Experimental and theoretical charge-density analysis of hippuric acid: insight into its binding with human serum albumin

In order to comprehend the binding of an important metabolite, hippuric acid, with human serum albumin and to understand its chemical and electronic nature, an experimental charge-density analysis has been carried out using high-resolution diffraction data collected under cryogenic conditions, and all the results have been compared with theoretical findings using the B3LYP/6-311++g(2d,2p) level of theory. The structure displays very strong classical hydrogen bonds as well as other noncovalent interactions, which have been fully characterized using Hirshfeld surface analysis and Bader's quantum theory of atoms in molecules. Contact analysis on the Hirshfeld surfaces shows that the O...H, C...H and C...N intermolecular interactions are enriched and gives their relative strengths. Topological analysis of the electron density shows the charge concentration/depletion of hippuric acid bonds in the crystal structure. Electrostatic parameters such as atomic charges and dipole moments were calculated. The mapping of atomic basins and the calculation of respective charges show the atomic volumes of each atom as well as their charge contributions in the hippuric acid crystal structure. The dipole-moment calculations show that the molecule is very polar in nature. Calculations of the electrostatic potential show that the chain part of the molecule has a higher concentration of negative charge than the ring, which might be instrumental in its strong binding with the polar residues of site II of human serum albumin.

Computing details

Program(s) used to refine structure: MoPro (J. Appl. Cryst. 2005, 38, 38-54) for Multipolar, IAM_MoPro; SHELXL2018/3 (Sheldrick, 2018) for IAM_shelx.

Benzoylaminoethanoic acid (Multipolar)

Crystal data

C9H9NO3	F(000) = 376
Mr = 179.17	Dx = 1.425 Mg m^−3
Orthorhombic, P212121	Mo Kα radiation, λ = 0.71073 Å
Hall symbol: P 2ac 2ab	Cell parameters from 599 reflections
a = 8.6793 (4) Å	θ = 3.0–55.5°
b = 9.0674 (4) Å	µ = 0.11 mm^−1
c = 10.6085 (4) Å	T = 100 K
V = 834.88 (6) Å^3	Block, colorless
Z = 4	0.28 × 0.21 × 0.18 mm

Data collection

Bruker D8 Venture with PHOTON II Detector (Bruker, 2016) diffractometer	13610 reflections with > 2.0σ(I)
ω and phi scan	Rint = 0.058
Absorption correction: multi-scan SADAB, Bruker, 2016	θmax = 62.7°, θmin = 3.0°
Tmin = 0.971, Tmax = 0.981	h = −21→21
118392 measured reflections	k = −22→22
13610 independent reflections	l = −26→26

Refinement

Refinement on F^2	Secondary atom site location: difference Fourier map
Least-squares matrix: full	Hydrogen site location: difference Fourier map
R[F^2 > 2σ(F^2)] = 0.028	Only H-atom coordinates refined
wR(F^2) = 0.043	w = 1/[σ^2(Fo^2) + (0.0059P)^2 + 0.0083P] where P = (Fo^2 + 2Fc^2)
S = 0.97	(Δ/σ)max < 0.001
13610 reflections	Δρmax = 0.27 e Å^−3
1493 parameters	Δρmin = −0.28 e Å^−3
0 restraints	Absolute structure: Flack x determined using 4564 quotients [(I+)-(I-)]/[(I+)+(I-)] (Parsons, Flack and Wagner, Acta Cryst. B69 (2013) 249-259)
Primary atom site location: structure-invariant direct methods	Absolute structure parameter: −0.05 (11)

Special details

Refinement. Refinement of F² against reflections. The threshold expression of F² > 2sigma(F²) is used for calculating R-factors(gt) and is not relevant to the choice of reflections for refinement. R-factors based on F² are statistically about twice as large as those based on F, and R-factors based on ALL data will be even larger.

