Sphingophospholipids/Sphingolipids Palmitic acid = 16:0 Sphingomyelin: present in nerves Sphingomyelinase: enzyme that degrades sphingomyelin PORLANTE, Jefferson “Tino” Aguado REVIEW ON PROFESSIONAL SUBJECTS 1 MODULE 2: BIOCHEMISTRY & PHARMACOGNOSY Niemann-Pick disease: if sphingomyelin will not Lipopolysaccharides be degraded; patient lacks sphingomyelinase; ▪ Polysaccharide-containing lipids the child may suffer from early childhood death, and mental retardation because of accumulation Sulfolipids of sphingomyelin ▪ Sulfur containing lipids (lipoproteins) Glycerophospholipids Precursor and Derived Lipids ▪ Phosphatidic acid + alcohol ▪ Terpenoids = terpenes Serine + PA: Phosphatidylcholine Precursor: isoprene units Ethanolamine + PA: ▪ Steroids = sterols Phosphatidylethanolamine Nucleus: CPPP Choline + PA: Phosphatidylcholine ▪ Autacoids (E.g. Prostaglandin) Inositol + PA: Phospatidylinositol Precursor: Arachidonic acid Glycerol + PA: Phosphatidylglycerol Parent: Prostonoic acid If 2 glycerol are present, 1st detection: seminal fluid diphosphatidylglycerol 4 Major Groups of Lipoproteins Cardiolipin: diphosphatidylglycerol ▪ Which of the mentioned sterols is similar to Chylomicron lecithin? Phosphatidylcholine ✓ Comes from diet ▪ Other name of Phosphatidylethanolamine: ✓ Hyperchylomicronemia: Elevated Cephalin chylomicron (Type I lipoproteinemia) Correlation ✓ ↑ VLDL (exogenous) ▪ Lecithin Deficiency Low Density Lipoprotein Lecithin is a lung surfactant ✓ Bad cholesterol Respiratory stress syndrome (common to ✓ Hypercholesterolemia: Elevated LDL premature babies who lack lecithin) (Type IIa) Sphingolipids ✓ Combined Hyperlipidemia (Type IIb) = ↑ LDL, ↑ IDL ▪ Ceramide = Sphingosine + FA Precursor of glycolipids Very Low Density Lipoprotein ▪ Sphingomyelin = Sphingosine + ✓ Endogenous (produced in the liver) Phosphorylcholine ✓ Hypertriglyceridemia (Type IV): ↑ Constituent of myelin sheaths Triglycerides; at risk of acute pancreatitis High Density Lipoprotein ✓ Good cholesterol ✓ ↑ HDL = decrease risk of coronary artery disease *Absence of HDL: Tangier disease Decrease level of LDL: statins Decrease level of VLDL: fibrates Decrease level of chylomicrons: no option; diet Glycolipids control; Ezetimibe may do ▪ Ceramide + Carbohydrates Dietary Lipid Metabolism 1. Cerebrosides = ceramide + glu/gal ▪ Digestion starts in the stomach Glucocerebroside ▪ Lingual lipase + gastric lipase = helps in Galactocerebroside increasing the metabolism of lipids 2. Gangliosides = cerebroside + sialic acid ▪ Lipase = TAGs, fats (n-acetyl-neuraminic acid) ▪ Emulsification occurs in the duodenum 3. Sulfatide = cerebroside + sulfate ▪ Bile salts (made in the liver and stored in the gall bladder) + peristalsis → emulsification PORLANTE, Jefferson “Tino” Aguado REVIEW ON PROFESSIONAL SUBJECTS 1 MODULE 2: BIOCHEMISTRY & PHARMACOGNOSY LIPID METABOLISM How many In 1 mol of a Subtotal was substance, ▪ β-oxidation: produced how many ✓ medium chain fatty acid ATP was produced (mitochondria) Acetyl CoA 5 10 50 ✓ Very long chain fatty acid NADH 4 2.5 10 (peroxisome) Number of Carbon FADH2 4 1.5 6 Capric 10 TOTAL 66 ATP Lauric 12 NET ATP TOTAL ATP – 2 ATP (need 64 Myristic 14 for activation of β-oxidation) Palmitic 16 2. Palmitic acid (16C) Stearic 18 In 16 Carbons of Palmitic acid, 8 Acetyl Arachidic 20 CoA was produced ▪ α-oxidation: Acetyl CoA: 8 ✓ branched fatty acid 1 mol Acetyl CoA = 12 ATP or 10 ATP How many ATPs are formed in the β-oxidation Determine number of rounds, how many of: times it was divided? Or simply: 1. Capric acid (10C) NADH or FADH2 = # of Acetyl CoA – 1 = 8–1=7 NADH: 7 