PHARMAOSMOSIS

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NMR
INTRODUCTION NMR is absorption spectrometry. Permits identification of atomic configuration. Radiofrequency waves are used in NMR.RF waves have long wavelength so less energy. Range of RF is from 30 KHZ TO 30 MHZ. Felix Bloch and Edward Purcell received Nobel Prize for NMR discovery. NMR mechanism includes change in spin orientation on exposure to magnetic field. Spin quantum number(I) describes angular momentum. Total spin quantum number depends on the number of nucleons and symmetry of charge distribution. Precessional frequency-No. of spin carried out by the nucleus in the presence of magnetic field per second. Thus when magnetic field applied the nucleus spins with precessional frequency. Before application of magnetic field it was moving with random frequency. When no magnetic field, only one energy level. but in presence of magnetic field 2 energy levels with aligned and opposite orientations of the nuclear spins. Precessional frequency is directly proportional to field strength() Bo RememberWhen mag.field=1.4 tesla,=60MHZ. When mag.field=2.3 tesla,=100MHZ. When mag.field=7.0tesla,=300MHZ. When mag.field=14.1Tesla,=600MHZ.

1 Tesla=104 Gauss.so 1.4 Tesla=14000 Gauss. Imp-Tesla is measure of magnetic flux density not field strength. Resonance occurs when, Applied frequency from RF oscillator=precissional frequency of spinning nucleus.

VARIOUS NUCLEUS When mass no. is odd,atomic no.odd/even spin quantum number(SQN) is in fraction e:g 1H=1/2,11B=3/2.,13C=19F=31P=1/2. When mass no. is even,atomic no.odd spin quantum number(SQN) is in integer; E;g 10B=3,14N=1.2D=1. rem = they exhibit quadrapole moment. When mass no. is even,atomic no.even spin quantum number(SQN) is 0. E:g 12C=0,16O=0. IMPORTANT POINTS (continued) No of possible orientations= 2I+1,so 1H will have 2 and 2D & 14N will have 3 orientations.

Imp.formula = Ho =larmor frequency. But =2,so ultimate equation,

Bo/2 = /I

=gyro magnetic ratio =nuclear magnetic moment I=nuclear angular momentum.

Remember = gyro magnetic ratio of 1H=268,while of 13C NMR is 67.so 13 C NMR is 4 times less sensitive than 1H NMR. so if the resonance frequency of particular nucleus in1HNMR is 300 MHZ, in 13CNMR it is 300/4 = 75MHZ.

Gyro magnetic ratio of 2H is 1/3 of 1H. There are some more nuclei in lower energy state which are responsible for absorption of RF. so NMR phenomenon occurs. This distribution is according to Boltzmanns law. frequency 20 40 60 80 Excess nuclei 3 6 9 12 frequency 100 200 300 600 Excess nuclei 16 32 48 96

Relaxation process of nucleus

Spin-lattice or longitudinal relaxation-gives energy to lattice so system gets heated. Spin-spin or transverse relaxation-give energy to nearby nuclei so no net change in energy of system. REMEMBER:spin-lattice relaxation time (T1)determines the rate at which absorption of RF can occur. Rememeber : relaxation process follow first order process and T1 is the reciprocal of first order rate constant. T2 is spin-spin relaxation time. Remember-if T1 and T2 small,broad absorption lines,if they r large sharp abs. lines. FOR non-viscous solids T1 is large so sharp lines in NMR spectrum. FOR 14N nucleus T1 is small due to quadrapole so broad signals in 14N NMR.

REMEMBER Solids are difficult to analyze by NMR so MAGIC ANGLE is used for solid samples which is 54.6 degree. Thus solid samples are oriented at this angle and observed.thus solids can be analyzed. INSTRUMENTATION Magnet RF oscillator Sweep generator RF receiver Recorder Internal standards TMS(tetramethyl silane)(0.01-1%) universally accepted,used only in organic solvents.TMS is used in 1H and 13C NMR. Sodium salt of 3-(trimethylsilyl)-propane sulfonic acid and DDS is used in aqueous solvents. CFCl3 for 19F NMR. H3PO4 for 31P NMR. NH3(absolute standard) and nitromathane(mostly used) for 14N NMR. Water for 17O NMR. CHEMICAL SHIFT Expressed in ppm (delta or tau) units. =10- High delta value=downfield Low delta value=upfield For tau it is reverse. Rem - is dimensionless number and it is field independent. Chemical shift values depend on shielding and deshielding. TMS has 12 protons to shielding of nucleus by protons so less i:e=0. If electronegative element, like Cl than removal of electrons near nucleus so deshielding so large . Chemical shift values range from 1-20. FACTORS AFFECTING CHEMICAL SHIFT Hybridization effect Electronegativity- Protons which are attatched to the most electronegative atoms are considered to be highly deshielded. It results in increase in chemical shift value. Van der walls deshielding-observed in rigid molecules.effect less than 1 ppm. Anisotropic effect-called space effect also.anisotopic means effects are not same in all directions (in some direction effect is paramagnetic,in some directions it is diamagnetic).(TABLETS are anisotropic in nature). Due to this effect value of proton is higher in ethane,lower than expected in ethyne,higher in aldehyde and acids.(for more refer pavia). Hydrogen bonding-increases .