Fractional atomic coordinates and isotropic or equivalent isotropic displacement parameters (Ų)

	x	y	z	Uiso*/Ueq
C1	0.39576 (2)	0.63962 (2)	0.584745 (17)	0.011137 (13)
C2	0.46750 (3)	0.52381 (2)	0.519655 (18)	0.013753 (15)
H2	0.5322 (7)	0.441 (10)	0.5714 (5)	0.03054
C3	0.45604 (3)	0.51495 (3)	0.38874 (2)	0.016263 (18)
H3	0.5117 (9)	0.425 (10)	0.3381 (6)	0.03547
C4	0.37192 (3)	0.62073 (3)	0.322251 (19)	0.016216 (17)
H4	0.3625 (8)	0.6133 (8)	0.220 (10)	0.03314
C5	0.29881 (3)	0.73531 (3)	0.38671 (2)	0.016165 (17)
H5	0.2334 (7)	0.818 (10)	0.3356 (5)	0.03444
C6	0.31024 (3)	0.74439 (2)	0.517446 (19)	0.014126 (16)
H6	0.2526 (7)	0.833 (10)	0.5680 (5)	0.03204
C7	0.40741 (2)	0.65979 (2)	0.724012 (18)	0.010815 (12)
C8	0.46949 (3)	0.55965 (2)	0.929786 (17)	0.013104 (14)
H8A	0.4586 (8)	0.449 (10)	0.9705 (5)	0.03131
H8B	0.371 (10)	0.6248 (7)	0.9637 (6)	0.02953
C9	0.61677 (2)	0.62936 (2)	0.978615 (17)	0.012395 (14)
N1	0.46293 (2)	0.54944 (2)	0.793978 (15)	0.012668 (13)
O1	0.36584 (2)	0.778888 (19)	0.773460 (15)	0.014240 (12)
O2	0.62219 (2)	0.61872 (2)	1.103204 (15)	0.018054 (16)
O3	0.71465 (2)	0.68682 (3)	0.913930 (17)	0.019887 (17)
H1	0.4998 (9)	0.455 (10)	0.7514 (6)	0.02774
H9	0.717 (10)	0.6676 (8)	1.1371 (6)	0.03066

Atomic displacement parameters (Ų)

	U^11	U^22	U^33	U^12	U^13	U^23
C1	0.01219 (6)	0.01149 (6)	0.00973 (5)	0.00055 (5)	−0.00049 (4)	−0.00014 (4)
C2	0.01638 (7)	0.01408 (7)	0.01080 (6)	0.00271 (6)	−0.00068 (5)	−0.00122 (5)
H2	0.03913	0.02807	0.02440	0.01306	−0.00441	0.00051
C3	0.01831 (8)	0.01947 (8)	0.01101 (6)	0.00269 (7)	0.00025 (5)	−0.00236 (5)
H3	0.04435	0.03732	0.02474	0.01410	0.00064	−0.00931
C4	0.01623 (7)	0.02233 (9)	0.01009 (6)	0.00002 (7)	−0.00024 (5)	0.00074 (6)
H4	0.03875	0.04484	0.01582	0.00277	−0.00218	−0.00011
C5	0.01695 (8)	0.01929 (8)	0.01225 (6)	0.00203 (6)	−0.00206 (5)	0.00269 (6)
H5	0.03977	0.03600	0.02755	0.01124	−0.00611	0.00789
C6	0.01552 (7)	0.01447 (7)	0.01239 (6)	0.00276 (5)	−0.00135 (5)	0.00071 (5)
H6	0.03932	0.02887	0.02794	0.01382	0.00045	−0.00289
C7	0.01198 (6)	0.01051 (5)	0.00995 (5)	−0.00011 (4)	0.00017 (4)	−0.00050 (4)
C8	0.01361 (6)	0.01551 (7)	0.01019 (5)	−0.00168 (5)	−0.00039 (5)	0.00110 (5)
H8A	0.04273	0.02366	0.02755	−0.00666	−0.00481	0.00656
H8B	0.02528	0.03588	0.02744	0.00354	0.00212	−0.00590
C9	0.01291 (6)	0.01396 (6)	0.01031 (5)	−0.00123 (5)	0.00060 (4)	0.00001 (4)
N1	0.01635 (6)	0.01195 (5)	0.00971 (5)	0.00077 (5)	−0.00094 (4)	0.00005 (4)
O1	0.01777 (6)	0.01220 (5)	0.01275 (5)	0.00163 (4)	0.00193 (4)	−0.00177 (4)
O2	0.01797 (7)	0.02607 (8)	0.01012 (5)	−0.00639 (6)	−0.00107 (4)	0.00000 (5)
O3	0.01776 (7)	0.02801 (9)	0.01389 (6)	−0.00815 (6)	0.00234 (5)	0.00120 (5)
H1	0.04033	0.02052	0.02237	0.00710	−0.00097	−0.00324
H9	0.02667	0.04373	0.02158	−0.01118	−0.00289	−0.00006

Geometric parameters (Å, º)