FADH2: 7 Acet = 2 carbons 1 mol NADH = 3 ATP or 2.5 ATP In 10 Carbons of Capric acid, 5 Acetyl 1 mol FADH2 = 2 ATP or 1.5 ATP CoA was produced How many In 1 mol of a Subtotal Acetyl CoA: 5 was substance, 1 mol Acetyl CoA = 12 ATP or 10 ATP produced how many ATP was produced Determine number of rounds, how many Acetyl CoA 8 12 96 times it was divided? Or simply: NADH 7 3 21 NADH or FADH2 = # of Acetyl CoA – 1 = FADH2 7 2 14 5–1=4 TOTAL 131 ATP NADH: 4 NET ATP TOTAL ATP – 2 ATP (need 129 FADH2: 4 for activation of β-oxidation) 1 mol NADH = 3 ATP or 2.5 ATP 1 mol FADH2 = 2 ATP or 1.5 ATP How many In 1 mol of a Subtotal was substance, How many In 1 mol of a Subtotal produced how many was substance, ATP was produced how many produced ATP was Acetyl CoA 8 10 80 produced NADH 7 2.5 17.5 Acetyl CoA 5 12 60 FADH2 7 1.5 10.5 NADH 4 3 12 TOTAL 108 FADH2 4 2 8 ATP TOTAL 80 NET ATP TOTAL ATP – 2 ATP (need 106 ATP for activation of β-oxidation) NET ATP TOTAL ATP – 2 ATP (need 78 for activation of β-oxidation) PORLANTE, Jefferson “Tino” Aguado REVIEW ON PROFESSIONAL SUBJECTS 1 MODULE 2: BIOCHEMISTRY & PHARMACOGNOSY Lipid Metabolism ▪ Storage and Transport Ferritin: storage form of Iron CATABOLISM Transferrin: transport form of Iron Triglycerides (TG) = glycerol + fatty acids Hemoglobin: oxygen transport Glycerol could undergo: ▪ Biological catalyst: Enzymes (increases 𝐺𝑙𝑢𝑐𝑜𝑛𝑒𝑜𝑔𝑒𝑛𝑒𝑠𝑖𝑠 chemical reaction) Glycerol → Glucose ▪ Muscular contraction: contractile proteins 𝐺𝑙𝑦𝑐𝑜𝑙𝑦𝑠𝑖𝑠 (actin and myosin) Glycerol → Pyruvate ▪ Structural Proteins: collagen (most abundant Glycerol → Glyceraldehyde → Pyruvate protein in the body), keratin (found in hair Fatty acids will undergo: and horns 𝛽 𝑜𝑥𝑖𝑑𝑎𝑡𝑖𝑜𝑛 𝐾𝑟𝑒𝑏 ′ 𝑠 𝑐𝑦𝑐𝑙𝑒 Fatty acid → Acetyl CoA → CO2 + Immunoglobulins = antibodies H2O 1. IgM = largest among immunoglobulins 2. IgA = secretory antibody Steroids → can be catabolized into Bile acids, 3. IgD = found in cell surface Vit. D, hormones (cortisol, estrogen, androgen) 4. IgG = smallest; can also be seen in the ✓ Not broken down completely placenta 5. IgE = responsible for allergic reaction ANABOLISM (lipogénesis – formation of Proteins lipids) Monomer: Amino acids ▪ Formation of FA: cytoplasm Properties: ▪ Unsaturation: mitochondria, smooth ER ▪ Chirality: 4 different substituents (except Chemical Tests (Lipids) Glycine, because R of glycine is replaced by H). Glycine is the only achiral amino ▪ Rosenheim Test – used to detect the acid. presence of choline ▪ Liebermann-Burchard test – most sensitive test to detect the presence of cholesterol Expected result: green, emerald green Calories Generate after certain chemical reaction ▪ Amphoteric: carboxylic acid is acidic, ▪ 1000 cal = 1 kcal Amino group is basic, hence amino acid is amphoteric (being an acid and base) ▪ 1 g CHO = 4 kcal ▪ Zwitterionic: presence of dipolar charge ▪ 1 g Fat = 9 kcal (has + and – charge) ▪ 1 g dextrose = 3.4 kcal (parenteral) ▪ 1 g ethanol = 7.1 kcal PROTEINS ▪ Most abundant and most functionally diverse biomolecules in living systems ▪ Building blocks: Amino acids ▪ Isoelectric (isoionic) point: pH at which ▪ Bond: peptide bond the amino acid is electrically neutral (0 charge) Biomolecules Type of bond/linkage Carbohydrates Glycosidic (ether) Isoelectric point Considerations (pI) Lipids Ester Average of 2 pKas Proteins Peptide ▪ Neutral side chain: 2 pKas are given, get the Nucleic acid Phosphodiester average Protein Functions ▪ Acidic: get the average of 2 near pKas ▪ Basic: get the average of 2 near pKas PORLANTE, Jefferson “Tino” Aguado REVIEW ON PROFESSIONAL SUBJECTS 1 MODULE 2: BIOCHEMISTRY & PHARMACOGNOSY ▪ Ionizable group: Cysteine, Tyrosine; get the Arginine Arg R average of 2 lowest pKa values Asparagine Asn N Ex. Aspartate Asp D Aspartate or Asx B (D,N) Asparagine Glutamate Glu E 1. Glutamine Gln Q A B C D Glutamate or Glx Z Glutamine x / x x Phenylalanine Phe F 𝑝𝐾𝑎1 + 𝑝𝐾𝑎3 1.88+3.65 pI = = = 2.77 Tyrosine Tyr Y 2 2 2. Tyrosine (ionizable group) Tryptophan Trp W Lysine Lys K Must-know info Aromatic amino acids F, Y, W Sulfur-containing M, C amino acids Involved in urea cycle D Amino acid A, Q transported to the liver 𝑝𝐾𝑎1 + 𝑝𝐾𝑎2 2.2+9.1 pI = = = 5.65 Only achiral amino G 2 2 acid 3. Cysteine (ionizable group) With imino group P * Cysteine + Cysteine = Cystine (formed by disulfide bonds) Some notes ▪ Each amino acid has carboxyl group, a primary amino group (except Proline) & side chain pI = 𝑝𝐾𝑎1 + 𝑝𝐾𝑎2 = 1.7+8.3 =5 Proline has imino group 2 2 ▪ Simplest amino acid: Glycine Amino acid Amino acid Classification Amino acid 3 Letter 1 Letter Abbrev Abbrev ▪ Polar: R-SH, R-CONH2, R-OH Cysteine Cys C NY ST QC (New York ST. Cubao, Quezon City) Histidine His H Methionine Met M Serine Ser S Valine Val V Alanine Ala A Glycine Gly G Proline Pro P Threonine The T ▪ Nonpolar: alkyl group, R-S-R Leucine Leu L Isoleucine Ile I Amino acid 3 Letter 1 Letter Abbrev Abbrev PORLANTE, Jefferson “Tino” Aguado REVIEW ON PROFESSIONAL SUBJECTS 1 MODULE 2: BIOCHEMISTRY & PHARMACOGNOSY Collagen (skin and tendons) Histones ✓ Soluble in Thymus histone H2O and hemoglobin ✓ Insoluble in dilute NH3 ✓ Basic in reaction Protamines ✓ Soluble in Salmin and ▪ Acidic: -COOH NH3, H2O, sturin (fish ac E D ic dilute acid sperm) ✓ Strongly basic in reaction ▪ Conjugated Proteins: protein + non- protein portion (prosthetic group) CONJUGATED PROSTHETIC EXAMPLES PROTEIN GROUP ▪ Basic: -NH2 Phosphoproteins Phosphoric acid Casein (milk) Ovovitellin (egg H A L kaline yolk) Nucleoproteins Nucleic acid Nuclein (cell nuclei) Glycoproteins Carbohydrates Mucins (Vitreous humor and saliva) Chromoproteins Colored Hemoglobin substance (blood) Flavoproteins Classification of Proteins Lipoproteins Lipid Chylomicron Lecithin ▪ Simple Proteins: converted to amino acids Metalloproteins Metal Enzymes only after hydrolysis (tyrosinase, Simple Characteristics Examples arginase, Proteins xanthine Albumins ✓ Soluble in Serum albumin oxidase) H2O Egg white ✓ Coagulated by heat ▪ Derived Proteins: Globulins ✓ Insoluble in Serum globulin 1. + Denaturation: Denatured proteins; H2O primary derived proteins ✓ Soluble in dilute salt 2. Progressive hydrolysis: secondary solutions derived proteins ✓ Coagulated by heat Primary Derived Proteins Glutelins ✓ Insoluble in Glutenin H2O or dilute (wheat) ▪ Aka as Denatured proteins salt solution Primary Denaturant Examples ✓ Soluble in Denatured dilute acid Protein or alkali Prolamines ✓ Insoluble in Zein (corn) Proteans Water, enzyme, Fibrin from neutral Gliadin (wheat) dilute acid fibrinogen solutions Myosan from ✓ Soluble in myosin 80% alcohol Albuminoids ✓ Dissolved Keratin (hair Metaproteins Acid/alkali Albuminates only by and horny Coagulated Heat, alcohol Coagulated egg boiling in tissue) Proteins albumin strong acids Elastin (tendons and arteries) Cooked meat PORLANTE, Jefferson “Tino” Aguado REVIEW ON PROFESSIONAL SUBJECTS 1 MODULE 2: BIOCHEMISTRY & PHARMACOGNOSY Secondary Derived Proteins NH3 is toxic to the brain; in order to prevent toxicity associated to NH3, it undergoes Urea cycle. NH3 must be ▪ Progressive hydrolysis of protein converted to urea for it to be eliminated in the kidney. Secondary Denatured Characteristics ↑ NH3 levels will invade the brain and will cause Protein encephalopathy Proteoses Highest MW group Peptones Intermediate MW Peptides Lowest MW group Structural Organization ▪ Why needed? 