Solvents for NMR CDCl3 mostly used. Should not contain hydrogen (aprotic solvents) For polar solutes-D2O,DMSO-d6,CCl2,CS2 ,CF3. Compound solubility should be atleast 10%.

TWO METHODS OF RECORDING NMR 1. CONTINUOUS WAVE METHOD RF remains constant and mag. Field is varied. This is called sweeping. 2. Fourier transform methods. In this, the sample is irradiated with strong pulse of radiofrequency energy containing all frequency of 1H range. It gives spectrum in frequency domain. Pulse duration is 10 microsecond and then pulse is switched off so nuclei get relaxed. Process of transfer of magnetization from RF to nuclei is called free induction decay. Higher sensitivity, can work in low concentrations. Used for sparingly soluble substances. GOLAY COILS/SHIMMING COILS These are used to increase the accuracy of the instrument. They are electromagnetic coils which induce small electromagnetic fields in order to improve the inhomogenity of the field.(sample can also be spun to remove inhomogenity) IMP-ringing phenomenon is produced in CW mode and not in FTNMR mode. REMEMBER Sensitivity is ratio of signal to noise. For 1HNMR,quartet of ethyl benzene spectrum is used for calculating S/N ratio and for 13CNMR,largest aromatic peak in ethyl benzene spectrum is used as standard. A VERY VERY IMPORTANT POINT Superconducting magnets are used for higher fields. A metal becomes a super conductor when it is cooled in liquid helium at 4K(Kelvin). Niobium and Titanium alloy is used as superconductor. For higher fields like 10 T, Nb3Sn can be used.

Chemical shift table (only imp)(for more refer pavia-but these are enough) structure Chem..shift structure Chemical shift alkane 0.5-1.5 Aliphatic ketone 2-2.7

alkene alkyne Benzene aromatics aldehyde acid

1.5-3.0 1.7-2.7 and 6-9 8-10 11-12

OCH3(methoxy) Benzylic proton Aliphatic ester Phenolic proton amide

3.82 2.3-2.7 3.5-5 4-7 5-9

Imp-CDCl3 solvent gives peak at 7-7.5, which is a solvent peak. IMPORTANT POINTS If protons are magnetically and chemically equivalent as in CH3, only one signal for that proton. Two protons are chemically equivalent if they are in same chemical environment and have same chemical shift value. Two protons are magnetically equivalent if each couples equally to neighboring proton. Chemical eq. means chemical shift equivalency. Magnetic eq. means coupling equivalency. Accidental equivalence- It is observed in 2,5-dichloro nitrobenzene. SPIN-SPIN COUPLING The signal splits according to the n+1 rule. This is due to coupling interaction between neighboring protons. E;g CH3CH2OH Singlet for OH (bcoz proton is not directly attached with carbon). Quartet for CH2 (bcoz in neighbor 3 protons so n+1=3+1=4). Triplet for CH3 (bcoz in neighbor 2 protons so n+1=2+1=3). It is also field independent. COUPLING COSTANT(J) Measure of interaction between nuclei. It is separation between the peaks in the multiplets. It is independent of magnetic field. Unit is HZ Cis form in alkenes J value is 4-12. Trans form in alkenes J value is 12-18. TYPES OF COUPING Germinal-10-18 Hz,strong and on same carbon. Vicinal-0-12 Hz in rigid systems,and 8 Hz in freely rotating carbons.it is between neighbouring protons. Long range-in alkanes,small coupling. Aromatic: ortho-7-10Hz,meta- 2-3 Hz,para 0-1 Hz. Allylic-in allyl chloride,four bond coupling and it is small(0-2 Hz)

RESTRICTED ROTATION Observed in N,N dimethyl formamide. Two signals for methyl proton instead of one at room temp. This is due to restricted rotation due to resonance. At 130 deg. C it gives one signal. REMEMBER Karpluss equation is related with vicinal coupling which explains variation in vicinal coupling constant with dihedral angle( ) Largest vicinal C.C when protons are in trans position ( =180 deg.) Smallest vicinal C.C when ( =90 dg.). For cis form ( =0 dg.) and C.C is large. Dueteriation technique is used to detect the presence of OH groups. Virtual coupling is observed in long alkyl chains as a result spectra gets distorted. THERE ARE 3 WAYS TO SIMPLIFY COMPLEX NMR SPECTRA. By increasing field strength By double irradiation one proton is irradiated with high energy so speedy transitions so no coupling with neighboring proton so no complex spectra. For this technique chemical shift positions for the coupling multiplets should be no closer than 1 ppm. By using shift reagents praseodymium (gives low shifts), europium, ytterbium, holmium, erbium are used (all give higher shifts). Lanthanide elements are used. All shift reagents are mild lewis acids. Shift reagents are paramagnetic. They are used in non-polar solvents. NUCLEAR OVERHAUSER EFFECT Space effect One proton irradiated and other neighbouring proton in space gives enhanced signal. Effect is upto shorter distance (3.5 angstrom).It tells about whether two protons are near to each other are not. Range for this effect is 2-4 angstrom. Isovanillin shows this effect. Many other compounds can also show this effect.