C1—C7	1.4921 (3)	C6—H6	1.09
C1—C6	1.4011 (3)	C7—O1	1.2536 (3)
C1—C2	1.4026 (3)	C7—N1	1.3358 (3)
C2—C3	1.3947 (3)	C8—N1	1.4448 (3)
C2—H2	1.09	C8—C9	1.5172 (3)
C3—C4	1.3966 (4)	C8—H8A	1.10
C3—H3	1.09	C8—H8B	1.10
C4—C5	1.3963 (4)	C9—O2	1.3261 (2)
C4—H4	1.09	C9—O3	1.2099 (3)
C5—C6	1.3929 (3)	N1—H1	1.02
C5—H5	1.09	O2—H9	1.00
C7—C1—C6	117.216 (15)	C5—C6—H6	120.1
C7—C1—C2	123.307 (15)	O1—C7—N1	121.104 (17)
C6—C1—C2	119.469 (17)	O1—C7—C1	120.026 (15)
C1—C2—C3	120.112 (17)	N1—C7—C1	118.869 (15)
C1—C2—H2	120	N1—C8—C9	113.602 (15)
C3—C2—H2	120.0	N1—C8—H8A	109.3
C2—C3—C4	120.042 (18)	N1—C8—H8B	109.3
C2—C3—H3	120.2	C9—C8—H8A	109
C4—C3—H3	120	C9—C8—H8B	109
C5—C4—C3	120.08 (2)	H8A—C8—H8B	107
C5—C4—H4	120	O2—C9—O3	124.871 (17)
C3—C4—H4	120	O2—C9—C8	109.868 (16)
C6—C5—C4	119.950 (18)	O3—C9—C8	125.260 (18)
C6—C5—H5	119.5	C7—N1—C8	121.351 (15)
C4—C5—H5	121	C7—N1—H1	120
C1—C6—C5	120.334 (17)	C8—N1—H1	118.8
C1—C6—H6	120	C9—O2—H9	110.7
C1—C7—N1—C8	−176.76 (3)	H4—C4—C5—C6	180 (1)
C1—C7—N1—H1	2 (2)	H4—C4—C5—H5	−1 (4)
C1—C6—C5—C4	0.46 (3)	C5—C6—C1—C7	177.82 (3)
C1—C6—C5—H5	−179 (4)	H5—C5—C6—H6	1 (6)
C1—C2—C3—C4	−0.56 (3)	C6—C1—C7—O1	−12.34 (3)
C1—C2—C3—H3	−180 (4)	C6—C1—C7—N1	167.85 (3)
C2—C1—C7—O1	166.63 (3)	H6—C6—C1—C7	−2 (5)
C2—C1—C7—N1	−13.19 (3)	C7—N1—C8—C9	−88.04 (3)
C2—C1—C6—C5	−1.18 (3)	C7—N1—C8—H8A	150 (2)
C2—C1—C6—H6	179 (4)	C7—N1—C8—H8B	33 (5)
C2—C3—C4—C5	−0.17 (4)	C8—N1—C7—O1	3.43 (3)
C2—C3—C4—H4	−179.6 (8)	C8—C9—O2—H9	−178 (3)
H2—C2—C1—C7	2 (5)	H8A—C8—N1—H1	−28 (1)
H2—C2—C1—C6	−179 (5)	H8A—C8—C9—O2	−51 (3)
H2—C2—C3—C4	179 (5)	H8A—C8—C9—O3	130 (4)
H2—C2—C3—H3	0 (6)	H8B—C8—N1—H1	−145 (7)
C3—C2—C1—C7	−177.71 (4)	H8B—C8—C9—O2	65.6 (7)
C3—C2—C1—C6	1.23 (4)	H8B—C8—C9—O3	−114.1 (8)
C3—C4—C5—C6	0.22 (4)	C9—C8—N1—H1	93 (6)
C3—C4—C5—H5	180 (4)	N1—C8—C9—O2	−172.56 (3)
H3—C3—C4—C5	179 (5)	N1—C8—C9—O3	7.76 (4)
H3—C3—C4—H4	0 (5)	O1—C7—N1—H1	−178 (3)
C4—C5—C6—H6	−180 (4)	O3—C9—O2—H9	2 (3)

Hydrogen-bond geometry (Å, º)