1. Primary structure ▪ Application: 1. Purine → uric acid 2. Protein → urea Protein misfolding (in secondary ▪ Amino acid sequence structure) ▪ Bonds: Peptide bond ▪ Degradation: Hydrolysis a) Alzheimer’s disease: Amyloid-β 2. Secondary structure b) Parkinson’s disease: α-synuclein Enzymes ▪ Role: catalysts (↑ rate of chemical reaction) ▪ Are all enzymes protein? No ▪ Describes folding/bending of polypeptide ✓ Generally, YES chain into helix/sheets ✓ Nucleic acid enzymes: Ribozymes ▪ Bond: Hydrogen bond Parts of Enzymes ▪ Destroyed via Denaturation process ▪ HOLOENZYME: active enzyme 3. Tertiary Structure ▪ APOENZYME: protein portion (inactive form ; active : simple enzyme) ▪ COFACTOR: non-protein portion (Vitamins act as cofactor) 1. Activator 2. Coenzyme: loosely bound 3. Prosthetic group: tightly bound ▪ 3-D folding pattern of a polypeptide chain ▪ ZYMOGEN: inactive enzyme; it will be ▪ Several bonds (hydrophobic, hydrophilic, activated once cofactor is added ionic, hydrogen bond, disulfide bond) +𝐻𝐶𝑙 Pepsinogen → Pepsin 4. Quaternary Structure +𝑁𝑎𝑂𝐻 Trypsinogen → Trypsin Cofactors COFACTOR ENZYME/PROTEIN Zn+2 Carbonic anhydrase Zn+2 Alcohol dehydrogenase ▪ Most complicated Fe +2 Cytochromes, hemoglobin ▪ Overall arrangement of polypeptide Cu Cytochrome oxidase chains K+, Mg+2 Pyruvate phosphokinase ▪ Several bonds Coenzymes Nitrogen Metabolism Vitamin Coenzyme Coenzyme Nitrogen must be eliminated from the body because it is function toxic Initial step: Deamination – remove amine PORLANTE, Jefferson “Tino” Aguado REVIEW ON PROFESSIONAL SUBJECTS 1 MODULE 2: BIOCHEMISTRY & PHARMACOGNOSY Vitamin B1 Thiamine Oxidative ▪ Glucogenic amino acids form glucose as pyrophosphate decarboxylation (Thiamine) needed via GLUCONEOGENESIS (TPP) Vitamin B2 Flavin Adenine Redox reaction Glucogenic, Ketogenic, or Both Dinucleotide (Hydrogen) (Riboflavin) (FAD+) /Flavin ▪ Glucogenic AA: the rest mononucleotide ▪ Ketogenic: Leucine, Lysine Vitamin B3 Nicotinamide Redox reaction ▪ Glucogenic and Ketogenic: Aromatic AA Adenine (Hydride) (F,Y,W), Isoleucine (Niacin) Dinucleotide (NAD+) Anabolism of amino acids Vitamin B5 Coenzyme A Carrier of acyl ▪ Amino acids are formed from the citric acid group (Pantothenic cycle intermediates acid) ▪ Non-essential Amino Acids (synthesized in Vitamin B6 Pyridoxal Transfer of the body): Ex. Tyrosine phosphate various group to (Pyridoxine) ▪ Essential Amino Acids (obtained from and form amino acids exogenous sources such as diet): Vitamin B9 Tetrahydrofolate Carrier of one- (Phenylalanine, Valine, Threonine), carbon unit (e.g. (Tryptophan, Isoleucine, Methionine), (Folic acid) Formyl group) (Histidine, Arginine*, Lysine, Leucine) Vitamin B12 Methylcobalamin/ Intramolecular *semi-essential Deoxyadenosyl- arrangement (Cobalamin) Amino Acid Disorders cobalamin Disease Amino acid Notes involvement Major classes of Enzymes Phenylketonuria 𝐹 ℎ𝑦𝑑𝑟𝑜𝑥𝑦𝑙𝑎𝑠𝑒 ↓ Phenylalanine 𝐹 → 𝑌 (PKU = most hydroxylase Oxidoreductases Involved in Redox common reaction clinically inborn Ex. Dehydrogenase, error of AA oxidases, peroxidases metabolism) Black Urine F, Y (more ↓ Transferases Transfer of functional Disease important) Homogentisate groups (PO4, NH2) (Alkaptonuria) 1,2-dioxygenase Ex. Kinase, transaminase (deposition of homogentisic Hydrolases Responsible in hydrolysis acid) Ex. Esterases, Lipases, Infantile Y (important ↓ thyroxine = amylases cretinism precursor in Hypothyroidism synthesis of Lyases Functional group removal thyroid