13
3

C NMR

Gyromagnetic ratio of 1 C NMR is 4 times less than that of proton NMR so 4 times less sensitive. Spin quantum no. is for 13C. Relative abundance of 13C is 1.1 %. Generally spectra is recorded by pulsed FTNMR mode. Chemical shift values range from 1-200.

Heteronuclear coupling occurs because SQN for H and 13C is same (i:e ) so decoupler is to be used in order to prevent coupling between 13C and 1H. To increase the line intensity in spectrum paramagnetic species like chromium acetylacetonate and shift reagents are added. OTHER IMPORTANT POINTS DEPT (Distortionless Enhancement by Polarization Transfer) SPECTRA It increases sensitivity by a factor of for 13C. In DEPT technique,sample is irradiated with a complex sequence of pulses in both 13C and 1H channels.The result of these pulse sequences is that 13C signal in the carbon atom will exhibit different phases depending on number of hydrogen atoms attached to it. Each type of carbon behaves slight differently, depending on duration of complex pulses. COSY (Correlation Spectroscopy) It is a 2D-NMR. It can be homonuclear or heteronuclear. This techniques enables correlation to be made between protons which are coupled to each other HMQC (Heteronuclear Multiple Quantum Coherence) and HMBC (Heteronuclear Multiple Bond Coherence) 2D inverse correlation techniques that allow for the determination of connectivity between two different nuclear species. MRI (Magnetic Resonance Imaging) Diagnostic technique which uses NMR principle. MAXIMUM OF 1.5 TESLA MAGNETIC FIELD CAN BE USED. CARDIAC PATIENTS NOT ELIGIBLE FOR MRI.

CIDNP (Chemically Induced Dynamic Nuclear Polarization) A technique which induces polarization by chemical reaction.

GPAT AND NIPER ASKED QUESTIONS FROM THE CHAPTER (Answers find yourself)
Chemical shift is expressed in? Magic angle in NMR is? MRI uses which technique? Difference between 13C NMR and 1H NMR in terms of sensitivity (GPAT-2010)-4 options were given and u have to find wrong answer. 5. Splitting related question i:e a compound was given and we have to find out the no of splits for that. 6. Standard in NMR? 7. Hydrogen bond can be detected by?(IR can also detect, bur most widely used is Raman so if RAMAN is given you tick that) 8. Methyl chloride cause ( shielding or deshielding)? 9. No. of peaks in 1,3 dichloropropane? 10. RF range? 11. 1 Tesla is equal to how many gauss? 12. Radio frequency oscillator is used in? 13. Unit of coupling constant? 14. NMR can give information about? 15. Shimming coils are used in which technique? 16. Solvent for NMR? Choose appropriate from options. 17. No of peaks in 1,4 dioxane? 1. 2. 3. 4.

EXTRA QUESTIONS 1. Nuclear Overhauser Effect is upto how much angstrom? 2. COSY is? 3. If applied frequency for 1H NMR is 600 MHZ, then what will be the applied frequency in 13C NMR? 4. Relative abundance of 13C? 5. Which moeity will give inverse peak in DEPT? 6. Full form of DEPT? 7. Karpluss equation is related with? 8. Unit of delta?(dont say ppm, it is dimensionless) 9. Cis form and trans form J value? 10. CDCl3 will give peak at? 11. Which spectroscopy will result in quadrapole?

12. In 60 MHZ instrument, no. of excess nuclei will be? 13. Aldehyde chemical shift? 14. Which effect is responsible of low delta value in alkyne although it is more electronegative than alkene and alkane? 15. Gyro magnetic ratio is the ratio of? PHARMAOSMOSIS BELIEVES NOT IN QUALITY BUT QUALITY WITH ADEQUACY! We are sure that there is no material series in the market which provides this kind of preparative material. SO BE WITH US BY JOINING US AND LEAD TOWARDS THE HIGHER BECAUSE NOT MUCH TIME IS LEFT. Ok friends, best of luck to all GPAT giving students. Dont worry, just prepare hard, the path is yours.!!! Everybody is welcome for guidance whether he is subscriber or not!!!