D—H···A	D—H	H···A	D···A	D—H···A
C8—H8B···O3^i	1.10	2.54	3.5953 (3)	161
C4—H4···O2^ii	1.09	2.57	3.1810 (3)	114
N1—H1···C2	1.02	2.55	2.9197 (3)	101
O2—H9···O1^iii	1.00	1.67	2.6544 (3)	165
C2—H2···O1^iv	1.09	2.38	3.4411 (3)	166
C8—H8A···C6^iv	1.10	2.74	3.4842 (4)	125
N1—H1···O1^iv	1.02	1.99	2.9561 (3)	156

Symmetry codes: (i) x−1/2, −y+3/2, −z+2; (ii) x, y, z−1; (iii) x+1/2, −y+3/2, −z+2; (iv) −x+1, y−1/2, −z+3/2.

Benzoylaminoethanoic acid (IAM_MoPro)

Crystal data

C9H9NO3	F(000) = 376
Mr = 179.17	Dx = 1.425 Mg m^−3
Orthorhombic, P212121	Mo Kα radiation, λ = 0.71073 Å
Hall symbol: P 2ac 2ab	Cell parameters from 599 reflections
a = 8.6793 (4) Å	θ = 3.0–55.5°
b = 9.0674 (4) Å	µ = 0.11 mm^−1
c = 10.6085 (4) Å	T = 100 K
V = 834.88 (6) Å^3	Block, colorless
Z = 4	0.28 × 0.21 × 0.18 mm

Data collection

Bruker D8 Venture with PHOTON II Detector (Bruker, 2016) diffractometer	13610 reflections with > 2.0σ(I)
ω and phi scan	Rint = 0.058
Absorption correction: multi-scan SADAB, Bruker, 2016	θmax = 62.7°, θmin = 3.0°
Tmin = 0.971, Tmax = 0.981	h = −21→21
118392 measured reflections	k = −22→22
13610 independent reflections	l = −26→26

Refinement

Refinement on F^2	Secondary atom site location: difference Fourier map
Least-squares matrix: full	Hydrogen site location: difference Fourier map
R[F^2 > 2σ(F^2)] = 0.041	Only H-atom coordinates refined
wR(F^2) = 0.101	w = 1/[σ^2(Fo^2) + (0.057P)^2 + 0.0088P] where P = (Fo^2 + 2Fc^2)/
S = 1.08	(Δ/σ)max < 0.001
11896 reflections	Δρmax = 0.55 e Å^−3
145 parameters	Δρmin = −0.38 e Å^−3
0 restraints	Absolute structure: Flack x determined using 4564 quotients [(I+)-(I-)]/[(I+)+(I-)] (Parsons, Flack and Wagner, Acta Cryst. B69 (2013) 249-259)
Primary atom site location: structure-invariant direct methods	Absolute structure parameter: −0.05 (11)

Special details

Refinement. Refinement of F² against reflections. The threshold expression of F² > 2sigma(F²) is used for calculating R-factors(gt) and is not relevant to the choice of reflections for refinement. R-factors based on F² are statistically about twice as large as those based on F, and R-factors based on ALL data will be even larger.

Fractional atomic coordinates and isotropic or equivalent isotropic displacement parameters (Ų)

	x	y	z	Uiso*/Ueq
C1	0.39580 (4)	0.63965 (4)	0.58472 (3)	0.01133 (2)
C2	0.46753 (5)	0.52385 (4)	0.51962 (3)	0.01392 (2)
H2	0.532 (2)	0.441 (11)	0.5714 (14)	0.03054
C3	0.45598 (5)	0.51498 (5)	0.38867 (4)	0.01642 (3)
H3	0.5116 (19)	0.425 (11)	0.3381 (16)	0.03547
C4	0.37199 (5)	0.62074 (5)	0.32223 (3)	0.01637 (3)
H4	0.363 (2)	0.613 (2)	0.220 (11)	0.03314
C5	0.29887 (5)	0.73527 (5)	0.38667 (4)	0.01632 (3)
H5	0.2334 (17)	0.818 (11)	0.3356 (14)	0.03444
C6	0.31017 (5)	0.74441 (4)	0.51749 (3)	0.01430 (2)
H6	0.253 (2)	0.833 (11)	0.5680 (13)	0.03204
C7	0.40745 (4)	0.65973 (4)	0.72401 (3)	0.01103 (2)
C8	0.46941 (4)	0.55963 (4)	0.92983 (3)	0.01333 (2)
H8A	0.459 (2)	0.449 (11)	0.9705 (15)	0.03131
H8B	0.371 (11)	0.6248 (18)	0.9638 (16)	0.02953
C9	0.61687 (4)	0.62933 (4)	0.97854 (3)	0.01254 (2)
N1	0.46286 (4)	0.54933 (3)	0.79392 (3)	0.01290 (2)
O1	0.36575 (4)	0.77897 (3)	0.77352 (3)	0.01449 (2)
O2	0.62203 (4)	0.61883 (5)	1.10332 (3)	0.01833 (3)
O3	0.71468 (4)	0.68671 (5)	0.91385 (3)	0.01999 (3)
H1	0.501 (2)	0.458 (11)	0.7466 (16)	0.02774
H9	0.720 (11)	0.664 (2)	1.1343 (15)	0.03066