Ex. Decarboxylase, hormone) Deaminases Albinism Y (could not be ↓ production in converted to melanin (white) Ligases Coupling melanin) Ex. DNA ligase, Maple syrup Problems with ↓ α-keto-acid urine disease branched chain dehydrogenase Isomerases Isomerization amino acids Ex. Isomerase enzyme, (L, I, V) mutase Hartnup disease Problem with ↓ neutral amino neutral amino acid transport acids Catabolism of Amino Acids (W) ▪ Amino group removal: Transamination (Deamination) ▪ The C skeleton is broken down to: Acetyl CoA (ketogenic amino acids) Citric cycle intermediates (glucogenic amino acids) – oxidized to CO2 and H2O for energy PORLANTE, Jefferson “Tino” Aguado REVIEW ON PROFESSIONAL SUBJECTS 1 MODULE 2: BIOCHEMISTRY & PHARMACOGNOSY ▪ Bond: Phosphodiester bond Phosphate Nitrogenous base Sugar Nucleotide Structural Organization Alkaptonuria manifestation = ear is somewhat color black Primary Thyroxine was not synthesized = Infantile ▪ Base or nucleotide sequence cretinism ▪ Phosphodiester bonds If patient has PKU, phenylalanine cannot be given. Instruct the patient not to eat Phenylalanine rich foods, instead use Tyrosine (non-essential; essential to patients having PKU) rich foods. Protein Digestion ▪ Start: stomach ▪ End: small intestine ▪ Enzyme peptidases (proteases) Secondary Qualitative Tests (Proteins) ▪ Helical structure ▪ H-bonds Test Amino acid Result Ninhydrin test Amino group Rubemann’s blue Biuret (general Peptide Violet test) Xanthoprotein Aromatic amino Yellow-orange acids (F,Y,W) Millon-Nasse Phenol (Y) Old rose Hopkins-Cole W Purple (detection of indole ring) Bromine water Purple Pauly Diazo His, Tyr Red Lead acetate Sulfur (M, C) Black (PbS) Sakaguchi Arginine Orange-red Shiff Lysine Pink *imino = Proline = yellow Base NUCLEIC ACID Purine: Guanine, Adenine ▪ Building blocks: Nucleotides Pyrimidine: Cytosine, Uracil, Thymine ▪ Components 1. Nitrogenous base Pairs (in DNA) 2. Sugar (Ribose, Deoxyribose) Adenine = Thymine (2 H bonds) 3. Phosphate Cytosine = Guanine (3 H bonds) Nucleoside = Nitrogenous base + Sugar Nucleotide = Nitrogenous base + Sugar Pairs (in RNA) + Phosphate Adenine = Uracil (2 H bonds) PORLANTE, Jefferson “Tino” Aguado REVIEW ON PROFESSIONAL SUBJECTS 1 MODULE 2: BIOCHEMISTRY & PHARMACOGNOSY Cytosine = Guanine (3 H bonds) Comparison of Types of Nucleic Acids Nucleic Acid DNA RNA Number of Double stranded Single stranded strands Pentose deoxyribose Ribose Purine A,G A,G Pyrimidine C,T C,U Cell Location Mostly nucleus Cytoplasm DNA Forms Parameter B-DNA A-DNA Z-DNA Strand antiparallel antiparallel Antiparallel Type of Right Right Left- Key Enzymes Helix handed Base pair 10 11 12 ▪ Helicases – unzipping enzyme per turn ▪ Primases – make “primer” (to know where to B-DNA = most abundant form start to work) A-DNA = dehydrated B-DNA ▪ DNA polymerase – act as builder, build new strand of DNA Central Dogma of Molecular Biology ▪ Ligase – gluer or joiner ▪ Replication DNA → DNA ✓ Production of new DNA ✓ Duplication ✓ Identical copies of DNA Follows Principle of Semi-conservative replication and Anti-parallelism Anti-parallel (3’ → 5’; 5’ → 3’) Semi-conservative replication – divide into 2 strands, the original strand will be used to 1 Helicase Unwinding the double helix synthesize new strands of DNA; most (by disrupting H bonds) accepted hypothesis. 