Atomic displacement parameters (Ų)

	U^11	U^22	U^33	U^12	U^13	U^23
C1	0.01234 (9)	0.01171 (9)	0.00995 (8)	0.00029 (7)	−0.00047 (7)	−0.00004 (6)
C2	0.01636 (11)	0.01423 (10)	0.01116 (9)	0.00241 (9)	−0.00062 (8)	−0.00107 (8)
H2	0.03913	0.02807	0.02440	0.01306	−0.00441	0.00051
C3	0.01812 (13)	0.01968 (13)	0.01146 (10)	0.00240 (10)	0.00024 (9)	−0.00227 (9)
H3	0.04435	0.03732	0.02474	0.01410	0.00064	−0.00931
C4	0.01600 (12)	0.02257 (14)	0.01055 (9)	−0.00048 (11)	−0.00019 (8)	0.00068 (9)
H4	0.03875	0.04484	0.01582	0.00277	−0.00218	−0.00011
C5	0.01667 (12)	0.01957 (13)	0.01271 (10)	0.00166 (10)	−0.00196 (9)	0.00281 (9)
H5	0.03977	0.03600	0.02755	0.01124	−0.00611	0.00789
C6	0.01543 (11)	0.01471 (11)	0.01277 (10)	0.00256 (8)	−0.00140 (8)	0.00075 (8)
H6	0.03932	0.02887	0.02794	0.01382	0.00045	−0.00289
C7	0.01175 (9)	0.01102 (8)	0.01032 (8)	−0.00038 (7)	0.00020 (7)	−0.00036 (7)
C8	0.01387 (10)	0.01577 (11)	0.01034 (9)	−0.00180 (8)	−0.00049 (7)	0.00119 (8)
H8A	0.04273	0.02366	0.02755	−0.00666	−0.00481	0.00656
H8B	0.02528	0.03588	0.02744	0.00354	0.00212	−0.00590
C9	0.01331 (10)	0.01364 (10)	0.01067 (9)	−0.00083 (8)	0.00047 (7)	−0.00012 (7)
N1	0.01653 (10)	0.01217 (8)	0.01001 (7)	0.00068 (7)	−0.00100 (7)	0.00003 (6)
O1	0.01801 (10)	0.01222 (8)	0.01324 (8)	0.00145 (7)	0.00206 (7)	−0.00197 (6)
O2	0.01860 (11)	0.02648 (14)	0.00991 (8)	−0.00617 (10)	−0.00103 (7)	−0.00012 (8)
O3	0.01795 (11)	0.02761 (15)	0.01439 (10)	−0.00763 (10)	0.00246 (8)	0.00123 (9)
H1	0.04033	0.02052	0.02237	0.00710	−0.00097	−0.00324
H9	0.02667	0.04373	0.02158	−0.01118	−0.00289	−0.00006

Geometric parameters (Å, º)

C1—C7	1.4923 (4)	C6—H6	1.09
C1—C6	1.4012 (5)	C7—O1	1.2553 (4)
C1—C2	1.4025 (5)	C7—N1	1.3355 (4)
C2—C3	1.3951 (5)	C8—N1	1.4459 (4)
C2—H2	1.09	C8—C9	1.5181 (5)
C3—C4	1.3956 (6)	C8—H8A	1.10
C3—H3	1.09	C8—H8B	1.10
C4—C5	1.3959 (6)	C9—O2	1.3279 (4)
C4—H4	1.09	C9—O3	1.2092 (5)
C5—C6	1.3937 (5)	N1—H1	1.02
C5—H5	1.09	O2—H9	1.00
C7—C1—C6	117.21 (3)	C5—C6—H6	120.1
C7—C1—C2	123.29 (3)	O1—C7—N1	121.14 (3)
C6—C1—C2	119.49 (3)	O1—C7—C1	119.99 (3)
C1—C2—C3	120.11 (3)	N1—C7—C1	118.87 (3)
C1—C2—H2	120	N1—C8—C9	113.54 (3)
C3—C2—H2	120.0	N1—C8—H8A	109.3
C2—C3—C4	120.05 (3)	N1—C8—H8B	109.4
C2—C3—H3	120.1	C9—C8—H8A	109
C4—C3—H3	120	C9—C8—H8B	109
C3—C4—C5	120.08 (3)	H8A—C8—H8B	107
C3—C4—H4	120	O2—C9—O3	124.95 (3)
C5—C4—H4	120	O2—C9—C8	109.75 (3)
C6—C5—C4	120.00 (3)	O3—C9—C8	125.30 (3)
C6—C5—H5	119.5	C7—N1—C8	121.30 (3)
C4—C5—H5	121	C7—N1—H1	117
C1—C6—C5	120.25 (3)	C8—N1—H1	122
C1—C6—H6	120	C9—O2—H9	109.0
C1—C7—N1—C8	−176.78 (4)	H4—C4—C5—C6	180 (2)
C1—C7—N1—H1	4 (3)	H4—C4—C5—H5	−1 (4)
C1—C6—C5—C4	0.52 (5)	C5—C6—C1—C7	177.81 (5)
C1—C6—C5—H5	−179 (4)	H5—C5—C6—H6	1 (6)
C1—C2—C3—C4	−0.50 (5)	C6—C1—C7—O1	−12.37 (5)
C1—C2—C3—H3	180 (4)	C6—C1—C7—N1	167.76 (5)
C2—C1—C7—O1	166.66 (5)	H6—C6—C1—C7	−2 (5)
C2—C1—C7—N1	−13.21 (5)	C7—N1—C8—C9	−88.03 (5)
C2—C1—C6—C5	−1.26 (5)	C7—N1—C8—H8A	151 (3)
C2—C1—C6—H6	179 (4)	C7—N1—C8—H8B	33 (5)
C2—C3—C4—C5	−0.24 (6)	C8—N1—C7—O1	3.35 (5)
C2—C3—C4—H4	−180 (1)	C8—C9—O2—H9	179 (3)
H2—C2—C1—C7	2 (5)	H8A—C8—N1—H1	−30 (2)
H2—C2—C1—C6	−179 (5)	H8A—C8—C9—O2	−51 (3)
H2—C2—C3—C4	179 (5)	H8A—C8—C9—O3	130 (4)
H2—C2—C3—H3	0 (6)	H8B—C8—N1—H1	−147 (7)
C3—C2—C1—C7	−177.76 (6)	H8B—C8—C9—O2	65 (2)
C3—C2—C1—C6	1.25 (6)	H8B—C8—C9—O3	−114 (2)
C3—C4—C5—C6	0.23 (6)	C9—C8—N1—H1	92 (6)
C3—C4—C5—H5	180 (4)	N1—C8—C9—O2	−172.64 (5)
H3—C3—C4—C5	179 (5)	N1—C8—C9—O3	7.80 (6)
H3—C3—C4—H4	0 (5)	O1—C7—N1—H1	−176 (3)
C4—C5—C6—H6	−180 (4)	O3—C9—O2—H9	−1 (3)

Hydrogen-bond geometry (Å, º)

D—H···A	D—H	H···A	D···A	D—H···A
C8—H8B···O3^i	1.10	2.54	3.5956 (6)	161
C4—H4···O2^ii	1.09	2.57	3.1786 (5)	114
N1—H1···C2	1.02	2.50	2.9193 (5)	104
O2—H9···O1^iii	1.00	1.68	2.6534 (5)	163
C2—H2···O1^iv	1.09	2.38	3.4409 (5)	166
C8—H8A···C6^iv	1.10	2.74	3.4845 (6)	125
N1—H1···O1^iv	1.02	2.01	2.9553 (5)	153

Symmetry codes: (i) x−1/2, −y+3/2, −z+2; (ii) x, y, z−1; (iii) x+1/2, −y+3/2, −z+2; (iv) −x+1, y−1/2, −z+3/2.