2 DNA gyrase Relieves any form of tension that is produced after DNA Conservative – the whole DNA is being unwinding replicated 3 SSBP (single Prevents the separated Dispersive – divided into parts and linked strand binding strands of DNA from together protein annealing 4 RNA primer Composed of multiple bases that attached to the template strand to initiate DNA replication (initates new complementary strand to be built) 5 Primase Build RNA primers and assembles them at the origin of replication site 6 DNA polymerase ▪ Creates III (builder) complementary strands of DNA (5’→3’) ▪ Attaches to an RNA primer ▪ Reads sequence in Antiparallel 3’→5’ ▪ Adds nucleotides from 5’→3’ 7 DNA polymerase I Removes RNA primer and replaces them with the appropriate DNA nucleotides PORLANTE, Jefferson “Tino” Aguado REVIEW ON PROFESSIONAL SUBJECTS 1 MODULE 2: BIOCHEMISTRY & PHARMACOGNOSY 8 Okazaki Short, discontinuous RNA → protein fragments stretches of DNA made on ✓ Protein synthesis from mRNA the lagging strand during replication (1) Activation 9 DNA ligase Joins DNA fragments (by (2) Initiation: ribosomal subunit attaches to the phosphodiester bond) mRNA at the initiation; start codon: AUG Leading strand – continuous strand (Met) (3) Elongation: creates amino acid sequence in order to become polypeptide Catalysis of bond at site P with the help of dipeptidylpeptidase translocation (4) Termination: stop the process using stop codons (UAG, UGA, UAA) (1) Helicase unwinds double stranded DNA (2) SSBPs stabilize unwound DNA (prevent annealing) (3) Leading strand is formed continuously (5’→3’) (4) Lagging strand is formed discontinuously ▪ RNA primer Start codon: AUG ▪ Gaps are fill out by DNA polymerase I (5) DNA ligase joins the Okazaki fragment Stop codon: UAG, UGA, UAA Major Events in Replication (Prokaryotes) Genetic code Table 1. Helicases unwind the DNA double helix 2. Primase creates a temporary RNA primer 3. DNA polymerase (III) at the replication fork synthesizes DNA in a 5’→3’ direction Leading strand (continuous) Lagging strand (discontinuous): Okazaki fragment 4. DNA polymerase (I) then removes the RNA primer and fills the gaps between the Okazaki fragments (short stretches of discontinuous DNA) Mutation 5. DNA ligase then joins DNA fragments of Involves a change in the shape, the lagging strand, creating a single DNA structure, and nucleotide sequence molecule The newly synthesized DNA is further modified by (1) Point mutation TOPOISOMERASES (remove supercoils in helix) Transitional mutation Semiconservative replication Purine → Purine ▪ Transcription Pyrimidine → Pyrimidine DNA → RNA Transversional mutation ✓ Controlled by the enzyme RNA Purine → Pyrimidines polymerase (controller and modifier) Pyrimidines → Purines Human (80s): 60S and 40S Ex. UCA → UCU (Transversional Bacteria (70S): 50S and 30S (target mutation) of aminoglycoside) ▪ Sickle cell anemia ✓ Creates 3 basic types of RNA ✓ Biconcave RBCs → crescent shape Comparison mRNA rRNA tRNA ✓ Decreased oxygen transport Description Genetic Provides Brings ✓ Enlarged spleen message physical specific carrier make up amino acid of to the ribosomes ribosme %total RNA cell 5% 80% 15% ▪ Translation PORLANTE, Jefferson “Tino” Aguado REVIEW ON PROFESSIONAL SUBJECTS 1 MODULE 2: BIOCHEMISTRY & PHARMACOGNOSY (Beta-carotene - Anti-oxidant - Xerophthalmia (pro-vitamin): (beta- (dryness of precursor) carotene) conjunctiva) D2 - Ca balance - Ricketts (ergocalciferol) - Osteomalacia D3 (colecalciferol) ▪ Results of Point mutation E (alpha- - Anti-oxidant - Neurologic ✓ Silent mutation tocopherol) dysfunctions (rare) Change in codon, mutation occurs - RBC facility but the outcome is same amino K1 - Blood clotting - Hemorrhage acid (phylloquinone) ✓ Missense mutation K2 (menaquinone) Different amino acid produced K33 ✓ Nonsense mutation (menadione) Facilitate stop codon K4 (menadiol) ▪ Silent, Missense, or Nonsense? ▪ Water-soluble vitamins (B & C) VITAMIN FUNCTION DEFICIENCY ✓ UCA to UCU (Ser to Ser): Silent STATES ✓ UUU to UUG (Phe to Leu): Missense B1 - Coenzyme in - Beri-beri ✓ UAC to UAG: Nonsense (thiamine) pyruvate and α- - Wernicke- (2) Frameshift mutation (Insertion, Deletion) ketoglutarate, Korsakoff dehydrogenation syndrome Insertion or Deletion mutations , and (nystagmus, A major alteration in the sequence of transketolase ataxia, polymerized amino acids → altered confusion) B2 - Coenzyme in - Cheilosis and reading frame in the mRNA (riboflavin) redox reactions seborrheic - FAD dermatitis B3 (niacin, - Coenzyme in - Pellagra nicotinamide redox reactions , nicotinic - NAD and NADP acid) B5 - Functional part - Burning Foot (Panthotenic of CoA Syndrome acid) Nucleic Acid Metabolism B6 - Coenzyme in - Peripheral Gout ↑ uric acid (Pyridoxine) transamination neuritis Deposit of monosodium urate B9 (Folic - Transfer of 1C - Megaloblastic crystals, deposit in the joints acid) fragments anemia which causes formation of tophi B12 (Cyano- - Transfer of 1C - Pernicious Kidney cobalamin) fragments anemia Lesch-Nyhan Decreased Hyperuricemia - Metabolism of (Megaloblasti syndrome HGPRT Folic acid c anemia + (self- spinal cord mutilation) degeneration) Xeroderma Deficiency of Sunburn, Vitamin H - Coenzyme in the - Impaired fat pigmentosum DNA repair freckles, dry, (Biotin) also carboxylation and CHO mechanisms scaly skin called as Vit. reactions in metabolism HGPRT = hypoxanthineguaninephosphoribosyl B7 gluconeogenesis - Dermatitis transferase and FA synthesis (3) Spontaneous mutation = natural error B17 (laetrile) B15 (Pangamic acid) Reverse transcription (see in viruses, e.g. HIV) C (Ascorbic - Coenzyme in - Scurvy RNA → DNA acid) hydroxylation of (impaired proline and wound VITAMINS lysine in collagen healing, loss synthesis of dental ▪ Essential nutrients that our body needs - Anti-oxidant cement, SQ to sustain life - Enhances Fe hemorrhage) ▪ Fat soluble vitamins (A,D,E,K) absorption VITAMIN FUNCTION DEFICIENCY STATES A (retinol) - Visual - Nyctalopia - END - pigment (night blindness) PORLANTE, Jefferson “Tino” Aguado REVIEW ON PROFESSIONAL SUBJECTS 1 MODULE 2: BIOCHEMISTRY & PHARMACOGNOSY PHARMACOGNOSY & ▪ Drying ✓ Removal of moisture or water PHYTOCHEMISTRY (prevention of microbes) PHARMACOGNOSY ✓ To protect the sample from microbial damage According to C.A. Seydler: ✓ ↓ enzyme activity ▪ Pharmakon = drug ✓ Ensure good characteristics of ▪ Gnosis = knowledge crude drug “Knowledge on drugs” ▪ Curing ✓ Specialized method of drying ✓ Natural drugs ✓ Fresh samples that are highly toxic Biological or irritating, these are subjected to Biochemical curing to lower down the toxicity Economic features and reduce the irritating effect of Fluckiger: Scope of Pharmacognosy fresh sample. ✓ Increase the yield/amount of ✓ Various scientific disciplines active ingredient that can be Simultaneous application obtained. Crude Drugs ▪ Garbling ✓ Final step of crude drug ▪ Vegetable or animal drugs that consist of preparation natural substances that have undergone ✓ Separation or removal of only the process of (1) collection & (2) extraneous matter (undesired drying plant parts or there are traces of Terms dirt present and/or soil) ✓ Natural substance – found in nature ▪ Packaging, storage and ✓ Extractives/Derivatives – active preservation (optional steps) ingredients (single or mixture) ✓ Packaging – provide physical ✓ Extraction protection, ↑ marketability Marc – residue left after extraction ✓ Storage – protect crude drug from Menstruum – solvent for animal invasion such as insects or extraction