Benzoylaminoethanoic acid (IAM_shelx)

Crystal data

C9H9NO3	Dx = 1.425 Mg m^−3
Mr = 179.17	Mo Kα radiation, λ = 0.71073 Å
Orthorhombic, P212121	Cell parameters from 599 reflections
a = 8.6793 (4) Å	θ = 3.0–55.5°
b = 9.0674 (4) Å	µ = 0.11 mm^−1
c = 10.6085 (4) Å	T = 100 K
V = 834.88 (6) Å^3	Block, colorless
Z = 4	0.28 × 0.21 × 0.18 mm
F(000) = 376

Data collection

Bruker D8 Venture with PHOTON II Detector (Bruker, 2016) diffractometer	11263 reflections with I > 2σ(I)
ω and phi scan	Rint = 0.058
Absorption correction: multi-scan SADABS, Bruker, 2016	θmax = 62.7°, θmin = 3.0°
Tmin = 0.971, Tmax = 0.981	h = −21→21
118392 measured reflections	k = −22→22
13635 independent reflections	l = −26→26

Refinement

Refinement on F^2	Hydrogen site location: mixed
Least-squares matrix: full	H atoms treated by a mixture of independent and constrained refinement
R[F^2 > 2σ(F^2)] = 0.036	w = 1/[σ^2(Fo^2) + (0.0572P)^2 + 0.0088P] where P = (Fo^2 + 2Fc^2)/3
wR(F^2) = 0.107	(Δ/σ)max = 0.001
S = 1.11	Δρmax = 0.51 e Å^−3
13635 reflections	Δρmin = −0.30 e Å^−3
126 parameters	Absolute structure: Flack x determined using 4564 quotients [(I+)-(I-)]/[(I+)+(I-)] (Parsons, Flack and Wagner, Acta Cryst. B69 (2013) 249-259).
0 restraints	Absolute structure parameter: −0.05 (11)

Special details

Geometry. All esds (except the esd in the dihedral angle between two l.s. planes) are estimated using the full covariance matrix. The cell esds are taken into account individually in the estimation of esds in distances, angles and torsion angles; correlations between esds in cell parameters are only used when they are defined by crystal symmetry. An approximate (isotropic) treatment of cell esds is used for estimating esds involving l.s. planes.

Fractional atomic coordinates and isotropic or equivalent isotropic displacement parameters (Ų)

	x	y	z	Uiso*/Ueq
C1	0.39576 (5)	0.63962 (4)	0.58464 (4)	0.01144 (5)
C2	0.46749 (6)	0.52386 (5)	0.51968 (4)	0.01397 (6)
H2	0.524053	0.451396	0.564858	0.017*
C3	0.45590 (6)	0.51492 (6)	0.38867 (5)	0.01645 (7)
H3	0.505176	0.436769	0.344712	0.020*
C4	0.37198 (6)	0.62069 (6)	0.32222 (4)	0.01646 (7)
H4	0.364544	0.614726	0.233023	0.020*
C5	0.29893 (6)	0.73527 (6)	0.38669 (5)	0.01636 (7)
H5	0.241555	0.807039	0.341378	0.020*
C6	0.31018 (6)	0.74439 (5)	0.51747 (4)	0.01434 (6)
H6	0.259629	0.821934	0.561238	0.017*
C7	0.40743 (5)	0.65975 (4)	0.72403 (4)	0.01117 (5)
C8	0.46938 (5)	0.55963 (5)	0.92978 (4)	0.01334 (5)
H8A	0.459587	0.459377	0.965858	0.016*
H8B	0.380515	0.618466	0.959681	0.016*
C9	0.61684 (5)	0.62938 (5)	0.97856 (4)	0.01261 (5)
N1	0.46273 (5)	0.54938 (4)	0.79395 (3)	0.01294 (5)
O1	0.36571 (5)	0.77896 (4)	0.77349 (3)	0.01455 (5)
O2	0.62201 (5)	0.61874 (6)	1.10332 (4)	0.01829 (6)
O3	0.71455 (5)	0.68679 (6)	0.91376 (4)	0.02006 (7)
H1	0.493 (2)	0.4700 (19)	0.7538 (19)	0.036 (4)*
H9	0.7006 (18)	0.6563 (18)	1.1289 (15)	0.029 (4)*

Atomic displacement parameters (Ų)

	U^11	U^22	U^33	U^12	U^13	U^23
C1	0.01241 (12)	0.01172 (11)	0.01018 (10)	0.00024 (9)	−0.00041 (8)	−0.00002 (8)
C2	0.01644 (14)	0.01424 (13)	0.01121 (11)	0.00236 (11)	−0.00072 (10)	−0.00111 (10)
C3	0.01823 (16)	0.01959 (17)	0.01153 (12)	0.00228 (13)	0.00043 (11)	−0.00225 (11)
C4	0.01601 (15)	0.02277 (18)	0.01060 (11)	−0.00047 (13)	−0.00019 (10)	0.00071 (12)
C5	0.01673 (15)	0.01956 (17)	0.01278 (13)	0.00167 (13)	−0.00205 (11)	0.00284 (12)
C6	0.01542 (14)	0.01484 (13)	0.01276 (12)	0.00258 (11)	−0.00143 (10)	0.00070 (10)
C7	0.01197 (11)	0.01117 (10)	0.01036 (10)	−0.00033 (8)	0.00022 (9)	−0.00026 (8)
C8	0.01390 (12)	0.01584 (13)	0.01029 (11)	−0.00165 (10)	−0.00045 (9)	0.00108 (10)
C9	0.01320 (12)	0.01387 (12)	0.01076 (11)	−0.00083 (10)	0.00046 (9)	0.00000 (9)
N1	0.01657 (12)	0.01214 (10)	0.01010 (9)	0.00067 (9)	−0.00099 (8)	0.00007 (8)
O1	0.01802 (12)	0.01238 (10)	0.01326 (10)	0.00146 (9)	0.00195 (9)	−0.00198 (8)
O2	0.01851 (14)	0.02632 (17)	0.01005 (9)	−0.00604 (12)	−0.00098 (9)	−0.00018 (10)
O3	0.01811 (14)	0.02761 (18)	0.01445 (12)	−0.00767 (13)	0.00250 (10)	0.00125 (12)

Geometric parameters (Å, º)

C1—C6	1.4007 (6)	C6—H6	0.9500
C1—C2	1.4015 (6)	C7—O1	1.2549 (5)
C1—C7	1.4933 (5)	C7—N1	1.3349 (5)
C2—C3	1.3958 (6)	C8—N1	1.4451 (5)
C2—H2	0.9500	C8—C9	1.5185 (6)
C3—C4	1.3955 (8)	C8—H8A	0.9900
C3—H3	0.9500	C8—H8B	0.9900
C4—C5	1.3961 (8)	C9—O3	1.2095 (6)
C4—H4	0.9500	C9—O2	1.3278 (5)
C5—C6	1.3933 (6)	N1—H1	0.876 (18)
C5—H5	0.9500	O2—H9	0.809 (16)
C6—C1—C2	119.56 (4)	C1—C6—H6	119.9
C6—C1—C7	117.18 (4)	O1—C7—N1	121.14 (4)
C2—C1—C7	123.25 (4)	O1—C7—C1	119.98 (4)
C3—C2—C1	120.08 (4)	N1—C7—C1	118.88 (3)
C3—C2—H2	120.0	N1—C8—C9	113.60 (4)
C1—C2—H2	120.0	N1—C8—H8A	108.8
C4—C3—C2	120.04 (4)	C9—C8—H8A	108.8
C4—C3—H3	120.0	N1—C8—H8B	108.8
C2—C3—H3	120.0	C9—C8—H8B	108.8
C3—C4—C5	120.07 (4)	H8A—C8—H8B	107.7
C3—C4—H4	120.0	O3—C9—O2	125.04 (4)
C5—C4—H4	120.0	O3—C9—C8	125.21 (4)
C6—C5—C4	120.01 (4)	O2—C9—C8	109.75 (4)
C6—C5—H5	120.0	C7—N1—C8	121.35 (4)
C4—C5—H5	120.0	C7—N1—H1	116.9 (13)
C5—C6—C1	120.23 (4)	C8—N1—H1	121.7 (13)
C5—C6—H6	119.9	C9—O2—H9	109.4 (11)

Hydrogen-bond geometry (Å, º)

D—H···A	D—H	H···A	D···A	D—H···A
C8—H8B···O3^i	0.99	2.64	3.5963 (7)	161
N1—H1···O1^ii	0.876 (18)	2.143 (19)	2.9566 (6)	154.2 (19)
O2—H9···O1^iii	0.809 (16)	1.863 (16)	2.6537 (6)	165.3 (16)

Symmetry codes: (i) x−1/2, −y+3/2, −z+2; (ii) −x+1, y−1/2, −z+3/2; (iii) x+1/2, −y+3/2, −z+2.