rodent. Should be in appropriate o Fats – hexane storage condition. o Resins – alcohol ✓ Preservation – maintain the o Chlorophyll – acetone quality of crude drug. o Chrysarobin – hot benzene Temperature incorporation is ✓ Naturalized Plants – foreign country needed. ✓ Indigenous Plants – native country Heating is needed to preserve the crude drug at Crude Drug Preparation (CHAD CurGa Pa) 65°C ▪ Collection Crude drug can be ✓ From wild plants subjected to fumigation ✓ Ensuring the true natural source using Methyl bromide as of drug fumigant. ▪ Harvesting Chloroform or Carbon ✓ Gathering samples from cultivated tetrachloride species Drug Evaluations ✓ Timing: Quality of substance/Quantity – amount of For identification purposes, know the quality active principle or ingredient must and purity of the crude drug be high enough to use for a ▪ Organoleptic certain purpose that has biological ✓ Use of sense organ activity ✓ Evaluation using the sense organ PORLANTE, Jefferson “Tino” Aguado REVIEW ON PROFESSIONAL SUBJECTS 1 MODULE 2: BIOCHEMISTRY & PHARMACOGNOSY ✓ Macroscopic evaluation belong to the group? Answer: ▪ Microscopic Atropine – anticholinergic drug) ✓ Use of microscope ▪ Chemical ✓ Involvement of anatomical ✓ Based on constituents or active examination ingredient present in crude drug ▪ Biologic ✓ Preferred method of drug ✓ Use of animals classification ✓ Perform “Bioassay” to know the ✓ Alkaloids – ephedrine, atropine, pharmacologic use or profile of pilocarpine, physostigmine crude drug CARBOHYDRATES ✓ Test the antipyretic profile of a certain drug = rabbit “Hydrates of carbon” ✓ Mydriatic effect = atropine ▪ Building blocks: Monosaccharides ▪ Chemical Bond formed: Glycosidic bond – ether ✓ Best method linkage ✓ Involvement of chemical Functional groups: R-OH, RCHO, RCOR treatment with the use of [Polyhydroxyaldehyde (C#1 – Carbonyl appropriate reagent group (terminus part of the structure), ✓ For qualitative (e.g. Iodine test for Polyhydroxyketone (C#2)] presence of starch; Benedict’s ▪ Reactions: test, Fehling’s test, Barfoed’s test ✓ Synthesis: occurrence of for detection of reducing sugar) glycosidic linkage, Dehydration and quantitative testing (e.g. reaction is needed to remove the titration for % purity water molecule determination, Saponification ✓ Degradation: opposite of value, Ester value, etc.) dehydration (Hydrolysis), add ✓ Best method in determining the water molecule, which allow potency destruction of molecule (a ▪ Physical polymer could be converted into ✓ Physical constants (Sp Gr, density, simpler forms) melting point, boiling point, optical Classes of Carbohydrates activity = with the use of polarimeter, refractive index = Units Type (how (number of many class) refractometer) sugar units) Drug Classifications Monosaccharides Always 1 Always 1 Similarity of available plant material Disaccharides Always 2 Up to 2 ▪ Morphologic Oligosaccharides 2-10 1 or more ✓ Depending on plant part used “few” C ≠ 11 ▪ Taxonomic Polysaccharides ✓ Phylogeny – evolutionary ✓ Aka Glycans > 10 1 or more relationship present among plants ✓ Homoglycans > 10 Always 1 and animals. Refers to natural ✓ Heteroglycans > 10 > 1 (2 or more) relationship exists among them. Monosaccharides ✓ Plants (botanic) and animals (zoologic classification) Name Aldose Ketose ▪ Pharmacologic/Therapeutic Diose (2 C) Hydroxy- - acetaldehyde ✓ Medicinal use of substance Triose (3 C) Glyceraldehyde Dihydroxy- depending on therapeutic acetone category Tetrose Erythrose Erythrulose Threose ✓ Ex. Atropine, Ach, Bethanechol, Pentose Ribose Ribulose Pilocarpine (which does not Arabinose Xylose